Research ArticleMechanism of Activating the ProprioceptiveNT-3TrkC Signalling Pathway by Reverse Intervention forthe Anterior Cruciate LigamentndashHamstring ReflexArc with Electroacupuncture

Lei Zhang12 Yan Zeng1 Ji Qi2 Taiyuan Guan1 Xin Zhou1 Yancheng He 1

GuoyouWang1 and Shijie Fu 1

1The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University Luzhou Sichuan 646600 China2School of Traditional Chinese Medicine Southern Medical University Guangzhou Guangdong Province 510515 China

Correspondence should be addressed to Shijie Fu fushijieggj126com

Received 30 July 2017 Revised 2 December 2017 Accepted 14 December 2017 Published 18 January 2018

Academic Editor Magali Cucchiarini

Copyright copy 2018 Lei Zhang et alThis is an open access article distributed under theCreative CommonsAttribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

The anterior cruciate ligament (ACL) is an important structure maintaining stability of the knee joints Deficits in physical stabilityand the proprioceptive capabilities of the knee joints are observed when the ACL is damaged Additionally a unilateral ACL injurycan affect bilateral knee proprioception therefore proprioception of the ACL may play a key role in stability Electroacupuncturetherapy has a definite effect nerve regeneration In this study cynomolgus monkeys were randomly divided into 4 groups themodel control group intervention of the injured knee with electroacupuncture (IIKE) group intervention of the bilateral kneeswith electroacupuncture (IBKE) group and the blank control group The unilateral ACL injury model was developed in IIKE andIBKE groups acupuncture points around the knees underwent intervention similarly in the IIKE and IBKE groups Then mRNAand protein expressions of NT-3 and TrkC in the dorsal root ganglion and of growth-associated protein-43 in the ACL increasedaccording to reverse-transcription quantitative polymerase chain reaction andWestern blotting results Decreased incubations andincreased amplitudes were found for somatosensory-evoked potentials and motor nerve conduction velocityThe finding indicatesthat electroacupuncture may play an important role in the recovery of proprioception in the ACL by activating the NT-3TrkCsignalling pathway

1 Introduction

The anterior cruciate ligament (ACL) is an important struc-ture that maintains knee joint stability [1ndash3] ACL tears mostcommonly occur in active people and athletes [4] and maycause proprioceptive degeneration which is the main factorleading to the instability of the knee joints Proprioceptionrefers to when proprioceptors of muscles tendons andjoints and the sense of position motion perception andvibration which mainly occur in the nerve tissue are ableto complete information transmission [5ndash8] However it isunclear whether an ACL injury in the unilateral knee hadimpacts on the contralateral proprioception Therefore howto effectively restore proprioception of the injured ACL isgaining attention

The ACLndashhamstring reflex arc is an important feedbackmechanism for reflexively regulating muscular tension andcoordination of the knee joints that can be expressed by pro-prioception [9]When the ACL is pulled by forward displace-ment of the tibia the contraction of the hamstring confrontsthe tibia translation through regulating the ACLndashhamstringreflex arc which protects the ACL from injury [10] Severalstudies have been conducted to prove the existence of theACLndashhamstring reflex arc and proprioceptive regeneration[11ndash13]

Many studies have found that neurotrophin-3 (NT-3)one of the neurotrophic factors and its specific receptortyrosine protein kinase C (TrkC) play important roles in theneural transmission of proprioception proprioceptive reha-bilitation and nerve regeneration [14ndash18] Proprioceptive

HindawiBioMed Research InternationalVolume 2018 Article ID 6348764 10 pageshttpsdoiorg10115520186348764

2 BioMed Research International

neurons located in the spinal dorsal root ganglion (DRG)express TrkC suggesting that NT-3 impacts the proprio-ceptive neurons [19] Growth associated protein-43 (GAP-43) is a specific protein that widely exists in the nervoussystem [20] and is highly expressed in neuronal axons[21 22] If the ACLs were injured then the expression ofNT-3 would decrease in the proprioceptors thus reduc-ing the activity of TrkC however GAP-43 in the ACLwould be activated thereby starting the self-healing sys-tem

Previous studies [23 24] have shown that electricityused to stimulate ACL could reflexively activate the relatedflexormuscles or extensormuscles participating in stabilisingactivities of the knee joints When knee joint stability isstrengthened proprioception could be well recovered pre-venting reinjury of the ACL [25 26]

Clinically electroacupuncture therapy for acupuncturepoints around the knee joints is efficient for the treat-ment and recovery of local injuries However thus farno relevant research has definitively clarified the treatmentmechanism of electroacupuncture especially for improvingthe proprioception function Therefore this study aimed toexplore the mechanism of activating the proprioceptive NT-3TrkC signalling pathway by reverse intervention for theACLndashhamstring reflex arc with electroacupuncture

2 Materials and Methods

21 Ethical Statement All procedures were approved by theEthical Inspection Committee of Animal Experiments ofYunnan Yinmore Biological Technology Co Ltd (number2016001) Animal cares were performed in accordance withthe Guide for the Care and Use of Laboratory Animals (Officeof Science andHealth Reports CPRRNIH 1996) And animalresearch facilities were in accordance with the Associationfor Assessment andAccreditation of Laboratory Animal CareInternational (AAALAC)

22 Experimental Animals and Feeding Procedures Thirtymale specific pathogen-free (SPF) cynomolgus monkeyswere bred for the purpose of this study and purchased fromYunnan Yinmore Biological Technology Co Ltd (age range40 to 50 years weight range 60 to 70 kg) All monkeys werehoused in several stable cages during periods of sleepingactivities feeding and resting at the Laboratory AnimalsBreeding Center of Yunnan Yinmore Biological TechnologyCo Ltd (SYXK2009-0003)

23 Grouping and Modelling Thirty cynomolgus monkeyswere randomly divided into 4 groups blank control group(119899 = 3) model control group (119899 = 9) interventionfor the injured knee with electroacupuncture (IIKE) group(119899 = 9) and intervention for the bilateral knees withelectroacupuncture (IBKE) group (119899 = 9) In the modelcontrol IIKE and IBKE groups the unilateral ACL injurymode was developed using arthroscopy

Arthroscopic instruments (Smith amp Nephew MemphisTN USA) were prepared before the procedure and the

equipment was rigorously sterilised by operators Underanaesthesia (5mgkg intramuscularly [IM] Zoletil 50 Vir-bac Carros France) the monkeys were placed in the supineposition and the surgical area was shaved Then incisionswere marked in order with a tourniquet (ZSGB-BIO Bei-jing China) Small incisions were made around the kneejoints where the scope and surgical instruments would beinserted After preparations were completed anterior medialand anterior lateral approaches to the knee joints wereperformed using 05 cm long incisions to explore the jointsThe scope was inserted in the knee joints Saline solution wasadministered through a tube and into the knees to expandthe joints and improve visualization The image was sent toa video monitor so that the surgeon could see inside thejoints Meticulous diagnostic arthroscopy was performed toevaluate the knee structures including the meniscus artic-ular cartilage ACL and posterior cruciate ligament (PCL)Approximately one-fourth of the ACL was transversely cutusing arthroscopy At the end of the procedure the surgicalinstrumentswere removed and the skin incisionswere closedwith sutures (ZSGB-BIO) A bandage was wrapped aroundthe knee joints and the monkey was taken to recoveryModelling procedures were completed by the same surgeonsSoft padded bandages were placed and maintained on theoperated limbs for 1 week On postoperative day 3 aninjection of levofloxacin hydrochloride and sodium chloride(8mgkg intravenously once every 12 hours Heng AoZhejiang China) was used to prevent infection Moreovermonkeys were monitored daily and tramadol hydrochloridewas injected (2mgkg onceday IM QiMaiTe China) torelieve pain as needed The incisions healed within 7 days

24 Electroacupuncture Intervention In the IIKE and IBKEgroups monkeys were subjected to intervention with elec-troacupuncture at 7 days postoperatively Acupuncture pointswere selected including Wei Yang (BL39) Yin Gu (KI10) XiYangguan (GB33) and Qu Quan (LR8) which are all locatedon the hamstring

While awake and after routine disinfection filiformneedles (Zhongyan Taihe Beijing China) that were 03mmin diameter and 15 inches in lengthwere slightly tilted inwardto a depth of approximately 05 to 08 inches Monkeys weresubjected to proper lifting thrusting and rotating techniquesto induce muscle contraction of the hamstring After inser-tion in the acupuncture points for approximately 15 minutesthe needles were connected to stimulator electrodes Thestimulation frequency was fixed at 2100Hz with alternatingdispersed and dense waves a current intensity of 3mA wasselected Electroacupuncture treatment was performed for 10minutes each time Intervention was performed every dayfor 4 weeks The entire process was performed by the sameacupuncturist

The blank control and model control groups didnot undergo any intervention In the IIKE group elec-troacupuncture intervention for the injured kneewas appliedthe same intervention was applied for the bilateral knees (theinjured knee and healthy knee) in the IBKE group (Figure 1)

At 4 8 and 12 weeks after electroacupuncture interven-tion 3 monkeys were separately and randomly chosen from

BioMed Research International 3

(a) (b)

Figure 1 Cynomolgus monkeys underwent electroacupuncture intervention while awake (a) The acupuncture point nerve stimulator (G91-DKangling Zhejiang China G91-D) (b) A cynomolgus monkeys was placed on the fixture and subjected to electroacupuncture

the model control group IIKE group and IBKE group andthe proprioception changes of the ACL were examined usingthe neural electrophysiological method (somatosensory-evoked potentials [SEPs] and motor nerve conduction veloc-ity [MCV]) reverse-transcription quantitative polymerasechain reaction (RT-qPCR) and Western blotting SEPs andMCV are currently used for monitoring of peripheral nerveinjuries their indicators are incubations and amplitudesExtended incubations and decreased amplitudes over timeindicated nerve injury and proprioceptive degeneration RT-qPCR andWestern blottingwere used to examinemRNAandprotein expressions of NT-3 TrkC and GAP-43 Examina-tions were performed formonkeys in the blank control groupafter group division

25 SEPs Monkeys were given Zoletil 50 (5mgkg IM) forgeneral anaesthesia after anaesthesia the heads and limbsof the monkeys were fixed and only the injured knee wasexamined Electrode points were 1 cm from the point ofintersection of the bilateral ear tips attachments the rootof the nose and the inion (lower limb cortex area) thereference electrode point was placed in the nasal root andthe ground wires were connected to the ears SEPs weremeasured at 26ndash28∘C and the bipolar surface electrodes wereused to stimulate the area of the body corresponding toboth sides of the ACL Stimulation parameters were constantvoltage and electrical stimulations of a single square wavewere used (wave width 01ms frequency 2Hz stimulationintensity range 15 to 20mA) Incubations and amplitudesof SEPs for cynomolgus monkeys were recorded by theevoked potentiometer (MEB-9402C Nihon kohden TokyoJapan) Eventually the information was entered in the micro-computer operation system and the graphics incubationsand amplitudes of SEPs were measured and analysed (Fig-ure 2(a))

26 MCV The stimulation electrode was placed in thepopliteal space for stimulation the recording electrode wasplaced on the muscle belly of the hamstring at a temperature

of 26ndash28∘C the reference electrode was placed 2 cm fromthe recording electrode and bipolar surface electrodes wereplaced on the ACL for stimulation Stimulation parameterswere constant voltage wave width of 02ms frequency of1Hz and stimulation intensity range of 25 to 30mA ThenMCV incubations and amplitudes were recorded by theevoked potentiometer Finally the informationwas entered inthemicrocomputer operation system and theMCVgraphicsincubations and amplitudes were measured and analysed(Figure 2(b)) Only the injured knee was examined

27 Specimen Extraction Monkeys were euthanised withZoletil 50 and the DRG and ACL were rapidly exposed andtaken out to be used as experimental materials (Figure 3)They were washed with physiological saline (ZSGB-BIO)without RNase destroyer (ABI Vernon CA USA) placed in5ml frozen pipes (Sangon Shanghai China) quickly plungedinto liquid nitrogen (ABI) and kept in the refrigerator atminus80∘C Bilateral DRGs were obtained from the 4 groupsbilateral ACLs were obtained from the blank control groupand injured ACLs were obtained from the model controlIIKE and IBKE groups

28 RT-qPCR Total RNA was isolated from the DRG usingTRIZOL reagent (Sangon) The concentration and purity ofthe extracted RNA were determined by ultramicronucleicacid protein detector (AlphaSpec Alpha Innotech San Lean-dro CA USA) The primers of the target gene and referencegene were provided by Sangon and designed and synthesisedusing Primer Premier 50 software Reference gene (120573-actin)primer sequences were R-120573-actin forward 51015840-GATCAA-GATCATTGCTCCTCCTG-31015840 5893 and R-120573-actin reverse51015840-GTCACAGTCCGCCTAGAAGC-31015840 6046 163 bp Tar-get gene primer sequences were R-NT-3 forward 51015840-TGC-CACGATCTTACAGGTGAAC-31015840 6061 R-NT-3 reverse51015840-TCCTTAACGTCCACCATCTGC-31015840 6007 198 bp R-TrkC forward 51015840-CAAATGCTCCACATCGCCAG-31015840 599R-TrkC reverse 51015840-CCTCCCACCCTGTAATAATCCG-31015840

4 BioMed Research International

(a) (b)

Figure 2Cynomolgus monkeys underwent electrophysiological examination involving somatosensory-evoked potentials (SEPs) andmotor nerveconduction velocity (MCV) under general anaesthesia (a) SEPs examination (b) MCV examination

(a) (b)

Figure 3 Obtaining experimental specimens (a) The white arrows indicate the dorsal root ganglion (DRG) (b) The red arrow indicates theanterior cruciate ligament (ACL) Bilateral DRGs were obtained from all groups Bilateral ACLs were obtained from the blank control groupInjured ACLs were obtained from the model control IIKE and IBKE groups

5996 179 bp R-GAP-43 forward 51015840-TCCACTGATAAC-TCGCCGTC-31015840 5955 and R-GAP-43 reverse 51015840-TCC-ACTGATAACTCGCCGTC-31015840 6044 98 bp cDNA wasgenerated by total RNA using a reverse-transcriptase cDNAsynthesis kit With SYBR Green (SYBR Premix Ex Taq IIAxygen Union City CA USA) as the fluorescent label and120573-actin as the internal control 1 120583l cDNA was added to the20120583l reaction system for 40 cycles for amplification Dataresults were analysed using the relative quantitative methodof 2minusΔΔCt

29 Western Blotting After grinding with liquid nitrogeneach 03 g sample was added to 1ml protein extractionpyrolysis liquid (Sangon) and placed in an ice box (PCRCooler Eppendorf Hamburg Germany) for ultrasonic

crushing Then the centrifuged supernatant fluid (MultifugeX3 Thermo Fisher Scientific Waltham MA USA) wasdetermined quantitatively using the bicinchoninic acid assayAfter the concentration was adjusted 20120583l protein samplebuffer was added to 20120583l sample and boiled for 10 minutesThe pretreated protein samples were separated by SDS-PAGEand transferred to PVDF membranes (Sangon) Then thePVDF membranes were moved to TBST solution (Sangon)for blockingThe membrane was incubated with the primaryantibody solution for 1 to 2 hours at 20∘C on a shakingtable After being washed with TBST solution repeatedly themembrane was incubated for 1 to 2 hours at 20∘C with theconjugated antibody (Abcam Cambridge MA USA) Thebands were visualized using ECL (Sangon) and the pictureswere conserved by a chemiluminescence imaging apparatus(SmartChemi 610 Sage Beijing China)

BioMed Research International 5

IIKE groupIBKE groupModel control groupBlank control group

0

10

20

30

40In

cuba

tion

of S

EPs (

ms)

8 124(Weeks)

abcde

bcde

cde

abdebde

dee

beabe

(a)

0

2

4

6

8

10

Am

plitu

de o

f SEP

s (

V)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde

dee

beabe

(b)

0

10

20

30

Incu

batio

n of

MCV

(ms)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abde

bde

de ebe

abe

(c)

0

4

8

12

Am

plitu

de o

f MCV

(mV

)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde cde

abdebde

dee

beabe

(d)

Figure 4 Comparison of incubations and amplitudes of SEPs and MCV among all groups at different time points (a) Incubations of SEPs (b)Amplitudes of SEPs (c) Incubations of MCV (d) Amplitudes of MCV a versus 8 weeks in the same group 119875 lt 005 b versus 12 weeks inthe same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 d versus the IBKE group at the same time 119875 lt 005 e versusthe blank control group 119875 lt 005

210 Statistical Analysis Measurement data were expressedasmeanplusmn SD RT-qPCRwas performed during three separateexperiments and data were collected Statistical differencesbetween the 4 groups were assessed by one-way analysis ofvariance (ANOVA) followed by the least significant difference(LSD) 119905-test In the same group differences in the repeated-measurement ANOVA and LSD 119905-test results were comparedamong three different time points All statistical analyseswereperformed using SPSS 200 software (IBM Corp ArmonkNY USA) 119875 lt 005 was considered statistically significant

3 Results

31 Neural Electrophysiological Indices At 4 8 and 12 weekscompared with the blank control group the incubationsof SEPs and MCV in the model control IIKE and IBKE

groups were extended but the amplitudes of SEPs and MCVwere decreased The changes of the IBKE group were betterthan those of the IIKE group and the changes of the IIKEgroup were better than those of the model control group Alldifferences were statistically significant (119875 lt 005)

In addition in the model control group the incubationsof the SEPs and MCV were extended over time but theamplitudes of SEPs and MCV were decreased In the IIKEand the IBKE groups the incubations of SEPs and MCVwere decreased but the amplitudes of SEPs and MCV wereextended Differences were statistically significant (119875 lt 005)(Figure 4)

32 RT-qPCR and Western Blotting At the same time pointregarding DRGs when comparing the mRNA and proteinexpressions of NT-3 and TrkC fewer changes occurred

6 BioMed Research International

00

05

10

15N

T-3

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde cde

abdebde de

ebeabe

(a)

00

05

10

15

TrkC

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcdecde

abde bde deebeabe

(b)

00

05

10

15

20

25

GA

P-43

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcde cde

abde bde de ebeabe

(c)

Figure 5 Expressions of mRNA of NT-3 TrkC and GAP-43 among all groups at different time points (a) Expression of NT-3 mRNA (b)Expression of TrkC mRNA (c) Expression of GAP-43 mRNA a versus 8 weeks in the same group 119875 lt 005 b versus 12 weeks in the samegroup 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 d versus the IBKE group at the same time 119875 lt 005 e versus the blankcontrol group 119875 lt 005

in the model control IIKE and IIKE groups than in theblank control group However more changes occurred inthe IBKE group than in the IIKE group More changes alsooccurred in the IIKE group than in the model control groupAll differences were statistically significant (119875 lt 005)Regarding the ACL when comparing the mRNA and proteinexpressions of GAP-43 more changes occurred in the modelcontrol group IIKE group and IBKE group than in the blankcontrol group more changes occurred in the IBKE groupthan in the IIKE group and more changes occurred in theIIKE group than in themodel control groupAll changes werestatistically significant (119875 lt 005)

At different time points in terms of mRNA and proteinexpressions of NT-3 and TrkC there were fewer changesin the model control IIKE and IBKE groups than in theblank control group however the expressions of the IIKE

and IBKE groups increased over time and the differenceswere statistically significant (119875 lt 005) In contrast theexpressions of the model control group decreased over timeand the differences were statistically significant (119875 lt 005) Interms of themRNA and protein expressions of GAP-43 thoseof the model control group decreased over time Howeverthe expressions of the IIKE and IBKE groups increased withincreased intervention time All changes were statisticallysignificant (119875 lt 005) (Figures 5 and 6)

4 Discussion

Proprioception of the ACL is closely related to knee jointstability which is realised through feedback regulation of thecentral nervous system however the key node on the path-way remains unknown [27ndash30] Many studies confirmed that

BioMed Research International 7

00

05

10

15

20N

T-3-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de e

beabe

(a)

00

05

10

15

20

TrkC

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebeabe

(b)

00

05

10

15

20

GA

P-43

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebe

abe

(c)

4 8 12

TrkCNT-3

GAP-43-actin

IIKE group IBKE group model control group blank control group

(Weeks)

(d)

Figure 6 Expressions of NT-3120573-actin TrkC120573-actin and GAP-43120573-actin proteins among the four groups at different time points (a)Expressions of NT-3120573-actin proteins (b) Expressions of TrkC120573-actin proteins (c) Expressions of GAP-43120573-actin proteins a versus 8weeks in the same group 119875 lt 005 b versus 12 weeks in the same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 dversus the IBKE group at the same time 119875 lt 005 e versus the blank control group 119875 lt 005 (d) The expression of protein of NT-3 TrkCand 120573-actin among the four groups in different time points

when the ACL proprioception function is decreased the neu-ral feedback regulation pathway is bound to be disorderedtherefore it could have further impact on the proprioceptionfunction [31 32] Some studies showed that NT-3 was the keyinduced signal of proprioceptive transduction when NT-3expression was normal the proprioceptive nerve fibres couldcontrol the target organs and have physiological roles [33ndash35] If the expressions of NT-3 and TrkC were decreasedthen apoptosis of proprioceptive neurons in the DRG couldoccur thus causing lower limb proprioception deficienciesNT-3 and TrkC could also accelerate the development andmaturation of neurons thus maintaining the survival ofmature neurons andpromoting the repair and regeneration ofinjured neurons When the proprioceptive nerve conductionpathway is injured proprioceptive neurons appear abnormal

however exogenous NT-3 could protect the proprioceptiveneurons and improve their function [36 37] ACL injuriescould damage peripheral nerve fibres When the nerve isdamaged expression of upregulation could be included andthe functional relationship could be decreased immediatelyafter reconstruction which is considered as a marker ofneuronal growth and repair [38]

During this study at 4 8 and 12 weeks in comparisonwith the blank control group incubations of SEPs and MCVin the model control group were significantly extended butthe amplitudes were significantly decreased (119875 lt 005)Comparisons of the three time points showed that incuba-tions of the SEPs and MCV in the model control group weresignificantly extended but the amplitudes were significantlydecreased over time (119875 lt 005) Furthermore at the same

8 BioMed Research International

time point in the DRG when comparing the relative mRNAand protein expressions of NT-3 and TrkC fewer changesoccurred in the model control group than in the blankcontrol group In the ACL when comparing the relativemRNA and protein expressions of GAP-43 more changesoccurred in the model control group than in the blankcontrol group all the differences were statistically significant(119875 lt 005) In the model control group at different timepoints the relative expression of mRNA and protein of NT-3 and TrkC decreased over time while the relative mRNAand protein expressions of GAP-43 increased over time allchanges were statistically significant (119875 lt 005) The resultsof this study illustrated that when the ACL was injured theexpressions of NT-3 and TrkC in neuronal cells of the DRGcould be reduced and the levels of phosphorylation of TrkCdecreased after activation thereby further reducing TrkCactivity When the ACL was injured GAP-43 was activatedin the proprioceptors which caused the increased expressionof GAP-43 Eventually changes in these cytokine expressionlevels can lead to decreased proprioceptive functions

Electroacupuncture is a form of acupuncture whereby asmall electric current is passed between pairs of acupunc-ture needles The use of electroacupuncture therapy (thecombination of nerve electrical stimulation and acupunc-ture) has gradually replaced other acupuncture therapiesElectroacupuncture therapy has been widely used to treatpain or injury in the hind limbs for many decades inChina Both human and animal experiments have confirmedthat electroacupuncture therapy has a definite effect on thetreatment of pain movement disturbance tissue regener-ation and so on [39ndash44] Some studies have found thatthe effects of acupuncture were mediated by the centralnervous system thus causing specific physiological effect thatachieved the purpose of treatment and improved the growthof neurons axonal regeneration synaptic reconstruction andthe functional connection of target organs [45 46]

When the ACL is pulled by the forward displacement ofthe tibia the contraction of the hamstring confronts tibialtranslation through regulating the ACLndashhamstring reflex arcwhich plays an important role in maintaining the stabilityof the knee joints The acupuncture points included in thisstudy were Wei Yang Yin Gu Xi Yangguan and Qu QuanThese points were chosen because they are all located nextto the hamstring When these acupuncture points werestimulated by electroacupuncture muscle contraction of thehamstring occurred and created the effect of reverse interven-tion Furthermore the ACL belongs to the arthromyodyniacategory of Traditional Chinese Medicine Among theseselected acupuncture points Wei Yang (BL19) is the Xiahetriple energiser acupuncture point that is mainly used totreat strong pain in the lumbar spine and pain and spasmsin the legs and feet Yin Gu (KI10) is mainly used to treatstrong pain in the knee joints Xi Yangguan (GB33) is mainlyused to treat swelling pain spasms in the knee joint andpopliteal area numbness in the leg and other diseases ofthe lower limbs and knee joints Qu Quan (LR8) is mainlyused to treat swelling pain in the knee joints and patellaand paralysis of lower limb Through the stimulation ofrelated points electroacupuncture transmits a large number

of proprioceptive and skin sensorial information to thecentral nervous system thereby participating in stabilisingthe activities of the knee joints and reconstruction of theproprioceptive function of the knee joints

In this study at 4 8 and 12 weeks compared withthe blank control group SEP and MCV incubations in themodel control group and the IIKE group were extendedbut the amplitudes were decreased The changes in the IIKEgroup were better than those in the model control group alldifferences were statistically significant (119875 lt 005) At thesame time point in the DRG when comparing the relativemRNA and protein expressions of NT-3 and TrkC therewere fewer changes in the model control group than in theblank control group however more changes occurred in theIIKE group than in the model control group in which alldifferences were statistically significant (119875 lt 005) It was alsoindicated that electroacupuncture could reversely intervenein the ACLndashhamstring reflex arc thereby activating the NT-3TrkC signal pathway and increasing the expression of NT-3 in the proprioceptors of hamstring NT-3 was reverselytransported to neurons in the DRG and then combined withthe corresponding TrkC receptor which promoted phospho-rylation of cytosolic TrkC after receptor activation whichcould enhance TrkC activity It could also activate GAP-43in the ACL which enhances the polymerisation ability ofGAP-43 in proprioceptors of ACL thereby increasing theactivity of ACL proprioceptionThe results found during ourexperiment may supplement certain potential explanationsabout the mechanisms of acupuncture treatment for ACLinjury to some extent

Some studies have found that unilateral ACL injury couldlead to proprioceptive degeneration of the bilateral kneejoints [47] In theory because of the characteristics of nerveconduction damage or intervention the proprioception ofone-side limb had a negative effect on the proprioceptionof the contralateral limb In this study the IIKE groupand the IBKE group were designed for the purpose ofproving that intervention for the healthy side can promote theproprioceptive recovery of the injured limb through centralregulation of the DRG

In terms of nerve electrophysiological examination andthe relativemRNA and protein expressions of NT-3 and TrkCin the DRG and of GAP-43 in the ACL the recovery effect ofthe IBKE group was more remarkable than that of the IIKEgroup (119875 lt 005) It showed that using the DRG as thenode the proprioception of bilateral ACLs could be regulatedand that the model of bilateral knee joint interventionwas better than the model of unilateral knee interventionAdditionally the effect of electroacupuncture interventionwas more significant with increased intervention time (119875 lt005)

This research had some limitations First all animalswere male and it is unclear whether that affected the resultsSecond because of the limitations of the objective conditionsthe period of observation during the experiment was only12 weeks therefore the long-term changes in proprioceptioncould not be observed Third without some antagonists ofthe NT-3TrkC signalling pathway it might be insufficientto check the role of the NT-3TrkC signalling pathway in

BioMed Research International 9

reverse intervention for the ACLndashhamstring reflex arc withelectroacupuncture Because these limitations existed thisstudy is considered basic All these issues remain to be solvedby further related studies

5 Conclusions

In this study the mechanism of reverse intervention for ACLproprioception with electroacupuncture which was basedon the ACLndashhamstring reflex arc and through mediationof the NT-3TrkC signalling pathway was explored Thisstudy helped establish the basis for the clinical application ofelectroacupuncture therapy and effective treatment methodsand patterns for proprioception disturbances caused by ACLinjuries thereby enriching and developing the scientificconnotation of electroacupuncture therapy guided by theory

Disclosure

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan are co-firstauthors

Conflicts of Interest

The authors declare that they have no conflicts of interest

Authorsrsquo Contributions

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan contributedequally to this work

Acknowledgments

This study was supported by the National Natural ScienceFoundation of China (no 81674095) Science-TechnologySupport Plan Project of Sichuan Province (no 2014SZ0185)and Science and Technology Strategic CooperationProject of Luzhou City-Southwest Medical University(no 2016LZXNYD-J08)

References

[1] H Zhang M Qiu A Zhou J Zhang and D Jiang ldquoAnatomicanterolateral ligament reconstruction improves postoperativeclinical outcomes combined with anatomic anterior cruciateligament reconstructionrdquo Journal of Sports Science andMedicine vol 15 no 4 pp 688ndash696 2016

[2] A A Rahnemai-Azar S Sabzevari S Irarrazaval T Chaoand F H Fu ldquoAnatomical individualized ACL reconstructionrdquoArchives of Bone and Joint Surgery vol 4 no 4 pp 291ndash2972016

[3] M R Sheth S R Tapasvi and S S Patil ldquoPrimary Repairof Tibial-Sided Avulsion of the Anterior Cruciate LigamentrdquoArthroscopy Techniques vol 5 no 4 pp e901ndashe906 2016

[4] F Sepulveda L Sanchez E Amy and W Micheo ldquoAnteriorCruciate Ligament InjuryrdquoCurrent SportsMedicine Reports vol16 no 3 pp 172ndash178 2017

[5] T M El-Gohary O A Khaled S R Ibrahim A M Alshenqitiand M I Ibrahim ldquoEffect of proprioception cross training on

repositioning accuracy and balance among healthy individualsrdquoJournal of PhysicalTherapy Science vol 28 no 11 pp 3178ndash31822016

[6] A Delle Vedove M Storbeck R Heller et al ldquoBiallelic loss ofproprioception-related PIEZO2 causes muscular atrophy withperinatal respiratory distress arthrogryposis and scoliosisrdquoAmerican Journal of Human Genetics vol 99 no 6 pp 1406ndash1408 2016

[7] W K Chang Y S Jung M Oh and K Kim ldquoQuantitativeassessment of proprioception using dynamometer in incom-plete spinal cord injury patients a preliminary studyrdquo Annalsof Rehabilitation Medicine vol 41 no 2 pp 218ndash224 2017

[8] G Haliloglu K Becker C Temucin et al ldquoRecessive PIEZO2stop mutation causes distal arthrogryposis with distal muscleweakness scoliosis and proprioception defectsrdquo Journal ofHuman Genetics vol 62 no 4 pp 497ndash501 2017

[9] J Li F Kong X Gao Y Shen and S Gao ldquoProspectiverandomized comparison of knee stability and proprioceptionfor posterior cruciate ligament reconstruction with autografthybrid graft and 120574-irradiated allograftrdquoArthroscopy vol 32 no12 pp 2548ndash2555 2016

[10] J F Abulhasan C M Anley M D Snow and M J GreyldquoHamstring stretch reflex could it be a reproducible objectivemeasure of functional knee stabilityrdquordquo Journal of ExperimentalOrthopaedics vol 3 no 1 2016

[11] M Solomonow R Baratta B H Zhou et al ldquoThe synergisticaction of the anterior cruciate ligament and thigh musclesin maintaining joint stabilityrdquo The American Journal of SportsMedicine vol 15 no 3 pp 207ndash213 1987

[12] P Dyhre-Poulsen and M R Krogsgaard ldquoMuscular reflexeselicited by electrical stimulation of the anterior cruciate liga-ment in humansrdquo Journal of Applied Physiology vol 89 no 6pp 2191ndash2195 2000

[13] E Tsuda Y Ishibashi Y Okamura and S Toh ldquoRestorationof anterior cruciate ligament-hamstring reflex arc after anteriorcruciate ligament reconstructionrdquoKnee Surgery Sports Trauma-tology Arthroscopy vol 11 no 2 pp 63ndash67 2003

[14] X-Y Wang P-Y Gu S-W Chen et al ldquoEndogenousneurotrophin-3 promotes neuronal sprouting from dorsalroot gangliardquo Neural Regeneration Research vol 10 no 11 pp1865ndash1868 2015

[15] K A Han D Woo S Kim et al ldquoNeurotrophin-3 regu-lates synapse development by modulating TrkC-PTP120590 synapticadhesion and intracellular signaling pathwaysrdquo The Journal ofNeuroscience vol 36 no 17 pp 4816ndash4831 2016

[16] M-H Yang K-C Chen P-W Chiang et al ldquoProteomicprofiling of neuroblastoma cells adhesion on hyaluronic acid-based surface for neural tissue engineeringrdquo BioMed ResearchInternational vol 2016 Article ID 1917394 2016

[17] S Rao M Martınez-Cengotitabengoa Y Yao et al ldquoPeripheralblood nerve growth factor levels inmajor psychiatric disordersrdquoJournal of Psychiatric Research vol 86 pp 39ndash45 2016

[18] M Yong S Kim and S Cheon ldquoEffects of skilled reach trainingwith affected forelimb and treadmill exercise on the expressionof neurotrophic factor following ischemia-induced brain injuryin ratsrdquo Journal of Physical Therapy Science vol 29 no 4 pp647ndash650 2017

[19] S Liu S Blanchard S Bigou S Vitry D Bohl and J-M HeardldquoNeurotrophin 3 improves delayed reconstruction of sensorypathways after cervical dorsal root injuryrdquoNeurosurgery vol 68no 2 pp 450ndash461 2011

10 BioMed Research International

[20] J Cui C Cui Y Cui et al ldquoBone marrow mesenchymal stemcell transplantation increases GAP-43 Expression via ERK12and PI3KAkt pathways in intracerebral hemorrhagerdquo CellularPhysiology and Biochemistry vol 42 no 1 pp 137ndash144 2017

[21] Y Li XGaoQWang et al ldquoRetinoic acid protects from experi-mental cerebral infarction by upregulating GAP-43 expressionrdquoBrazilian Journal of Medical and Biological Research vol 50 no4 2017

[22] H B Kim and B S Yoo ldquoPropolis inhibits neurite outgrowthin differentiating SH-SY5Y human neuroblastoma cellsrdquo Toxi-cological Research vol 32 no 3 pp 239ndash243 2016

[23] J Iwasa M Ochi Y Uchio N Adachi and K KawasakildquoDecrease in anterior knee laxity by electrical stimulation ofnormal and reconstructed anterior cruciate ligamentsrdquo TheJournal of Boneamp Joint Surgery vol 88 no 4 pp 477ndash483 2006

[24] A Biscarini F M Botti and V E Pettorossi ldquoSelectivecontribution of each hamstring muscle to anterior cruciateligament protection and tibiofemoral joint stability in leg-extension exercise A simulation studyrdquo European Journal ofApplied Physiology vol 113 no 9 pp 2263ndash2273 2013

[25] D Sugimoto E Alentorn-Geli J Mendiguchıa K SamuelssonJ Karlsson andGDMyer ldquoBiomechanical and neuromuscularcharacteristics of male athletes implications for the develop-ment of anterior cruciate ligament injury prevention programsrdquoSports Medicine vol 45 no 6 pp 809ndash822 2015

[26] M G Barcellona M C Morrissey P Milligan M Clinton andA A Amis ldquoThe effect of knee extensor open kinetic chainresistance training in the ACL-injured kneerdquo Knee SurgerySports Traumatology Arthroscopy vol 23 no 11 pp 3168ndash31772015

[27] T S Furlanetto L A Peyre-Tartaruga A S Do Pinho E SDa Bernardes and M A Zaro ldquoProprioception body balanceand functionality in individuals with ACL reconstructionrdquoActaOrtopedica Brasileira vol 24 no 2 pp 67ndash72 2016

[28] A J Wodowski C W Swigler H Liu K M Nord P C ToyandW M Mihalko ldquoProprioception and Knee Arthroplasty ALiterature ReviewrdquoOrthopedic Clinics of North America vol 47no 2 pp 301ndash309 2016

[29] I Dallo J Chahla J JMitchell C Pascual-Garrido J A Feaginand R F LaPrade ldquoBiologic Approaches for the Treatment ofPartial Tears of the Anterior Cruciate Ligament A CurrentConcepts Reviewrdquo Orthopaedic Journal of Sports Medicine vol5 no 1 2017

[30] W R Lowe R J Warth E P Davis and L Bailey ldquoFunc-tional bracing after anterior cruciate ligament reconstructionA systematic reviewrdquo Journal of the American Academy ofOrthopaedicSurgeons vol 25 no 3 pp 239ndash249 2017

[31] Y Ma M Deie D Iwaki et al ldquoBalance Ability and Pro-prioception after Single-Bundle Single-Bundle Augmentationand Double-Bundle ACL ReconstructionrdquoThe Scientific WorldJournal vol 2014 Article ID 342012 8 pages 2014

[32] N Relph L Herrington and S Tyson ldquoThe effects of ACLinjury on knee proprioception Ameta-analysisrdquo Physiotherapyvol 100 no 3 pp 187ndash195 2014

[33] B P Livingston and T R Nichols ldquoEffects of reinnervation ofthe triceps brachii on joint kinematics and electromyographicpatterns of the feline forelimb during level and upslope walk-ingrdquo Cells Tissues Organs vol 199 no 5-6 pp 405ndash422 2014

[34] E Appel S Weissmann Y Salzberg et al ldquoAn ensemble ofregulatory elements controls Runx3 spatiotemporal expressionin subsets of dorsal root ganglia proprioceptive neuronsrdquoGenesamp Development vol 30 no 23 pp 2607ndash2622 2016

[35] S M OrsquoToole M M Ferrer J Mekonnen et al ldquoDicermaintains the identity and function of proprioceptive sensoryneuronsrdquo Journal of Neurophysiology vol 117 no 3 pp 1057ndash1069 2017

[36] Y Liu L Kelamangalath H Kim et al ldquoNT-3 promotes pro-prioceptive axon regeneration when combined with activationof the mTor intrinsic growth pathway but not with reduction ofmyelin extrinsic inhibitorsrdquo Experimental Neurology vol 283pp 73ndash84 2016

[37] S Li T Shen Y Liang Y Zhang and B Bai ldquoEffects ofminiscalpel-needle release on chronic neck pain A retrospec-tive analysis with 12-month follow-uprdquo PLoS ONE vol 10 no8 Article ID e0137033 pp 133ndash137 2015

[38] W-S Juan S-Y Huang C-C Chang et al ldquoMelatoninimproves neuroplasticity by upregulating the growth-associatedprotein-43 (GAP-43) and NMDAR postsynaptic density-95(PSD-95) proteins in cultured neurons exposed to glutamateexcitotoxicity and in rats subjected to transient focal cerebralischemia even during a long-term recovery periodrdquo Journal ofPineal Research vol 56 no 2 pp 213ndash223 2014

[39] Y-HGaoC-W Li J-YWang et al ldquoActivation of hippocampalMEK1 contributes to the cumulative antinociceptive effect ofelectroacupuncture in neuropathic pain ratsrdquo BMC Comple-mentary and Alternative Medicine vol 16 no 1 article no 5172016

[40] K-Y Huang S Liang M-L Yu S-P Fu X Chen and S-F Lu ldquoA systematic review and meta-analysis of acupuncturefor improving learning and memory ability in animalsrdquo BMCComplementary and Alternative Medicine vol 16 no 1 articleno 297 2016

[41] J Zhou S Li Y Wang et al ldquoEffects and mechanisms ofauricular electroacupuncture on gastric hypersensitivity in arodent model of functional dyspepsiardquo PLoS ONE vol 12 no3 Article ID e0174568 2017

[42] Y H Gao C W Li J Y Wang et al ldquoEffect of electroacupunc-ture on the cervicospinal P2X7 receptorfractalkineCX3CR1signaling pathway in a rat neck-incision painmodelrdquo PurinergicSignalling vol 13 no 2 pp 215ndash225 2017

[43] JH LeeDGoWKimet al ldquoInvolvement of spinalmuscarinicand serotonergic receptors in the anti-allodynic effect of elec-troacupuncture in rats with oxaliplatin-induced neuropathicpainrdquoKorean Journal of Physiology amp Pharmacology vol 20 no4 pp 407ndash414 2016

[44] S P Chen Y Kan J L Zhang et al ldquoInvolvement of hippocam-pal acetylcholinergic receptors in electroacupuncture analgesiain neuropathic pain ratsrdquo Behavioral and Brain Functions vol12 no 1 article no 13 2016

[45] M Villarreal Santiago S Tumilty A Mącznik and R ManildquoDoes acupuncture alter pain-related functional connectivity ofthe central nervous system a systematic reviewrdquo JAMS Journalof Acupuncture and Meridian Studies vol 9 no 4 pp 167ndash1772016

[46] T E Salazar M R Richardson E Beli et al ldquoElectroacupunc-ture Promotes Central Nervous System-Dependent Release ofMesenchymal Stem Cellsrdquo Stem Cells vol 35 no 5 pp 1303ndash1315 2017

[47] D-H Lee J-H Lee S-E Ahn and M-J Park ldquoEffect of timeafter anterior cruciate ligament tears on proprioception andpostural stabilityrdquo PLoS ONE vol 10 no 9 Article ID e01390382015

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

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Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

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Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

BioMed Research International 3

(a) (b)

Figure 1 Cynomolgus monkeys underwent electroacupuncture intervention while awake (a) The acupuncture point nerve stimulator (G91-DKangling Zhejiang China G91-D) (b) A cynomolgus monkeys was placed on the fixture and subjected to electroacupuncture

the model control group IIKE group and IBKE group andthe proprioception changes of the ACL were examined usingthe neural electrophysiological method (somatosensory-evoked potentials [SEPs] and motor nerve conduction veloc-ity [MCV]) reverse-transcription quantitative polymerasechain reaction (RT-qPCR) and Western blotting SEPs andMCV are currently used for monitoring of peripheral nerveinjuries their indicators are incubations and amplitudesExtended incubations and decreased amplitudes over timeindicated nerve injury and proprioceptive degeneration RT-qPCR andWestern blottingwere used to examinemRNAandprotein expressions of NT-3 TrkC and GAP-43 Examina-tions were performed formonkeys in the blank control groupafter group division

25 SEPs Monkeys were given Zoletil 50 (5mgkg IM) forgeneral anaesthesia after anaesthesia the heads and limbsof the monkeys were fixed and only the injured knee wasexamined Electrode points were 1 cm from the point ofintersection of the bilateral ear tips attachments the rootof the nose and the inion (lower limb cortex area) thereference electrode point was placed in the nasal root andthe ground wires were connected to the ears SEPs weremeasured at 26ndash28∘C and the bipolar surface electrodes wereused to stimulate the area of the body corresponding toboth sides of the ACL Stimulation parameters were constantvoltage and electrical stimulations of a single square wavewere used (wave width 01ms frequency 2Hz stimulationintensity range 15 to 20mA) Incubations and amplitudesof SEPs for cynomolgus monkeys were recorded by theevoked potentiometer (MEB-9402C Nihon kohden TokyoJapan) Eventually the information was entered in the micro-computer operation system and the graphics incubationsand amplitudes of SEPs were measured and analysed (Fig-ure 2(a))

26 MCV The stimulation electrode was placed in thepopliteal space for stimulation the recording electrode wasplaced on the muscle belly of the hamstring at a temperature

of 26ndash28∘C the reference electrode was placed 2 cm fromthe recording electrode and bipolar surface electrodes wereplaced on the ACL for stimulation Stimulation parameterswere constant voltage wave width of 02ms frequency of1Hz and stimulation intensity range of 25 to 30mA ThenMCV incubations and amplitudes were recorded by theevoked potentiometer Finally the informationwas entered inthemicrocomputer operation system and theMCVgraphicsincubations and amplitudes were measured and analysed(Figure 2(b)) Only the injured knee was examined

27 Specimen Extraction Monkeys were euthanised withZoletil 50 and the DRG and ACL were rapidly exposed andtaken out to be used as experimental materials (Figure 3)They were washed with physiological saline (ZSGB-BIO)without RNase destroyer (ABI Vernon CA USA) placed in5ml frozen pipes (Sangon Shanghai China) quickly plungedinto liquid nitrogen (ABI) and kept in the refrigerator atminus80∘C Bilateral DRGs were obtained from the 4 groupsbilateral ACLs were obtained from the blank control groupand injured ACLs were obtained from the model controlIIKE and IBKE groups

28 RT-qPCR Total RNA was isolated from the DRG usingTRIZOL reagent (Sangon) The concentration and purity ofthe extracted RNA were determined by ultramicronucleicacid protein detector (AlphaSpec Alpha Innotech San Lean-dro CA USA) The primers of the target gene and referencegene were provided by Sangon and designed and synthesisedusing Primer Premier 50 software Reference gene (120573-actin)primer sequences were R-120573-actin forward 51015840-GATCAA-GATCATTGCTCCTCCTG-31015840 5893 and R-120573-actin reverse51015840-GTCACAGTCCGCCTAGAAGC-31015840 6046 163 bp Tar-get gene primer sequences were R-NT-3 forward 51015840-TGC-CACGATCTTACAGGTGAAC-31015840 6061 R-NT-3 reverse51015840-TCCTTAACGTCCACCATCTGC-31015840 6007 198 bp R-TrkC forward 51015840-CAAATGCTCCACATCGCCAG-31015840 599R-TrkC reverse 51015840-CCTCCCACCCTGTAATAATCCG-31015840

4 BioMed Research International

(a) (b)

Figure 2Cynomolgus monkeys underwent electrophysiological examination involving somatosensory-evoked potentials (SEPs) andmotor nerveconduction velocity (MCV) under general anaesthesia (a) SEPs examination (b) MCV examination

(a) (b)

Figure 3 Obtaining experimental specimens (a) The white arrows indicate the dorsal root ganglion (DRG) (b) The red arrow indicates theanterior cruciate ligament (ACL) Bilateral DRGs were obtained from all groups Bilateral ACLs were obtained from the blank control groupInjured ACLs were obtained from the model control IIKE and IBKE groups

5996 179 bp R-GAP-43 forward 51015840-TCCACTGATAAC-TCGCCGTC-31015840 5955 and R-GAP-43 reverse 51015840-TCC-ACTGATAACTCGCCGTC-31015840 6044 98 bp cDNA wasgenerated by total RNA using a reverse-transcriptase cDNAsynthesis kit With SYBR Green (SYBR Premix Ex Taq IIAxygen Union City CA USA) as the fluorescent label and120573-actin as the internal control 1 120583l cDNA was added to the20120583l reaction system for 40 cycles for amplification Dataresults were analysed using the relative quantitative methodof 2minusΔΔCt

29 Western Blotting After grinding with liquid nitrogeneach 03 g sample was added to 1ml protein extractionpyrolysis liquid (Sangon) and placed in an ice box (PCRCooler Eppendorf Hamburg Germany) for ultrasonic

crushing Then the centrifuged supernatant fluid (MultifugeX3 Thermo Fisher Scientific Waltham MA USA) wasdetermined quantitatively using the bicinchoninic acid assayAfter the concentration was adjusted 20120583l protein samplebuffer was added to 20120583l sample and boiled for 10 minutesThe pretreated protein samples were separated by SDS-PAGEand transferred to PVDF membranes (Sangon) Then thePVDF membranes were moved to TBST solution (Sangon)for blockingThe membrane was incubated with the primaryantibody solution for 1 to 2 hours at 20∘C on a shakingtable After being washed with TBST solution repeatedly themembrane was incubated for 1 to 2 hours at 20∘C with theconjugated antibody (Abcam Cambridge MA USA) Thebands were visualized using ECL (Sangon) and the pictureswere conserved by a chemiluminescence imaging apparatus(SmartChemi 610 Sage Beijing China)

BioMed Research International 5

IIKE groupIBKE groupModel control groupBlank control group

0

10

20

30

40In

cuba

tion

of S

EPs (

ms)

8 124(Weeks)

abcde

bcde

cde

abdebde

dee

beabe

(a)

0

2

4

6

8

10

Am

plitu

de o

f SEP

s (

V)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde

dee

beabe

(b)

0

10

20

30

Incu

batio

n of

MCV

(ms)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abde

bde

de ebe

abe

(c)

0

4

8

12

Am

plitu

de o

f MCV

(mV

)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde cde

abdebde

dee

beabe

(d)

Figure 4 Comparison of incubations and amplitudes of SEPs and MCV among all groups at different time points (a) Incubations of SEPs (b)Amplitudes of SEPs (c) Incubations of MCV (d) Amplitudes of MCV a versus 8 weeks in the same group 119875 lt 005 b versus 12 weeks inthe same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 d versus the IBKE group at the same time 119875 lt 005 e versusthe blank control group 119875 lt 005

210 Statistical Analysis Measurement data were expressedasmeanplusmn SD RT-qPCRwas performed during three separateexperiments and data were collected Statistical differencesbetween the 4 groups were assessed by one-way analysis ofvariance (ANOVA) followed by the least significant difference(LSD) 119905-test In the same group differences in the repeated-measurement ANOVA and LSD 119905-test results were comparedamong three different time points All statistical analyseswereperformed using SPSS 200 software (IBM Corp ArmonkNY USA) 119875 lt 005 was considered statistically significant

3 Results

31 Neural Electrophysiological Indices At 4 8 and 12 weekscompared with the blank control group the incubationsof SEPs and MCV in the model control IIKE and IBKE

groups were extended but the amplitudes of SEPs and MCVwere decreased The changes of the IBKE group were betterthan those of the IIKE group and the changes of the IIKEgroup were better than those of the model control group Alldifferences were statistically significant (119875 lt 005)

In addition in the model control group the incubationsof the SEPs and MCV were extended over time but theamplitudes of SEPs and MCV were decreased In the IIKEand the IBKE groups the incubations of SEPs and MCVwere decreased but the amplitudes of SEPs and MCV wereextended Differences were statistically significant (119875 lt 005)(Figure 4)

32 RT-qPCR and Western Blotting At the same time pointregarding DRGs when comparing the mRNA and proteinexpressions of NT-3 and TrkC fewer changes occurred

6 BioMed Research International

00

05

10

15N

T-3

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde cde

abdebde de

ebeabe

(a)

00

05

10

15

TrkC

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcdecde

abde bde deebeabe

(b)

00

05

10

15

20

25

GA

P-43

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcde cde

abde bde de ebeabe

(c)

Figure 5 Expressions of mRNA of NT-3 TrkC and GAP-43 among all groups at different time points (a) Expression of NT-3 mRNA (b)Expression of TrkC mRNA (c) Expression of GAP-43 mRNA a versus 8 weeks in the same group 119875 lt 005 b versus 12 weeks in the samegroup 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 d versus the IBKE group at the same time 119875 lt 005 e versus the blankcontrol group 119875 lt 005

in the model control IIKE and IIKE groups than in theblank control group However more changes occurred inthe IBKE group than in the IIKE group More changes alsooccurred in the IIKE group than in the model control groupAll differences were statistically significant (119875 lt 005)Regarding the ACL when comparing the mRNA and proteinexpressions of GAP-43 more changes occurred in the modelcontrol group IIKE group and IBKE group than in the blankcontrol group more changes occurred in the IBKE groupthan in the IIKE group and more changes occurred in theIIKE group than in themodel control groupAll changes werestatistically significant (119875 lt 005)

At different time points in terms of mRNA and proteinexpressions of NT-3 and TrkC there were fewer changesin the model control IIKE and IBKE groups than in theblank control group however the expressions of the IIKE

and IBKE groups increased over time and the differenceswere statistically significant (119875 lt 005) In contrast theexpressions of the model control group decreased over timeand the differences were statistically significant (119875 lt 005) Interms of themRNA and protein expressions of GAP-43 thoseof the model control group decreased over time Howeverthe expressions of the IIKE and IBKE groups increased withincreased intervention time All changes were statisticallysignificant (119875 lt 005) (Figures 5 and 6)

4 Discussion

Proprioception of the ACL is closely related to knee jointstability which is realised through feedback regulation of thecentral nervous system however the key node on the path-way remains unknown [27ndash30] Many studies confirmed that

BioMed Research International 7

00

05

10

15

20N

T-3-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de e

beabe

(a)

00

05

10

15

20

TrkC

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebeabe

(b)

00

05

10

15

20

GA

P-43

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebe

abe

(c)

4 8 12

TrkCNT-3

GAP-43-actin

IIKE group IBKE group model control group blank control group

(Weeks)

(d)

Figure 6 Expressions of NT-3120573-actin TrkC120573-actin and GAP-43120573-actin proteins among the four groups at different time points (a)Expressions of NT-3120573-actin proteins (b) Expressions of TrkC120573-actin proteins (c) Expressions of GAP-43120573-actin proteins a versus 8weeks in the same group 119875 lt 005 b versus 12 weeks in the same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 dversus the IBKE group at the same time 119875 lt 005 e versus the blank control group 119875 lt 005 (d) The expression of protein of NT-3 TrkCand 120573-actin among the four groups in different time points

when the ACL proprioception function is decreased the neu-ral feedback regulation pathway is bound to be disorderedtherefore it could have further impact on the proprioceptionfunction [31 32] Some studies showed that NT-3 was the keyinduced signal of proprioceptive transduction when NT-3expression was normal the proprioceptive nerve fibres couldcontrol the target organs and have physiological roles [33ndash35] If the expressions of NT-3 and TrkC were decreasedthen apoptosis of proprioceptive neurons in the DRG couldoccur thus causing lower limb proprioception deficienciesNT-3 and TrkC could also accelerate the development andmaturation of neurons thus maintaining the survival ofmature neurons andpromoting the repair and regeneration ofinjured neurons When the proprioceptive nerve conductionpathway is injured proprioceptive neurons appear abnormal

however exogenous NT-3 could protect the proprioceptiveneurons and improve their function [36 37] ACL injuriescould damage peripheral nerve fibres When the nerve isdamaged expression of upregulation could be included andthe functional relationship could be decreased immediatelyafter reconstruction which is considered as a marker ofneuronal growth and repair [38]

During this study at 4 8 and 12 weeks in comparisonwith the blank control group incubations of SEPs and MCVin the model control group were significantly extended butthe amplitudes were significantly decreased (119875 lt 005)Comparisons of the three time points showed that incuba-tions of the SEPs and MCV in the model control group weresignificantly extended but the amplitudes were significantlydecreased over time (119875 lt 005) Furthermore at the same

8 BioMed Research International

time point in the DRG when comparing the relative mRNAand protein expressions of NT-3 and TrkC fewer changesoccurred in the model control group than in the blankcontrol group In the ACL when comparing the relativemRNA and protein expressions of GAP-43 more changesoccurred in the model control group than in the blankcontrol group all the differences were statistically significant(119875 lt 005) In the model control group at different timepoints the relative expression of mRNA and protein of NT-3 and TrkC decreased over time while the relative mRNAand protein expressions of GAP-43 increased over time allchanges were statistically significant (119875 lt 005) The resultsof this study illustrated that when the ACL was injured theexpressions of NT-3 and TrkC in neuronal cells of the DRGcould be reduced and the levels of phosphorylation of TrkCdecreased after activation thereby further reducing TrkCactivity When the ACL was injured GAP-43 was activatedin the proprioceptors which caused the increased expressionof GAP-43 Eventually changes in these cytokine expressionlevels can lead to decreased proprioceptive functions

Electroacupuncture is a form of acupuncture whereby asmall electric current is passed between pairs of acupunc-ture needles The use of electroacupuncture therapy (thecombination of nerve electrical stimulation and acupunc-ture) has gradually replaced other acupuncture therapiesElectroacupuncture therapy has been widely used to treatpain or injury in the hind limbs for many decades inChina Both human and animal experiments have confirmedthat electroacupuncture therapy has a definite effect on thetreatment of pain movement disturbance tissue regener-ation and so on [39ndash44] Some studies have found thatthe effects of acupuncture were mediated by the centralnervous system thus causing specific physiological effect thatachieved the purpose of treatment and improved the growthof neurons axonal regeneration synaptic reconstruction andthe functional connection of target organs [45 46]

When the ACL is pulled by the forward displacement ofthe tibia the contraction of the hamstring confronts tibialtranslation through regulating the ACLndashhamstring reflex arcwhich plays an important role in maintaining the stabilityof the knee joints The acupuncture points included in thisstudy were Wei Yang Yin Gu Xi Yangguan and Qu QuanThese points were chosen because they are all located nextto the hamstring When these acupuncture points werestimulated by electroacupuncture muscle contraction of thehamstring occurred and created the effect of reverse interven-tion Furthermore the ACL belongs to the arthromyodyniacategory of Traditional Chinese Medicine Among theseselected acupuncture points Wei Yang (BL19) is the Xiahetriple energiser acupuncture point that is mainly used totreat strong pain in the lumbar spine and pain and spasmsin the legs and feet Yin Gu (KI10) is mainly used to treatstrong pain in the knee joints Xi Yangguan (GB33) is mainlyused to treat swelling pain spasms in the knee joint andpopliteal area numbness in the leg and other diseases ofthe lower limbs and knee joints Qu Quan (LR8) is mainlyused to treat swelling pain in the knee joints and patellaand paralysis of lower limb Through the stimulation ofrelated points electroacupuncture transmits a large number

of proprioceptive and skin sensorial information to thecentral nervous system thereby participating in stabilisingthe activities of the knee joints and reconstruction of theproprioceptive function of the knee joints

In this study at 4 8 and 12 weeks compared withthe blank control group SEP and MCV incubations in themodel control group and the IIKE group were extendedbut the amplitudes were decreased The changes in the IIKEgroup were better than those in the model control group alldifferences were statistically significant (119875 lt 005) At thesame time point in the DRG when comparing the relativemRNA and protein expressions of NT-3 and TrkC therewere fewer changes in the model control group than in theblank control group however more changes occurred in theIIKE group than in the model control group in which alldifferences were statistically significant (119875 lt 005) It was alsoindicated that electroacupuncture could reversely intervenein the ACLndashhamstring reflex arc thereby activating the NT-3TrkC signal pathway and increasing the expression of NT-3 in the proprioceptors of hamstring NT-3 was reverselytransported to neurons in the DRG and then combined withthe corresponding TrkC receptor which promoted phospho-rylation of cytosolic TrkC after receptor activation whichcould enhance TrkC activity It could also activate GAP-43in the ACL which enhances the polymerisation ability ofGAP-43 in proprioceptors of ACL thereby increasing theactivity of ACL proprioceptionThe results found during ourexperiment may supplement certain potential explanationsabout the mechanisms of acupuncture treatment for ACLinjury to some extent

Some studies have found that unilateral ACL injury couldlead to proprioceptive degeneration of the bilateral kneejoints [47] In theory because of the characteristics of nerveconduction damage or intervention the proprioception ofone-side limb had a negative effect on the proprioceptionof the contralateral limb In this study the IIKE groupand the IBKE group were designed for the purpose ofproving that intervention for the healthy side can promote theproprioceptive recovery of the injured limb through centralregulation of the DRG

In terms of nerve electrophysiological examination andthe relativemRNA and protein expressions of NT-3 and TrkCin the DRG and of GAP-43 in the ACL the recovery effect ofthe IBKE group was more remarkable than that of the IIKEgroup (119875 lt 005) It showed that using the DRG as thenode the proprioception of bilateral ACLs could be regulatedand that the model of bilateral knee joint interventionwas better than the model of unilateral knee interventionAdditionally the effect of electroacupuncture interventionwas more significant with increased intervention time (119875 lt005)

This research had some limitations First all animalswere male and it is unclear whether that affected the resultsSecond because of the limitations of the objective conditionsthe period of observation during the experiment was only12 weeks therefore the long-term changes in proprioceptioncould not be observed Third without some antagonists ofthe NT-3TrkC signalling pathway it might be insufficientto check the role of the NT-3TrkC signalling pathway in

BioMed Research International 9

reverse intervention for the ACLndashhamstring reflex arc withelectroacupuncture Because these limitations existed thisstudy is considered basic All these issues remain to be solvedby further related studies

5 Conclusions

In this study the mechanism of reverse intervention for ACLproprioception with electroacupuncture which was basedon the ACLndashhamstring reflex arc and through mediationof the NT-3TrkC signalling pathway was explored Thisstudy helped establish the basis for the clinical application ofelectroacupuncture therapy and effective treatment methodsand patterns for proprioception disturbances caused by ACLinjuries thereby enriching and developing the scientificconnotation of electroacupuncture therapy guided by theory

Disclosure

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan are co-firstauthors

Conflicts of Interest

The authors declare that they have no conflicts of interest

Authorsrsquo Contributions

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan contributedequally to this work

Acknowledgments

This study was supported by the National Natural ScienceFoundation of China (no 81674095) Science-TechnologySupport Plan Project of Sichuan Province (no 2014SZ0185)and Science and Technology Strategic CooperationProject of Luzhou City-Southwest Medical University(no 2016LZXNYD-J08)

References

[1] H Zhang M Qiu A Zhou J Zhang and D Jiang ldquoAnatomicanterolateral ligament reconstruction improves postoperativeclinical outcomes combined with anatomic anterior cruciateligament reconstructionrdquo Journal of Sports Science andMedicine vol 15 no 4 pp 688ndash696 2016

[2] A A Rahnemai-Azar S Sabzevari S Irarrazaval T Chaoand F H Fu ldquoAnatomical individualized ACL reconstructionrdquoArchives of Bone and Joint Surgery vol 4 no 4 pp 291ndash2972016

[3] M R Sheth S R Tapasvi and S S Patil ldquoPrimary Repairof Tibial-Sided Avulsion of the Anterior Cruciate LigamentrdquoArthroscopy Techniques vol 5 no 4 pp e901ndashe906 2016

[4] F Sepulveda L Sanchez E Amy and W Micheo ldquoAnteriorCruciate Ligament InjuryrdquoCurrent SportsMedicine Reports vol16 no 3 pp 172ndash178 2017

[5] T M El-Gohary O A Khaled S R Ibrahim A M Alshenqitiand M I Ibrahim ldquoEffect of proprioception cross training on

repositioning accuracy and balance among healthy individualsrdquoJournal of PhysicalTherapy Science vol 28 no 11 pp 3178ndash31822016

[6] A Delle Vedove M Storbeck R Heller et al ldquoBiallelic loss ofproprioception-related PIEZO2 causes muscular atrophy withperinatal respiratory distress arthrogryposis and scoliosisrdquoAmerican Journal of Human Genetics vol 99 no 6 pp 1406ndash1408 2016

[7] W K Chang Y S Jung M Oh and K Kim ldquoQuantitativeassessment of proprioception using dynamometer in incom-plete spinal cord injury patients a preliminary studyrdquo Annalsof Rehabilitation Medicine vol 41 no 2 pp 218ndash224 2017

[8] G Haliloglu K Becker C Temucin et al ldquoRecessive PIEZO2stop mutation causes distal arthrogryposis with distal muscleweakness scoliosis and proprioception defectsrdquo Journal ofHuman Genetics vol 62 no 4 pp 497ndash501 2017

[9] J Li F Kong X Gao Y Shen and S Gao ldquoProspectiverandomized comparison of knee stability and proprioceptionfor posterior cruciate ligament reconstruction with autografthybrid graft and 120574-irradiated allograftrdquoArthroscopy vol 32 no12 pp 2548ndash2555 2016

[10] J F Abulhasan C M Anley M D Snow and M J GreyldquoHamstring stretch reflex could it be a reproducible objectivemeasure of functional knee stabilityrdquordquo Journal of ExperimentalOrthopaedics vol 3 no 1 2016

[11] M Solomonow R Baratta B H Zhou et al ldquoThe synergisticaction of the anterior cruciate ligament and thigh musclesin maintaining joint stabilityrdquo The American Journal of SportsMedicine vol 15 no 3 pp 207ndash213 1987

[12] P Dyhre-Poulsen and M R Krogsgaard ldquoMuscular reflexeselicited by electrical stimulation of the anterior cruciate liga-ment in humansrdquo Journal of Applied Physiology vol 89 no 6pp 2191ndash2195 2000

[13] E Tsuda Y Ishibashi Y Okamura and S Toh ldquoRestorationof anterior cruciate ligament-hamstring reflex arc after anteriorcruciate ligament reconstructionrdquoKnee Surgery Sports Trauma-tology Arthroscopy vol 11 no 2 pp 63ndash67 2003

[14] X-Y Wang P-Y Gu S-W Chen et al ldquoEndogenousneurotrophin-3 promotes neuronal sprouting from dorsalroot gangliardquo Neural Regeneration Research vol 10 no 11 pp1865ndash1868 2015

[15] K A Han D Woo S Kim et al ldquoNeurotrophin-3 regu-lates synapse development by modulating TrkC-PTP120590 synapticadhesion and intracellular signaling pathwaysrdquo The Journal ofNeuroscience vol 36 no 17 pp 4816ndash4831 2016

[16] M-H Yang K-C Chen P-W Chiang et al ldquoProteomicprofiling of neuroblastoma cells adhesion on hyaluronic acid-based surface for neural tissue engineeringrdquo BioMed ResearchInternational vol 2016 Article ID 1917394 2016

[17] S Rao M Martınez-Cengotitabengoa Y Yao et al ldquoPeripheralblood nerve growth factor levels inmajor psychiatric disordersrdquoJournal of Psychiatric Research vol 86 pp 39ndash45 2016

[18] M Yong S Kim and S Cheon ldquoEffects of skilled reach trainingwith affected forelimb and treadmill exercise on the expressionof neurotrophic factor following ischemia-induced brain injuryin ratsrdquo Journal of Physical Therapy Science vol 29 no 4 pp647ndash650 2017

[19] S Liu S Blanchard S Bigou S Vitry D Bohl and J-M HeardldquoNeurotrophin 3 improves delayed reconstruction of sensorypathways after cervical dorsal root injuryrdquoNeurosurgery vol 68no 2 pp 450ndash461 2011

10 BioMed Research International

[20] J Cui C Cui Y Cui et al ldquoBone marrow mesenchymal stemcell transplantation increases GAP-43 Expression via ERK12and PI3KAkt pathways in intracerebral hemorrhagerdquo CellularPhysiology and Biochemistry vol 42 no 1 pp 137ndash144 2017

[21] Y Li XGaoQWang et al ldquoRetinoic acid protects from experi-mental cerebral infarction by upregulating GAP-43 expressionrdquoBrazilian Journal of Medical and Biological Research vol 50 no4 2017

[22] H B Kim and B S Yoo ldquoPropolis inhibits neurite outgrowthin differentiating SH-SY5Y human neuroblastoma cellsrdquo Toxi-cological Research vol 32 no 3 pp 239ndash243 2016

[23] J Iwasa M Ochi Y Uchio N Adachi and K KawasakildquoDecrease in anterior knee laxity by electrical stimulation ofnormal and reconstructed anterior cruciate ligamentsrdquo TheJournal of Boneamp Joint Surgery vol 88 no 4 pp 477ndash483 2006

[24] A Biscarini F M Botti and V E Pettorossi ldquoSelectivecontribution of each hamstring muscle to anterior cruciateligament protection and tibiofemoral joint stability in leg-extension exercise A simulation studyrdquo European Journal ofApplied Physiology vol 113 no 9 pp 2263ndash2273 2013

[25] D Sugimoto E Alentorn-Geli J Mendiguchıa K SamuelssonJ Karlsson andGDMyer ldquoBiomechanical and neuromuscularcharacteristics of male athletes implications for the develop-ment of anterior cruciate ligament injury prevention programsrdquoSports Medicine vol 45 no 6 pp 809ndash822 2015

[26] M G Barcellona M C Morrissey P Milligan M Clinton andA A Amis ldquoThe effect of knee extensor open kinetic chainresistance training in the ACL-injured kneerdquo Knee SurgerySports Traumatology Arthroscopy vol 23 no 11 pp 3168ndash31772015

[27] T S Furlanetto L A Peyre-Tartaruga A S Do Pinho E SDa Bernardes and M A Zaro ldquoProprioception body balanceand functionality in individuals with ACL reconstructionrdquoActaOrtopedica Brasileira vol 24 no 2 pp 67ndash72 2016

[28] A J Wodowski C W Swigler H Liu K M Nord P C ToyandW M Mihalko ldquoProprioception and Knee Arthroplasty ALiterature ReviewrdquoOrthopedic Clinics of North America vol 47no 2 pp 301ndash309 2016

[29] I Dallo J Chahla J JMitchell C Pascual-Garrido J A Feaginand R F LaPrade ldquoBiologic Approaches for the Treatment ofPartial Tears of the Anterior Cruciate Ligament A CurrentConcepts Reviewrdquo Orthopaedic Journal of Sports Medicine vol5 no 1 2017

[30] W R Lowe R J Warth E P Davis and L Bailey ldquoFunc-tional bracing after anterior cruciate ligament reconstructionA systematic reviewrdquo Journal of the American Academy ofOrthopaedicSurgeons vol 25 no 3 pp 239ndash249 2017

[31] Y Ma M Deie D Iwaki et al ldquoBalance Ability and Pro-prioception after Single-Bundle Single-Bundle Augmentationand Double-Bundle ACL ReconstructionrdquoThe Scientific WorldJournal vol 2014 Article ID 342012 8 pages 2014

[32] N Relph L Herrington and S Tyson ldquoThe effects of ACLinjury on knee proprioception Ameta-analysisrdquo Physiotherapyvol 100 no 3 pp 187ndash195 2014

[33] B P Livingston and T R Nichols ldquoEffects of reinnervation ofthe triceps brachii on joint kinematics and electromyographicpatterns of the feline forelimb during level and upslope walk-ingrdquo Cells Tissues Organs vol 199 no 5-6 pp 405ndash422 2014

[34] E Appel S Weissmann Y Salzberg et al ldquoAn ensemble ofregulatory elements controls Runx3 spatiotemporal expressionin subsets of dorsal root ganglia proprioceptive neuronsrdquoGenesamp Development vol 30 no 23 pp 2607ndash2622 2016

[35] S M OrsquoToole M M Ferrer J Mekonnen et al ldquoDicermaintains the identity and function of proprioceptive sensoryneuronsrdquo Journal of Neurophysiology vol 117 no 3 pp 1057ndash1069 2017

[36] Y Liu L Kelamangalath H Kim et al ldquoNT-3 promotes pro-prioceptive axon regeneration when combined with activationof the mTor intrinsic growth pathway but not with reduction ofmyelin extrinsic inhibitorsrdquo Experimental Neurology vol 283pp 73ndash84 2016

[37] S Li T Shen Y Liang Y Zhang and B Bai ldquoEffects ofminiscalpel-needle release on chronic neck pain A retrospec-tive analysis with 12-month follow-uprdquo PLoS ONE vol 10 no8 Article ID e0137033 pp 133ndash137 2015

[38] W-S Juan S-Y Huang C-C Chang et al ldquoMelatoninimproves neuroplasticity by upregulating the growth-associatedprotein-43 (GAP-43) and NMDAR postsynaptic density-95(PSD-95) proteins in cultured neurons exposed to glutamateexcitotoxicity and in rats subjected to transient focal cerebralischemia even during a long-term recovery periodrdquo Journal ofPineal Research vol 56 no 2 pp 213ndash223 2014

[39] Y-HGaoC-W Li J-YWang et al ldquoActivation of hippocampalMEK1 contributes to the cumulative antinociceptive effect ofelectroacupuncture in neuropathic pain ratsrdquo BMC Comple-mentary and Alternative Medicine vol 16 no 1 article no 5172016

[40] K-Y Huang S Liang M-L Yu S-P Fu X Chen and S-F Lu ldquoA systematic review and meta-analysis of acupuncturefor improving learning and memory ability in animalsrdquo BMCComplementary and Alternative Medicine vol 16 no 1 articleno 297 2016

[41] J Zhou S Li Y Wang et al ldquoEffects and mechanisms ofauricular electroacupuncture on gastric hypersensitivity in arodent model of functional dyspepsiardquo PLoS ONE vol 12 no3 Article ID e0174568 2017

[42] Y H Gao C W Li J Y Wang et al ldquoEffect of electroacupunc-ture on the cervicospinal P2X7 receptorfractalkineCX3CR1signaling pathway in a rat neck-incision painmodelrdquo PurinergicSignalling vol 13 no 2 pp 215ndash225 2017

[43] JH LeeDGoWKimet al ldquoInvolvement of spinalmuscarinicand serotonergic receptors in the anti-allodynic effect of elec-troacupuncture in rats with oxaliplatin-induced neuropathicpainrdquoKorean Journal of Physiology amp Pharmacology vol 20 no4 pp 407ndash414 2016

[44] S P Chen Y Kan J L Zhang et al ldquoInvolvement of hippocam-pal acetylcholinergic receptors in electroacupuncture analgesiain neuropathic pain ratsrdquo Behavioral and Brain Functions vol12 no 1 article no 13 2016

[45] M Villarreal Santiago S Tumilty A Mącznik and R ManildquoDoes acupuncture alter pain-related functional connectivity ofthe central nervous system a systematic reviewrdquo JAMS Journalof Acupuncture and Meridian Studies vol 9 no 4 pp 167ndash1772016

[46] T E Salazar M R Richardson E Beli et al ldquoElectroacupunc-ture Promotes Central Nervous System-Dependent Release ofMesenchymal Stem Cellsrdquo Stem Cells vol 35 no 5 pp 1303ndash1315 2017

[47] D-H Lee J-H Lee S-E Ahn and M-J Park ldquoEffect of timeafter anterior cruciate ligament tears on proprioception andpostural stabilityrdquo PLoS ONE vol 10 no 9 Article ID e01390382015

Stem Cells International

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Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

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Disease Markers

Hindawiwwwhindawicom Volume 2018

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Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

4 BioMed Research International

(a) (b)

Figure 2Cynomolgus monkeys underwent electrophysiological examination involving somatosensory-evoked potentials (SEPs) andmotor nerveconduction velocity (MCV) under general anaesthesia (a) SEPs examination (b) MCV examination

(a) (b)

Figure 3 Obtaining experimental specimens (a) The white arrows indicate the dorsal root ganglion (DRG) (b) The red arrow indicates theanterior cruciate ligament (ACL) Bilateral DRGs were obtained from all groups Bilateral ACLs were obtained from the blank control groupInjured ACLs were obtained from the model control IIKE and IBKE groups

5996 179 bp R-GAP-43 forward 51015840-TCCACTGATAAC-TCGCCGTC-31015840 5955 and R-GAP-43 reverse 51015840-TCC-ACTGATAACTCGCCGTC-31015840 6044 98 bp cDNA wasgenerated by total RNA using a reverse-transcriptase cDNAsynthesis kit With SYBR Green (SYBR Premix Ex Taq IIAxygen Union City CA USA) as the fluorescent label and120573-actin as the internal control 1 120583l cDNA was added to the20120583l reaction system for 40 cycles for amplification Dataresults were analysed using the relative quantitative methodof 2minusΔΔCt

29 Western Blotting After grinding with liquid nitrogeneach 03 g sample was added to 1ml protein extractionpyrolysis liquid (Sangon) and placed in an ice box (PCRCooler Eppendorf Hamburg Germany) for ultrasonic

crushing Then the centrifuged supernatant fluid (MultifugeX3 Thermo Fisher Scientific Waltham MA USA) wasdetermined quantitatively using the bicinchoninic acid assayAfter the concentration was adjusted 20120583l protein samplebuffer was added to 20120583l sample and boiled for 10 minutesThe pretreated protein samples were separated by SDS-PAGEand transferred to PVDF membranes (Sangon) Then thePVDF membranes were moved to TBST solution (Sangon)for blockingThe membrane was incubated with the primaryantibody solution for 1 to 2 hours at 20∘C on a shakingtable After being washed with TBST solution repeatedly themembrane was incubated for 1 to 2 hours at 20∘C with theconjugated antibody (Abcam Cambridge MA USA) Thebands were visualized using ECL (Sangon) and the pictureswere conserved by a chemiluminescence imaging apparatus(SmartChemi 610 Sage Beijing China)

BioMed Research International 5

IIKE groupIBKE groupModel control groupBlank control group

0

10

20

30

40In

cuba

tion

of S

EPs (

ms)

8 124(Weeks)

abcde

bcde

cde

abdebde

dee

beabe

(a)

0

2

4

6

8

10

Am

plitu

de o

f SEP

s (

V)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde

dee

beabe

(b)

0

10

20

30

Incu

batio

n of

MCV

(ms)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abde

bde

de ebe

abe

(c)

0

4

8

12

Am

plitu

de o

f MCV

(mV

)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde cde

abdebde

dee

beabe

(d)

Figure 4 Comparison of incubations and amplitudes of SEPs and MCV among all groups at different time points (a) Incubations of SEPs (b)Amplitudes of SEPs (c) Incubations of MCV (d) Amplitudes of MCV a versus 8 weeks in the same group 119875 lt 005 b versus 12 weeks inthe same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 d versus the IBKE group at the same time 119875 lt 005 e versusthe blank control group 119875 lt 005

210 Statistical Analysis Measurement data were expressedasmeanplusmn SD RT-qPCRwas performed during three separateexperiments and data were collected Statistical differencesbetween the 4 groups were assessed by one-way analysis ofvariance (ANOVA) followed by the least significant difference(LSD) 119905-test In the same group differences in the repeated-measurement ANOVA and LSD 119905-test results were comparedamong three different time points All statistical analyseswereperformed using SPSS 200 software (IBM Corp ArmonkNY USA) 119875 lt 005 was considered statistically significant

3 Results

31 Neural Electrophysiological Indices At 4 8 and 12 weekscompared with the blank control group the incubationsof SEPs and MCV in the model control IIKE and IBKE

groups were extended but the amplitudes of SEPs and MCVwere decreased The changes of the IBKE group were betterthan those of the IIKE group and the changes of the IIKEgroup were better than those of the model control group Alldifferences were statistically significant (119875 lt 005)

In addition in the model control group the incubationsof the SEPs and MCV were extended over time but theamplitudes of SEPs and MCV were decreased In the IIKEand the IBKE groups the incubations of SEPs and MCVwere decreased but the amplitudes of SEPs and MCV wereextended Differences were statistically significant (119875 lt 005)(Figure 4)

32 RT-qPCR and Western Blotting At the same time pointregarding DRGs when comparing the mRNA and proteinexpressions of NT-3 and TrkC fewer changes occurred

6 BioMed Research International

00

05

10

15N

T-3

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde cde

abdebde de

ebeabe

(a)

00

05

10

15

TrkC

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcdecde

abde bde deebeabe

(b)

00

05

10

15

20

25

GA

P-43

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcde cde

abde bde de ebeabe

(c)

Figure 5 Expressions of mRNA of NT-3 TrkC and GAP-43 among all groups at different time points (a) Expression of NT-3 mRNA (b)Expression of TrkC mRNA (c) Expression of GAP-43 mRNA a versus 8 weeks in the same group 119875 lt 005 b versus 12 weeks in the samegroup 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 d versus the IBKE group at the same time 119875 lt 005 e versus the blankcontrol group 119875 lt 005

in the model control IIKE and IIKE groups than in theblank control group However more changes occurred inthe IBKE group than in the IIKE group More changes alsooccurred in the IIKE group than in the model control groupAll differences were statistically significant (119875 lt 005)Regarding the ACL when comparing the mRNA and proteinexpressions of GAP-43 more changes occurred in the modelcontrol group IIKE group and IBKE group than in the blankcontrol group more changes occurred in the IBKE groupthan in the IIKE group and more changes occurred in theIIKE group than in themodel control groupAll changes werestatistically significant (119875 lt 005)

At different time points in terms of mRNA and proteinexpressions of NT-3 and TrkC there were fewer changesin the model control IIKE and IBKE groups than in theblank control group however the expressions of the IIKE

and IBKE groups increased over time and the differenceswere statistically significant (119875 lt 005) In contrast theexpressions of the model control group decreased over timeand the differences were statistically significant (119875 lt 005) Interms of themRNA and protein expressions of GAP-43 thoseof the model control group decreased over time Howeverthe expressions of the IIKE and IBKE groups increased withincreased intervention time All changes were statisticallysignificant (119875 lt 005) (Figures 5 and 6)

4 Discussion

Proprioception of the ACL is closely related to knee jointstability which is realised through feedback regulation of thecentral nervous system however the key node on the path-way remains unknown [27ndash30] Many studies confirmed that

BioMed Research International 7

00

05

10

15

20N

T-3-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de e

beabe

(a)

00

05

10

15

20

TrkC

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebeabe

(b)

00

05

10

15

20

GA

P-43

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebe

abe

(c)

4 8 12

TrkCNT-3

GAP-43-actin

IIKE group IBKE group model control group blank control group

(Weeks)

(d)

Figure 6 Expressions of NT-3120573-actin TrkC120573-actin and GAP-43120573-actin proteins among the four groups at different time points (a)Expressions of NT-3120573-actin proteins (b) Expressions of TrkC120573-actin proteins (c) Expressions of GAP-43120573-actin proteins a versus 8weeks in the same group 119875 lt 005 b versus 12 weeks in the same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 dversus the IBKE group at the same time 119875 lt 005 e versus the blank control group 119875 lt 005 (d) The expression of protein of NT-3 TrkCand 120573-actin among the four groups in different time points

when the ACL proprioception function is decreased the neu-ral feedback regulation pathway is bound to be disorderedtherefore it could have further impact on the proprioceptionfunction [31 32] Some studies showed that NT-3 was the keyinduced signal of proprioceptive transduction when NT-3expression was normal the proprioceptive nerve fibres couldcontrol the target organs and have physiological roles [33ndash35] If the expressions of NT-3 and TrkC were decreasedthen apoptosis of proprioceptive neurons in the DRG couldoccur thus causing lower limb proprioception deficienciesNT-3 and TrkC could also accelerate the development andmaturation of neurons thus maintaining the survival ofmature neurons andpromoting the repair and regeneration ofinjured neurons When the proprioceptive nerve conductionpathway is injured proprioceptive neurons appear abnormal

however exogenous NT-3 could protect the proprioceptiveneurons and improve their function [36 37] ACL injuriescould damage peripheral nerve fibres When the nerve isdamaged expression of upregulation could be included andthe functional relationship could be decreased immediatelyafter reconstruction which is considered as a marker ofneuronal growth and repair [38]

During this study at 4 8 and 12 weeks in comparisonwith the blank control group incubations of SEPs and MCVin the model control group were significantly extended butthe amplitudes were significantly decreased (119875 lt 005)Comparisons of the three time points showed that incuba-tions of the SEPs and MCV in the model control group weresignificantly extended but the amplitudes were significantlydecreased over time (119875 lt 005) Furthermore at the same

8 BioMed Research International

time point in the DRG when comparing the relative mRNAand protein expressions of NT-3 and TrkC fewer changesoccurred in the model control group than in the blankcontrol group In the ACL when comparing the relativemRNA and protein expressions of GAP-43 more changesoccurred in the model control group than in the blankcontrol group all the differences were statistically significant(119875 lt 005) In the model control group at different timepoints the relative expression of mRNA and protein of NT-3 and TrkC decreased over time while the relative mRNAand protein expressions of GAP-43 increased over time allchanges were statistically significant (119875 lt 005) The resultsof this study illustrated that when the ACL was injured theexpressions of NT-3 and TrkC in neuronal cells of the DRGcould be reduced and the levels of phosphorylation of TrkCdecreased after activation thereby further reducing TrkCactivity When the ACL was injured GAP-43 was activatedin the proprioceptors which caused the increased expressionof GAP-43 Eventually changes in these cytokine expressionlevels can lead to decreased proprioceptive functions

Electroacupuncture is a form of acupuncture whereby asmall electric current is passed between pairs of acupunc-ture needles The use of electroacupuncture therapy (thecombination of nerve electrical stimulation and acupunc-ture) has gradually replaced other acupuncture therapiesElectroacupuncture therapy has been widely used to treatpain or injury in the hind limbs for many decades inChina Both human and animal experiments have confirmedthat electroacupuncture therapy has a definite effect on thetreatment of pain movement disturbance tissue regener-ation and so on [39ndash44] Some studies have found thatthe effects of acupuncture were mediated by the centralnervous system thus causing specific physiological effect thatachieved the purpose of treatment and improved the growthof neurons axonal regeneration synaptic reconstruction andthe functional connection of target organs [45 46]

When the ACL is pulled by the forward displacement ofthe tibia the contraction of the hamstring confronts tibialtranslation through regulating the ACLndashhamstring reflex arcwhich plays an important role in maintaining the stabilityof the knee joints The acupuncture points included in thisstudy were Wei Yang Yin Gu Xi Yangguan and Qu QuanThese points were chosen because they are all located nextto the hamstring When these acupuncture points werestimulated by electroacupuncture muscle contraction of thehamstring occurred and created the effect of reverse interven-tion Furthermore the ACL belongs to the arthromyodyniacategory of Traditional Chinese Medicine Among theseselected acupuncture points Wei Yang (BL19) is the Xiahetriple energiser acupuncture point that is mainly used totreat strong pain in the lumbar spine and pain and spasmsin the legs and feet Yin Gu (KI10) is mainly used to treatstrong pain in the knee joints Xi Yangguan (GB33) is mainlyused to treat swelling pain spasms in the knee joint andpopliteal area numbness in the leg and other diseases ofthe lower limbs and knee joints Qu Quan (LR8) is mainlyused to treat swelling pain in the knee joints and patellaand paralysis of lower limb Through the stimulation ofrelated points electroacupuncture transmits a large number

of proprioceptive and skin sensorial information to thecentral nervous system thereby participating in stabilisingthe activities of the knee joints and reconstruction of theproprioceptive function of the knee joints

In this study at 4 8 and 12 weeks compared withthe blank control group SEP and MCV incubations in themodel control group and the IIKE group were extendedbut the amplitudes were decreased The changes in the IIKEgroup were better than those in the model control group alldifferences were statistically significant (119875 lt 005) At thesame time point in the DRG when comparing the relativemRNA and protein expressions of NT-3 and TrkC therewere fewer changes in the model control group than in theblank control group however more changes occurred in theIIKE group than in the model control group in which alldifferences were statistically significant (119875 lt 005) It was alsoindicated that electroacupuncture could reversely intervenein the ACLndashhamstring reflex arc thereby activating the NT-3TrkC signal pathway and increasing the expression of NT-3 in the proprioceptors of hamstring NT-3 was reverselytransported to neurons in the DRG and then combined withthe corresponding TrkC receptor which promoted phospho-rylation of cytosolic TrkC after receptor activation whichcould enhance TrkC activity It could also activate GAP-43in the ACL which enhances the polymerisation ability ofGAP-43 in proprioceptors of ACL thereby increasing theactivity of ACL proprioceptionThe results found during ourexperiment may supplement certain potential explanationsabout the mechanisms of acupuncture treatment for ACLinjury to some extent

Some studies have found that unilateral ACL injury couldlead to proprioceptive degeneration of the bilateral kneejoints [47] In theory because of the characteristics of nerveconduction damage or intervention the proprioception ofone-side limb had a negative effect on the proprioceptionof the contralateral limb In this study the IIKE groupand the IBKE group were designed for the purpose ofproving that intervention for the healthy side can promote theproprioceptive recovery of the injured limb through centralregulation of the DRG

In terms of nerve electrophysiological examination andthe relativemRNA and protein expressions of NT-3 and TrkCin the DRG and of GAP-43 in the ACL the recovery effect ofthe IBKE group was more remarkable than that of the IIKEgroup (119875 lt 005) It showed that using the DRG as thenode the proprioception of bilateral ACLs could be regulatedand that the model of bilateral knee joint interventionwas better than the model of unilateral knee interventionAdditionally the effect of electroacupuncture interventionwas more significant with increased intervention time (119875 lt005)

This research had some limitations First all animalswere male and it is unclear whether that affected the resultsSecond because of the limitations of the objective conditionsthe period of observation during the experiment was only12 weeks therefore the long-term changes in proprioceptioncould not be observed Third without some antagonists ofthe NT-3TrkC signalling pathway it might be insufficientto check the role of the NT-3TrkC signalling pathway in

BioMed Research International 9

reverse intervention for the ACLndashhamstring reflex arc withelectroacupuncture Because these limitations existed thisstudy is considered basic All these issues remain to be solvedby further related studies

5 Conclusions

In this study the mechanism of reverse intervention for ACLproprioception with electroacupuncture which was basedon the ACLndashhamstring reflex arc and through mediationof the NT-3TrkC signalling pathway was explored Thisstudy helped establish the basis for the clinical application ofelectroacupuncture therapy and effective treatment methodsand patterns for proprioception disturbances caused by ACLinjuries thereby enriching and developing the scientificconnotation of electroacupuncture therapy guided by theory

Disclosure

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan are co-firstauthors

Conflicts of Interest

The authors declare that they have no conflicts of interest

Authorsrsquo Contributions

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan contributedequally to this work

Acknowledgments

This study was supported by the National Natural ScienceFoundation of China (no 81674095) Science-TechnologySupport Plan Project of Sichuan Province (no 2014SZ0185)and Science and Technology Strategic CooperationProject of Luzhou City-Southwest Medical University(no 2016LZXNYD-J08)

References

[1] H Zhang M Qiu A Zhou J Zhang and D Jiang ldquoAnatomicanterolateral ligament reconstruction improves postoperativeclinical outcomes combined with anatomic anterior cruciateligament reconstructionrdquo Journal of Sports Science andMedicine vol 15 no 4 pp 688ndash696 2016

[2] A A Rahnemai-Azar S Sabzevari S Irarrazaval T Chaoand F H Fu ldquoAnatomical individualized ACL reconstructionrdquoArchives of Bone and Joint Surgery vol 4 no 4 pp 291ndash2972016

[3] M R Sheth S R Tapasvi and S S Patil ldquoPrimary Repairof Tibial-Sided Avulsion of the Anterior Cruciate LigamentrdquoArthroscopy Techniques vol 5 no 4 pp e901ndashe906 2016

[4] F Sepulveda L Sanchez E Amy and W Micheo ldquoAnteriorCruciate Ligament InjuryrdquoCurrent SportsMedicine Reports vol16 no 3 pp 172ndash178 2017

[5] T M El-Gohary O A Khaled S R Ibrahim A M Alshenqitiand M I Ibrahim ldquoEffect of proprioception cross training on

repositioning accuracy and balance among healthy individualsrdquoJournal of PhysicalTherapy Science vol 28 no 11 pp 3178ndash31822016

[6] A Delle Vedove M Storbeck R Heller et al ldquoBiallelic loss ofproprioception-related PIEZO2 causes muscular atrophy withperinatal respiratory distress arthrogryposis and scoliosisrdquoAmerican Journal of Human Genetics vol 99 no 6 pp 1406ndash1408 2016

[7] W K Chang Y S Jung M Oh and K Kim ldquoQuantitativeassessment of proprioception using dynamometer in incom-plete spinal cord injury patients a preliminary studyrdquo Annalsof Rehabilitation Medicine vol 41 no 2 pp 218ndash224 2017

[8] G Haliloglu K Becker C Temucin et al ldquoRecessive PIEZO2stop mutation causes distal arthrogryposis with distal muscleweakness scoliosis and proprioception defectsrdquo Journal ofHuman Genetics vol 62 no 4 pp 497ndash501 2017

[9] J Li F Kong X Gao Y Shen and S Gao ldquoProspectiverandomized comparison of knee stability and proprioceptionfor posterior cruciate ligament reconstruction with autografthybrid graft and 120574-irradiated allograftrdquoArthroscopy vol 32 no12 pp 2548ndash2555 2016

[10] J F Abulhasan C M Anley M D Snow and M J GreyldquoHamstring stretch reflex could it be a reproducible objectivemeasure of functional knee stabilityrdquordquo Journal of ExperimentalOrthopaedics vol 3 no 1 2016

[11] M Solomonow R Baratta B H Zhou et al ldquoThe synergisticaction of the anterior cruciate ligament and thigh musclesin maintaining joint stabilityrdquo The American Journal of SportsMedicine vol 15 no 3 pp 207ndash213 1987

[12] P Dyhre-Poulsen and M R Krogsgaard ldquoMuscular reflexeselicited by electrical stimulation of the anterior cruciate liga-ment in humansrdquo Journal of Applied Physiology vol 89 no 6pp 2191ndash2195 2000

[13] E Tsuda Y Ishibashi Y Okamura and S Toh ldquoRestorationof anterior cruciate ligament-hamstring reflex arc after anteriorcruciate ligament reconstructionrdquoKnee Surgery Sports Trauma-tology Arthroscopy vol 11 no 2 pp 63ndash67 2003

[14] X-Y Wang P-Y Gu S-W Chen et al ldquoEndogenousneurotrophin-3 promotes neuronal sprouting from dorsalroot gangliardquo Neural Regeneration Research vol 10 no 11 pp1865ndash1868 2015

[15] K A Han D Woo S Kim et al ldquoNeurotrophin-3 regu-lates synapse development by modulating TrkC-PTP120590 synapticadhesion and intracellular signaling pathwaysrdquo The Journal ofNeuroscience vol 36 no 17 pp 4816ndash4831 2016

[16] M-H Yang K-C Chen P-W Chiang et al ldquoProteomicprofiling of neuroblastoma cells adhesion on hyaluronic acid-based surface for neural tissue engineeringrdquo BioMed ResearchInternational vol 2016 Article ID 1917394 2016

[17] S Rao M Martınez-Cengotitabengoa Y Yao et al ldquoPeripheralblood nerve growth factor levels inmajor psychiatric disordersrdquoJournal of Psychiatric Research vol 86 pp 39ndash45 2016

[18] M Yong S Kim and S Cheon ldquoEffects of skilled reach trainingwith affected forelimb and treadmill exercise on the expressionof neurotrophic factor following ischemia-induced brain injuryin ratsrdquo Journal of Physical Therapy Science vol 29 no 4 pp647ndash650 2017

[19] S Liu S Blanchard S Bigou S Vitry D Bohl and J-M HeardldquoNeurotrophin 3 improves delayed reconstruction of sensorypathways after cervical dorsal root injuryrdquoNeurosurgery vol 68no 2 pp 450ndash461 2011

10 BioMed Research International

[20] J Cui C Cui Y Cui et al ldquoBone marrow mesenchymal stemcell transplantation increases GAP-43 Expression via ERK12and PI3KAkt pathways in intracerebral hemorrhagerdquo CellularPhysiology and Biochemistry vol 42 no 1 pp 137ndash144 2017

[21] Y Li XGaoQWang et al ldquoRetinoic acid protects from experi-mental cerebral infarction by upregulating GAP-43 expressionrdquoBrazilian Journal of Medical and Biological Research vol 50 no4 2017

[22] H B Kim and B S Yoo ldquoPropolis inhibits neurite outgrowthin differentiating SH-SY5Y human neuroblastoma cellsrdquo Toxi-cological Research vol 32 no 3 pp 239ndash243 2016

[23] J Iwasa M Ochi Y Uchio N Adachi and K KawasakildquoDecrease in anterior knee laxity by electrical stimulation ofnormal and reconstructed anterior cruciate ligamentsrdquo TheJournal of Boneamp Joint Surgery vol 88 no 4 pp 477ndash483 2006

[24] A Biscarini F M Botti and V E Pettorossi ldquoSelectivecontribution of each hamstring muscle to anterior cruciateligament protection and tibiofemoral joint stability in leg-extension exercise A simulation studyrdquo European Journal ofApplied Physiology vol 113 no 9 pp 2263ndash2273 2013

[25] D Sugimoto E Alentorn-Geli J Mendiguchıa K SamuelssonJ Karlsson andGDMyer ldquoBiomechanical and neuromuscularcharacteristics of male athletes implications for the develop-ment of anterior cruciate ligament injury prevention programsrdquoSports Medicine vol 45 no 6 pp 809ndash822 2015

[26] M G Barcellona M C Morrissey P Milligan M Clinton andA A Amis ldquoThe effect of knee extensor open kinetic chainresistance training in the ACL-injured kneerdquo Knee SurgerySports Traumatology Arthroscopy vol 23 no 11 pp 3168ndash31772015

[27] T S Furlanetto L A Peyre-Tartaruga A S Do Pinho E SDa Bernardes and M A Zaro ldquoProprioception body balanceand functionality in individuals with ACL reconstructionrdquoActaOrtopedica Brasileira vol 24 no 2 pp 67ndash72 2016

[28] A J Wodowski C W Swigler H Liu K M Nord P C ToyandW M Mihalko ldquoProprioception and Knee Arthroplasty ALiterature ReviewrdquoOrthopedic Clinics of North America vol 47no 2 pp 301ndash309 2016

[29] I Dallo J Chahla J JMitchell C Pascual-Garrido J A Feaginand R F LaPrade ldquoBiologic Approaches for the Treatment ofPartial Tears of the Anterior Cruciate Ligament A CurrentConcepts Reviewrdquo Orthopaedic Journal of Sports Medicine vol5 no 1 2017

[30] W R Lowe R J Warth E P Davis and L Bailey ldquoFunc-tional bracing after anterior cruciate ligament reconstructionA systematic reviewrdquo Journal of the American Academy ofOrthopaedicSurgeons vol 25 no 3 pp 239ndash249 2017

[31] Y Ma M Deie D Iwaki et al ldquoBalance Ability and Pro-prioception after Single-Bundle Single-Bundle Augmentationand Double-Bundle ACL ReconstructionrdquoThe Scientific WorldJournal vol 2014 Article ID 342012 8 pages 2014

[32] N Relph L Herrington and S Tyson ldquoThe effects of ACLinjury on knee proprioception Ameta-analysisrdquo Physiotherapyvol 100 no 3 pp 187ndash195 2014

[33] B P Livingston and T R Nichols ldquoEffects of reinnervation ofthe triceps brachii on joint kinematics and electromyographicpatterns of the feline forelimb during level and upslope walk-ingrdquo Cells Tissues Organs vol 199 no 5-6 pp 405ndash422 2014

[34] E Appel S Weissmann Y Salzberg et al ldquoAn ensemble ofregulatory elements controls Runx3 spatiotemporal expressionin subsets of dorsal root ganglia proprioceptive neuronsrdquoGenesamp Development vol 30 no 23 pp 2607ndash2622 2016

[35] S M OrsquoToole M M Ferrer J Mekonnen et al ldquoDicermaintains the identity and function of proprioceptive sensoryneuronsrdquo Journal of Neurophysiology vol 117 no 3 pp 1057ndash1069 2017

[36] Y Liu L Kelamangalath H Kim et al ldquoNT-3 promotes pro-prioceptive axon regeneration when combined with activationof the mTor intrinsic growth pathway but not with reduction ofmyelin extrinsic inhibitorsrdquo Experimental Neurology vol 283pp 73ndash84 2016

[37] S Li T Shen Y Liang Y Zhang and B Bai ldquoEffects ofminiscalpel-needle release on chronic neck pain A retrospec-tive analysis with 12-month follow-uprdquo PLoS ONE vol 10 no8 Article ID e0137033 pp 133ndash137 2015

[38] W-S Juan S-Y Huang C-C Chang et al ldquoMelatoninimproves neuroplasticity by upregulating the growth-associatedprotein-43 (GAP-43) and NMDAR postsynaptic density-95(PSD-95) proteins in cultured neurons exposed to glutamateexcitotoxicity and in rats subjected to transient focal cerebralischemia even during a long-term recovery periodrdquo Journal ofPineal Research vol 56 no 2 pp 213ndash223 2014

[39] Y-HGaoC-W Li J-YWang et al ldquoActivation of hippocampalMEK1 contributes to the cumulative antinociceptive effect ofelectroacupuncture in neuropathic pain ratsrdquo BMC Comple-mentary and Alternative Medicine vol 16 no 1 article no 5172016

[40] K-Y Huang S Liang M-L Yu S-P Fu X Chen and S-F Lu ldquoA systematic review and meta-analysis of acupuncturefor improving learning and memory ability in animalsrdquo BMCComplementary and Alternative Medicine vol 16 no 1 articleno 297 2016

[41] J Zhou S Li Y Wang et al ldquoEffects and mechanisms ofauricular electroacupuncture on gastric hypersensitivity in arodent model of functional dyspepsiardquo PLoS ONE vol 12 no3 Article ID e0174568 2017

[42] Y H Gao C W Li J Y Wang et al ldquoEffect of electroacupunc-ture on the cervicospinal P2X7 receptorfractalkineCX3CR1signaling pathway in a rat neck-incision painmodelrdquo PurinergicSignalling vol 13 no 2 pp 215ndash225 2017

[43] JH LeeDGoWKimet al ldquoInvolvement of spinalmuscarinicand serotonergic receptors in the anti-allodynic effect of elec-troacupuncture in rats with oxaliplatin-induced neuropathicpainrdquoKorean Journal of Physiology amp Pharmacology vol 20 no4 pp 407ndash414 2016

[44] S P Chen Y Kan J L Zhang et al ldquoInvolvement of hippocam-pal acetylcholinergic receptors in electroacupuncture analgesiain neuropathic pain ratsrdquo Behavioral and Brain Functions vol12 no 1 article no 13 2016

[45] M Villarreal Santiago S Tumilty A Mącznik and R ManildquoDoes acupuncture alter pain-related functional connectivity ofthe central nervous system a systematic reviewrdquo JAMS Journalof Acupuncture and Meridian Studies vol 9 no 4 pp 167ndash1772016

[46] T E Salazar M R Richardson E Beli et al ldquoElectroacupunc-ture Promotes Central Nervous System-Dependent Release ofMesenchymal Stem Cellsrdquo Stem Cells vol 35 no 5 pp 1303ndash1315 2017

[47] D-H Lee J-H Lee S-E Ahn and M-J Park ldquoEffect of timeafter anterior cruciate ligament tears on proprioception andpostural stabilityrdquo PLoS ONE vol 10 no 9 Article ID e01390382015

Stem Cells International

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Disease Markers

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Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

BioMed Research International 5

IIKE groupIBKE groupModel control groupBlank control group

0

10

20

30

40In

cuba

tion

of S

EPs (

ms)

8 124(Weeks)

abcde

bcde

cde

abdebde

dee

beabe

(a)

0

2

4

6

8

10

Am

plitu

de o

f SEP

s (

V)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde

dee

beabe

(b)

0

10

20

30

Incu

batio

n of

MCV

(ms)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abde

bde

de ebe

abe

(c)

0

4

8

12

Am

plitu

de o

f MCV

(mV

)

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde cde

abdebde

dee

beabe

(d)

Figure 4 Comparison of incubations and amplitudes of SEPs and MCV among all groups at different time points (a) Incubations of SEPs (b)Amplitudes of SEPs (c) Incubations of MCV (d) Amplitudes of MCV a versus 8 weeks in the same group 119875 lt 005 b versus 12 weeks inthe same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 d versus the IBKE group at the same time 119875 lt 005 e versusthe blank control group 119875 lt 005

210 Statistical Analysis Measurement data were expressedasmeanplusmn SD RT-qPCRwas performed during three separateexperiments and data were collected Statistical differencesbetween the 4 groups were assessed by one-way analysis ofvariance (ANOVA) followed by the least significant difference(LSD) 119905-test In the same group differences in the repeated-measurement ANOVA and LSD 119905-test results were comparedamong three different time points All statistical analyseswereperformed using SPSS 200 software (IBM Corp ArmonkNY USA) 119875 lt 005 was considered statistically significant

3 Results

31 Neural Electrophysiological Indices At 4 8 and 12 weekscompared with the blank control group the incubationsof SEPs and MCV in the model control IIKE and IBKE

groups were extended but the amplitudes of SEPs and MCVwere decreased The changes of the IBKE group were betterthan those of the IIKE group and the changes of the IIKEgroup were better than those of the model control group Alldifferences were statistically significant (119875 lt 005)

In addition in the model control group the incubationsof the SEPs and MCV were extended over time but theamplitudes of SEPs and MCV were decreased In the IIKEand the IBKE groups the incubations of SEPs and MCVwere decreased but the amplitudes of SEPs and MCV wereextended Differences were statistically significant (119875 lt 005)(Figure 4)

32 RT-qPCR and Western Blotting At the same time pointregarding DRGs when comparing the mRNA and proteinexpressions of NT-3 and TrkC fewer changes occurred

6 BioMed Research International

00

05

10

15N

T-3

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde cde

abdebde de

ebeabe

(a)

00

05

10

15

TrkC

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcdecde

abde bde deebeabe

(b)

00

05

10

15

20

25

GA

P-43

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcde cde

abde bde de ebeabe

(c)

Figure 5 Expressions of mRNA of NT-3 TrkC and GAP-43 among all groups at different time points (a) Expression of NT-3 mRNA (b)Expression of TrkC mRNA (c) Expression of GAP-43 mRNA a versus 8 weeks in the same group 119875 lt 005 b versus 12 weeks in the samegroup 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 d versus the IBKE group at the same time 119875 lt 005 e versus the blankcontrol group 119875 lt 005

in the model control IIKE and IIKE groups than in theblank control group However more changes occurred inthe IBKE group than in the IIKE group More changes alsooccurred in the IIKE group than in the model control groupAll differences were statistically significant (119875 lt 005)Regarding the ACL when comparing the mRNA and proteinexpressions of GAP-43 more changes occurred in the modelcontrol group IIKE group and IBKE group than in the blankcontrol group more changes occurred in the IBKE groupthan in the IIKE group and more changes occurred in theIIKE group than in themodel control groupAll changes werestatistically significant (119875 lt 005)

At different time points in terms of mRNA and proteinexpressions of NT-3 and TrkC there were fewer changesin the model control IIKE and IBKE groups than in theblank control group however the expressions of the IIKE

and IBKE groups increased over time and the differenceswere statistically significant (119875 lt 005) In contrast theexpressions of the model control group decreased over timeand the differences were statistically significant (119875 lt 005) Interms of themRNA and protein expressions of GAP-43 thoseof the model control group decreased over time Howeverthe expressions of the IIKE and IBKE groups increased withincreased intervention time All changes were statisticallysignificant (119875 lt 005) (Figures 5 and 6)

4 Discussion

Proprioception of the ACL is closely related to knee jointstability which is realised through feedback regulation of thecentral nervous system however the key node on the path-way remains unknown [27ndash30] Many studies confirmed that

BioMed Research International 7

00

05

10

15

20N

T-3-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de e

beabe

(a)

00

05

10

15

20

TrkC

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebeabe

(b)

00

05

10

15

20

GA

P-43

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebe

abe

(c)

4 8 12

TrkCNT-3

GAP-43-actin

IIKE group IBKE group model control group blank control group

(Weeks)

(d)

Figure 6 Expressions of NT-3120573-actin TrkC120573-actin and GAP-43120573-actin proteins among the four groups at different time points (a)Expressions of NT-3120573-actin proteins (b) Expressions of TrkC120573-actin proteins (c) Expressions of GAP-43120573-actin proteins a versus 8weeks in the same group 119875 lt 005 b versus 12 weeks in the same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 dversus the IBKE group at the same time 119875 lt 005 e versus the blank control group 119875 lt 005 (d) The expression of protein of NT-3 TrkCand 120573-actin among the four groups in different time points

when the ACL proprioception function is decreased the neu-ral feedback regulation pathway is bound to be disorderedtherefore it could have further impact on the proprioceptionfunction [31 32] Some studies showed that NT-3 was the keyinduced signal of proprioceptive transduction when NT-3expression was normal the proprioceptive nerve fibres couldcontrol the target organs and have physiological roles [33ndash35] If the expressions of NT-3 and TrkC were decreasedthen apoptosis of proprioceptive neurons in the DRG couldoccur thus causing lower limb proprioception deficienciesNT-3 and TrkC could also accelerate the development andmaturation of neurons thus maintaining the survival ofmature neurons andpromoting the repair and regeneration ofinjured neurons When the proprioceptive nerve conductionpathway is injured proprioceptive neurons appear abnormal

however exogenous NT-3 could protect the proprioceptiveneurons and improve their function [36 37] ACL injuriescould damage peripheral nerve fibres When the nerve isdamaged expression of upregulation could be included andthe functional relationship could be decreased immediatelyafter reconstruction which is considered as a marker ofneuronal growth and repair [38]

During this study at 4 8 and 12 weeks in comparisonwith the blank control group incubations of SEPs and MCVin the model control group were significantly extended butthe amplitudes were significantly decreased (119875 lt 005)Comparisons of the three time points showed that incuba-tions of the SEPs and MCV in the model control group weresignificantly extended but the amplitudes were significantlydecreased over time (119875 lt 005) Furthermore at the same

8 BioMed Research International

time point in the DRG when comparing the relative mRNAand protein expressions of NT-3 and TrkC fewer changesoccurred in the model control group than in the blankcontrol group In the ACL when comparing the relativemRNA and protein expressions of GAP-43 more changesoccurred in the model control group than in the blankcontrol group all the differences were statistically significant(119875 lt 005) In the model control group at different timepoints the relative expression of mRNA and protein of NT-3 and TrkC decreased over time while the relative mRNAand protein expressions of GAP-43 increased over time allchanges were statistically significant (119875 lt 005) The resultsof this study illustrated that when the ACL was injured theexpressions of NT-3 and TrkC in neuronal cells of the DRGcould be reduced and the levels of phosphorylation of TrkCdecreased after activation thereby further reducing TrkCactivity When the ACL was injured GAP-43 was activatedin the proprioceptors which caused the increased expressionof GAP-43 Eventually changes in these cytokine expressionlevels can lead to decreased proprioceptive functions

Electroacupuncture is a form of acupuncture whereby asmall electric current is passed between pairs of acupunc-ture needles The use of electroacupuncture therapy (thecombination of nerve electrical stimulation and acupunc-ture) has gradually replaced other acupuncture therapiesElectroacupuncture therapy has been widely used to treatpain or injury in the hind limbs for many decades inChina Both human and animal experiments have confirmedthat electroacupuncture therapy has a definite effect on thetreatment of pain movement disturbance tissue regener-ation and so on [39ndash44] Some studies have found thatthe effects of acupuncture were mediated by the centralnervous system thus causing specific physiological effect thatachieved the purpose of treatment and improved the growthof neurons axonal regeneration synaptic reconstruction andthe functional connection of target organs [45 46]

When the ACL is pulled by the forward displacement ofthe tibia the contraction of the hamstring confronts tibialtranslation through regulating the ACLndashhamstring reflex arcwhich plays an important role in maintaining the stabilityof the knee joints The acupuncture points included in thisstudy were Wei Yang Yin Gu Xi Yangguan and Qu QuanThese points were chosen because they are all located nextto the hamstring When these acupuncture points werestimulated by electroacupuncture muscle contraction of thehamstring occurred and created the effect of reverse interven-tion Furthermore the ACL belongs to the arthromyodyniacategory of Traditional Chinese Medicine Among theseselected acupuncture points Wei Yang (BL19) is the Xiahetriple energiser acupuncture point that is mainly used totreat strong pain in the lumbar spine and pain and spasmsin the legs and feet Yin Gu (KI10) is mainly used to treatstrong pain in the knee joints Xi Yangguan (GB33) is mainlyused to treat swelling pain spasms in the knee joint andpopliteal area numbness in the leg and other diseases ofthe lower limbs and knee joints Qu Quan (LR8) is mainlyused to treat swelling pain in the knee joints and patellaand paralysis of lower limb Through the stimulation ofrelated points electroacupuncture transmits a large number

of proprioceptive and skin sensorial information to thecentral nervous system thereby participating in stabilisingthe activities of the knee joints and reconstruction of theproprioceptive function of the knee joints

In this study at 4 8 and 12 weeks compared withthe blank control group SEP and MCV incubations in themodel control group and the IIKE group were extendedbut the amplitudes were decreased The changes in the IIKEgroup were better than those in the model control group alldifferences were statistically significant (119875 lt 005) At thesame time point in the DRG when comparing the relativemRNA and protein expressions of NT-3 and TrkC therewere fewer changes in the model control group than in theblank control group however more changes occurred in theIIKE group than in the model control group in which alldifferences were statistically significant (119875 lt 005) It was alsoindicated that electroacupuncture could reversely intervenein the ACLndashhamstring reflex arc thereby activating the NT-3TrkC signal pathway and increasing the expression of NT-3 in the proprioceptors of hamstring NT-3 was reverselytransported to neurons in the DRG and then combined withthe corresponding TrkC receptor which promoted phospho-rylation of cytosolic TrkC after receptor activation whichcould enhance TrkC activity It could also activate GAP-43in the ACL which enhances the polymerisation ability ofGAP-43 in proprioceptors of ACL thereby increasing theactivity of ACL proprioceptionThe results found during ourexperiment may supplement certain potential explanationsabout the mechanisms of acupuncture treatment for ACLinjury to some extent

Some studies have found that unilateral ACL injury couldlead to proprioceptive degeneration of the bilateral kneejoints [47] In theory because of the characteristics of nerveconduction damage or intervention the proprioception ofone-side limb had a negative effect on the proprioceptionof the contralateral limb In this study the IIKE groupand the IBKE group were designed for the purpose ofproving that intervention for the healthy side can promote theproprioceptive recovery of the injured limb through centralregulation of the DRG

In terms of nerve electrophysiological examination andthe relativemRNA and protein expressions of NT-3 and TrkCin the DRG and of GAP-43 in the ACL the recovery effect ofthe IBKE group was more remarkable than that of the IIKEgroup (119875 lt 005) It showed that using the DRG as thenode the proprioception of bilateral ACLs could be regulatedand that the model of bilateral knee joint interventionwas better than the model of unilateral knee interventionAdditionally the effect of electroacupuncture interventionwas more significant with increased intervention time (119875 lt005)

This research had some limitations First all animalswere male and it is unclear whether that affected the resultsSecond because of the limitations of the objective conditionsthe period of observation during the experiment was only12 weeks therefore the long-term changes in proprioceptioncould not be observed Third without some antagonists ofthe NT-3TrkC signalling pathway it might be insufficientto check the role of the NT-3TrkC signalling pathway in

BioMed Research International 9

reverse intervention for the ACLndashhamstring reflex arc withelectroacupuncture Because these limitations existed thisstudy is considered basic All these issues remain to be solvedby further related studies

5 Conclusions

In this study the mechanism of reverse intervention for ACLproprioception with electroacupuncture which was basedon the ACLndashhamstring reflex arc and through mediationof the NT-3TrkC signalling pathway was explored Thisstudy helped establish the basis for the clinical application ofelectroacupuncture therapy and effective treatment methodsand patterns for proprioception disturbances caused by ACLinjuries thereby enriching and developing the scientificconnotation of electroacupuncture therapy guided by theory

Disclosure

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan are co-firstauthors

Conflicts of Interest

The authors declare that they have no conflicts of interest

Authorsrsquo Contributions

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan contributedequally to this work

Acknowledgments

This study was supported by the National Natural ScienceFoundation of China (no 81674095) Science-TechnologySupport Plan Project of Sichuan Province (no 2014SZ0185)and Science and Technology Strategic CooperationProject of Luzhou City-Southwest Medical University(no 2016LZXNYD-J08)

References

[1] H Zhang M Qiu A Zhou J Zhang and D Jiang ldquoAnatomicanterolateral ligament reconstruction improves postoperativeclinical outcomes combined with anatomic anterior cruciateligament reconstructionrdquo Journal of Sports Science andMedicine vol 15 no 4 pp 688ndash696 2016

[2] A A Rahnemai-Azar S Sabzevari S Irarrazaval T Chaoand F H Fu ldquoAnatomical individualized ACL reconstructionrdquoArchives of Bone and Joint Surgery vol 4 no 4 pp 291ndash2972016

[3] M R Sheth S R Tapasvi and S S Patil ldquoPrimary Repairof Tibial-Sided Avulsion of the Anterior Cruciate LigamentrdquoArthroscopy Techniques vol 5 no 4 pp e901ndashe906 2016

[4] F Sepulveda L Sanchez E Amy and W Micheo ldquoAnteriorCruciate Ligament InjuryrdquoCurrent SportsMedicine Reports vol16 no 3 pp 172ndash178 2017

[5] T M El-Gohary O A Khaled S R Ibrahim A M Alshenqitiand M I Ibrahim ldquoEffect of proprioception cross training on

repositioning accuracy and balance among healthy individualsrdquoJournal of PhysicalTherapy Science vol 28 no 11 pp 3178ndash31822016

[6] A Delle Vedove M Storbeck R Heller et al ldquoBiallelic loss ofproprioception-related PIEZO2 causes muscular atrophy withperinatal respiratory distress arthrogryposis and scoliosisrdquoAmerican Journal of Human Genetics vol 99 no 6 pp 1406ndash1408 2016

[7] W K Chang Y S Jung M Oh and K Kim ldquoQuantitativeassessment of proprioception using dynamometer in incom-plete spinal cord injury patients a preliminary studyrdquo Annalsof Rehabilitation Medicine vol 41 no 2 pp 218ndash224 2017

[8] G Haliloglu K Becker C Temucin et al ldquoRecessive PIEZO2stop mutation causes distal arthrogryposis with distal muscleweakness scoliosis and proprioception defectsrdquo Journal ofHuman Genetics vol 62 no 4 pp 497ndash501 2017

[9] J Li F Kong X Gao Y Shen and S Gao ldquoProspectiverandomized comparison of knee stability and proprioceptionfor posterior cruciate ligament reconstruction with autografthybrid graft and 120574-irradiated allograftrdquoArthroscopy vol 32 no12 pp 2548ndash2555 2016

[10] J F Abulhasan C M Anley M D Snow and M J GreyldquoHamstring stretch reflex could it be a reproducible objectivemeasure of functional knee stabilityrdquordquo Journal of ExperimentalOrthopaedics vol 3 no 1 2016

[11] M Solomonow R Baratta B H Zhou et al ldquoThe synergisticaction of the anterior cruciate ligament and thigh musclesin maintaining joint stabilityrdquo The American Journal of SportsMedicine vol 15 no 3 pp 207ndash213 1987

[12] P Dyhre-Poulsen and M R Krogsgaard ldquoMuscular reflexeselicited by electrical stimulation of the anterior cruciate liga-ment in humansrdquo Journal of Applied Physiology vol 89 no 6pp 2191ndash2195 2000

[13] E Tsuda Y Ishibashi Y Okamura and S Toh ldquoRestorationof anterior cruciate ligament-hamstring reflex arc after anteriorcruciate ligament reconstructionrdquoKnee Surgery Sports Trauma-tology Arthroscopy vol 11 no 2 pp 63ndash67 2003

[14] X-Y Wang P-Y Gu S-W Chen et al ldquoEndogenousneurotrophin-3 promotes neuronal sprouting from dorsalroot gangliardquo Neural Regeneration Research vol 10 no 11 pp1865ndash1868 2015

[15] K A Han D Woo S Kim et al ldquoNeurotrophin-3 regu-lates synapse development by modulating TrkC-PTP120590 synapticadhesion and intracellular signaling pathwaysrdquo The Journal ofNeuroscience vol 36 no 17 pp 4816ndash4831 2016

[16] M-H Yang K-C Chen P-W Chiang et al ldquoProteomicprofiling of neuroblastoma cells adhesion on hyaluronic acid-based surface for neural tissue engineeringrdquo BioMed ResearchInternational vol 2016 Article ID 1917394 2016

[17] S Rao M Martınez-Cengotitabengoa Y Yao et al ldquoPeripheralblood nerve growth factor levels inmajor psychiatric disordersrdquoJournal of Psychiatric Research vol 86 pp 39ndash45 2016

[18] M Yong S Kim and S Cheon ldquoEffects of skilled reach trainingwith affected forelimb and treadmill exercise on the expressionof neurotrophic factor following ischemia-induced brain injuryin ratsrdquo Journal of Physical Therapy Science vol 29 no 4 pp647ndash650 2017

[19] S Liu S Blanchard S Bigou S Vitry D Bohl and J-M HeardldquoNeurotrophin 3 improves delayed reconstruction of sensorypathways after cervical dorsal root injuryrdquoNeurosurgery vol 68no 2 pp 450ndash461 2011

10 BioMed Research International

[20] J Cui C Cui Y Cui et al ldquoBone marrow mesenchymal stemcell transplantation increases GAP-43 Expression via ERK12and PI3KAkt pathways in intracerebral hemorrhagerdquo CellularPhysiology and Biochemistry vol 42 no 1 pp 137ndash144 2017

[21] Y Li XGaoQWang et al ldquoRetinoic acid protects from experi-mental cerebral infarction by upregulating GAP-43 expressionrdquoBrazilian Journal of Medical and Biological Research vol 50 no4 2017

[22] H B Kim and B S Yoo ldquoPropolis inhibits neurite outgrowthin differentiating SH-SY5Y human neuroblastoma cellsrdquo Toxi-cological Research vol 32 no 3 pp 239ndash243 2016

[23] J Iwasa M Ochi Y Uchio N Adachi and K KawasakildquoDecrease in anterior knee laxity by electrical stimulation ofnormal and reconstructed anterior cruciate ligamentsrdquo TheJournal of Boneamp Joint Surgery vol 88 no 4 pp 477ndash483 2006

[24] A Biscarini F M Botti and V E Pettorossi ldquoSelectivecontribution of each hamstring muscle to anterior cruciateligament protection and tibiofemoral joint stability in leg-extension exercise A simulation studyrdquo European Journal ofApplied Physiology vol 113 no 9 pp 2263ndash2273 2013

[25] D Sugimoto E Alentorn-Geli J Mendiguchıa K SamuelssonJ Karlsson andGDMyer ldquoBiomechanical and neuromuscularcharacteristics of male athletes implications for the develop-ment of anterior cruciate ligament injury prevention programsrdquoSports Medicine vol 45 no 6 pp 809ndash822 2015

[26] M G Barcellona M C Morrissey P Milligan M Clinton andA A Amis ldquoThe effect of knee extensor open kinetic chainresistance training in the ACL-injured kneerdquo Knee SurgerySports Traumatology Arthroscopy vol 23 no 11 pp 3168ndash31772015

[27] T S Furlanetto L A Peyre-Tartaruga A S Do Pinho E SDa Bernardes and M A Zaro ldquoProprioception body balanceand functionality in individuals with ACL reconstructionrdquoActaOrtopedica Brasileira vol 24 no 2 pp 67ndash72 2016

[28] A J Wodowski C W Swigler H Liu K M Nord P C ToyandW M Mihalko ldquoProprioception and Knee Arthroplasty ALiterature ReviewrdquoOrthopedic Clinics of North America vol 47no 2 pp 301ndash309 2016

[29] I Dallo J Chahla J JMitchell C Pascual-Garrido J A Feaginand R F LaPrade ldquoBiologic Approaches for the Treatment ofPartial Tears of the Anterior Cruciate Ligament A CurrentConcepts Reviewrdquo Orthopaedic Journal of Sports Medicine vol5 no 1 2017

[30] W R Lowe R J Warth E P Davis and L Bailey ldquoFunc-tional bracing after anterior cruciate ligament reconstructionA systematic reviewrdquo Journal of the American Academy ofOrthopaedicSurgeons vol 25 no 3 pp 239ndash249 2017

[31] Y Ma M Deie D Iwaki et al ldquoBalance Ability and Pro-prioception after Single-Bundle Single-Bundle Augmentationand Double-Bundle ACL ReconstructionrdquoThe Scientific WorldJournal vol 2014 Article ID 342012 8 pages 2014

[32] N Relph L Herrington and S Tyson ldquoThe effects of ACLinjury on knee proprioception Ameta-analysisrdquo Physiotherapyvol 100 no 3 pp 187ndash195 2014

[33] B P Livingston and T R Nichols ldquoEffects of reinnervation ofthe triceps brachii on joint kinematics and electromyographicpatterns of the feline forelimb during level and upslope walk-ingrdquo Cells Tissues Organs vol 199 no 5-6 pp 405ndash422 2014

[34] E Appel S Weissmann Y Salzberg et al ldquoAn ensemble ofregulatory elements controls Runx3 spatiotemporal expressionin subsets of dorsal root ganglia proprioceptive neuronsrdquoGenesamp Development vol 30 no 23 pp 2607ndash2622 2016

[35] S M OrsquoToole M M Ferrer J Mekonnen et al ldquoDicermaintains the identity and function of proprioceptive sensoryneuronsrdquo Journal of Neurophysiology vol 117 no 3 pp 1057ndash1069 2017

[36] Y Liu L Kelamangalath H Kim et al ldquoNT-3 promotes pro-prioceptive axon regeneration when combined with activationof the mTor intrinsic growth pathway but not with reduction ofmyelin extrinsic inhibitorsrdquo Experimental Neurology vol 283pp 73ndash84 2016

[37] S Li T Shen Y Liang Y Zhang and B Bai ldquoEffects ofminiscalpel-needle release on chronic neck pain A retrospec-tive analysis with 12-month follow-uprdquo PLoS ONE vol 10 no8 Article ID e0137033 pp 133ndash137 2015

[38] W-S Juan S-Y Huang C-C Chang et al ldquoMelatoninimproves neuroplasticity by upregulating the growth-associatedprotein-43 (GAP-43) and NMDAR postsynaptic density-95(PSD-95) proteins in cultured neurons exposed to glutamateexcitotoxicity and in rats subjected to transient focal cerebralischemia even during a long-term recovery periodrdquo Journal ofPineal Research vol 56 no 2 pp 213ndash223 2014

[39] Y-HGaoC-W Li J-YWang et al ldquoActivation of hippocampalMEK1 contributes to the cumulative antinociceptive effect ofelectroacupuncture in neuropathic pain ratsrdquo BMC Comple-mentary and Alternative Medicine vol 16 no 1 article no 5172016

[40] K-Y Huang S Liang M-L Yu S-P Fu X Chen and S-F Lu ldquoA systematic review and meta-analysis of acupuncturefor improving learning and memory ability in animalsrdquo BMCComplementary and Alternative Medicine vol 16 no 1 articleno 297 2016

[41] J Zhou S Li Y Wang et al ldquoEffects and mechanisms ofauricular electroacupuncture on gastric hypersensitivity in arodent model of functional dyspepsiardquo PLoS ONE vol 12 no3 Article ID e0174568 2017

[42] Y H Gao C W Li J Y Wang et al ldquoEffect of electroacupunc-ture on the cervicospinal P2X7 receptorfractalkineCX3CR1signaling pathway in a rat neck-incision painmodelrdquo PurinergicSignalling vol 13 no 2 pp 215ndash225 2017

[43] JH LeeDGoWKimet al ldquoInvolvement of spinalmuscarinicand serotonergic receptors in the anti-allodynic effect of elec-troacupuncture in rats with oxaliplatin-induced neuropathicpainrdquoKorean Journal of Physiology amp Pharmacology vol 20 no4 pp 407ndash414 2016

[44] S P Chen Y Kan J L Zhang et al ldquoInvolvement of hippocam-pal acetylcholinergic receptors in electroacupuncture analgesiain neuropathic pain ratsrdquo Behavioral and Brain Functions vol12 no 1 article no 13 2016

[45] M Villarreal Santiago S Tumilty A Mącznik and R ManildquoDoes acupuncture alter pain-related functional connectivity ofthe central nervous system a systematic reviewrdquo JAMS Journalof Acupuncture and Meridian Studies vol 9 no 4 pp 167ndash1772016

[46] T E Salazar M R Richardson E Beli et al ldquoElectroacupunc-ture Promotes Central Nervous System-Dependent Release ofMesenchymal Stem Cellsrdquo Stem Cells vol 35 no 5 pp 1303ndash1315 2017

[47] D-H Lee J-H Lee S-E Ahn and M-J Park ldquoEffect of timeafter anterior cruciate ligament tears on proprioception andpostural stabilityrdquo PLoS ONE vol 10 no 9 Article ID e01390382015

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

OncologyJournal of

Hindawiwwwhindawicom Volume 2013

Hindawiwwwhindawicom Volume 2018

Oxidative Medicine and Cellular Longevity

Hindawiwwwhindawicom Volume 2018

PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of

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Hindawiwwwhindawicom Volume 2018

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Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

6 BioMed Research International

00

05

10

15N

T-3

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde cde

abdebde de

ebeabe

(a)

00

05

10

15

TrkC

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcdecde

abde bde deebeabe

(b)

00

05

10

15

20

25

GA

P-43

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcde bcde cde

abde bde de ebeabe

(c)

Figure 5 Expressions of mRNA of NT-3 TrkC and GAP-43 among all groups at different time points (a) Expression of NT-3 mRNA (b)Expression of TrkC mRNA (c) Expression of GAP-43 mRNA a versus 8 weeks in the same group 119875 lt 005 b versus 12 weeks in the samegroup 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 d versus the IBKE group at the same time 119875 lt 005 e versus the blankcontrol group 119875 lt 005

in the model control IIKE and IIKE groups than in theblank control group However more changes occurred inthe IBKE group than in the IIKE group More changes alsooccurred in the IIKE group than in the model control groupAll differences were statistically significant (119875 lt 005)Regarding the ACL when comparing the mRNA and proteinexpressions of GAP-43 more changes occurred in the modelcontrol group IIKE group and IBKE group than in the blankcontrol group more changes occurred in the IBKE groupthan in the IIKE group and more changes occurred in theIIKE group than in themodel control groupAll changes werestatistically significant (119875 lt 005)

At different time points in terms of mRNA and proteinexpressions of NT-3 and TrkC there were fewer changesin the model control IIKE and IBKE groups than in theblank control group however the expressions of the IIKE

and IBKE groups increased over time and the differenceswere statistically significant (119875 lt 005) In contrast theexpressions of the model control group decreased over timeand the differences were statistically significant (119875 lt 005) Interms of themRNA and protein expressions of GAP-43 thoseof the model control group decreased over time Howeverthe expressions of the IIKE and IBKE groups increased withincreased intervention time All changes were statisticallysignificant (119875 lt 005) (Figures 5 and 6)

4 Discussion

Proprioception of the ACL is closely related to knee jointstability which is realised through feedback regulation of thecentral nervous system however the key node on the path-way remains unknown [27ndash30] Many studies confirmed that

BioMed Research International 7

00

05

10

15

20N

T-3-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de e

beabe

(a)

00

05

10

15

20

TrkC

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebeabe

(b)

00

05

10

15

20

GA

P-43

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebe

abe

(c)

4 8 12

TrkCNT-3

GAP-43-actin

IIKE group IBKE group model control group blank control group

(Weeks)

(d)

Figure 6 Expressions of NT-3120573-actin TrkC120573-actin and GAP-43120573-actin proteins among the four groups at different time points (a)Expressions of NT-3120573-actin proteins (b) Expressions of TrkC120573-actin proteins (c) Expressions of GAP-43120573-actin proteins a versus 8weeks in the same group 119875 lt 005 b versus 12 weeks in the same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 dversus the IBKE group at the same time 119875 lt 005 e versus the blank control group 119875 lt 005 (d) The expression of protein of NT-3 TrkCand 120573-actin among the four groups in different time points

when the ACL proprioception function is decreased the neu-ral feedback regulation pathway is bound to be disorderedtherefore it could have further impact on the proprioceptionfunction [31 32] Some studies showed that NT-3 was the keyinduced signal of proprioceptive transduction when NT-3expression was normal the proprioceptive nerve fibres couldcontrol the target organs and have physiological roles [33ndash35] If the expressions of NT-3 and TrkC were decreasedthen apoptosis of proprioceptive neurons in the DRG couldoccur thus causing lower limb proprioception deficienciesNT-3 and TrkC could also accelerate the development andmaturation of neurons thus maintaining the survival ofmature neurons andpromoting the repair and regeneration ofinjured neurons When the proprioceptive nerve conductionpathway is injured proprioceptive neurons appear abnormal

however exogenous NT-3 could protect the proprioceptiveneurons and improve their function [36 37] ACL injuriescould damage peripheral nerve fibres When the nerve isdamaged expression of upregulation could be included andthe functional relationship could be decreased immediatelyafter reconstruction which is considered as a marker ofneuronal growth and repair [38]

During this study at 4 8 and 12 weeks in comparisonwith the blank control group incubations of SEPs and MCVin the model control group were significantly extended butthe amplitudes were significantly decreased (119875 lt 005)Comparisons of the three time points showed that incuba-tions of the SEPs and MCV in the model control group weresignificantly extended but the amplitudes were significantlydecreased over time (119875 lt 005) Furthermore at the same

8 BioMed Research International

time point in the DRG when comparing the relative mRNAand protein expressions of NT-3 and TrkC fewer changesoccurred in the model control group than in the blankcontrol group In the ACL when comparing the relativemRNA and protein expressions of GAP-43 more changesoccurred in the model control group than in the blankcontrol group all the differences were statistically significant(119875 lt 005) In the model control group at different timepoints the relative expression of mRNA and protein of NT-3 and TrkC decreased over time while the relative mRNAand protein expressions of GAP-43 increased over time allchanges were statistically significant (119875 lt 005) The resultsof this study illustrated that when the ACL was injured theexpressions of NT-3 and TrkC in neuronal cells of the DRGcould be reduced and the levels of phosphorylation of TrkCdecreased after activation thereby further reducing TrkCactivity When the ACL was injured GAP-43 was activatedin the proprioceptors which caused the increased expressionof GAP-43 Eventually changes in these cytokine expressionlevels can lead to decreased proprioceptive functions

Electroacupuncture is a form of acupuncture whereby asmall electric current is passed between pairs of acupunc-ture needles The use of electroacupuncture therapy (thecombination of nerve electrical stimulation and acupunc-ture) has gradually replaced other acupuncture therapiesElectroacupuncture therapy has been widely used to treatpain or injury in the hind limbs for many decades inChina Both human and animal experiments have confirmedthat electroacupuncture therapy has a definite effect on thetreatment of pain movement disturbance tissue regener-ation and so on [39ndash44] Some studies have found thatthe effects of acupuncture were mediated by the centralnervous system thus causing specific physiological effect thatachieved the purpose of treatment and improved the growthof neurons axonal regeneration synaptic reconstruction andthe functional connection of target organs [45 46]

When the ACL is pulled by the forward displacement ofthe tibia the contraction of the hamstring confronts tibialtranslation through regulating the ACLndashhamstring reflex arcwhich plays an important role in maintaining the stabilityof the knee joints The acupuncture points included in thisstudy were Wei Yang Yin Gu Xi Yangguan and Qu QuanThese points were chosen because they are all located nextto the hamstring When these acupuncture points werestimulated by electroacupuncture muscle contraction of thehamstring occurred and created the effect of reverse interven-tion Furthermore the ACL belongs to the arthromyodyniacategory of Traditional Chinese Medicine Among theseselected acupuncture points Wei Yang (BL19) is the Xiahetriple energiser acupuncture point that is mainly used totreat strong pain in the lumbar spine and pain and spasmsin the legs and feet Yin Gu (KI10) is mainly used to treatstrong pain in the knee joints Xi Yangguan (GB33) is mainlyused to treat swelling pain spasms in the knee joint andpopliteal area numbness in the leg and other diseases ofthe lower limbs and knee joints Qu Quan (LR8) is mainlyused to treat swelling pain in the knee joints and patellaand paralysis of lower limb Through the stimulation ofrelated points electroacupuncture transmits a large number

of proprioceptive and skin sensorial information to thecentral nervous system thereby participating in stabilisingthe activities of the knee joints and reconstruction of theproprioceptive function of the knee joints

In this study at 4 8 and 12 weeks compared withthe blank control group SEP and MCV incubations in themodel control group and the IIKE group were extendedbut the amplitudes were decreased The changes in the IIKEgroup were better than those in the model control group alldifferences were statistically significant (119875 lt 005) At thesame time point in the DRG when comparing the relativemRNA and protein expressions of NT-3 and TrkC therewere fewer changes in the model control group than in theblank control group however more changes occurred in theIIKE group than in the model control group in which alldifferences were statistically significant (119875 lt 005) It was alsoindicated that electroacupuncture could reversely intervenein the ACLndashhamstring reflex arc thereby activating the NT-3TrkC signal pathway and increasing the expression of NT-3 in the proprioceptors of hamstring NT-3 was reverselytransported to neurons in the DRG and then combined withthe corresponding TrkC receptor which promoted phospho-rylation of cytosolic TrkC after receptor activation whichcould enhance TrkC activity It could also activate GAP-43in the ACL which enhances the polymerisation ability ofGAP-43 in proprioceptors of ACL thereby increasing theactivity of ACL proprioceptionThe results found during ourexperiment may supplement certain potential explanationsabout the mechanisms of acupuncture treatment for ACLinjury to some extent

Some studies have found that unilateral ACL injury couldlead to proprioceptive degeneration of the bilateral kneejoints [47] In theory because of the characteristics of nerveconduction damage or intervention the proprioception ofone-side limb had a negative effect on the proprioceptionof the contralateral limb In this study the IIKE groupand the IBKE group were designed for the purpose ofproving that intervention for the healthy side can promote theproprioceptive recovery of the injured limb through centralregulation of the DRG

In terms of nerve electrophysiological examination andthe relativemRNA and protein expressions of NT-3 and TrkCin the DRG and of GAP-43 in the ACL the recovery effect ofthe IBKE group was more remarkable than that of the IIKEgroup (119875 lt 005) It showed that using the DRG as thenode the proprioception of bilateral ACLs could be regulatedand that the model of bilateral knee joint interventionwas better than the model of unilateral knee interventionAdditionally the effect of electroacupuncture interventionwas more significant with increased intervention time (119875 lt005)

This research had some limitations First all animalswere male and it is unclear whether that affected the resultsSecond because of the limitations of the objective conditionsthe period of observation during the experiment was only12 weeks therefore the long-term changes in proprioceptioncould not be observed Third without some antagonists ofthe NT-3TrkC signalling pathway it might be insufficientto check the role of the NT-3TrkC signalling pathway in

BioMed Research International 9

reverse intervention for the ACLndashhamstring reflex arc withelectroacupuncture Because these limitations existed thisstudy is considered basic All these issues remain to be solvedby further related studies

5 Conclusions

In this study the mechanism of reverse intervention for ACLproprioception with electroacupuncture which was basedon the ACLndashhamstring reflex arc and through mediationof the NT-3TrkC signalling pathway was explored Thisstudy helped establish the basis for the clinical application ofelectroacupuncture therapy and effective treatment methodsand patterns for proprioception disturbances caused by ACLinjuries thereby enriching and developing the scientificconnotation of electroacupuncture therapy guided by theory

Disclosure

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan are co-firstauthors

Conflicts of Interest

The authors declare that they have no conflicts of interest

Authorsrsquo Contributions

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan contributedequally to this work

Acknowledgments

This study was supported by the National Natural ScienceFoundation of China (no 81674095) Science-TechnologySupport Plan Project of Sichuan Province (no 2014SZ0185)and Science and Technology Strategic CooperationProject of Luzhou City-Southwest Medical University(no 2016LZXNYD-J08)

References

[1] H Zhang M Qiu A Zhou J Zhang and D Jiang ldquoAnatomicanterolateral ligament reconstruction improves postoperativeclinical outcomes combined with anatomic anterior cruciateligament reconstructionrdquo Journal of Sports Science andMedicine vol 15 no 4 pp 688ndash696 2016

[2] A A Rahnemai-Azar S Sabzevari S Irarrazaval T Chaoand F H Fu ldquoAnatomical individualized ACL reconstructionrdquoArchives of Bone and Joint Surgery vol 4 no 4 pp 291ndash2972016

[3] M R Sheth S R Tapasvi and S S Patil ldquoPrimary Repairof Tibial-Sided Avulsion of the Anterior Cruciate LigamentrdquoArthroscopy Techniques vol 5 no 4 pp e901ndashe906 2016

[4] F Sepulveda L Sanchez E Amy and W Micheo ldquoAnteriorCruciate Ligament InjuryrdquoCurrent SportsMedicine Reports vol16 no 3 pp 172ndash178 2017

[5] T M El-Gohary O A Khaled S R Ibrahim A M Alshenqitiand M I Ibrahim ldquoEffect of proprioception cross training on

repositioning accuracy and balance among healthy individualsrdquoJournal of PhysicalTherapy Science vol 28 no 11 pp 3178ndash31822016

[6] A Delle Vedove M Storbeck R Heller et al ldquoBiallelic loss ofproprioception-related PIEZO2 causes muscular atrophy withperinatal respiratory distress arthrogryposis and scoliosisrdquoAmerican Journal of Human Genetics vol 99 no 6 pp 1406ndash1408 2016

[7] W K Chang Y S Jung M Oh and K Kim ldquoQuantitativeassessment of proprioception using dynamometer in incom-plete spinal cord injury patients a preliminary studyrdquo Annalsof Rehabilitation Medicine vol 41 no 2 pp 218ndash224 2017

[8] G Haliloglu K Becker C Temucin et al ldquoRecessive PIEZO2stop mutation causes distal arthrogryposis with distal muscleweakness scoliosis and proprioception defectsrdquo Journal ofHuman Genetics vol 62 no 4 pp 497ndash501 2017

[9] J Li F Kong X Gao Y Shen and S Gao ldquoProspectiverandomized comparison of knee stability and proprioceptionfor posterior cruciate ligament reconstruction with autografthybrid graft and 120574-irradiated allograftrdquoArthroscopy vol 32 no12 pp 2548ndash2555 2016

[10] J F Abulhasan C M Anley M D Snow and M J GreyldquoHamstring stretch reflex could it be a reproducible objectivemeasure of functional knee stabilityrdquordquo Journal of ExperimentalOrthopaedics vol 3 no 1 2016

[11] M Solomonow R Baratta B H Zhou et al ldquoThe synergisticaction of the anterior cruciate ligament and thigh musclesin maintaining joint stabilityrdquo The American Journal of SportsMedicine vol 15 no 3 pp 207ndash213 1987

[12] P Dyhre-Poulsen and M R Krogsgaard ldquoMuscular reflexeselicited by electrical stimulation of the anterior cruciate liga-ment in humansrdquo Journal of Applied Physiology vol 89 no 6pp 2191ndash2195 2000

[13] E Tsuda Y Ishibashi Y Okamura and S Toh ldquoRestorationof anterior cruciate ligament-hamstring reflex arc after anteriorcruciate ligament reconstructionrdquoKnee Surgery Sports Trauma-tology Arthroscopy vol 11 no 2 pp 63ndash67 2003

[14] X-Y Wang P-Y Gu S-W Chen et al ldquoEndogenousneurotrophin-3 promotes neuronal sprouting from dorsalroot gangliardquo Neural Regeneration Research vol 10 no 11 pp1865ndash1868 2015

[15] K A Han D Woo S Kim et al ldquoNeurotrophin-3 regu-lates synapse development by modulating TrkC-PTP120590 synapticadhesion and intracellular signaling pathwaysrdquo The Journal ofNeuroscience vol 36 no 17 pp 4816ndash4831 2016

[16] M-H Yang K-C Chen P-W Chiang et al ldquoProteomicprofiling of neuroblastoma cells adhesion on hyaluronic acid-based surface for neural tissue engineeringrdquo BioMed ResearchInternational vol 2016 Article ID 1917394 2016

[17] S Rao M Martınez-Cengotitabengoa Y Yao et al ldquoPeripheralblood nerve growth factor levels inmajor psychiatric disordersrdquoJournal of Psychiatric Research vol 86 pp 39ndash45 2016

[18] M Yong S Kim and S Cheon ldquoEffects of skilled reach trainingwith affected forelimb and treadmill exercise on the expressionof neurotrophic factor following ischemia-induced brain injuryin ratsrdquo Journal of Physical Therapy Science vol 29 no 4 pp647ndash650 2017

[19] S Liu S Blanchard S Bigou S Vitry D Bohl and J-M HeardldquoNeurotrophin 3 improves delayed reconstruction of sensorypathways after cervical dorsal root injuryrdquoNeurosurgery vol 68no 2 pp 450ndash461 2011

10 BioMed Research International

[20] J Cui C Cui Y Cui et al ldquoBone marrow mesenchymal stemcell transplantation increases GAP-43 Expression via ERK12and PI3KAkt pathways in intracerebral hemorrhagerdquo CellularPhysiology and Biochemistry vol 42 no 1 pp 137ndash144 2017

[21] Y Li XGaoQWang et al ldquoRetinoic acid protects from experi-mental cerebral infarction by upregulating GAP-43 expressionrdquoBrazilian Journal of Medical and Biological Research vol 50 no4 2017

[22] H B Kim and B S Yoo ldquoPropolis inhibits neurite outgrowthin differentiating SH-SY5Y human neuroblastoma cellsrdquo Toxi-cological Research vol 32 no 3 pp 239ndash243 2016

[23] J Iwasa M Ochi Y Uchio N Adachi and K KawasakildquoDecrease in anterior knee laxity by electrical stimulation ofnormal and reconstructed anterior cruciate ligamentsrdquo TheJournal of Boneamp Joint Surgery vol 88 no 4 pp 477ndash483 2006

[24] A Biscarini F M Botti and V E Pettorossi ldquoSelectivecontribution of each hamstring muscle to anterior cruciateligament protection and tibiofemoral joint stability in leg-extension exercise A simulation studyrdquo European Journal ofApplied Physiology vol 113 no 9 pp 2263ndash2273 2013

[25] D Sugimoto E Alentorn-Geli J Mendiguchıa K SamuelssonJ Karlsson andGDMyer ldquoBiomechanical and neuromuscularcharacteristics of male athletes implications for the develop-ment of anterior cruciate ligament injury prevention programsrdquoSports Medicine vol 45 no 6 pp 809ndash822 2015

[26] M G Barcellona M C Morrissey P Milligan M Clinton andA A Amis ldquoThe effect of knee extensor open kinetic chainresistance training in the ACL-injured kneerdquo Knee SurgerySports Traumatology Arthroscopy vol 23 no 11 pp 3168ndash31772015

[27] T S Furlanetto L A Peyre-Tartaruga A S Do Pinho E SDa Bernardes and M A Zaro ldquoProprioception body balanceand functionality in individuals with ACL reconstructionrdquoActaOrtopedica Brasileira vol 24 no 2 pp 67ndash72 2016

[28] A J Wodowski C W Swigler H Liu K M Nord P C ToyandW M Mihalko ldquoProprioception and Knee Arthroplasty ALiterature ReviewrdquoOrthopedic Clinics of North America vol 47no 2 pp 301ndash309 2016

[29] I Dallo J Chahla J JMitchell C Pascual-Garrido J A Feaginand R F LaPrade ldquoBiologic Approaches for the Treatment ofPartial Tears of the Anterior Cruciate Ligament A CurrentConcepts Reviewrdquo Orthopaedic Journal of Sports Medicine vol5 no 1 2017

[30] W R Lowe R J Warth E P Davis and L Bailey ldquoFunc-tional bracing after anterior cruciate ligament reconstructionA systematic reviewrdquo Journal of the American Academy ofOrthopaedicSurgeons vol 25 no 3 pp 239ndash249 2017

[31] Y Ma M Deie D Iwaki et al ldquoBalance Ability and Pro-prioception after Single-Bundle Single-Bundle Augmentationand Double-Bundle ACL ReconstructionrdquoThe Scientific WorldJournal vol 2014 Article ID 342012 8 pages 2014

[32] N Relph L Herrington and S Tyson ldquoThe effects of ACLinjury on knee proprioception Ameta-analysisrdquo Physiotherapyvol 100 no 3 pp 187ndash195 2014

[33] B P Livingston and T R Nichols ldquoEffects of reinnervation ofthe triceps brachii on joint kinematics and electromyographicpatterns of the feline forelimb during level and upslope walk-ingrdquo Cells Tissues Organs vol 199 no 5-6 pp 405ndash422 2014

[34] E Appel S Weissmann Y Salzberg et al ldquoAn ensemble ofregulatory elements controls Runx3 spatiotemporal expressionin subsets of dorsal root ganglia proprioceptive neuronsrdquoGenesamp Development vol 30 no 23 pp 2607ndash2622 2016

[35] S M OrsquoToole M M Ferrer J Mekonnen et al ldquoDicermaintains the identity and function of proprioceptive sensoryneuronsrdquo Journal of Neurophysiology vol 117 no 3 pp 1057ndash1069 2017

[36] Y Liu L Kelamangalath H Kim et al ldquoNT-3 promotes pro-prioceptive axon regeneration when combined with activationof the mTor intrinsic growth pathway but not with reduction ofmyelin extrinsic inhibitorsrdquo Experimental Neurology vol 283pp 73ndash84 2016

[37] S Li T Shen Y Liang Y Zhang and B Bai ldquoEffects ofminiscalpel-needle release on chronic neck pain A retrospec-tive analysis with 12-month follow-uprdquo PLoS ONE vol 10 no8 Article ID e0137033 pp 133ndash137 2015

[38] W-S Juan S-Y Huang C-C Chang et al ldquoMelatoninimproves neuroplasticity by upregulating the growth-associatedprotein-43 (GAP-43) and NMDAR postsynaptic density-95(PSD-95) proteins in cultured neurons exposed to glutamateexcitotoxicity and in rats subjected to transient focal cerebralischemia even during a long-term recovery periodrdquo Journal ofPineal Research vol 56 no 2 pp 213ndash223 2014

[39] Y-HGaoC-W Li J-YWang et al ldquoActivation of hippocampalMEK1 contributes to the cumulative antinociceptive effect ofelectroacupuncture in neuropathic pain ratsrdquo BMC Comple-mentary and Alternative Medicine vol 16 no 1 article no 5172016

[40] K-Y Huang S Liang M-L Yu S-P Fu X Chen and S-F Lu ldquoA systematic review and meta-analysis of acupuncturefor improving learning and memory ability in animalsrdquo BMCComplementary and Alternative Medicine vol 16 no 1 articleno 297 2016

[41] J Zhou S Li Y Wang et al ldquoEffects and mechanisms ofauricular electroacupuncture on gastric hypersensitivity in arodent model of functional dyspepsiardquo PLoS ONE vol 12 no3 Article ID e0174568 2017

[42] Y H Gao C W Li J Y Wang et al ldquoEffect of electroacupunc-ture on the cervicospinal P2X7 receptorfractalkineCX3CR1signaling pathway in a rat neck-incision painmodelrdquo PurinergicSignalling vol 13 no 2 pp 215ndash225 2017

[43] JH LeeDGoWKimet al ldquoInvolvement of spinalmuscarinicand serotonergic receptors in the anti-allodynic effect of elec-troacupuncture in rats with oxaliplatin-induced neuropathicpainrdquoKorean Journal of Physiology amp Pharmacology vol 20 no4 pp 407ndash414 2016

[44] S P Chen Y Kan J L Zhang et al ldquoInvolvement of hippocam-pal acetylcholinergic receptors in electroacupuncture analgesiain neuropathic pain ratsrdquo Behavioral and Brain Functions vol12 no 1 article no 13 2016

[45] M Villarreal Santiago S Tumilty A Mącznik and R ManildquoDoes acupuncture alter pain-related functional connectivity ofthe central nervous system a systematic reviewrdquo JAMS Journalof Acupuncture and Meridian Studies vol 9 no 4 pp 167ndash1772016

[46] T E Salazar M R Richardson E Beli et al ldquoElectroacupunc-ture Promotes Central Nervous System-Dependent Release ofMesenchymal Stem Cellsrdquo Stem Cells vol 35 no 5 pp 1303ndash1315 2017

[47] D-H Lee J-H Lee S-E Ahn and M-J Park ldquoEffect of timeafter anterior cruciate ligament tears on proprioception andpostural stabilityrdquo PLoS ONE vol 10 no 9 Article ID e01390382015

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

OncologyJournal of

Hindawiwwwhindawicom Volume 2013

Hindawiwwwhindawicom Volume 2018

Oxidative Medicine and Cellular Longevity

Hindawiwwwhindawicom Volume 2018

PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

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Hindawiwwwhindawicom Volume 2018

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Computational and Mathematical Methods in Medicine

Hindawiwwwhindawicom Volume 2018

Behavioural Neurology

OphthalmologyJournal of

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Research and TreatmentAIDS

Hindawiwwwhindawicom Volume 2018

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Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

BioMed Research International 7

00

05

10

15

20N

T-3-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de e

beabe

(a)

00

05

10

15

20

TrkC

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebeabe

(b)

00

05

10

15

20

GA

P-43

-a

ctin

IIKE groupIBKE groupModel control groupBlank control group

8 124(Weeks)

abcdebcde

cde

abdebde de

ebe

abe

(c)

4 8 12

TrkCNT-3

GAP-43-actin

IIKE group IBKE group model control group blank control group

(Weeks)

(d)

Figure 6 Expressions of NT-3120573-actin TrkC120573-actin and GAP-43120573-actin proteins among the four groups at different time points (a)Expressions of NT-3120573-actin proteins (b) Expressions of TrkC120573-actin proteins (c) Expressions of GAP-43120573-actin proteins a versus 8weeks in the same group 119875 lt 005 b versus 12 weeks in the same group 119875 lt 005 c versus the IIKE group at the same time 119875 lt 005 dversus the IBKE group at the same time 119875 lt 005 e versus the blank control group 119875 lt 005 (d) The expression of protein of NT-3 TrkCand 120573-actin among the four groups in different time points

when the ACL proprioception function is decreased the neu-ral feedback regulation pathway is bound to be disorderedtherefore it could have further impact on the proprioceptionfunction [31 32] Some studies showed that NT-3 was the keyinduced signal of proprioceptive transduction when NT-3expression was normal the proprioceptive nerve fibres couldcontrol the target organs and have physiological roles [33ndash35] If the expressions of NT-3 and TrkC were decreasedthen apoptosis of proprioceptive neurons in the DRG couldoccur thus causing lower limb proprioception deficienciesNT-3 and TrkC could also accelerate the development andmaturation of neurons thus maintaining the survival ofmature neurons andpromoting the repair and regeneration ofinjured neurons When the proprioceptive nerve conductionpathway is injured proprioceptive neurons appear abnormal

however exogenous NT-3 could protect the proprioceptiveneurons and improve their function [36 37] ACL injuriescould damage peripheral nerve fibres When the nerve isdamaged expression of upregulation could be included andthe functional relationship could be decreased immediatelyafter reconstruction which is considered as a marker ofneuronal growth and repair [38]

During this study at 4 8 and 12 weeks in comparisonwith the blank control group incubations of SEPs and MCVin the model control group were significantly extended butthe amplitudes were significantly decreased (119875 lt 005)Comparisons of the three time points showed that incuba-tions of the SEPs and MCV in the model control group weresignificantly extended but the amplitudes were significantlydecreased over time (119875 lt 005) Furthermore at the same

8 BioMed Research International

time point in the DRG when comparing the relative mRNAand protein expressions of NT-3 and TrkC fewer changesoccurred in the model control group than in the blankcontrol group In the ACL when comparing the relativemRNA and protein expressions of GAP-43 more changesoccurred in the model control group than in the blankcontrol group all the differences were statistically significant(119875 lt 005) In the model control group at different timepoints the relative expression of mRNA and protein of NT-3 and TrkC decreased over time while the relative mRNAand protein expressions of GAP-43 increased over time allchanges were statistically significant (119875 lt 005) The resultsof this study illustrated that when the ACL was injured theexpressions of NT-3 and TrkC in neuronal cells of the DRGcould be reduced and the levels of phosphorylation of TrkCdecreased after activation thereby further reducing TrkCactivity When the ACL was injured GAP-43 was activatedin the proprioceptors which caused the increased expressionof GAP-43 Eventually changes in these cytokine expressionlevels can lead to decreased proprioceptive functions

Electroacupuncture is a form of acupuncture whereby asmall electric current is passed between pairs of acupunc-ture needles The use of electroacupuncture therapy (thecombination of nerve electrical stimulation and acupunc-ture) has gradually replaced other acupuncture therapiesElectroacupuncture therapy has been widely used to treatpain or injury in the hind limbs for many decades inChina Both human and animal experiments have confirmedthat electroacupuncture therapy has a definite effect on thetreatment of pain movement disturbance tissue regener-ation and so on [39ndash44] Some studies have found thatthe effects of acupuncture were mediated by the centralnervous system thus causing specific physiological effect thatachieved the purpose of treatment and improved the growthof neurons axonal regeneration synaptic reconstruction andthe functional connection of target organs [45 46]

When the ACL is pulled by the forward displacement ofthe tibia the contraction of the hamstring confronts tibialtranslation through regulating the ACLndashhamstring reflex arcwhich plays an important role in maintaining the stabilityof the knee joints The acupuncture points included in thisstudy were Wei Yang Yin Gu Xi Yangguan and Qu QuanThese points were chosen because they are all located nextto the hamstring When these acupuncture points werestimulated by electroacupuncture muscle contraction of thehamstring occurred and created the effect of reverse interven-tion Furthermore the ACL belongs to the arthromyodyniacategory of Traditional Chinese Medicine Among theseselected acupuncture points Wei Yang (BL19) is the Xiahetriple energiser acupuncture point that is mainly used totreat strong pain in the lumbar spine and pain and spasmsin the legs and feet Yin Gu (KI10) is mainly used to treatstrong pain in the knee joints Xi Yangguan (GB33) is mainlyused to treat swelling pain spasms in the knee joint andpopliteal area numbness in the leg and other diseases ofthe lower limbs and knee joints Qu Quan (LR8) is mainlyused to treat swelling pain in the knee joints and patellaand paralysis of lower limb Through the stimulation ofrelated points electroacupuncture transmits a large number

of proprioceptive and skin sensorial information to thecentral nervous system thereby participating in stabilisingthe activities of the knee joints and reconstruction of theproprioceptive function of the knee joints

In this study at 4 8 and 12 weeks compared withthe blank control group SEP and MCV incubations in themodel control group and the IIKE group were extendedbut the amplitudes were decreased The changes in the IIKEgroup were better than those in the model control group alldifferences were statistically significant (119875 lt 005) At thesame time point in the DRG when comparing the relativemRNA and protein expressions of NT-3 and TrkC therewere fewer changes in the model control group than in theblank control group however more changes occurred in theIIKE group than in the model control group in which alldifferences were statistically significant (119875 lt 005) It was alsoindicated that electroacupuncture could reversely intervenein the ACLndashhamstring reflex arc thereby activating the NT-3TrkC signal pathway and increasing the expression of NT-3 in the proprioceptors of hamstring NT-3 was reverselytransported to neurons in the DRG and then combined withthe corresponding TrkC receptor which promoted phospho-rylation of cytosolic TrkC after receptor activation whichcould enhance TrkC activity It could also activate GAP-43in the ACL which enhances the polymerisation ability ofGAP-43 in proprioceptors of ACL thereby increasing theactivity of ACL proprioceptionThe results found during ourexperiment may supplement certain potential explanationsabout the mechanisms of acupuncture treatment for ACLinjury to some extent

Some studies have found that unilateral ACL injury couldlead to proprioceptive degeneration of the bilateral kneejoints [47] In theory because of the characteristics of nerveconduction damage or intervention the proprioception ofone-side limb had a negative effect on the proprioceptionof the contralateral limb In this study the IIKE groupand the IBKE group were designed for the purpose ofproving that intervention for the healthy side can promote theproprioceptive recovery of the injured limb through centralregulation of the DRG

In terms of nerve electrophysiological examination andthe relativemRNA and protein expressions of NT-3 and TrkCin the DRG and of GAP-43 in the ACL the recovery effect ofthe IBKE group was more remarkable than that of the IIKEgroup (119875 lt 005) It showed that using the DRG as thenode the proprioception of bilateral ACLs could be regulatedand that the model of bilateral knee joint interventionwas better than the model of unilateral knee interventionAdditionally the effect of electroacupuncture interventionwas more significant with increased intervention time (119875 lt005)

This research had some limitations First all animalswere male and it is unclear whether that affected the resultsSecond because of the limitations of the objective conditionsthe period of observation during the experiment was only12 weeks therefore the long-term changes in proprioceptioncould not be observed Third without some antagonists ofthe NT-3TrkC signalling pathway it might be insufficientto check the role of the NT-3TrkC signalling pathway in

BioMed Research International 9

reverse intervention for the ACLndashhamstring reflex arc withelectroacupuncture Because these limitations existed thisstudy is considered basic All these issues remain to be solvedby further related studies

5 Conclusions

In this study the mechanism of reverse intervention for ACLproprioception with electroacupuncture which was basedon the ACLndashhamstring reflex arc and through mediationof the NT-3TrkC signalling pathway was explored Thisstudy helped establish the basis for the clinical application ofelectroacupuncture therapy and effective treatment methodsand patterns for proprioception disturbances caused by ACLinjuries thereby enriching and developing the scientificconnotation of electroacupuncture therapy guided by theory

Disclosure

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan are co-firstauthors

Conflicts of Interest

The authors declare that they have no conflicts of interest

Authorsrsquo Contributions

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan contributedequally to this work

Acknowledgments

This study was supported by the National Natural ScienceFoundation of China (no 81674095) Science-TechnologySupport Plan Project of Sichuan Province (no 2014SZ0185)and Science and Technology Strategic CooperationProject of Luzhou City-Southwest Medical University(no 2016LZXNYD-J08)

References

[1] H Zhang M Qiu A Zhou J Zhang and D Jiang ldquoAnatomicanterolateral ligament reconstruction improves postoperativeclinical outcomes combined with anatomic anterior cruciateligament reconstructionrdquo Journal of Sports Science andMedicine vol 15 no 4 pp 688ndash696 2016

[2] A A Rahnemai-Azar S Sabzevari S Irarrazaval T Chaoand F H Fu ldquoAnatomical individualized ACL reconstructionrdquoArchives of Bone and Joint Surgery vol 4 no 4 pp 291ndash2972016

[3] M R Sheth S R Tapasvi and S S Patil ldquoPrimary Repairof Tibial-Sided Avulsion of the Anterior Cruciate LigamentrdquoArthroscopy Techniques vol 5 no 4 pp e901ndashe906 2016

[4] F Sepulveda L Sanchez E Amy and W Micheo ldquoAnteriorCruciate Ligament InjuryrdquoCurrent SportsMedicine Reports vol16 no 3 pp 172ndash178 2017

[5] T M El-Gohary O A Khaled S R Ibrahim A M Alshenqitiand M I Ibrahim ldquoEffect of proprioception cross training on

repositioning accuracy and balance among healthy individualsrdquoJournal of PhysicalTherapy Science vol 28 no 11 pp 3178ndash31822016

[6] A Delle Vedove M Storbeck R Heller et al ldquoBiallelic loss ofproprioception-related PIEZO2 causes muscular atrophy withperinatal respiratory distress arthrogryposis and scoliosisrdquoAmerican Journal of Human Genetics vol 99 no 6 pp 1406ndash1408 2016

[7] W K Chang Y S Jung M Oh and K Kim ldquoQuantitativeassessment of proprioception using dynamometer in incom-plete spinal cord injury patients a preliminary studyrdquo Annalsof Rehabilitation Medicine vol 41 no 2 pp 218ndash224 2017

[8] G Haliloglu K Becker C Temucin et al ldquoRecessive PIEZO2stop mutation causes distal arthrogryposis with distal muscleweakness scoliosis and proprioception defectsrdquo Journal ofHuman Genetics vol 62 no 4 pp 497ndash501 2017

[9] J Li F Kong X Gao Y Shen and S Gao ldquoProspectiverandomized comparison of knee stability and proprioceptionfor posterior cruciate ligament reconstruction with autografthybrid graft and 120574-irradiated allograftrdquoArthroscopy vol 32 no12 pp 2548ndash2555 2016

[10] J F Abulhasan C M Anley M D Snow and M J GreyldquoHamstring stretch reflex could it be a reproducible objectivemeasure of functional knee stabilityrdquordquo Journal of ExperimentalOrthopaedics vol 3 no 1 2016

[11] M Solomonow R Baratta B H Zhou et al ldquoThe synergisticaction of the anterior cruciate ligament and thigh musclesin maintaining joint stabilityrdquo The American Journal of SportsMedicine vol 15 no 3 pp 207ndash213 1987

[12] P Dyhre-Poulsen and M R Krogsgaard ldquoMuscular reflexeselicited by electrical stimulation of the anterior cruciate liga-ment in humansrdquo Journal of Applied Physiology vol 89 no 6pp 2191ndash2195 2000

[13] E Tsuda Y Ishibashi Y Okamura and S Toh ldquoRestorationof anterior cruciate ligament-hamstring reflex arc after anteriorcruciate ligament reconstructionrdquoKnee Surgery Sports Trauma-tology Arthroscopy vol 11 no 2 pp 63ndash67 2003

[14] X-Y Wang P-Y Gu S-W Chen et al ldquoEndogenousneurotrophin-3 promotes neuronal sprouting from dorsalroot gangliardquo Neural Regeneration Research vol 10 no 11 pp1865ndash1868 2015

[15] K A Han D Woo S Kim et al ldquoNeurotrophin-3 regu-lates synapse development by modulating TrkC-PTP120590 synapticadhesion and intracellular signaling pathwaysrdquo The Journal ofNeuroscience vol 36 no 17 pp 4816ndash4831 2016

[16] M-H Yang K-C Chen P-W Chiang et al ldquoProteomicprofiling of neuroblastoma cells adhesion on hyaluronic acid-based surface for neural tissue engineeringrdquo BioMed ResearchInternational vol 2016 Article ID 1917394 2016

[17] S Rao M Martınez-Cengotitabengoa Y Yao et al ldquoPeripheralblood nerve growth factor levels inmajor psychiatric disordersrdquoJournal of Psychiatric Research vol 86 pp 39ndash45 2016

[18] M Yong S Kim and S Cheon ldquoEffects of skilled reach trainingwith affected forelimb and treadmill exercise on the expressionof neurotrophic factor following ischemia-induced brain injuryin ratsrdquo Journal of Physical Therapy Science vol 29 no 4 pp647ndash650 2017

[19] S Liu S Blanchard S Bigou S Vitry D Bohl and J-M HeardldquoNeurotrophin 3 improves delayed reconstruction of sensorypathways after cervical dorsal root injuryrdquoNeurosurgery vol 68no 2 pp 450ndash461 2011

10 BioMed Research International

[20] J Cui C Cui Y Cui et al ldquoBone marrow mesenchymal stemcell transplantation increases GAP-43 Expression via ERK12and PI3KAkt pathways in intracerebral hemorrhagerdquo CellularPhysiology and Biochemistry vol 42 no 1 pp 137ndash144 2017

[21] Y Li XGaoQWang et al ldquoRetinoic acid protects from experi-mental cerebral infarction by upregulating GAP-43 expressionrdquoBrazilian Journal of Medical and Biological Research vol 50 no4 2017

[22] H B Kim and B S Yoo ldquoPropolis inhibits neurite outgrowthin differentiating SH-SY5Y human neuroblastoma cellsrdquo Toxi-cological Research vol 32 no 3 pp 239ndash243 2016

[23] J Iwasa M Ochi Y Uchio N Adachi and K KawasakildquoDecrease in anterior knee laxity by electrical stimulation ofnormal and reconstructed anterior cruciate ligamentsrdquo TheJournal of Boneamp Joint Surgery vol 88 no 4 pp 477ndash483 2006

[24] A Biscarini F M Botti and V E Pettorossi ldquoSelectivecontribution of each hamstring muscle to anterior cruciateligament protection and tibiofemoral joint stability in leg-extension exercise A simulation studyrdquo European Journal ofApplied Physiology vol 113 no 9 pp 2263ndash2273 2013

[25] D Sugimoto E Alentorn-Geli J Mendiguchıa K SamuelssonJ Karlsson andGDMyer ldquoBiomechanical and neuromuscularcharacteristics of male athletes implications for the develop-ment of anterior cruciate ligament injury prevention programsrdquoSports Medicine vol 45 no 6 pp 809ndash822 2015

[26] M G Barcellona M C Morrissey P Milligan M Clinton andA A Amis ldquoThe effect of knee extensor open kinetic chainresistance training in the ACL-injured kneerdquo Knee SurgerySports Traumatology Arthroscopy vol 23 no 11 pp 3168ndash31772015

[27] T S Furlanetto L A Peyre-Tartaruga A S Do Pinho E SDa Bernardes and M A Zaro ldquoProprioception body balanceand functionality in individuals with ACL reconstructionrdquoActaOrtopedica Brasileira vol 24 no 2 pp 67ndash72 2016

[28] A J Wodowski C W Swigler H Liu K M Nord P C ToyandW M Mihalko ldquoProprioception and Knee Arthroplasty ALiterature ReviewrdquoOrthopedic Clinics of North America vol 47no 2 pp 301ndash309 2016

[29] I Dallo J Chahla J JMitchell C Pascual-Garrido J A Feaginand R F LaPrade ldquoBiologic Approaches for the Treatment ofPartial Tears of the Anterior Cruciate Ligament A CurrentConcepts Reviewrdquo Orthopaedic Journal of Sports Medicine vol5 no 1 2017

[30] W R Lowe R J Warth E P Davis and L Bailey ldquoFunc-tional bracing after anterior cruciate ligament reconstructionA systematic reviewrdquo Journal of the American Academy ofOrthopaedicSurgeons vol 25 no 3 pp 239ndash249 2017

[31] Y Ma M Deie D Iwaki et al ldquoBalance Ability and Pro-prioception after Single-Bundle Single-Bundle Augmentationand Double-Bundle ACL ReconstructionrdquoThe Scientific WorldJournal vol 2014 Article ID 342012 8 pages 2014

[32] N Relph L Herrington and S Tyson ldquoThe effects of ACLinjury on knee proprioception Ameta-analysisrdquo Physiotherapyvol 100 no 3 pp 187ndash195 2014

[33] B P Livingston and T R Nichols ldquoEffects of reinnervation ofthe triceps brachii on joint kinematics and electromyographicpatterns of the feline forelimb during level and upslope walk-ingrdquo Cells Tissues Organs vol 199 no 5-6 pp 405ndash422 2014

[34] E Appel S Weissmann Y Salzberg et al ldquoAn ensemble ofregulatory elements controls Runx3 spatiotemporal expressionin subsets of dorsal root ganglia proprioceptive neuronsrdquoGenesamp Development vol 30 no 23 pp 2607ndash2622 2016

[35] S M OrsquoToole M M Ferrer J Mekonnen et al ldquoDicermaintains the identity and function of proprioceptive sensoryneuronsrdquo Journal of Neurophysiology vol 117 no 3 pp 1057ndash1069 2017

[36] Y Liu L Kelamangalath H Kim et al ldquoNT-3 promotes pro-prioceptive axon regeneration when combined with activationof the mTor intrinsic growth pathway but not with reduction ofmyelin extrinsic inhibitorsrdquo Experimental Neurology vol 283pp 73ndash84 2016

[37] S Li T Shen Y Liang Y Zhang and B Bai ldquoEffects ofminiscalpel-needle release on chronic neck pain A retrospec-tive analysis with 12-month follow-uprdquo PLoS ONE vol 10 no8 Article ID e0137033 pp 133ndash137 2015

[38] W-S Juan S-Y Huang C-C Chang et al ldquoMelatoninimproves neuroplasticity by upregulating the growth-associatedprotein-43 (GAP-43) and NMDAR postsynaptic density-95(PSD-95) proteins in cultured neurons exposed to glutamateexcitotoxicity and in rats subjected to transient focal cerebralischemia even during a long-term recovery periodrdquo Journal ofPineal Research vol 56 no 2 pp 213ndash223 2014

[39] Y-HGaoC-W Li J-YWang et al ldquoActivation of hippocampalMEK1 contributes to the cumulative antinociceptive effect ofelectroacupuncture in neuropathic pain ratsrdquo BMC Comple-mentary and Alternative Medicine vol 16 no 1 article no 5172016

[40] K-Y Huang S Liang M-L Yu S-P Fu X Chen and S-F Lu ldquoA systematic review and meta-analysis of acupuncturefor improving learning and memory ability in animalsrdquo BMCComplementary and Alternative Medicine vol 16 no 1 articleno 297 2016

[41] J Zhou S Li Y Wang et al ldquoEffects and mechanisms ofauricular electroacupuncture on gastric hypersensitivity in arodent model of functional dyspepsiardquo PLoS ONE vol 12 no3 Article ID e0174568 2017

[42] Y H Gao C W Li J Y Wang et al ldquoEffect of electroacupunc-ture on the cervicospinal P2X7 receptorfractalkineCX3CR1signaling pathway in a rat neck-incision painmodelrdquo PurinergicSignalling vol 13 no 2 pp 215ndash225 2017

[43] JH LeeDGoWKimet al ldquoInvolvement of spinalmuscarinicand serotonergic receptors in the anti-allodynic effect of elec-troacupuncture in rats with oxaliplatin-induced neuropathicpainrdquoKorean Journal of Physiology amp Pharmacology vol 20 no4 pp 407ndash414 2016

[44] S P Chen Y Kan J L Zhang et al ldquoInvolvement of hippocam-pal acetylcholinergic receptors in electroacupuncture analgesiain neuropathic pain ratsrdquo Behavioral and Brain Functions vol12 no 1 article no 13 2016

[45] M Villarreal Santiago S Tumilty A Mącznik and R ManildquoDoes acupuncture alter pain-related functional connectivity ofthe central nervous system a systematic reviewrdquo JAMS Journalof Acupuncture and Meridian Studies vol 9 no 4 pp 167ndash1772016

[46] T E Salazar M R Richardson E Beli et al ldquoElectroacupunc-ture Promotes Central Nervous System-Dependent Release ofMesenchymal Stem Cellsrdquo Stem Cells vol 35 no 5 pp 1303ndash1315 2017

[47] D-H Lee J-H Lee S-E Ahn and M-J Park ldquoEffect of timeafter anterior cruciate ligament tears on proprioception andpostural stabilityrdquo PLoS ONE vol 10 no 9 Article ID e01390382015

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

OncologyJournal of

Hindawiwwwhindawicom Volume 2013

Hindawiwwwhindawicom Volume 2018

Oxidative Medicine and Cellular Longevity

Hindawiwwwhindawicom Volume 2018

PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Computational and Mathematical Methods in Medicine

Hindawiwwwhindawicom Volume 2018

Behavioural Neurology

OphthalmologyJournal of

Hindawiwwwhindawicom Volume 2018

Diabetes ResearchJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Research and TreatmentAIDS

Hindawiwwwhindawicom Volume 2018

Gastroenterology Research and Practice

Hindawiwwwhindawicom Volume 2018

Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

8 BioMed Research International

time point in the DRG when comparing the relative mRNAand protein expressions of NT-3 and TrkC fewer changesoccurred in the model control group than in the blankcontrol group In the ACL when comparing the relativemRNA and protein expressions of GAP-43 more changesoccurred in the model control group than in the blankcontrol group all the differences were statistically significant(119875 lt 005) In the model control group at different timepoints the relative expression of mRNA and protein of NT-3 and TrkC decreased over time while the relative mRNAand protein expressions of GAP-43 increased over time allchanges were statistically significant (119875 lt 005) The resultsof this study illustrated that when the ACL was injured theexpressions of NT-3 and TrkC in neuronal cells of the DRGcould be reduced and the levels of phosphorylation of TrkCdecreased after activation thereby further reducing TrkCactivity When the ACL was injured GAP-43 was activatedin the proprioceptors which caused the increased expressionof GAP-43 Eventually changes in these cytokine expressionlevels can lead to decreased proprioceptive functions

Electroacupuncture is a form of acupuncture whereby asmall electric current is passed between pairs of acupunc-ture needles The use of electroacupuncture therapy (thecombination of nerve electrical stimulation and acupunc-ture) has gradually replaced other acupuncture therapiesElectroacupuncture therapy has been widely used to treatpain or injury in the hind limbs for many decades inChina Both human and animal experiments have confirmedthat electroacupuncture therapy has a definite effect on thetreatment of pain movement disturbance tissue regener-ation and so on [39ndash44] Some studies have found thatthe effects of acupuncture were mediated by the centralnervous system thus causing specific physiological effect thatachieved the purpose of treatment and improved the growthof neurons axonal regeneration synaptic reconstruction andthe functional connection of target organs [45 46]

When the ACL is pulled by the forward displacement ofthe tibia the contraction of the hamstring confronts tibialtranslation through regulating the ACLndashhamstring reflex arcwhich plays an important role in maintaining the stabilityof the knee joints The acupuncture points included in thisstudy were Wei Yang Yin Gu Xi Yangguan and Qu QuanThese points were chosen because they are all located nextto the hamstring When these acupuncture points werestimulated by electroacupuncture muscle contraction of thehamstring occurred and created the effect of reverse interven-tion Furthermore the ACL belongs to the arthromyodyniacategory of Traditional Chinese Medicine Among theseselected acupuncture points Wei Yang (BL19) is the Xiahetriple energiser acupuncture point that is mainly used totreat strong pain in the lumbar spine and pain and spasmsin the legs and feet Yin Gu (KI10) is mainly used to treatstrong pain in the knee joints Xi Yangguan (GB33) is mainlyused to treat swelling pain spasms in the knee joint andpopliteal area numbness in the leg and other diseases ofthe lower limbs and knee joints Qu Quan (LR8) is mainlyused to treat swelling pain in the knee joints and patellaand paralysis of lower limb Through the stimulation ofrelated points electroacupuncture transmits a large number

of proprioceptive and skin sensorial information to thecentral nervous system thereby participating in stabilisingthe activities of the knee joints and reconstruction of theproprioceptive function of the knee joints

In this study at 4 8 and 12 weeks compared withthe blank control group SEP and MCV incubations in themodel control group and the IIKE group were extendedbut the amplitudes were decreased The changes in the IIKEgroup were better than those in the model control group alldifferences were statistically significant (119875 lt 005) At thesame time point in the DRG when comparing the relativemRNA and protein expressions of NT-3 and TrkC therewere fewer changes in the model control group than in theblank control group however more changes occurred in theIIKE group than in the model control group in which alldifferences were statistically significant (119875 lt 005) It was alsoindicated that electroacupuncture could reversely intervenein the ACLndashhamstring reflex arc thereby activating the NT-3TrkC signal pathway and increasing the expression of NT-3 in the proprioceptors of hamstring NT-3 was reverselytransported to neurons in the DRG and then combined withthe corresponding TrkC receptor which promoted phospho-rylation of cytosolic TrkC after receptor activation whichcould enhance TrkC activity It could also activate GAP-43in the ACL which enhances the polymerisation ability ofGAP-43 in proprioceptors of ACL thereby increasing theactivity of ACL proprioceptionThe results found during ourexperiment may supplement certain potential explanationsabout the mechanisms of acupuncture treatment for ACLinjury to some extent

Some studies have found that unilateral ACL injury couldlead to proprioceptive degeneration of the bilateral kneejoints [47] In theory because of the characteristics of nerveconduction damage or intervention the proprioception ofone-side limb had a negative effect on the proprioceptionof the contralateral limb In this study the IIKE groupand the IBKE group were designed for the purpose ofproving that intervention for the healthy side can promote theproprioceptive recovery of the injured limb through centralregulation of the DRG

In terms of nerve electrophysiological examination andthe relativemRNA and protein expressions of NT-3 and TrkCin the DRG and of GAP-43 in the ACL the recovery effect ofthe IBKE group was more remarkable than that of the IIKEgroup (119875 lt 005) It showed that using the DRG as thenode the proprioception of bilateral ACLs could be regulatedand that the model of bilateral knee joint interventionwas better than the model of unilateral knee interventionAdditionally the effect of electroacupuncture interventionwas more significant with increased intervention time (119875 lt005)

This research had some limitations First all animalswere male and it is unclear whether that affected the resultsSecond because of the limitations of the objective conditionsthe period of observation during the experiment was only12 weeks therefore the long-term changes in proprioceptioncould not be observed Third without some antagonists ofthe NT-3TrkC signalling pathway it might be insufficientto check the role of the NT-3TrkC signalling pathway in

BioMed Research International 9

reverse intervention for the ACLndashhamstring reflex arc withelectroacupuncture Because these limitations existed thisstudy is considered basic All these issues remain to be solvedby further related studies

5 Conclusions

In this study the mechanism of reverse intervention for ACLproprioception with electroacupuncture which was basedon the ACLndashhamstring reflex arc and through mediationof the NT-3TrkC signalling pathway was explored Thisstudy helped establish the basis for the clinical application ofelectroacupuncture therapy and effective treatment methodsand patterns for proprioception disturbances caused by ACLinjuries thereby enriching and developing the scientificconnotation of electroacupuncture therapy guided by theory

Disclosure

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan are co-firstauthors

Conflicts of Interest

The authors declare that they have no conflicts of interest

Authorsrsquo Contributions

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan contributedequally to this work

Acknowledgments

This study was supported by the National Natural ScienceFoundation of China (no 81674095) Science-TechnologySupport Plan Project of Sichuan Province (no 2014SZ0185)and Science and Technology Strategic CooperationProject of Luzhou City-Southwest Medical University(no 2016LZXNYD-J08)

References

[1] H Zhang M Qiu A Zhou J Zhang and D Jiang ldquoAnatomicanterolateral ligament reconstruction improves postoperativeclinical outcomes combined with anatomic anterior cruciateligament reconstructionrdquo Journal of Sports Science andMedicine vol 15 no 4 pp 688ndash696 2016

[2] A A Rahnemai-Azar S Sabzevari S Irarrazaval T Chaoand F H Fu ldquoAnatomical individualized ACL reconstructionrdquoArchives of Bone and Joint Surgery vol 4 no 4 pp 291ndash2972016

[3] M R Sheth S R Tapasvi and S S Patil ldquoPrimary Repairof Tibial-Sided Avulsion of the Anterior Cruciate LigamentrdquoArthroscopy Techniques vol 5 no 4 pp e901ndashe906 2016

[4] F Sepulveda L Sanchez E Amy and W Micheo ldquoAnteriorCruciate Ligament InjuryrdquoCurrent SportsMedicine Reports vol16 no 3 pp 172ndash178 2017

[5] T M El-Gohary O A Khaled S R Ibrahim A M Alshenqitiand M I Ibrahim ldquoEffect of proprioception cross training on

repositioning accuracy and balance among healthy individualsrdquoJournal of PhysicalTherapy Science vol 28 no 11 pp 3178ndash31822016

[6] A Delle Vedove M Storbeck R Heller et al ldquoBiallelic loss ofproprioception-related PIEZO2 causes muscular atrophy withperinatal respiratory distress arthrogryposis and scoliosisrdquoAmerican Journal of Human Genetics vol 99 no 6 pp 1406ndash1408 2016

[7] W K Chang Y S Jung M Oh and K Kim ldquoQuantitativeassessment of proprioception using dynamometer in incom-plete spinal cord injury patients a preliminary studyrdquo Annalsof Rehabilitation Medicine vol 41 no 2 pp 218ndash224 2017

[8] G Haliloglu K Becker C Temucin et al ldquoRecessive PIEZO2stop mutation causes distal arthrogryposis with distal muscleweakness scoliosis and proprioception defectsrdquo Journal ofHuman Genetics vol 62 no 4 pp 497ndash501 2017

[9] J Li F Kong X Gao Y Shen and S Gao ldquoProspectiverandomized comparison of knee stability and proprioceptionfor posterior cruciate ligament reconstruction with autografthybrid graft and 120574-irradiated allograftrdquoArthroscopy vol 32 no12 pp 2548ndash2555 2016

[10] J F Abulhasan C M Anley M D Snow and M J GreyldquoHamstring stretch reflex could it be a reproducible objectivemeasure of functional knee stabilityrdquordquo Journal of ExperimentalOrthopaedics vol 3 no 1 2016

[11] M Solomonow R Baratta B H Zhou et al ldquoThe synergisticaction of the anterior cruciate ligament and thigh musclesin maintaining joint stabilityrdquo The American Journal of SportsMedicine vol 15 no 3 pp 207ndash213 1987

[12] P Dyhre-Poulsen and M R Krogsgaard ldquoMuscular reflexeselicited by electrical stimulation of the anterior cruciate liga-ment in humansrdquo Journal of Applied Physiology vol 89 no 6pp 2191ndash2195 2000

[13] E Tsuda Y Ishibashi Y Okamura and S Toh ldquoRestorationof anterior cruciate ligament-hamstring reflex arc after anteriorcruciate ligament reconstructionrdquoKnee Surgery Sports Trauma-tology Arthroscopy vol 11 no 2 pp 63ndash67 2003

[14] X-Y Wang P-Y Gu S-W Chen et al ldquoEndogenousneurotrophin-3 promotes neuronal sprouting from dorsalroot gangliardquo Neural Regeneration Research vol 10 no 11 pp1865ndash1868 2015

[15] K A Han D Woo S Kim et al ldquoNeurotrophin-3 regu-lates synapse development by modulating TrkC-PTP120590 synapticadhesion and intracellular signaling pathwaysrdquo The Journal ofNeuroscience vol 36 no 17 pp 4816ndash4831 2016

[16] M-H Yang K-C Chen P-W Chiang et al ldquoProteomicprofiling of neuroblastoma cells adhesion on hyaluronic acid-based surface for neural tissue engineeringrdquo BioMed ResearchInternational vol 2016 Article ID 1917394 2016

[17] S Rao M Martınez-Cengotitabengoa Y Yao et al ldquoPeripheralblood nerve growth factor levels inmajor psychiatric disordersrdquoJournal of Psychiatric Research vol 86 pp 39ndash45 2016

[18] M Yong S Kim and S Cheon ldquoEffects of skilled reach trainingwith affected forelimb and treadmill exercise on the expressionof neurotrophic factor following ischemia-induced brain injuryin ratsrdquo Journal of Physical Therapy Science vol 29 no 4 pp647ndash650 2017

[19] S Liu S Blanchard S Bigou S Vitry D Bohl and J-M HeardldquoNeurotrophin 3 improves delayed reconstruction of sensorypathways after cervical dorsal root injuryrdquoNeurosurgery vol 68no 2 pp 450ndash461 2011

10 BioMed Research International

[20] J Cui C Cui Y Cui et al ldquoBone marrow mesenchymal stemcell transplantation increases GAP-43 Expression via ERK12and PI3KAkt pathways in intracerebral hemorrhagerdquo CellularPhysiology and Biochemistry vol 42 no 1 pp 137ndash144 2017

[21] Y Li XGaoQWang et al ldquoRetinoic acid protects from experi-mental cerebral infarction by upregulating GAP-43 expressionrdquoBrazilian Journal of Medical and Biological Research vol 50 no4 2017

[22] H B Kim and B S Yoo ldquoPropolis inhibits neurite outgrowthin differentiating SH-SY5Y human neuroblastoma cellsrdquo Toxi-cological Research vol 32 no 3 pp 239ndash243 2016

[23] J Iwasa M Ochi Y Uchio N Adachi and K KawasakildquoDecrease in anterior knee laxity by electrical stimulation ofnormal and reconstructed anterior cruciate ligamentsrdquo TheJournal of Boneamp Joint Surgery vol 88 no 4 pp 477ndash483 2006

[24] A Biscarini F M Botti and V E Pettorossi ldquoSelectivecontribution of each hamstring muscle to anterior cruciateligament protection and tibiofemoral joint stability in leg-extension exercise A simulation studyrdquo European Journal ofApplied Physiology vol 113 no 9 pp 2263ndash2273 2013

[25] D Sugimoto E Alentorn-Geli J Mendiguchıa K SamuelssonJ Karlsson andGDMyer ldquoBiomechanical and neuromuscularcharacteristics of male athletes implications for the develop-ment of anterior cruciate ligament injury prevention programsrdquoSports Medicine vol 45 no 6 pp 809ndash822 2015

[26] M G Barcellona M C Morrissey P Milligan M Clinton andA A Amis ldquoThe effect of knee extensor open kinetic chainresistance training in the ACL-injured kneerdquo Knee SurgerySports Traumatology Arthroscopy vol 23 no 11 pp 3168ndash31772015

[27] T S Furlanetto L A Peyre-Tartaruga A S Do Pinho E SDa Bernardes and M A Zaro ldquoProprioception body balanceand functionality in individuals with ACL reconstructionrdquoActaOrtopedica Brasileira vol 24 no 2 pp 67ndash72 2016

[28] A J Wodowski C W Swigler H Liu K M Nord P C ToyandW M Mihalko ldquoProprioception and Knee Arthroplasty ALiterature ReviewrdquoOrthopedic Clinics of North America vol 47no 2 pp 301ndash309 2016

[29] I Dallo J Chahla J JMitchell C Pascual-Garrido J A Feaginand R F LaPrade ldquoBiologic Approaches for the Treatment ofPartial Tears of the Anterior Cruciate Ligament A CurrentConcepts Reviewrdquo Orthopaedic Journal of Sports Medicine vol5 no 1 2017

[30] W R Lowe R J Warth E P Davis and L Bailey ldquoFunc-tional bracing after anterior cruciate ligament reconstructionA systematic reviewrdquo Journal of the American Academy ofOrthopaedicSurgeons vol 25 no 3 pp 239ndash249 2017

[31] Y Ma M Deie D Iwaki et al ldquoBalance Ability and Pro-prioception after Single-Bundle Single-Bundle Augmentationand Double-Bundle ACL ReconstructionrdquoThe Scientific WorldJournal vol 2014 Article ID 342012 8 pages 2014

[32] N Relph L Herrington and S Tyson ldquoThe effects of ACLinjury on knee proprioception Ameta-analysisrdquo Physiotherapyvol 100 no 3 pp 187ndash195 2014

[33] B P Livingston and T R Nichols ldquoEffects of reinnervation ofthe triceps brachii on joint kinematics and electromyographicpatterns of the feline forelimb during level and upslope walk-ingrdquo Cells Tissues Organs vol 199 no 5-6 pp 405ndash422 2014

[34] E Appel S Weissmann Y Salzberg et al ldquoAn ensemble ofregulatory elements controls Runx3 spatiotemporal expressionin subsets of dorsal root ganglia proprioceptive neuronsrdquoGenesamp Development vol 30 no 23 pp 2607ndash2622 2016

[35] S M OrsquoToole M M Ferrer J Mekonnen et al ldquoDicermaintains the identity and function of proprioceptive sensoryneuronsrdquo Journal of Neurophysiology vol 117 no 3 pp 1057ndash1069 2017

[36] Y Liu L Kelamangalath H Kim et al ldquoNT-3 promotes pro-prioceptive axon regeneration when combined with activationof the mTor intrinsic growth pathway but not with reduction ofmyelin extrinsic inhibitorsrdquo Experimental Neurology vol 283pp 73ndash84 2016

[37] S Li T Shen Y Liang Y Zhang and B Bai ldquoEffects ofminiscalpel-needle release on chronic neck pain A retrospec-tive analysis with 12-month follow-uprdquo PLoS ONE vol 10 no8 Article ID e0137033 pp 133ndash137 2015

[38] W-S Juan S-Y Huang C-C Chang et al ldquoMelatoninimproves neuroplasticity by upregulating the growth-associatedprotein-43 (GAP-43) and NMDAR postsynaptic density-95(PSD-95) proteins in cultured neurons exposed to glutamateexcitotoxicity and in rats subjected to transient focal cerebralischemia even during a long-term recovery periodrdquo Journal ofPineal Research vol 56 no 2 pp 213ndash223 2014

[39] Y-HGaoC-W Li J-YWang et al ldquoActivation of hippocampalMEK1 contributes to the cumulative antinociceptive effect ofelectroacupuncture in neuropathic pain ratsrdquo BMC Comple-mentary and Alternative Medicine vol 16 no 1 article no 5172016

[40] K-Y Huang S Liang M-L Yu S-P Fu X Chen and S-F Lu ldquoA systematic review and meta-analysis of acupuncturefor improving learning and memory ability in animalsrdquo BMCComplementary and Alternative Medicine vol 16 no 1 articleno 297 2016

[41] J Zhou S Li Y Wang et al ldquoEffects and mechanisms ofauricular electroacupuncture on gastric hypersensitivity in arodent model of functional dyspepsiardquo PLoS ONE vol 12 no3 Article ID e0174568 2017

[42] Y H Gao C W Li J Y Wang et al ldquoEffect of electroacupunc-ture on the cervicospinal P2X7 receptorfractalkineCX3CR1signaling pathway in a rat neck-incision painmodelrdquo PurinergicSignalling vol 13 no 2 pp 215ndash225 2017

[43] JH LeeDGoWKimet al ldquoInvolvement of spinalmuscarinicand serotonergic receptors in the anti-allodynic effect of elec-troacupuncture in rats with oxaliplatin-induced neuropathicpainrdquoKorean Journal of Physiology amp Pharmacology vol 20 no4 pp 407ndash414 2016

[44] S P Chen Y Kan J L Zhang et al ldquoInvolvement of hippocam-pal acetylcholinergic receptors in electroacupuncture analgesiain neuropathic pain ratsrdquo Behavioral and Brain Functions vol12 no 1 article no 13 2016

[45] M Villarreal Santiago S Tumilty A Mącznik and R ManildquoDoes acupuncture alter pain-related functional connectivity ofthe central nervous system a systematic reviewrdquo JAMS Journalof Acupuncture and Meridian Studies vol 9 no 4 pp 167ndash1772016

[46] T E Salazar M R Richardson E Beli et al ldquoElectroacupunc-ture Promotes Central Nervous System-Dependent Release ofMesenchymal Stem Cellsrdquo Stem Cells vol 35 no 5 pp 1303ndash1315 2017

[47] D-H Lee J-H Lee S-E Ahn and M-J Park ldquoEffect of timeafter anterior cruciate ligament tears on proprioception andpostural stabilityrdquo PLoS ONE vol 10 no 9 Article ID e01390382015

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

OncologyJournal of

Hindawiwwwhindawicom Volume 2013

Hindawiwwwhindawicom Volume 2018

Oxidative Medicine and Cellular Longevity

Hindawiwwwhindawicom Volume 2018

PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Computational and Mathematical Methods in Medicine

Hindawiwwwhindawicom Volume 2018

Behavioural Neurology

OphthalmologyJournal of

Hindawiwwwhindawicom Volume 2018

Diabetes ResearchJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Research and TreatmentAIDS

Hindawiwwwhindawicom Volume 2018

Gastroenterology Research and Practice

Hindawiwwwhindawicom Volume 2018

Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

BioMed Research International 9

reverse intervention for the ACLndashhamstring reflex arc withelectroacupuncture Because these limitations existed thisstudy is considered basic All these issues remain to be solvedby further related studies

5 Conclusions

In this study the mechanism of reverse intervention for ACLproprioception with electroacupuncture which was basedon the ACLndashhamstring reflex arc and through mediationof the NT-3TrkC signalling pathway was explored Thisstudy helped establish the basis for the clinical application ofelectroacupuncture therapy and effective treatment methodsand patterns for proprioception disturbances caused by ACLinjuries thereby enriching and developing the scientificconnotation of electroacupuncture therapy guided by theory

Disclosure

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan are co-firstauthors

Conflicts of Interest

The authors declare that they have no conflicts of interest

Authorsrsquo Contributions

Lei Zhang Yan Zeng Ji Qi and Taiyuan Guan contributedequally to this work

Acknowledgments

This study was supported by the National Natural ScienceFoundation of China (no 81674095) Science-TechnologySupport Plan Project of Sichuan Province (no 2014SZ0185)and Science and Technology Strategic CooperationProject of Luzhou City-Southwest Medical University(no 2016LZXNYD-J08)

References

[1] H Zhang M Qiu A Zhou J Zhang and D Jiang ldquoAnatomicanterolateral ligament reconstruction improves postoperativeclinical outcomes combined with anatomic anterior cruciateligament reconstructionrdquo Journal of Sports Science andMedicine vol 15 no 4 pp 688ndash696 2016

[2] A A Rahnemai-Azar S Sabzevari S Irarrazaval T Chaoand F H Fu ldquoAnatomical individualized ACL reconstructionrdquoArchives of Bone and Joint Surgery vol 4 no 4 pp 291ndash2972016

[3] M R Sheth S R Tapasvi and S S Patil ldquoPrimary Repairof Tibial-Sided Avulsion of the Anterior Cruciate LigamentrdquoArthroscopy Techniques vol 5 no 4 pp e901ndashe906 2016

[4] F Sepulveda L Sanchez E Amy and W Micheo ldquoAnteriorCruciate Ligament InjuryrdquoCurrent SportsMedicine Reports vol16 no 3 pp 172ndash178 2017

[5] T M El-Gohary O A Khaled S R Ibrahim A M Alshenqitiand M I Ibrahim ldquoEffect of proprioception cross training on

repositioning accuracy and balance among healthy individualsrdquoJournal of PhysicalTherapy Science vol 28 no 11 pp 3178ndash31822016

[6] A Delle Vedove M Storbeck R Heller et al ldquoBiallelic loss ofproprioception-related PIEZO2 causes muscular atrophy withperinatal respiratory distress arthrogryposis and scoliosisrdquoAmerican Journal of Human Genetics vol 99 no 6 pp 1406ndash1408 2016

[7] W K Chang Y S Jung M Oh and K Kim ldquoQuantitativeassessment of proprioception using dynamometer in incom-plete spinal cord injury patients a preliminary studyrdquo Annalsof Rehabilitation Medicine vol 41 no 2 pp 218ndash224 2017

[8] G Haliloglu K Becker C Temucin et al ldquoRecessive PIEZO2stop mutation causes distal arthrogryposis with distal muscleweakness scoliosis and proprioception defectsrdquo Journal ofHuman Genetics vol 62 no 4 pp 497ndash501 2017

[9] J Li F Kong X Gao Y Shen and S Gao ldquoProspectiverandomized comparison of knee stability and proprioceptionfor posterior cruciate ligament reconstruction with autografthybrid graft and 120574-irradiated allograftrdquoArthroscopy vol 32 no12 pp 2548ndash2555 2016

[10] J F Abulhasan C M Anley M D Snow and M J GreyldquoHamstring stretch reflex could it be a reproducible objectivemeasure of functional knee stabilityrdquordquo Journal of ExperimentalOrthopaedics vol 3 no 1 2016

[11] M Solomonow R Baratta B H Zhou et al ldquoThe synergisticaction of the anterior cruciate ligament and thigh musclesin maintaining joint stabilityrdquo The American Journal of SportsMedicine vol 15 no 3 pp 207ndash213 1987

[12] P Dyhre-Poulsen and M R Krogsgaard ldquoMuscular reflexeselicited by electrical stimulation of the anterior cruciate liga-ment in humansrdquo Journal of Applied Physiology vol 89 no 6pp 2191ndash2195 2000

[13] E Tsuda Y Ishibashi Y Okamura and S Toh ldquoRestorationof anterior cruciate ligament-hamstring reflex arc after anteriorcruciate ligament reconstructionrdquoKnee Surgery Sports Trauma-tology Arthroscopy vol 11 no 2 pp 63ndash67 2003

[14] X-Y Wang P-Y Gu S-W Chen et al ldquoEndogenousneurotrophin-3 promotes neuronal sprouting from dorsalroot gangliardquo Neural Regeneration Research vol 10 no 11 pp1865ndash1868 2015

[15] K A Han D Woo S Kim et al ldquoNeurotrophin-3 regu-lates synapse development by modulating TrkC-PTP120590 synapticadhesion and intracellular signaling pathwaysrdquo The Journal ofNeuroscience vol 36 no 17 pp 4816ndash4831 2016

[16] M-H Yang K-C Chen P-W Chiang et al ldquoProteomicprofiling of neuroblastoma cells adhesion on hyaluronic acid-based surface for neural tissue engineeringrdquo BioMed ResearchInternational vol 2016 Article ID 1917394 2016

[17] S Rao M Martınez-Cengotitabengoa Y Yao et al ldquoPeripheralblood nerve growth factor levels inmajor psychiatric disordersrdquoJournal of Psychiatric Research vol 86 pp 39ndash45 2016

[18] M Yong S Kim and S Cheon ldquoEffects of skilled reach trainingwith affected forelimb and treadmill exercise on the expressionof neurotrophic factor following ischemia-induced brain injuryin ratsrdquo Journal of Physical Therapy Science vol 29 no 4 pp647ndash650 2017

[19] S Liu S Blanchard S Bigou S Vitry D Bohl and J-M HeardldquoNeurotrophin 3 improves delayed reconstruction of sensorypathways after cervical dorsal root injuryrdquoNeurosurgery vol 68no 2 pp 450ndash461 2011

10 BioMed Research International

[20] J Cui C Cui Y Cui et al ldquoBone marrow mesenchymal stemcell transplantation increases GAP-43 Expression via ERK12and PI3KAkt pathways in intracerebral hemorrhagerdquo CellularPhysiology and Biochemistry vol 42 no 1 pp 137ndash144 2017

[21] Y Li XGaoQWang et al ldquoRetinoic acid protects from experi-mental cerebral infarction by upregulating GAP-43 expressionrdquoBrazilian Journal of Medical and Biological Research vol 50 no4 2017

[22] H B Kim and B S Yoo ldquoPropolis inhibits neurite outgrowthin differentiating SH-SY5Y human neuroblastoma cellsrdquo Toxi-cological Research vol 32 no 3 pp 239ndash243 2016

[23] J Iwasa M Ochi Y Uchio N Adachi and K KawasakildquoDecrease in anterior knee laxity by electrical stimulation ofnormal and reconstructed anterior cruciate ligamentsrdquo TheJournal of Boneamp Joint Surgery vol 88 no 4 pp 477ndash483 2006

[24] A Biscarini F M Botti and V E Pettorossi ldquoSelectivecontribution of each hamstring muscle to anterior cruciateligament protection and tibiofemoral joint stability in leg-extension exercise A simulation studyrdquo European Journal ofApplied Physiology vol 113 no 9 pp 2263ndash2273 2013

[25] D Sugimoto E Alentorn-Geli J Mendiguchıa K SamuelssonJ Karlsson andGDMyer ldquoBiomechanical and neuromuscularcharacteristics of male athletes implications for the develop-ment of anterior cruciate ligament injury prevention programsrdquoSports Medicine vol 45 no 6 pp 809ndash822 2015

[26] M G Barcellona M C Morrissey P Milligan M Clinton andA A Amis ldquoThe effect of knee extensor open kinetic chainresistance training in the ACL-injured kneerdquo Knee SurgerySports Traumatology Arthroscopy vol 23 no 11 pp 3168ndash31772015

[27] T S Furlanetto L A Peyre-Tartaruga A S Do Pinho E SDa Bernardes and M A Zaro ldquoProprioception body balanceand functionality in individuals with ACL reconstructionrdquoActaOrtopedica Brasileira vol 24 no 2 pp 67ndash72 2016

[28] A J Wodowski C W Swigler H Liu K M Nord P C ToyandW M Mihalko ldquoProprioception and Knee Arthroplasty ALiterature ReviewrdquoOrthopedic Clinics of North America vol 47no 2 pp 301ndash309 2016

[29] I Dallo J Chahla J JMitchell C Pascual-Garrido J A Feaginand R F LaPrade ldquoBiologic Approaches for the Treatment ofPartial Tears of the Anterior Cruciate Ligament A CurrentConcepts Reviewrdquo Orthopaedic Journal of Sports Medicine vol5 no 1 2017

[30] W R Lowe R J Warth E P Davis and L Bailey ldquoFunc-tional bracing after anterior cruciate ligament reconstructionA systematic reviewrdquo Journal of the American Academy ofOrthopaedicSurgeons vol 25 no 3 pp 239ndash249 2017

[31] Y Ma M Deie D Iwaki et al ldquoBalance Ability and Pro-prioception after Single-Bundle Single-Bundle Augmentationand Double-Bundle ACL ReconstructionrdquoThe Scientific WorldJournal vol 2014 Article ID 342012 8 pages 2014

[32] N Relph L Herrington and S Tyson ldquoThe effects of ACLinjury on knee proprioception Ameta-analysisrdquo Physiotherapyvol 100 no 3 pp 187ndash195 2014

[33] B P Livingston and T R Nichols ldquoEffects of reinnervation ofthe triceps brachii on joint kinematics and electromyographicpatterns of the feline forelimb during level and upslope walk-ingrdquo Cells Tissues Organs vol 199 no 5-6 pp 405ndash422 2014

[34] E Appel S Weissmann Y Salzberg et al ldquoAn ensemble ofregulatory elements controls Runx3 spatiotemporal expressionin subsets of dorsal root ganglia proprioceptive neuronsrdquoGenesamp Development vol 30 no 23 pp 2607ndash2622 2016

[35] S M OrsquoToole M M Ferrer J Mekonnen et al ldquoDicermaintains the identity and function of proprioceptive sensoryneuronsrdquo Journal of Neurophysiology vol 117 no 3 pp 1057ndash1069 2017

[36] Y Liu L Kelamangalath H Kim et al ldquoNT-3 promotes pro-prioceptive axon regeneration when combined with activationof the mTor intrinsic growth pathway but not with reduction ofmyelin extrinsic inhibitorsrdquo Experimental Neurology vol 283pp 73ndash84 2016

[37] S Li T Shen Y Liang Y Zhang and B Bai ldquoEffects ofminiscalpel-needle release on chronic neck pain A retrospec-tive analysis with 12-month follow-uprdquo PLoS ONE vol 10 no8 Article ID e0137033 pp 133ndash137 2015

[38] W-S Juan S-Y Huang C-C Chang et al ldquoMelatoninimproves neuroplasticity by upregulating the growth-associatedprotein-43 (GAP-43) and NMDAR postsynaptic density-95(PSD-95) proteins in cultured neurons exposed to glutamateexcitotoxicity and in rats subjected to transient focal cerebralischemia even during a long-term recovery periodrdquo Journal ofPineal Research vol 56 no 2 pp 213ndash223 2014

[39] Y-HGaoC-W Li J-YWang et al ldquoActivation of hippocampalMEK1 contributes to the cumulative antinociceptive effect ofelectroacupuncture in neuropathic pain ratsrdquo BMC Comple-mentary and Alternative Medicine vol 16 no 1 article no 5172016

[40] K-Y Huang S Liang M-L Yu S-P Fu X Chen and S-F Lu ldquoA systematic review and meta-analysis of acupuncturefor improving learning and memory ability in animalsrdquo BMCComplementary and Alternative Medicine vol 16 no 1 articleno 297 2016

[41] J Zhou S Li Y Wang et al ldquoEffects and mechanisms ofauricular electroacupuncture on gastric hypersensitivity in arodent model of functional dyspepsiardquo PLoS ONE vol 12 no3 Article ID e0174568 2017

[42] Y H Gao C W Li J Y Wang et al ldquoEffect of electroacupunc-ture on the cervicospinal P2X7 receptorfractalkineCX3CR1signaling pathway in a rat neck-incision painmodelrdquo PurinergicSignalling vol 13 no 2 pp 215ndash225 2017

[43] JH LeeDGoWKimet al ldquoInvolvement of spinalmuscarinicand serotonergic receptors in the anti-allodynic effect of elec-troacupuncture in rats with oxaliplatin-induced neuropathicpainrdquoKorean Journal of Physiology amp Pharmacology vol 20 no4 pp 407ndash414 2016

[44] S P Chen Y Kan J L Zhang et al ldquoInvolvement of hippocam-pal acetylcholinergic receptors in electroacupuncture analgesiain neuropathic pain ratsrdquo Behavioral and Brain Functions vol12 no 1 article no 13 2016

[45] M Villarreal Santiago S Tumilty A Mącznik and R ManildquoDoes acupuncture alter pain-related functional connectivity ofthe central nervous system a systematic reviewrdquo JAMS Journalof Acupuncture and Meridian Studies vol 9 no 4 pp 167ndash1772016

[46] T E Salazar M R Richardson E Beli et al ldquoElectroacupunc-ture Promotes Central Nervous System-Dependent Release ofMesenchymal Stem Cellsrdquo Stem Cells vol 35 no 5 pp 1303ndash1315 2017

[47] D-H Lee J-H Lee S-E Ahn and M-J Park ldquoEffect of timeafter anterior cruciate ligament tears on proprioception andpostural stabilityrdquo PLoS ONE vol 10 no 9 Article ID e01390382015

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

OncologyJournal of

Hindawiwwwhindawicom Volume 2013

Hindawiwwwhindawicom Volume 2018

Oxidative Medicine and Cellular Longevity

Hindawiwwwhindawicom Volume 2018

PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Computational and Mathematical Methods in Medicine

Hindawiwwwhindawicom Volume 2018

Behavioural Neurology

OphthalmologyJournal of

Hindawiwwwhindawicom Volume 2018

Diabetes ResearchJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Research and TreatmentAIDS

Hindawiwwwhindawicom Volume 2018

Gastroenterology Research and Practice

Hindawiwwwhindawicom Volume 2018

Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

10 BioMed Research International

[20] J Cui C Cui Y Cui et al ldquoBone marrow mesenchymal stemcell transplantation increases GAP-43 Expression via ERK12and PI3KAkt pathways in intracerebral hemorrhagerdquo CellularPhysiology and Biochemistry vol 42 no 1 pp 137ndash144 2017

[21] Y Li XGaoQWang et al ldquoRetinoic acid protects from experi-mental cerebral infarction by upregulating GAP-43 expressionrdquoBrazilian Journal of Medical and Biological Research vol 50 no4 2017

[22] H B Kim and B S Yoo ldquoPropolis inhibits neurite outgrowthin differentiating SH-SY5Y human neuroblastoma cellsrdquo Toxi-cological Research vol 32 no 3 pp 239ndash243 2016

[23] J Iwasa M Ochi Y Uchio N Adachi and K KawasakildquoDecrease in anterior knee laxity by electrical stimulation ofnormal and reconstructed anterior cruciate ligamentsrdquo TheJournal of Boneamp Joint Surgery vol 88 no 4 pp 477ndash483 2006

[24] A Biscarini F M Botti and V E Pettorossi ldquoSelectivecontribution of each hamstring muscle to anterior cruciateligament protection and tibiofemoral joint stability in leg-extension exercise A simulation studyrdquo European Journal ofApplied Physiology vol 113 no 9 pp 2263ndash2273 2013

[25] D Sugimoto E Alentorn-Geli J Mendiguchıa K SamuelssonJ Karlsson andGDMyer ldquoBiomechanical and neuromuscularcharacteristics of male athletes implications for the develop-ment of anterior cruciate ligament injury prevention programsrdquoSports Medicine vol 45 no 6 pp 809ndash822 2015

[26] M G Barcellona M C Morrissey P Milligan M Clinton andA A Amis ldquoThe effect of knee extensor open kinetic chainresistance training in the ACL-injured kneerdquo Knee SurgerySports Traumatology Arthroscopy vol 23 no 11 pp 3168ndash31772015

[27] T S Furlanetto L A Peyre-Tartaruga A S Do Pinho E SDa Bernardes and M A Zaro ldquoProprioception body balanceand functionality in individuals with ACL reconstructionrdquoActaOrtopedica Brasileira vol 24 no 2 pp 67ndash72 2016

[28] A J Wodowski C W Swigler H Liu K M Nord P C ToyandW M Mihalko ldquoProprioception and Knee Arthroplasty ALiterature ReviewrdquoOrthopedic Clinics of North America vol 47no 2 pp 301ndash309 2016

[29] I Dallo J Chahla J JMitchell C Pascual-Garrido J A Feaginand R F LaPrade ldquoBiologic Approaches for the Treatment ofPartial Tears of the Anterior Cruciate Ligament A CurrentConcepts Reviewrdquo Orthopaedic Journal of Sports Medicine vol5 no 1 2017

[30] W R Lowe R J Warth E P Davis and L Bailey ldquoFunc-tional bracing after anterior cruciate ligament reconstructionA systematic reviewrdquo Journal of the American Academy ofOrthopaedicSurgeons vol 25 no 3 pp 239ndash249 2017

[31] Y Ma M Deie D Iwaki et al ldquoBalance Ability and Pro-prioception after Single-Bundle Single-Bundle Augmentationand Double-Bundle ACL ReconstructionrdquoThe Scientific WorldJournal vol 2014 Article ID 342012 8 pages 2014

[32] N Relph L Herrington and S Tyson ldquoThe effects of ACLinjury on knee proprioception Ameta-analysisrdquo Physiotherapyvol 100 no 3 pp 187ndash195 2014

[33] B P Livingston and T R Nichols ldquoEffects of reinnervation ofthe triceps brachii on joint kinematics and electromyographicpatterns of the feline forelimb during level and upslope walk-ingrdquo Cells Tissues Organs vol 199 no 5-6 pp 405ndash422 2014

[34] E Appel S Weissmann Y Salzberg et al ldquoAn ensemble ofregulatory elements controls Runx3 spatiotemporal expressionin subsets of dorsal root ganglia proprioceptive neuronsrdquoGenesamp Development vol 30 no 23 pp 2607ndash2622 2016

[35] S M OrsquoToole M M Ferrer J Mekonnen et al ldquoDicermaintains the identity and function of proprioceptive sensoryneuronsrdquo Journal of Neurophysiology vol 117 no 3 pp 1057ndash1069 2017

[36] Y Liu L Kelamangalath H Kim et al ldquoNT-3 promotes pro-prioceptive axon regeneration when combined with activationof the mTor intrinsic growth pathway but not with reduction ofmyelin extrinsic inhibitorsrdquo Experimental Neurology vol 283pp 73ndash84 2016

[37] S Li T Shen Y Liang Y Zhang and B Bai ldquoEffects ofminiscalpel-needle release on chronic neck pain A retrospec-tive analysis with 12-month follow-uprdquo PLoS ONE vol 10 no8 Article ID e0137033 pp 133ndash137 2015

[38] W-S Juan S-Y Huang C-C Chang et al ldquoMelatoninimproves neuroplasticity by upregulating the growth-associatedprotein-43 (GAP-43) and NMDAR postsynaptic density-95(PSD-95) proteins in cultured neurons exposed to glutamateexcitotoxicity and in rats subjected to transient focal cerebralischemia even during a long-term recovery periodrdquo Journal ofPineal Research vol 56 no 2 pp 213ndash223 2014

[39] Y-HGaoC-W Li J-YWang et al ldquoActivation of hippocampalMEK1 contributes to the cumulative antinociceptive effect ofelectroacupuncture in neuropathic pain ratsrdquo BMC Comple-mentary and Alternative Medicine vol 16 no 1 article no 5172016

[40] K-Y Huang S Liang M-L Yu S-P Fu X Chen and S-F Lu ldquoA systematic review and meta-analysis of acupuncturefor improving learning and memory ability in animalsrdquo BMCComplementary and Alternative Medicine vol 16 no 1 articleno 297 2016

[41] J Zhou S Li Y Wang et al ldquoEffects and mechanisms ofauricular electroacupuncture on gastric hypersensitivity in arodent model of functional dyspepsiardquo PLoS ONE vol 12 no3 Article ID e0174568 2017

[42] Y H Gao C W Li J Y Wang et al ldquoEffect of electroacupunc-ture on the cervicospinal P2X7 receptorfractalkineCX3CR1signaling pathway in a rat neck-incision painmodelrdquo PurinergicSignalling vol 13 no 2 pp 215ndash225 2017

[43] JH LeeDGoWKimet al ldquoInvolvement of spinalmuscarinicand serotonergic receptors in the anti-allodynic effect of elec-troacupuncture in rats with oxaliplatin-induced neuropathicpainrdquoKorean Journal of Physiology amp Pharmacology vol 20 no4 pp 407ndash414 2016

[44] S P Chen Y Kan J L Zhang et al ldquoInvolvement of hippocam-pal acetylcholinergic receptors in electroacupuncture analgesiain neuropathic pain ratsrdquo Behavioral and Brain Functions vol12 no 1 article no 13 2016

[45] M Villarreal Santiago S Tumilty A Mącznik and R ManildquoDoes acupuncture alter pain-related functional connectivity ofthe central nervous system a systematic reviewrdquo JAMS Journalof Acupuncture and Meridian Studies vol 9 no 4 pp 167ndash1772016

[46] T E Salazar M R Richardson E Beli et al ldquoElectroacupunc-ture Promotes Central Nervous System-Dependent Release ofMesenchymal Stem Cellsrdquo Stem Cells vol 35 no 5 pp 1303ndash1315 2017

[47] D-H Lee J-H Lee S-E Ahn and M-J Park ldquoEffect of timeafter anterior cruciate ligament tears on proprioception andpostural stabilityrdquo PLoS ONE vol 10 no 9 Article ID e01390382015

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

OncologyJournal of

Hindawiwwwhindawicom Volume 2013

Hindawiwwwhindawicom Volume 2018

Oxidative Medicine and Cellular Longevity

Hindawiwwwhindawicom Volume 2018

PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Computational and Mathematical Methods in Medicine

Hindawiwwwhindawicom Volume 2018

Behavioural Neurology

OphthalmologyJournal of

Hindawiwwwhindawicom Volume 2018

Diabetes ResearchJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Research and TreatmentAIDS

Hindawiwwwhindawicom Volume 2018

Gastroenterology Research and Practice

Hindawiwwwhindawicom Volume 2018

Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom

Stem Cells International

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

MEDIATORSINFLAMMATION

of

EndocrinologyInternational Journal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Disease Markers

Hindawiwwwhindawicom Volume 2018

BioMed Research International

OncologyJournal of

Hindawiwwwhindawicom Volume 2013

Hindawiwwwhindawicom Volume 2018

Oxidative Medicine and Cellular Longevity

Hindawiwwwhindawicom Volume 2018

PPAR Research

Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Immunology ResearchHindawiwwwhindawicom Volume 2018

Journal of

ObesityJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Computational and Mathematical Methods in Medicine

Hindawiwwwhindawicom Volume 2018

Behavioural Neurology

OphthalmologyJournal of

Hindawiwwwhindawicom Volume 2018

Diabetes ResearchJournal of

Hindawiwwwhindawicom Volume 2018

Hindawiwwwhindawicom Volume 2018

Research and TreatmentAIDS

Hindawiwwwhindawicom Volume 2018

Gastroenterology Research and Practice

Hindawiwwwhindawicom Volume 2018

Parkinsonrsquos Disease

Evidence-Based Complementary andAlternative Medicine

Volume 2018Hindawiwwwhindawicom

Submit your manuscripts atwwwhindawicom