Motor neuron-specific RhoA knockout delays degeneration and promotes regeneration of dendrites in spinal ventral horn after brachial plexus injury

Dendrites play irreplaceable roles in the nerve conduction pathway and are vulnerable to various insults.Peripheral axotomy of motor neurons results in the retraction of dendritic arbors,and the dendritic arbor can be re-expanded when reinnervation is allowed.RhoA is a target that regulates the cytoskeleton and promotes neuronal survival and axon regeneration.However,the role of RhoA in dendrite degeneration and regeneration is unknown.In this study,we explored the potential role of RhoA in dendrites.A line of motor neuronal conditional knockout mice was developed by crossbreeding HB9~(Cre+)mice with RhoA~(flox/flox)mice.We established two models for assaying dendrite degeneration and regeneration,in which the brachial plexus was transection or crush injured,respectively.We found that at 28 days after brachial plexus transection,the density,complexity,and structural integrity of dendrites in the ventral horn of the spinal cord of RhoA conditional knockout mice were slightly decreased compared with that in Cre mice.Dendrites underwent degeneration at 7 and 14 days after brachial plexus transection and recovered at 28–56 days.The density,complexity,and structural integrity of dendrites in the ventral horn of the spinal cord of RhoA conditional knockout mice recovered compared with results in Cre mice.These findings suggest that RhoA knockout in motor neurons attenuates dendrite degeneration and promotes dendrite regeneration after peripheral nerve injury.

Hemodynamic instability following intravenous dexmedetomidine infusion for sedation under brachial plexus block: Two case reports

BACKGROUND Dexmedetomidine(DMED)is frequently used as a sedative in several medical fields.The benefits of DMED include enhanced quality of regional anesthesia,prolonged analgesia,and postoperative opioid-sparing when administered intravenously or perineurally in combination with regional anesthesia.Severe hemodynamic complications,such as profound bradycardia and hypotension,can occur after DMED administration in critically ill patients or overdosage;however,there are few reports of complications with DMED administration following brachial plexus block(BPB).CASE SUMMARY We present two cases of hemodynamic instability that occurred following the initial loading of DMED under supraclavicular BPB.A healthy 29-year-old man without any medical history showed profound bradycardia after receiving a loading dose of DMED 0.9μg/kg for 9 min.DMED administration was promptly stopped,and after receiving a second dose of atropine,the heart rate recovered.A 62-year-old woman with a history of cardiomyopathy became hypotensive abruptly,requiring the administration of inotrope and vasopressors after receiving a reduced loading dose of 0.5μg/kg for 10 min.Half of the recommended loading dose of DMED was administered due to the underlying heart dysfunction.Decrea-sed blood pressure was maintained despite the intravenous administration of ephedrine.With continuous infusion of dopamine and norepinephrine,the vital signs were maintained within normal ranges.Inotropic and vasopressor support was required for over 6 h after the initial loading dose of DMED.CONCLUSION DMED administration following BPB could trigger hemodynamic instability in patients with decreased cardiac function as well as in healthy individuals.

Axillary Vessels and Brachial Plexus Traumas in Abidjan: Lesional Aspects and Surgical Difficulties

Introduction-Objectives: Through the presentation of epidemiological, anatomo-clinical and surgical aspects, we report our experience in the management of traumatic axillary lesions. Materials and Methods: A descriptive retrospective study was based on the medical records of patients who suffered vascular axillary and/or brachial plexus trauma and who underwent surgical repair at the Abidjan Cardiology Institute from January 2008 to June 2022. Epidemiological, anatomo-clinical and surgical data were studied. Results: Thirty-four medical files belonging to 33 men and one woman, aged 32 on average, were collected. The circumstances of occurrence were dominated by the stab wound (n = 22). The combinations of injuries were as follows: associated involvement of the axillary artery and vein (n = 4);isolated involvement of axillary artery (n = 3);isolated involvement of the axillary vein (n = 2);associated involvement of the axillary artery and brachial plexus (n = 17);associated involvement of the axillary artery and vein and brachial plexus (n = 08). Anatomic lesions included acute arterial lesions (n = 29) and arteriovenous fistula (n = 1) and false aneurysms (n = 4). All patients were operated on under general anesthesia;vascular repair included direct suturing (n = 16), arterial and venous bypass using a long saphenous graft (n = 9), prosthetic arterial bypass (n = 5) and prosthetic flattening-graft (n = 4). Brachial plexus surgery consisted of an end-to-end anastomosis of each transected bundle in all cases (n = 25). The medium-term postoperative course was marked by success without functional sequelae in 88.24% of cases (n = 30) and by the persistence of distal paralysis of the thoracic limb after 6 months in 05.88% (n = 2) of all patients, i.e., 8% of patients who presented with brachial plexus injury. Conclusion: The concomitant surgical treatment of these axillary vascular and nerve lesions has given good results. However, if paralysis of the thoracic limb persists after 6 to 12 months, the patient should be referred to a specialist in brachial plexus surgery.

Electroacupuncture attenuates neuropathic pain after brachial plexus injury

Electroacupuncture has traditionally been used to treat pain, but its effect on pain following brachial plexus injury is still unknown. In this study, rat models of an avulsion injury to the left brachial plexus root （associated with upper-limb chronic neuropathic pain） were given electroacupuncture stimulation at bilateral Quchi （LIll）, Hegu （LI04）, Zusanli （ST36） and Yanglingquan （GB34）. After electroacupuncture therapy, chronic neuropathic pain in the rats＇ upper limbs was significantly attenuated. Immunofluorescence staining showed that the expression of β-endorphins in the arcuate nucleus was significantly increased after therapy. Thus, experimental findings indi- cate that electroacupuncture can attenuate neuropathic pain after brachial plexus injury through upregulating β-endorphin expression.

Structure of the brachial plexus root and adjacent regions displayed by ultrasound imaging

Brachial plexuses of 110 healthy volunteers were examined using high resolution color Doppler ultrasound. Ultrasonic characteristics and anatomic variation in the intervertebral foramen, interscalene, supraclavicular and infraclavicular, as well as the axillary brachial plexus were investigated. Results confirmed that the normal brachial plexus on cross section exhibited round or elliptic hypoechoic texture. Longitudinal section imaging showed many parallel linear hypo-moderate echoes, with hypo-echo. The transverse processes of the seventh cervical vertebra, the scalene space, the subclavian artery and the deep cervical artery are important markers in an examination. The display rates for the interscalene, and supraclavicular and axillary brachial plexuses were 100% each, while that for the infraclavicular brachial plexus was 97%. The region where the normal brachial plexus root traversed the intervertebral foramen exhibited a regular hypo-echo. The display rate for the C5-7 nerve roots was 100%, while those for C8 and T1 were 83% and 68%, respectively. A total of 20 of the 110 subjects underwent cervical CT scan. High-frequency ultrasound can clearly display the outline of the transverse processes of the vertebrae, which were consistent with CT results. These results indicate that high-frequency ultrasound provides a new method for observing the morphology of the brachial plexus. The C~ vertebra is a marker for identifying the position of brachial plexus nerve roots.

Human amniotic epithelial cell transplantation for the repair of injured brachial plexus nerve: evaluation of nerve viscoelastic properties

The transplantation of embryonic stem cells can effectively improve the creeping strength of nerves near an injury site in animals. Amniotic epithelial cells have similar biological properties as em-bryonic stem cells; therefore, we hypothesized that transplantation of amniotic epithelial cells can repair peripheral nerve injury and recover the creeping strength of the brachial plexus nerve. In the present study, a brachial plexus injury model was established in rabbits using the C6root avulsion method. A suspension of human amniotic epithelial cells was repeatedly injected over an area 4.0 mm lateral to the cephal and caudal ends of the C6 brachial plexus injury site （1 × 106 cells/mL, 3μL/injection, 25 injections） immediately after the injury. The results showed that the decrease in stress and increase in strain at 7,200 seconds in the injured rabbit C6 brachial plexus nerve were mitigated by the cell transplantation, restoring the viscoelastic stress relaxation and creep properties of the brachial plexus nerve. The forepaw functions were also signiifcantly improved at 26 weeks after injury. These data indicate that transplantation of human amniotic epithelial cells can effec-tively restore the mechanical properties of the brachial plexus nerve after injury in rabbits and that viscoelasticity may be an important index for the evaluation of brachial plexus injury in animals.

Magnetic resonance imaging with three-dimensional fast imaging employing steady-state acquisition with phase-cycled and short T1 inversion recovery pulse sequence for evaluating brachial plexus injury

There is a large amount of fat in the postganglionic segment of the brachial plexus nerve.The use of short T1 inversion recovery pulse sequence may improve signal strength of the brachial plexus postganglionic segment.The present study revealed that the combination of three-dimensional fast imaging employing steady-state acquisition with phase-cycled and short T1 inversion recovery pulse sequence clearly displayed the anatomical morphology and structure of the brachial plexus nerve,together with maximum intensity projection,volume rendering and other three-dimensional reconstruction techniques.Our results suggested that this method is also suitable for providing accurate assessment and diagnosis of the site,severity and scope of brachial plexus injury.

Small-worldness of brain networks after brachial plexus injury: a resting-state functional magnetic resonance imaging study

Research on brain function after brachial plexus injury focuses on local cortical functional reorganization,and few studies have focused on brain networks after brachial plexus injury.Changes in brain networks may help understanding of brain plasticity at the global level.We hypothesized that topology of the global cerebral resting-state functional network changes after unilateral brachial plexus injury.Thus,in this cross-sectional study,we recruited eight male patients with unilateral brachial plexus injury（right handedness,mean age of 27.9±5.4years old）and eight male healthy controls（right handedness,mean age of 28.6±3.2）.After acquiring and preprocessing resting-state magnetic resonance imaging data,the cerebrum was divided into 90 regions and Pearson’s correlation coefficient calculated between regions.These correlation matrices were then converted into a binary matrix with affixed sparsity values of 0.1–0.46.Under sparsity conditions,both groups satisfied this small-world property.The clustering coefficient was markedly lower,while average shortest path remarkably higher in patients compared with healthy controls.These findings confirm that cerebral functional networks in patients still show smallworld characteristics,which are highly effective in information transmission in the brain,as well as normal controls.Alternatively,varied small-worldness suggests that capacity of information transmission and integration in different brain regions in brachial plexus injury patients is damaged.

Total brachial plexus injury： contralateral C7 root transfer to the lower trunk versus the median nerve

Contralateral C7（cC7） root transfer to the healthy side is the main method for the treatment of brachial plexus root injury. A relatively new modification of this method involves cC7 root transfer to the lower trunk via the prespinal route. In the current study, we examined the effectiveness of this method using electrophysiological and histological analyses. To this end, we used a rat model of total brachial plexus injury, and cC7 root transfer was performed to either the lower trunk via the prespinal route or the median nerve via a subcutaneous tunnel to repair the injury. At 4, 8 and 12 weeks, the grasping test was used to measure the changes in grasp strength of the injured forepaw. Electrophysiological changes were examined in the flexor digitorum superficialis muscle. The change in the wet weight of the forearm flexor was also measured. Atrophy of the flexor digitorum superficialis muscle was assessed by hematoxylin-eosin staining. Toluidine blue staining was used to count the number of myelinated nerve fibers in the injured nerves. Compared with the traditional method, cC7 root transfer to the lower trunk via the prespinal route increased grasp strength of the injured forepaw, increased the compound muscle action potential maximum amplitude, shortened latency, substantially restored tetanic contraction of the forearm flexor muscles, increased the wet weight of the muscle, reduced atrophy of the flexor digitorum superficialis muscle, and increased the number of myelinated nerve fibers. These findings demonstrate that for finger flexion functional recovery in rats with total brachial plexus injury, transfer of the cC7 root to the lower trunk via the prespinal route is more effective than transfer to the median nerve via subcutaneous tunnel.

Nerve transfer helps repair brachial plexus injury by increasing cerebral cortical plasticity

In the treatment of brachial plexus injury, nerves that are functionally less important are transferred onto the distal ends of damaged crucial nerves to help recover neuromuscular function in the target region. For example, intercostal nerves are transferred onto axillary nerves, and accessory nerves are transferred onto suprascapular nerves, the phrenic nerve is transferred onto the musculocutaneous nerves, and the contralateral C7 nerve is transferred onto the median or radial nerves. Nerve transfer has become a major method for reconstructing the brachial plexus after avulsion injury. Many experiments have shown that nerve transfers for treatment of brachial plexus injury can help reconstruct cerebral cortical function and increase cortical plasticity. In this review article, we summarize the recent progress in the use of diverse nerve transfer methods for the repair of brachial plexus injury, and we discuss the impact of nerve transfer on cerebral cortical plasticity after brachial plexus injury.

Transplantation of human amniotic epithelial cells repairs brachial plexus injury:pathological and biomechanical analyses

A brachial plexus injury model was established in rabbits by stretching the C6 nerve root. Imme- diately after the stretching, a suspension of human amniotic epithelial cells was injected into the injured brachial plexus. The results of tensile mechanical testing of the brachial plexus showed that the tensile elastic limit strain, elastic limit stress, maximum stress, and maximum strain of the injured brachial plexuses were significantly increased at 24 weeks after the injection. The treatment clearly improved the pathological morphology of the injured brachial plexus nerve, as seen by hematoxylin eosin staining, and the functions of the rabbit forepaw were restored. These data indicate that the injection of human amniotic epithelial cells contributed to the repair of brachial plexus injury, and that this technique may transform into current clinical treatment strategies.

Effect of Acupuncture Signal after Brachial Plexus Blockade on Cerebral Blood Perfusion and Brain Cell Function

Objective: Using single photon emission computed tomography (SPECT) to observe the influence of the up-transmitting of acupuncture signal into the brain in health volunteers whose nerve trunk was blocked by anesthetics. Methods: Thirty-one healthy volunteers were divided into two groups, the control group of 20 cases, and the brachial plexus blockade (BPB) group of 11 cases, with supraclavicular BPB route adopted. With the control group 2 acupoints were randomly selected (Hegu and Quchi of both sides), while with the BPB group Hegu and Quchi of anesthetic arm side were selected. Siemens ECAM/ ICON SPECT system was used to conduct brain imaging using double imaging assay before acupuncture and 99mTc-ECD imaging agent during acupuncture for cerebral perfusion. The data were quantitatively analyzed by blood functional changing rate (BFCR%) mathematics model. Results: Before acupuncture, the control and BPB groups showed insignificant change by SPECT, but after electro-acupuncture (EA), the control group displayed improved motor and sensory cortex excitability in basal nuclei, contra-lateral thal-amus, parietal and frontal lobe; while BPB group was characterized with reduction of the blood perfusion and cell function of contra-lateral thalamus of anesthetized arm. The difference between the two groups was significant (P<0.01) . Conclusion: (1) After BPB, the up-transmitting of the acupuncture signal via upper limb into the brain, and its strength was impaired or blocked; (2) After BPB, the effect of acupuncture on cerebral perfusion and brain cell function of contra-lateral thalamus was impaired or blocked.

Phrenic nerve transfer to the musculocutaneous nerve for the repair of brachial plexus injury: electrophysiological characteristics

Phrenic nerve transfer is a major dynamic treatment used to repair brachial plexus root avulsion. We analyzed 72 relevant articles on phrenic nerve transfer to repair injured brachial plexus that were indexed by Science Citation Index. The keywords searched were brachial plexus injury, phrenic nerve, repair, surgery, protection, nerve transfer, and nerve graft. In addition, we per-formed neurophysiological analysis of the preoperative condition and prognosis of 10 patients undergoing ipsilateral phrenic nerve transfer to the musculocutaneous nerve in our hospital from 2008 to 201 3 and observed the electromyograms of the biceps brachii and motor conduc-tion function of the musculocutaneous nerve. Clinically, approximately 28% of patients had brachial plexus injury combined with phrenic nerve injury, and injured phrenic nerve cannot be used as a nerve graft. After phrenic nerve transfer to the musculocutaneous nerve, the regener-ated potentials ifrst appeared at 3 months. Recovery of motor unit action potential occurred 6 months later and became more apparent at 12 months. The percent of patients recovering ‘ex-cellent’ and ‘good’ muscle strength in the biceps brachii was 80% after 18 months. At 12 months after surgery, motor nerve conduction potential appeared in the musculocutaneous nerve in seven cases. These data suggest that preoperative evaluation of phrenic nerve function may help identify the most appropriate nerve graft in patients with an injured brachial plexus. The func-tional recovery of a transplanted nerve can be dynamically observed after the surgery.

Valproic acid protects neurons and promotes neuronal regeneration after brachial plexus avulsion

Valproic acid has been shown to exert neuroprotective effects and promote neurite outgrowth in several peripheral nerve injury models. However, whether valproic acid can exert its beneficial effect on neurons after brachial plexus avulsion injury is currently unknown. In this study, brachial plexus root avulsion models, established in Wistar rats, were administered daily with valproic acid dis-solved in drinking water （300 mg/kg） or normal water. On days 1, 2, 3, 7, 14 and 28 after avulsion injury, tissues of the C 5-T 1 spinal cord segments of the avulsion injured side were harvested to in-vestigate the expression of Bcl-2, c-Jun and growth associated protein 43 by real-time PCR and western blot assay. Results showed that valproic acid significantly increased the expression of Bcl-2 and growth associated protein 43, and reduced the c-Jun expression after brachial plexus avulsion. Our findings indicate that valproic acid can protect neurons in the spinal cord and enhance neuronal regeneration fol owing brachial plexus root avulsion.

Electroacupuncture stimulation of the brachial plexus trunk on the healthy side promotes brain-derived neurotrophic factor mRNA expression in the ischemic cerebral cortex of a rat model of cerebral ischemia/reperfusion injury

A rat model of cerebral ischemia/reperfusion was established by suture occlusion of the left middle cerebral artery. In situ hybridization results showed that the number of brain-derived neurotrophic factor mRNA-positive cells in the ischemic rat cerebral cortex increased after cerebral ischemia/ reperfusion injury. Low frequency continuous wave electroacupuncture （frequency 2-6 Hz, current intensity 2 mA） stimulation of the brachial plexus trunk on the healthy （right） side increased the number of brain-derived neurotrophic factor mRNA-positive cells in the ischemic cerebral cortex 14 days after cerebral ischemia/reperfusion injury. At the same time, electroacupuncture stimulation of the healthy brachial plexus truck significantly decreased neurological function scores and alleviated neurological function deficits. These findings suggest that electroacupuncture stimulation of the brachial plexus trunk on the healthy （right） side can greatly increase brain-derived neurotrophic factor mRNA expression and improve neurological function.

Clinical application of ultrasound-guided selective proximal and distal brachial plexus block in rapid rehabilitation surgery for hand trauma

BACKGROUND Anesthesia plays an essential role in the successful surgical procedures for hand trauma.Compared with general anesthesia,brachial plexus block shows lots of benefits for the upper extremity.Specifically,ultrasound-guided selective proximal and distal brachial plexus block may overcome the issues of incomplete block and failure of anesthesia in such circumstances.The present study assessed the efficacy of ultrasound-guided selective proximal and distal brachial plexus block in clinical practice.AIM To explore the efficacy of ultrasound-guided selective proximal and distal brachial plexus block in rapid recovery surgery for hand trauma.METHODS A total of 68 patients with traumatic hand injuries treated at our hospital from January 2018 to June 2019 were selected.They were divided into an observation group and a control group with 34 patients in each group.The observation group underwent selective brachial plexus block guided by ultrasound and the control group underwent conventional brachial plexus block.The analgesic efficacy,intraoperative complications,wound healing time,and length of hospital stay were compared between the two groups under different anesthesia.RESULTS The percentage of cases with analgesia graded as“excellent or very good”was higher in the observation group than in the control group(P<0.05).Moreover,the incidence of intraoperative complications was lower and the wound healing time and length of hospital stay were shorter in the observation group than in the control group(P<0.05).CONCLUSION Ultrasound-guided selective proximal and distal brachial plexus block has remarkable analgesic efficacy in patients with traumatic hand injuries.It can reduce the incidence of intraoperative complications,promote wound healing,and shorten the length of hospital stay.

Restoration and protection of brachial plexus injury: hot topics in the last decade

Brachial plexus injury is frequently induced by injuries, accidents or birth trauma. Upper limb function may be partially or totally lost after injury, or left permanently disabled. With the de- velopment of various medical technologies, different types of interventions are used, but their effectiveness is wide ranging. Many repair methods have phasic characteristics, i.e., repairs are done in different phases. This study explored research progress and hot topic methods for pro- tection after brachial plexus injury, by analyzing 1,797 articles concerning the repair of brachial plexus injuries, published between 2004 and 2013 and indexed by the Science Citation Index database. Results revealed that there are many methods used to repair brachial plexus injury, and their effects are varied. Intervention methods include nerve transfer surgery, electrical stimula- tion, cell transplantation, neurotrophic factor therapy and drug treatment. Therapeutic methods in this field change according to the hot topic of research.

Effects of brachial plexus injury anastomosis simulation on biomechanical properties of adult brachial plexus

BACKGROUND： Previous studies of peripheral nerve mechanical properties in animals have utilized one-dimensional drawing methods. OBJECTIVE： To analyze the effects of brachial plexus injury anastomosis simulation on biomechanical properties of adult brachial plexus by observing tensile mechanical properties, stress relaxation, and creep deformation of the brachial plexus in normal human cadavers and brachial plexus from simulated brachial plexus injury anastomosis samples. DESIGN, TIME AND SETTING： The in vitro experiment was performed at the Mechanics Experimental Center, Jilin University, China from April to May 2007. MATERIALS： A total of six adult, male cadavers, who had died from acute trauma, and were aged 20-29 years, were supplied by the Research Room of Anatomy, Medical Department, Jilin University, China. AG-10TA Universal Material Testing Machine （Shimadzu, Japan） was used in this study. METHODS： A total of 36 samples of fresh brachial plexus were collected from the cadavers, comprising 12 C5 nerve roots, 12 C6 nerve roots at the left and right sides of the superior truck, and 12 C7 nerve roots at the middle truck. The C5 and C6 nerve roots were processed into 50 samples and the C7 nerve roots into 24 samples. A total of 36 C5 and C6 nerve root samples were randomly assigned to a non-surgery control group （n = 18） and brachial plexus injury anastomosis simulation group （n = 18）. Brachial plexus injury simulation anastomosis samples underwent an incision in the middle, and then received anastomosis. Samples in both groups underwent a tension test at 5 mm/min on the AG-10TA universal material testing machine. A total of 24 samples from the C6 superior trunk and C7 middle trunk of the brachial plexus were subjected to stress relaxation and creep tests. Test duration was 7 200 seconds. A total of 100 data points were collected and analyzed using a normalization method. MAIN OUTCOME MEASURES： The following parameters were measured： tension maximum displacement, maximum load, maximum stress, maximum strain and stress-strain curve, stress relaxation at 7 200 seconds, creep deformation at 7 200 seconds, stress relaxation, and creep curve in the non-surgery control group and brachial plexus injury simulation anastomosis group. RESULTS： The tension maximum load of brachial plexus was （140.36 ± 30.50） N, maximum stress was （10.67 ± 2.52） MPa, maximum displacement was （7.78 ± 1.48） mm, and maximum strain was （31.64 ± 5.32）% in the non-surgery control group. The tension maximum load of brachial plexus was （93.23 ± 20.65） N, maximum stress was （7.09 ± 1.57） MPa, maximum displacement was （6.13 ± 0.86） mm,and maximum strain was （24.55 ± 3.45）% in the brachial plexus injury simulation anastomosis group. The above-mentioned indices were greater in the non-surgery control group than in the brachial plexus injury simulation anastomosis group （P 〈 0.01）. Stress relaxation at 7 200 seconds was 2.07 MPa and 2.11 MPa, respectively, in the non-surgery control and brachial plexus injury simulation anastomosis groups. Creep deformation at 7 200 seconds was 4.68% and 3.52%, respectively, in the non-surgery control and brachial plexus injury simulation anastomosis groups. CONCLUSION： Decreased tension maximum load, maximum displacement, maximum stress, maximum strain, and creep deformation at 7 200 seconds affected the biomechanical properties of the brachial plexus following brachial plexus injury.

Stability of motor endplates is greater in the biceps than in the interossei in a rat model of obstetric brachial plexus palsy

The time window for repair of the lower trunk is shorter than that of the upper trunk in patients with obstetric brachial plexus palsy. The denervated intrinsic muscles of the hand become irreversibly atrophic much faster than the denervated biceps. However, it is unclear whether the motor endplates of the denervated interosseous muscles degenerate more rapidly than those of the denervated biceps. In this study, we used a rat model of obstetric brachial plexus palsy of the right upper limb. C5–6 was lacerated distal to the intervertebral foramina, with concurrent avulsion of C7–8 and T1, with the left upper limb used as the control. Bilateral interossei and biceps were collected at 5 and 7 weeks. Immunofluorescence was used to assess the morphology of the motor endplates. Real-time quantitative polymerase chain reaction and western blot assay were used to assess mRNA and protein expression levels of acetylcholine receptor subunits(α, β and δ), rapsyn and β-catenin. Immunofluorescence microscopy showed that motor endplates in the denervated interossei were fragmented, while those in the denervated biceps were morphologically intact with little fragmentation. The number and area of motor endplates, relative to the control side, were significantly lower in the denervated interossei compared with the denervated biceps. mRNA and protein expression levels of acetylcholine receptor subunits(α, β and δ) were significantly lower, whereas β-catenin protein expression was higher, in the denervated interossei compared with the denervated biceps. The protein expression of rapsyn was higher in the denervated biceps than in the denervated interossei at 7 weeks. Our findings demonstrate that motor endplates of interossei are destabilized, whereas those of the biceps remain stable, in the rat model of obstetric brachial plexus palsy. All procedures were approved by the Experimental Animal Ethics Committee of Fudan University, China(approval No. DF-187) in January 2016.

Calcitonin gene-related peptide in anterior and posterior horns of spinal cord after brachial plexus injury

BACKGROUND： The changes of calcitonin gene-related peptide （CGRP） expression are closely associated with peripheral nerve injury, whereas it should be further investigated whether the damage of central nerve can lead to the changes of CGRP expression, and whether it is associated with the neural regeneration and repair. OBJECTIVE： To observe the changing law of CGRP expression in the anterior and posterior horns of spinal cord following brachial plexus injury. DESIGN： A randomized controlled trial. SETTINGS： Department of Anatomy, Yunyang Medical College; Department of Anatomy, Basic Medical College, Sun Yat-sen University. MATERIALS： Sixty-five adult male SD rats of clean degree, weighing 180 - 220 g, provided by the experimental animal center of the Basic Medical College, Sun Yat-sen University, were randomly divided into control group （n =5） and experimental group （n =60）, and the latter was subdivided into three damage groups： avulsion of anterior root group （n =20）, disjunction of posterior root group （n =20） and transection of spinal cord group （n =20）. Diaminobenzidine （DAB） chromogen, rabbit anti-CGRP polyclonal antibody were the products of Sigma Company; Leica image analytical apparatus was produced by QUIN Company （Germany）; Histotome by Sigma Company. METHODS： The experiments were carried out in the Department of Anatomy, Basic Medical College, Sun Yat-sen University from September 2004 to March 2005. Three kinds of models of brachial plexus injury were established： In the avulsion of anterior root group, right C7 anterior root was avulsed, and the distal nerve residual root was transected. In the disjunction of posterior root group, right C7 anterior root was avulsed and right C5 - T1 posterior horns were cut to block the sensory afferent pathway. In the transection of spinal cord group, right C7 anterior root was avulsed and C5-6 segments of right spinal cord were semi-transected to block the cortical descending pathway. In the control group, C5 - T1 vertebral plates were prayed open, and then the skin was sutured. The C7 segments of spinal cord were removed on the 1^st, 3^rd, 7^th and 14^th days postoperatively respectively, and the CGRP expressions in the anterior and posterior horns of spinal cord were determined and analyzed using immunohistochemical method and image analysis. MAIN OUTCOME MEASURES： ① Number of CGRP immuno-positive motor neurons in the anterior horn of spinal cord; ② Total area ofCGRP immuno-positive fibers in the posterior horn of spinal cord. RESULTS： All the 65 rats were involved in the analysis of results. ① Number of CGRP immuno-positive motor neurons in the anterior horn of spinal cord： CGRP immuno-positive motor neurons could be observed in the anterior horns of C7 spinal cord in the control group and damage groups, the neurons had big cell body with stained cytoplasm, appeared as brown granules, and mainly distributed in the ventral lateral anterior horn of spinal cord. On the Ist day postoperatively, the number of CGRP positive neurons was obviously higher in the in the avulsion of anterior root group than in the control group （P 〈 0.01）, whereas obviously lower in the disjunction of posterior root group than in the control group （P 〈 0.01）, and there was no obvious difference between the transection of spinal cord group and the control group （P 〉 0.05）. On the 7^th day, the numbers of CGRP positive neurons in the damage groups were obviously higher than that in the control group （P 〈 0.01）, also obviously different from those on the 1^st day in the same group respectively （P 〈 0.01）. On the 14^th day, the number of CGRP positive neurons in the disjunction of posterior root group was decreased, but there was no obvious difference as compared with that in the control group, whereas those in the avulsion of anterior root group and transection of spinal cord group were still obviously higher than that in the control group （P 〈 0.01）. The number of CGRP positive neurons was the most in the avulsion of anterior root group, followed by the transection of spinal cord group, and the least in the disjunction of posterior root group, and there were significant differences among them （P 〈 0.01）. ② Total area of CGRP immuno-positive fibers in the posterior horn of spinal cord： Dense CGRP immuno-positive nerve fibers distributed in the layers Ⅰ and Ⅱ of the C7 posterior horn of spinal cord in the control group. On the 1^st day postoperatively, the total areas of CGRP positive fibers in the avulsion of anterior root group and transection of spinal cord group were obviously larger than that in the control group （P 〈 0.01）, whereas there was no obvious difference between the disjunction of posterior root group and control group. On the 7^th day, the CGRP expression in the posterior horn of spinal cord decreased to the lowest level in the disjunction of posterior root group, whereas there were no obvious differences in the avulsion of anterior root group and transection of spinal cord group as compared with that in the control group （P 〉 0.05）. On the 14^th day, the area continued to decrease in the avulsion of anterior root group and transection of spinal cord group, and it was obviously lower in the transection of spinal cord group than in the control group （P 〈 0.01）, and it was slightly increased in the disjunction of posterior root group as compared with that on 7^th day, but still obviously lower than that in the control group （P 〈 0.01）. CONCLUSION： The expression and role of CGRP are in discrepancy in the anterior and posterior horns of spinal cord after brachial plexus injury. The CGRP in anterior horn of spinal cord are derived from the cell body of motor neurons, and may be involved in the repairing mechanism of nerve injury regeneration; Whereas those in the posterior horn are mainly derived from posterior root ganglion, and may be associated with the conduction of noxious stimulations.