The dorsal root ganglion as a target for neurorestoration in neuropathic pain

Neuropathic pain is a severe and chronic condition widely found in the general population.The reason for this is the extensive variety of damage or diseases that can spark this unpleasant constant feeling in patients.During the processing of pain,the dorsal root ganglia constitute an important region where dorsal root ganglion neurons play a crucial role in the transmission and propagation of sensory electrical stimulation.Furthermore,the dorsal root ganglia have recently exhibited a regenerative capacity that should not be neglected in the understanding of the development and resolution of neuropathic pain and in the elucidation of innovative therapies.Here,we will review the complex interplay between cells(satellite glial cells and inflammatory cells)and factors(cytokines,neurotrophic factors and genetic factors)that takes place within the dorsal root ganglia and accounts for the generation of the aberrant excitation of primary sensory neurons occurring in neuropathic pain.More importantly,we will summarize an updated view of the current pharmacologic and nonpharmacologic therapies targeting the dorsal root ganglia for the treatment of neuropathic pain.

Prediction of cell-cell communication patierns of dorsal root ganglion cells:single-cell RNA sequencing data analysis

Dorsal root ganglion neurons transmit peripheral somatic information to the central nervous system,and dorsal root ganglion neuron excitability affects pain perception.Dorsal root ganglion stimulation is a new approach for managing pain sensation.Knowledge of the cell-cell communication among dorsal root ganglion cells may help in the development of new pain and itch management strategies.Here,we used the single-cell RNA-sequencing(scRNA-seq)database to investigate intercellular communication networks among dorsal root ganglion cells.We collected scRNA-seq data from six samples from three studies,yielding data on a total of 17,766 cells.Based on genetic profiles,we identified satellite glial cells,Schwann cells,neurons,vascular endothelial cells,immune cells,fibroblasts,and vascular smooth muscle cells.Further analysis revealed that eight types of dorsal root ganglion neurons mediated proprioceptive,itch,touch,mechanical,heat,and cold sensations.Moreover,we predicted several distinct forms of intercellular communication among dorsal root ganglion cells,including cell-cell contact,secreted signals,extracellular matrix,and neurotransmitter-mediated signals.The data mining predicted that Mrgpra3-positive neurons robustly express the genes encoding the adenosine Adora2b(A2B)receptor and glial cell line-derived neurotrophic factor family receptor alpha 1(GFRα-1).Our immunohistochemistry results confirmed the coexpression of the A2B receptor and GFRα-1.Intrathecal injection of the A2B receptor antagonist PSB-603 effectively prevented histamine-induced scratching behaviour in a dose-dependent manner.Our results demonstrate the involvement of the A2B receptor in the modulation of itch sensation.Furthermore,our findings provide insight into dorsal root ganglion cell-cell communication patterns and mechanisms.Our results should contribute to the development of new strategies for the regulation of dorsal root ganglion excitability.

Surgical Repair of Dorsal Capsulo-Ligamentous Complex for Dislocation of the Thumb Carpometacarpal Joint

Background: Acute dislocation of the thumb carpometacarpal (CMC) joint is a rare and challenging injury, and its treatment remains controversial. The purpose of the study was to evaluate the clinical outcome after surgical repair of the dorsal capsulo-ligamentous complex. Materials and methods: Eight patients with a mean age of 39.8 years (range, 23 - 56 years) treated for the thumb CMC joint were treated in our hospital between 2008 and 2018. The ruptured dorsal capsulo-ligamentous complex was repaired using suture anchors in all patients, and the joint was immobilized with a Kirschner wire pinning and splint for 3 weeks. The clinical outcome was assessed by measuring the range of motion of the thumb CMC joint and the Japanese Society for Surgery of the Hand version of the Quick Disability of the Arm, Shoulder, and Hand (Q-DASH-JSSH) score. Results: The mean radial abduction of the thumb CMC joint was 66°, and the volar abduction was 63°. The mean Q-DASH-JSSH score was 4.8 (range, 0 - 15.6). There were no cases of infection, nerve disturbance, or osteoarthritis. Anatomical reduction was observed in seven patients at the final follow-up and in a rugby football player at 6 months of follow-up;however, subluxation of the thumb CMC joint at 12 months of follow-up due to reinjury was observed. Conclusion: Surgical repair of the dorsal capsulo-ligamentous complex with suture anchors is a reliable and simple treatment for acute unstable dislocation of the thumb CMC joint.

Polyethylene glycol fusion repair of severed rat sciatic nerves reestablishes axonal continuity and reorganizes sensory terminal fields in the spinal cord

Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene glycol(PEG),to immediately fuse closely apposed open ends of severed proximal and distal axons in rat sciatic nerves.We have previously reported that sciatic nerve axons repaired by PEG-fusion do not undergo Wallerian degeneration,and PEG-fused animals exhibit rapid(within 2–6 weeks)and extensive locomotor recovery.Furthermore,our previous report showed that PEG-fusion of severed sciatic motor axons was non-specific,i.e.,spinal motoneurons in PEG-fused animals were found to project to appropriate as well as inappropriate target muscles.In this study,we examined the consequences of PEG-fusion for sensory axons of the sciatic nerve.Young adult male and female rats(Sprague–Dawley)received either a unilateral single cut or ablation injury to the sciatic nerve and subsequent repair with or without(Negative Control)the application of PEG.Compound action potentials recorded immediately after PEG-fusion repair confirmed conduction across the injury site.The success of PEG-fusion was confirmed through Sciatic Functional Index testing with PEG-fused animals showing improvement in locomotor function beginning at 35 days postoperatively.At 2–42 days postoperatively,we anterogradely labeled sensory afferents from the dorsal aspect of the hindpaw following bilateral intradermal injection of wheat germ agglutinin conjugated horseradish peroxidase.PEG-fusion repair reestablished axonal continuity.Compared to unoperated animals,labeled sensory afferents ipsilateral to the injury in PEG-fused animals were found in the appropriate area of the dorsal horn,as well as inappropriate mediolateral and rostrocaudal areas.Unexpectedly,despite having intact peripheral nerves,similar reorganizations of labeled sensory afferents were also observed contralateral to the injury and repair.This central reorganization may contribute to the improved behavioral recovery seen after PEG-fusion repair,supporting the use of this novel repair methodology over currently available treatments.

Outcomes following minimally invasive dorsal cheilectomy for hallux rigidus:A systematic review

BACKGROUND Cheilectomy of the 1^(st)metatarsophalangeal joint(MTPJ)is one of the most common procedures for the management of hallux rigidus.However,there is no consensus regarding outcomes following minimally invasive dorsal cheilectomy(MIDC)for the management of hallux rigidus.AIM To evaluate outcomes following MIDC for the management of hallux rigidus.METHODS During November 2023,the PubMed,EMBASE and Cochrane Library databases were systematically reviewed to identify clinical studies examining outcomes following MIDC for the management of hallux rigidus.RESULTS Six studies were included.In total,348 patients(370 feet)underwent MIDC for hallux rigidus at a weighted mean follow-up of 37.9±16.5 months.The distribution of patients by Coughlin and Shurna's classification was recorded in 4 studies as follows:Ⅰ(58 patients,27.1%),Ⅱ(112 patients,52.3%),Ⅲ(44 patients,20.6%).Three studies performed an additional 1^(st)MTPJ arthroscopy and debridement following MIDC.Retained intra-articular bone debris was observed in 100%of patients in 1 study.The weighted mean American orthopedic foot and ankle society score improved from a preoperative score of 68.9±3.2 to a postoperative score of 87.1.The complication rate was 8.4%,the most common of which was persistent joint pain and stiffness.Thirty-two failures(8.7%)were observed.Thirty-three secondary procedures(8.9%)were performed at a weighted mean time of 8.6±3.2 months following the index procedure.CONCLUSION This systematic review demonstrated improvements in subjective clinical outcomes together with a moderate complication rate following MIDC for the management of hallux rigidus at short-term follow-up.A moderate reoperation rate at short-term follow-up was recorded.The marked heterogeneity between included studies and paucity of high quality comparative studies limits the generation of any robust conclusions.

Electroacupuncture Alleviates Memory Deficits in APP/PS1 Mice by Targeting Serotonergic Neurons in Dorsal Raphe Nucleus

Objective Alzheimer’s disease(AD)has become a significant global concern,but effective drugs able to slow down AD progression is still lacked.Electroacupuncture(EA)has been demonstrated to ameliorate cognitive impairment in individuals with AD.However,the underlying mechanisms remains poorly understood.This study aimed at examining the neuroprotective properties of EA and its potential mechanism of action against AD.Methods APP/PS1 transgenic mice were employed to evaluate the protective effects of EA on Shenshu(BL 23)and Baihui(GV 20).Chemogenetic manipulation was used to activate or inhibit serotonergic neurons within the dorsal raphe nucleus(DRN).Learning and memory abilities were assessed by the novel object recognition and Morris water maze tests.Golgi staining,western blot,and immunostaining were utilized to determine EA-induced neuroprotection.Results EA at Shenshu(BL 23)and Baihui(GV 20)effectively ameliorated learning and memory impairments in APP/PS1 mice.EA attenuated dendritic spine loss,increased the expression levels of PSD95,synaptophysin,and brain-derived neurotrophic factor in hippocampus.Activation of serotonergic neurons within the DRN can ameliorate cognitive deficits in AD by activating glutamatergic neurons mediated by 5-HT1B.Chemogenetic inhibition of serotonergic neurons in the DRN reversed the effects of EA on synaptic plasticity and memory.Conclusion EA can alleviate cognitive dysfunction in APP/PS1 mice by activating serotonergic neurons in the DRN.Further study is necessary to better understand how the serotonergic neurons-related neural circuits involves in EA-induced memory improvement in AD.

Dorsal approach for isolated volar fracture-dislocation of the base of the second metacarpal:A case report

BACKGROUND We report a case with the displacement of an articular fracture fragment of the base of the second metacarpal from the ulnar to the volar side,treated via the dorsal approach.The dorsal approach can be a good option not only because it allows direct observation of ligament damage and fixation of bone fragments but also because the thin subcutaneous tissue makes the approach easier.CASE SUMMARY A 45-year-old man with a right hand injury visited the hospital.A small bone fragment was identified using plain radiography.Lateral radiography revealed the fragment as lying over the volar aspect of the carpometacarpal(CMC)joint.Computed tomography revealed that approximately one-third of the CMC joint surface of the second metacarpal was damaged.We provisionally diagnosed an intra-articular fracture with significant CMC joint instability and performed open reduction and internal fixation.We made a dorsal longitudinal incision over the CMC joint between the second and third metacarpals.The dorsal ligament of the third CMC joint was torn.We thought it had been dislocated to the volar side and spontaneously reduced to that position.There are only few reports of volar dislocation of CMC joint fractures,particularly of the second and third metacarpals;our report is unique as our patient had an intact interosseous ligament between the second and third metacarpals.CONCLUSION Although past reports have used a palmar approach,the dorsal approach is a good option for these cases.

Protein arginine methyltransferase-6 regulates heterogeneous nuclear ribonucleoprotein-F expression and is a potential target for the treatment of neuropathic pain

Protein arginine methyltransferase-6 participates in a range of biological functions,particularly RNA processing,transcription,chromatin remodeling,and endosomal trafficking.However,it remains unclear whether protein arginine methyl transferase-6 modifies neuropathic pain and,if so,what the mechanisms of this effect.In this study,protein arginine methyltransferase-6 expression levels and its effect on neuropathic pain were investigated in the spared nerve injury model,chronic constriction injury model and bone cancer pain model,using immunohistochemistry,western blotting,immunoprecipitation,and label-free proteomic analysis.The results showed that protein arginine methyltransferase-6 mostly co-localized withβ-tubulinⅢin the dorsal root ganglion,and that its expression decreased following spared nerve injury,chronic constriction injury and bone cancer pain.In addition,PRMT6 knockout(Prmt6~(-/-))mice exhibited pain hypersensitivity.Furthermore,the development of spared nerve injury-induced hypersensitivity to mechanical pain was attenuated by blocking the decrease in protein arginine methyltransferase-6 expression.Moreover,when protein arginine methyltransferase-6 expression was downregulated in the dorsal root ganglion in mice without spared nerve injury,increased levels of phosphorylated extracellular signal-regulated kinases were observed in the ipsilateral dorsal horn,and the response to mechanical stimuli was enhanced.Mechanistically,protein arginine methyltransferase-6 appeared to contribute to spared nerve injury-induced neuropathic pain by regulating the expression of heterogeneous nuclear ribonucleoprotein-F.Additionally,protein arginine methyltransfe rase-6-mediated modulation of hete rogeneous nuclear ribonucleoprotein-F expression required amino atids 319 to 388,but not classical H3R2 methylation.These findings indicated that protein arginine methyltransferase-6 is a potential therapeutic target fo r the treatment of peripheral neuro pathic pain.

Bidirectional regulation of the brain-gut-microbiota axis following traumatic brain injury

Traumatic brain injury is a prevalent disorder of the central nervous system.In addition to primary brain parenchymal damage,the enduring biological consequences of traumatic brain injury pose long-term risks for patients with traumatic brain injury;however,the underlying pathogenesis remains unclear,and effective intervention methods are lacking.Intestinal dysfunction is a significant consequence of traumatic brain injury.Being the most densely innervated peripheral tissue in the body,the gut possesses multiple pathways for the establishment of a bidirectional“brain-gut axis”with the central nervous system.The gut harbors a vast microbial community,and alterations of the gut niche contribute to the progression of traumatic brain injury and its unfavorable prognosis through neuronal,hormonal,and immune pathways.A comprehensive understanding of microbiota-mediated peripheral neuroimmunomodulation mechanisms is needed to enhance treatment strategies for traumatic brain injury and its associated complications.We comprehensively reviewed alterations in the gut microecological environment following traumatic brain injury,with a specific focus on the complex biological processes of peripheral nerves,immunity,and microbes triggered by traumatic brain injury,encompassing autonomic dysfunction,neuroendocrine disturbances,peripheral immunosuppression,increased intestinal barrier permeability,compromised responses of sensory nerves to microorganisms,and potential effector nuclei in the central nervous system influenced by gut microbiota.Additionally,we reviewed the mechanisms underlying secondary biological injury and the dynamic pathological responses that occur following injury to enhance our current understanding of how peripheral pathways impact the outcome of patients with traumatic brain injury.This review aimed to propose a conceptual model for future risk assessment of central nervous system-related diseases while elucidating novel insights into the bidirectional effects of the“brain-gut-microbiota axis.”

Unusual capitate fracture with dorsal shearing pattern and concomitant carpometacarpal dislocation with a 6-year follow-up:A case report

BACKGROUND Isolated capitate fractures are rare carpal fractures.Following high-energy injuries,capitate fractures are usually associated with other carpal fractures or ligament injuries.The management of capitate fractures depends on the fracture pattern.Here,we report an unusual capitate fracture with a dorsal shearing pattern and concomitant carpometacarpal dislocation,with a 6-year follow-up.To the best of our knowledge,this fracture pattern and surgical management have not been previously reported.CASE SUMMARY A 28-year-old man presented with left-hand volar tenderness and decreased grip strength that persisted for one month after a traffic accident.Radiography showed a distal capitate fracture with carpometacarpal joint incongruence.Computed tomography(CT)revealed a distal capitate fracture with carpometacarpal joint dislocation.The distal fragment was rotated by 90°in the sagittal plane,and an oblique shearing fracture pattern was noted.Open reduction and internal fixation(ORIF)with a locking plate were performed using the dorsal approach.The imaging studies performed 3 mo and 6 years following surgery revealed a healed fracture,and the Disabilities of the Arm,Shoulder,and Hand and visual analog scale scores were significantly improved.CONCLUSION CT can detect capitate fractures with dorsal shearing pattern and concomitant carpometacarpal dislocation.ORIF using a locking plate are possible.

The Effect of Fascial Pedicled Island Flap of the First Metacarpal Radial Dorsal Artery Skin Branch Chain on the Repair of Thumb Tissue Defect

Objective To analyze the effect of fascial pedicled island flap of the first metacarpal radial dorsal artery skin branch chain on the repair of thumb tissue defect.Methods take the 100 patients with thumb tissue defects treated in our hospital from 2017.01 to 2019.01 as the research objects,and divide them into control group and observation group randomly,50 cases in each group.In the observation group,the fascial pedicled island flap of the first metacarpal radial dorsal artery skin branch chain is used for repair,and in the control group,the thumb tissue is repaired with abdominal flap.Compare the DASH scores and outcomes of two groups.Results After treatment,the DASH score in the observation group is significantly lower than that in the control group,which is statistically significant(P<0.05);The operation time and intraoperative blood loss of Observation Group is significantly lower than Control Group,which is statistically significant(P<0.05).Conclusion Repairing defected thumb with the fascial pedicled island flap of the first metacarpal radial dorsal artery skin branch chaint has the advantages of simple operation and easy mastery,and has significant curative effect on patients.It is an effective way to repair partial thumb defect,and worth popularizing.

Complete Dorsal Carpo-Metacarpal Spatular Dislocation: A Case Report

Carpo-metacarpal dislocations are rare traumatic injuries resulting from violent trauma;they are mostly observed in young subjects. The spatular form is often accompanied by metacarpal fractures. We report a clinical case of complete dorsal carpo-metacarpal spatular dislocation. This was a 34-year-old patient, admitted for a closed trauma of the right hand, in whom the clinical examination noted edema of the hand. The radiological workup showed a complete dorsal spatular dislocation associated with a fracture of the base of the 2nd metacarpal. The reduction of these lesions in closed focus followed by stabilization by metacarpal broaching associated with an intermetacarpal transverse broach was done under scopic control in emergency. Functional rehabilitation was started in the 3rd week. The broaches were ablated at the 6th week. The functional result at 3 months was satisfying with good muscle strength. Resumption of work was effective at 8 weeks. Carpo-metacarpal dislocations are rare injuries, often unrecognized, especially in polytrauma patients, and have a good functional prognosis if they are managed correctly in an emergency.

Clinical application of dorsal carpometacarpal reversed island flap with dorsal metacarpal nerve to reconstruct finger

To investigate the clinical effect of dorsal carpometacarpal reversed island flap with dorsal metacarpal nerve to reconstruct finger.Methods We designed the dorsal reverse carpal and metacarpal island flaps with nerve by using the adjacent two dorsal matacarpal arteries as blood-supply and applied the stand of bone and tendon in waste finger or the free iliac transplantation to reconstruct the every sensory finger.Results Eighten cases were survived completely,and the skin degloving injuries of the finger in 3 cases.The maximum of the flap was 9 cm by 8 cm.Patients were followed up 3 months to 2 years,7 weeks later pain sense of reconstructed finger was recovered.Sensation over S3 amounts to 89% of the digits.Two-point-discrimination of the digits was 5～10 mm.After the operation,the reconstructed finger obtained good appearance,the sensory recovery approach normally.The patients have ability to fulfil daily activities.Conclusion This methods has advantages as follow:simple and practical,high survival rate,low impairment,sensible and good appearance.7 refs,3 figs.

Regulatory effect of nerve growth factor on release of substance P in cultured dorsal root ganglion neurons of rat

Objective To investigate the regulatory effects of nerve growth factor （NGF） on basal and capsaicin-induced release of neuropeptide substance P （SP） in primary cultured embryonic rat dorsal root ganglion （DRG） neurons. Methods DRGs were dissected from 15-day-old embryonic Wistar rats. DRG neurons were dissociated and cultured, and then exposed to different concentrations of NGF （10 ng/mL, 30 ng/mL, or 100 ng/mL） for 72 h. The neurons cultured in media without NGF served as control. RT-PCR were used for detecting the mRNAs of SP and vanilloid receptor 1 （VR1） in the DRG neurons. The SP basal and capsaicin （100 nmol/L）-induced release in the culture were measured by radioimmunoassay （RIA）. Results SP mRNA and VR1 mRNA expression increased in primary cultured DRG neurons in a dose-dependent manner of NGF. Both basal release and capsaicin-evoked release of SP increased in NGF-treated DRG neurons compared with in control group. The capsaicin-evoked release of SP also increased in a dose-dependent manner of NGF. Conclusion NGF may promote both basal release and capsaicin-evoked release of SP. NGF might increase the sensitivity of nociceptors by increasing the SP mRNA or VR1 mRNA.

Identification of differentially expressed genes in dorsal root ganglion in early diabetic rats

Objective To screen and identify differentially expressed genes in the dorsal root ganglion （DRG） in early experimental diabetic rats. Methods Diabetic model rats were induced by single intraperitoneal injection of streptozotocin （STZ）. At the second week after STZ injection, the sensory nerve conduction velocities （SNCV） of sciatic nerve were measured as an indicator of neuropathy. The technique of silver-staining mRNA differential display polymerase chain reaction （DD-PCR） was used to detect the levels of differentially expressed genes in rat DRG. The cDNA fragments that displayed differentially were identified by reverse-hybridization, cloned and sequenced subsequently, and then confirmed by Northern blot. Results The SNCV in the diabetic model group [n = 9, （45.25±10.38） m/s] reduced obviously compared with the control group [n = 8, （60.10± 11.92） m/s] （P 〈 0.05）. Seven distinct cDNA clones, one was up-regulated gene and the others were downregulated ones, were isolated by silver-staining mRNA differential display method and confirmed by Northern blot. According to the results of sequence alignment with GenBank data, majority of the clones had no significant sequence similarity to previously reported genes except only one that showed high homology to 6-pyruvoyl-tetrahydropterin synthase mRNA （accession No., BC059140）, which had not been reported to relate to diabetic neuropathy. Conclusion These differentially expressed genes in the diabetic DRG may contribute to the pathogenesis of diabetic peripheral neuropathy.

葱蝇Dorsal基因的克隆·表达及生物信息学分析

[目的]探讨Dorsal与葱蝇滞育发育的关系。[方法]以葱蝇(Delia antiqua)为试虫,采用RACE方法克隆了葱蝇Dorsal的全长cDNA序列;将该序列推导的氨基酸序列在GenBank搜寻同源序列,与搜寻到的14个代表性昆虫Dorsal序列进行相似性比较,并进行系统进化分析;采用半定量RT-PCR方法分析了Dorsal基因在葱蝇非滞育、夏滞育、冬滞育蛹中的表达情况。[结果]克隆了葱蝇Dorsal基因的cDNA序列,全长2 412 bp,开放阅读框1 974 bp,编码657个氨基酸,等电点PI约8.5,分子量72.9 kDa。与GenBank中的其他14个昆虫代表性Dorsal序列相似性比较和进化分析显示葱蝇的Dorsal与黑腹果蝇(Drosophila melanogaster)、拟暗果蝇(Drosophlia pseudoob-scura)的Dorsal序列相似性最大。半定量分析显示葱蝇Dorsal基因在葱蝇冬滞育、夏滞育、非滞育蛹滞育发育关键期的表达量均明显升高,尤其在滞育后期表达量最高,初步判断Dorsal基因与葱蝇滞育发育相关。[结论]为进一步开展Dorsal基因在昆虫滞育发育过程中的功能研究奠定了基础。

Early Detection of Lesions of Dorsal Artery of Foot in Patients with Type 2 Diabetes Mellitus by High-frequency Ultrasonography

This study evaluated the value of high-frequency ultrasonograpy for early detection of dorsal artery of foot in patients with type 2 diabetes mellitus （MD）. Eighty subjects including 40 patients with type 2 MD （T2DM group） and 40 healthy volunteers （NC group） were recruited. The intima-media thickness （IMT）, the inner diameter and the perfusion of dorsal artery of foot were measured by using high-frequency ultrasonograpy. Meanwhile, the parameters of vascular elasticity, including stiffness parameter （]3）, pressure-strain elastic modulus （Ep）, arterial compliance （AC）, augment index （AI）, and pulse wave conducting velocity （PWV]3） were detected by means of echo-tracking technique. The results showed that no significant difference was found in the IMT, systolic diameter （Ds）, diastolic diameter （Dd） and peak systolic velocity （PSV） between T2DM and NC groups. Ep and PWVβ were increased, and AC was decreased in T2DM group as compared with those in NC group with the differences being significant （P〈0.05 for all）. There was no significant difference in β and AI between T2DM and NC groups. It was concluded that high-frequency ultra- sonography in combination with echo-tracking technique is sensitive and non-invasive, and can be used for early detection of sclerosis of the lower extremity artery in patients with type 2 MD.

Neural mechanism of gastric motility regulation by electroacupuncture at RN12 and BL21: A paraventricular hypothalamic nucleus-dorsal vagal complex-vagus nervegastric channel pathway

AIM: To study the neural mechanism by which electroacupuncture(EA) at RN12(Zhongwan) and BL21(Weishu) regulates gastric motility.METHODS: One hundred and forty-four adult Sprague Dawley rats were studied in four separate experiments. Intragastric pressure was measured using custommade rubber balloons, and extracellular neuron firing activity, which is sensitive to gastric distention in the dorsal vagal complex(DVC), was recorded by an electrophysiological technique. The expression levels of c-fos, motilin(MTL) and gastrin(GAS) in the paraventricular hypothalamic nucleus(PVN) were assayed by immunohistochemistry, and the expression levels of motilin receptor(MTL-R) and gastrin receptor(GAS-R) in both the PVN and the gastric antrum were assayed by western blotting.RESULTS: EA at RN12 + BL21(gastric Shu and Mu points), BL21(gastric Back-Shu point), RN12(gastric Front-Mu point), resulted in increased neuron-activating frequency in the DVC(2.08 ± 0.050, 1.17 ± 0.023, 1.55 ± 0.079 vs 0.75 ± 0.046, P < 0.001) compared with a model group. The expression of c-fos(36.24 ± 1.67, 29.41 ± 2.55, 31.79 ± 3.00 vs 5.73 ± 2.18, P < 0.001), MTL(22.48 ± 2.66, 20.76 ± 2.41, 19.17 ± 1.71 vs 11.68 ± 2.52, P < 0.001), GAS(24.99 ± 2.95, 21.69 ± 3.24, 23.03 ± 3.09 vs 12.53 ± 2.15, P < 0.001), MTL-R(1.39 ± 0.05, 1.22 ± 0.05, 1.17 ± 0.12 vs 0.84 ± 0.06, P < 0.001), and GAS-R(1.07 ± 0.07, 0.91 ± 0.06, 0.78 ± 0.05 vs 0.45 ± 0.04, P < 0.001) increased in the PVN after EA compared with the model group. The expression of MTL-R(1.46 ± 0.14, 1.26 ± 0.11, 0.99 ± 0.07 vs 0.65 ± 0.03, P < 0.001), and GAS-R(1.63 ± 0.11, 1.26 ± 0.16, 1.13 ± 0.02 vs 0.80 ± 0.11, P < 0.001) increased in the gastric antrum after EA compared with the model group. Damaging the PVN resulted in reduced intragastric pressure(13.67 ± 3.72 vs 4.27 ± 1.48, P < 0.001). These data demonstrate that the signals induced by EA stimulation of acupoints RN12 and BL21 are detectable in the DVC and the PVN, and increase the levels of gastrointestinal hormones and their receptors in the PVN and gastric antrum to regulate gastric motility. CONCLUSION: EA at RN12 and BL21 regulates gastric motility, which may be achieved through the PVN-DVCvagus-gastric neural pathway.

Proteomic analysis of the dorsal spinal cord in the mouse model of spared nerve injury-induced neuropathic pain

Peripheral nerve injury often causes neuropathic pain and is associated with changes in the expression of numerous proteins in the dorsal horn of the spinal cord. To date, proteomic analysis method has been used to simultaneously analyze hundreds or thousands of proteins differentially expressed in the dorsal horn of the spinal cord in rats or dorsal root ganglion of rats with certain type of peripheral nerve injury. However, a proteomic study using a mouse model of neuropathic pain could be attempted because of abundant protein database and the availability of transgenic mice. In this study, whole proteins were extracted from the ipsilateral dorsal half of the 4th-6th lumbar spinal cord in a mouse model of spared nerve injury（SNI）-induced neuropathic pain. In-gel digests of the proteins size-separated on a polyacrylamide gel were subjected to reverse-phase liquid-chromatography coupled with electrospray ionization ion trap tandem mass spectrometry（MS/MS）. After identifying proteins, the data were analyzed with subtractive proteomics using ProtAn, an in-house analytic program. Consequently, 15 downregulated and 35 upregulated proteins were identified in SNI mice. The identified proteins may contribute to the maintenance of neuropathic pain,and may provide new or valuable information in the discovery of new therapeutic targets for neuropathic pain.

Expression and effect of sodium-potassium-chloride cotransporter on dorsal root ganglion neurons in a rat model of chronic constriction injury

Sodium-potassium-chloride cotransporter 1 (NKCC1) and potassium-chloride cotransporter 2 (KCC2) are associated with the transmission of peripheral pain.We investigated whether the increase of NKCC1 and KCC2 is associated with peripheral pain transmission in dorsal root ganglion neurons.To this aim,rats with persistent hyperalgesia were randomly divided into four groups.Rats in the control group received no treatment,and the rat sciatic nerve was only exposed in the sham group.Rats in the chronic constriction injury group were established into chronic constriction injury models by ligating sciatic nerve and rats were given bumetanide,an inhibitor of NKCC1,based on chronic constriction injury modeling in the chronic constriction injury + bumetanide group.In the experiment measuring thermal withdrawal latency,bumetanide (15 mg/kg) was intravenously administered.In the patch clamp experiment,bumetanide (10 μg/μL) and acutely isolated dorsal root ganglion neurons (on day 14) were incubated for 1 hour,or bumetanide (5 μg/μL) was intrathecally injected.The Hargreaves test was conducted to detect changes in thermal hyperalgesia in rats.We found that the thermal withdrawal latency of rats was significantly decreased on days 7,14,and 21 after model establishment.After intravenous injection of bumetanide,the reduction in thermal retraction latency caused by model establishment was significantly inhibited.Immunohistochemistry and western blot assay results revealed that the immune response and protein expression of NKCC1 in dorsal root ganglion neurons of the chronic constriction injury group increased significantly on days 7,14,and 21 after model establishment.No immune response or protein expression of KCC2 was observed in dorsal root ganglion neurons before and after model establishment.The Cl^– (chloride ion) fluorescent probe technique was used to evaluate the change of Cl^– concentration in dorsal root ganglion neurons of chronic constriction injury model rats.We found that the relative optical density of N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (a Cl^– fluorescent probe whose fluorescence Cenintensity decreases as Cl– concentration increases) in the dorsal root ganglion neurons of the chronic constriction injury group was significantly decreased on days 7 and 14 after model establishment.The whole-cell patch clamp technique revealed that the resting potential and action potential frequency of dorsal root ganglion neurons increased,and the threshold and rheobase of action potentials decreased in the chronic constriction injury group on day 14 after model establishment.After bumetanide administration,the above indicators were significantly suppressed.These results confirm that CCI can induce abnormal overexpression of NKCC1,thereby increasing the Cl^– concentration in dorsal root ganglion neurons;this then enhances the excitability of dorsal root ganglion neurons and ultimately promotes hyperalgesia and allodynia.In addition,bumetanide can achieve analgesic effects.All experiments were approved by the Institutional Ethics Review Board at the First Affiliated Hospital,College of Medicine,Shihezi University,China on February 22,2017 (approval No.A2017-169-01).