Objective:To observe the therapeutic effects of electroacupuncture(EA)stimulation on chronic pain and associated negative emotions,and to investigate the activity changes of glutamatergic neurons in the primary motor cortex(M1)following EA stimulation.Methods:Male C57BL/6J mice were randomly divided into four groups:sham surgery(Sham),pure electroacupuncture stimulation(EA),nerve injury(SNI)and electroacupuncture treatment of nerve injury(SNI+EA).Thirty days after the SNI model establishment,EA stimulation was administered bilaterally to the Zusanli(ST36)acupoints in mice.von Frey filaments and Hargreaves heat sensitivity testing were used to detect mechanical and thermal hyperalgesia.The elevated plus maze test was conducted to assess the impact on anxiety-like behaviors in mice.Dual immunofluorescence staining was employed to observe changes in c-Fos expression in M1 gluta-matergic neurons.Results:As compared to Sham group,SNI-treated mice exhibited significant mechanical and thermal hyperalgesia in bilateral hindpaws at 30 days post-modeling(P＜0.01)and displayed obvious anxiety-like behaviors(P＜0.01).The SNI+EA group showed significant relief in pain and anxiety-like behaviors(P＜0.01).c-Fos expression in M1 glutamatergic neurons was significantly decreased in SNI mice.Conversely,after electroacupuncture(EA)treatment,compared to the SNI group,M1 glutamatergic neurons in the SNI+EA group showed significantly in-creased c-Fos expression(P＜0.01).Conclusion:Electroacupuncture significantly alleviated chronic pain and associ-ated anxiety-like behaviors induced by SNI,which might involve the activation of glutamatergic neurons in the M1 in this process.

Objective:To investigate the effects of spinal cord injury(SCI)on neuropathic pain(NP)and cognitive dysfunction in mice,as well as the activation of neurons in the anterior cingulate cortex(ACC),in order to provide experimental evidence for the connection between NP and cognitive dysfunction following SCI.Methods:Ten 8-week-old male C57BL/6J mice were used to prepare a model of spinal cord hemisection(SCI).Pain behavior and cognitive function of mice after SCI were assessed using von Frey,thermal stimulation,and Morris water maze behavioral experi-ments.Immunofluorescence staining were used to analyze the expression of c-Fos and GAD67/VGLUT2 positive cells in the anterior cingulate cortex(ACC)of the SCI model mice.Results:Compared with the Sham group,the SCI group of mice showed a significant decrease in mechanical threshold and thermal pain threshold(P＜0.05).The Morris water maze results indicated a significantly prolonged escape latency(P＜0.05)and a significant reduction in the use of search strategies for locating the hidden platform.Immunofluorescence results showed a significant increase in the num-ber of c-Fos positive cells in the ACC(P＜0.05),which were mainly co-labeled with excitatory neuron marker VGLUT2 positive cells.Conclusion:SCI leads to abnormal pain behavior and cognitive dysfunction in mice,and it induces the activation of neurons in the ACC,with a predominant activation of excitatory neurons.This study provides morphological evidence for the involvement of excitatory neurons in the ACC in the comorbidity of NP and cognitive impairment following SCI.

Objective:To explore the effects of peripheral nerve injury(PNI)on neuropathic pain(NP)and memo-ry function in mice,as well as the activation of neurons in the paraventricular nucleus(PVT)of the thalamus,so as to provide a basis for studying the relationship between NP and memory impairment.Methods:Twenty one 8-week-old male C57 BL/6J mice were randomly divided into sham group and experimental group,and the routine spared nerve inju-ry(SNI)was constructed in the mice of experimental group.The pain behavior and memory impairment of mice after SNI were evaluated with hot plate and eight-arm maze behavioral tests.Pearson correlation analysis was performed to an-alyze the correlation between pain behavior and memory impairment.The c-FOS expression in PVT was detected with immuno-staining.Results:Compared with the sham group,the heat pain threshold of mice in the experimental group was significantly reduced(P＜0.001).The results of the eight-arm maze test showed that the total rest time was signifi-cantly increased(P＜0.001),and the working memory error was increased from 1 to 4 days after SNI(P＜0.01).Correlation analysis indicated that early working memory errors were negatively correlated with heat pain threshold after SNI(P＜0.001).The immunofluorescence revealed that the number of c-FOS positive cells in PVT increased signifi-cantly(P＜0.001).Conclusion:SNI can cause abnormal pain behavior and memory impairment in mice,and cause neuronal activation in PVT.This study provides a basis for neurons in PVT to participate in the regulation of memory impairment in the context of NP.

Objective:To investigate the effects of the novel oral edaravone(EDA)on rats with diabetic encepha-lopathy(DE).Methods:The network pharmacology research methodology was employed to elucidate the mechanism of action of oral EDA in the treatment of diabetes mellitus,identify intersecting targets,and conduct initial validation of these findings in vivo.Thirty male SD rats were randomly assigned to three groups:A normal control(control)group,a diabetic encephalopathy DE(DE)group,and an oral edaravone treatment(DE+EDA)group.Diabetic encephalop-athy was induced in both the DE and DE+EDA groups using the streptozotocin(STZ)method.After successful model-ing,the DE+EDA group received oral administration of EDA,while the other two groups were administered equal doses of saline as controls.Serum samples were examined for lipid release rate,and the protein expression levels of oxidative stress factor 3-nitrotyrosine(3-NT)and apoptotic factor cysteinyl aspartate specific proteinase-3(caspase-3)in brain tissues were detected by Western blot.Brain samples were stained with HE staining to observe the pathological changes.Histopathological changes were observed through hematoxylin-eosin(HE)staining.Results:Network pharmacological analysis yielded 27 core targets,and functional annotation of gene bioprocesses showed that the intersecting targets were mainly enriched in response to oxidative stress and neuronal apoptosis.Serum-related lipid assay showed that the DE+EDA group had significantly improved lipid metabolism disorders compared with the DE group.Additionally,expression levels of 3-NT and caspase-3 were significantly higher in the DE group when compared with controls(P＜0.05);How-ever,both markers exhibited a significant decrease within the DE+EDA treatment cohort as opposed to their counter-parts in the DE group(P＜0.05).HE staining showed that in DE group the cellular arrangement was disordered,the cells were shrunk with intact plasma membrane,and the nuclei were condensed showing karyopyknosis,fragmented and dissolved.Compared with the DE group,the brain tissue in the DE+EDA group was relatively dense and neatly ar-ranged,and the cell karyopyknosis,fragmentation and lysis were significantly improved.Conclusion:Both network pharmacology and in vivo experiments provide preliminary evidence that oral EDA reduces damage in diabetic encepha-lopathy rats.

Objective:To investigate the neuroprotective effect of non-invasive vagus nerve stimulation(nVNS)on traumatic brain injury(TBI).Methods:Male SD rats were used to prepare controlled cortical impact(CCI)rat mod-el.nVNS treatment was performed.The motor function of rats was detected by beam walking test.The water content of rats was evaluated by the dry-wet ratio of rat brain tissue.The content of lactate dehydrogenase(LDH)in cerebrospinal fluid was detected by commercial kit.The expression of bain-derived neurotrophic factor(BDNF)and nerve growth fac-tor(NGF)mRNA in rat cortex was detected by RT-qPCR.Results:After nVNS stimulation,CCI rats significantly im-proved the neurological function deficit,improved the motor ability,decreased the cerebral water content,decreased the level of LDH in cerebrospinal fluid,and upregulated the expression of BDNF and NGF mRNA in cortex.Conclusion:nVNS has a neuroprotective effect on TBI rats,and its mechanism may be related to the increased expression of BDNF and NGF.

Objective:To explore the effects three of anesthetics(tribromoethanol,isoflurane,and pentobarbital so-dium)on the outcome of mice with post-stroke dysphagia(PSD)induced by photothrombosis(PT)method,and to e-valuate which anesthetic is more suitable for the preparation of this model.Methods:Sixty-six male C57BL/6J mice were divided into Tribromoethanol group,Isoflurane group,Pentobarbital sodium group and Sham group.The post-stroke dysphagia model was established by PT.Before and 5 min after modeling,a laser speckle imager was used to measure the regional cerebral blood flow(rCBF)decrease rate of mice and record the wake-up time of mice.Forty-eight hours after modeling,the mortality rate of PSD mice in three groups was recorded and the rCBF decrease rate was meas-ured again.The neurological function of mice was evaluated using the neurological deficit score,the water intake of mice was recorded using the 4-min drinking test,the infarct volume ratio was measured using the TTC staining method,and the swallowing counts induced by water administration was recorded using the multichannel physiological recorder MP160 and the myoelectric area of the swallowing muscle was calculated.Results:There was no statistical difference in the percentage of decrease in rCBF,infarct volume ratio,neurological deficit score,water intake,swallowing counts,and myoelectric area of swallowing muscle among the three groups of PSD mice 48 h after modeling(P＞0.05).Com-pared with the Tribromoethanol group and the Pentobarbital sodium group,the rCBF of the mice in the Isoflurane group decreased rapidly within 5 min(P＜0.05),and the mortality rate of the mice was lower and the awakening time was shorter.(P＜0.05).Conclusion:The use of different anesthesia will affect the mortality rate,wake-up time and the downward trend of rCBF within 48 h after modeling of PSD mice.Among the three anesthetics,isoflurane is more suit-able as an anesthetic for the PSD mouse model.

Objective:Peripheral nerve injury(PNI)is the most common clinical condition.In this study,the current hotspot and future trends of the therapeutic role of microRNA(miRNA)in posterior axon regeneration were evaluated,through bibliometric analysis of peripheral nerves,to provide a reliable reference for later related studies.Methods:Published data from 2009 to 2024 were collected from the Web of Science database by key words TS=mi-croRNA OR TS=miRNA AND TS=peripheral nerve injury OR TS=microRNA OR TS=miRNA AND TS=axonal re-generation screening.The visualization analysis software CiteSpace was used to evaluate the author,country,keywords,outbreak words,institutions,etc.Results:A total of 301 documents were retrieved from the Web of Science database,and 288 documents finally fit the study post eliminating and screening.The 288 articles used in this study have a total of 1513 authors from 34 countries,and 1513 authors from 358 institutions published in 148 journals,citing 13941 articles from 2038 journals.In addition,this study lists the top 5 authors of the annual publication,national distribution,5 arti-cles with the most cited times and the top ten institutions of articles.Moreover,among them,China is the country with the largest number of articles,but there is still a significant gap in the quality of articles when compared with European and American regions.The analysis of institutions found that domestic institutions account for the largest proportion of the research in this field,with close mutual cooperation,but lack of cooperation with foreign institutions.The most cited article in research is"Let-7 microRNAs Regenerate Peripheral Nerve Regeneration by Targeting Nerve Growth Factor"which was cited 34 times.The institution with the largest publication volume is 31 articles from Nantong University.Conclusion:Although the research on therapeutic role of tiny RNA in axon regeneration after peripheral nerve injury started late,the number of papers is relatively small,but it has great potential.As the most popular topic today,tiny RNA has been inuetigated regarding its mechanistic action in the fields of nerve inflammation,nerve cell repair and re-generation,neuroprotection and functional recovery.With the investment in scientific research in this field,more high-quality articles will be published in excellent journals at home and abroad,ushering in a new era in the treatment of pe-ripheral nerve injury.

Objective:To investigate the effect of riboflavin on cerebral infarction volume and the possible mecha-nism of apoptotic factors with cerebral ischemic injury in mice.Methods:Eighteen C57BL/6J male mice were divided into the sham group,middle cerebral artery occlusion(MCAO)group and riboflavin intervention group(MCAO+RF)randomly.TTC staining was used to observe the infarction of the cerebral tissues;Quantitative real-time PCR(RT-qPCR)was used to detect the mRNA expression of tumor protein p53(p53),cytochrome C(CytC),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X(Bax),cysteinyl aspartate specific proteinase-3(caspase-3),poly ADP-ribose poly-merase(PARP),cysteinyl aspartate specific proteinase-6(caspase-6)and apoptosis inducing factor(AIF)in different groups,to study the possible mechanism of riboflavin inhibiting neuronal apoptosis.The proteins expression of acetyl-p53(AC-p53),caspase-3 and PARP were analyzed by Western blot.Results:Compared with the MCAO group,the cerebral infarct volume of the MCAO+RF group was obviously reduced(P＜0.01);The relative expression of p53,CytC,caspase-3,PARP,caspase-6 and AIF were significantly lower in the MCAO+RF group(P＜0.05).Addition-ally,significant differences were observed in the proteins expression of AC-p53,caspase-3 and PARP between the MCAO group and MCAO+RF group.Conclusion:Riboflavin has a protective effect against cerebral ischemic injury,which is possibly realized by inhibiting neuronal apoptosis through multiple pathways.

Objective:To investigate the possible mechanism of treadmill exercise(TR)in improving cognitive function of rats with chronic unpredictable mild stress(CUMS)by detecting pathological changes and the expression of sigma-1 receptor(SIR),endoplasmic reticulum stress-related proteins in hippocampus.Methods:30 SD rats were ran-domly allocated into control group(n=10),CUMS group(n=10)and CUMS exercise group(CUMS+TR,n=10).Subsequently,depressive model was established in rats of CUMS group and CUMS+TR group,meanwhile,CUMS+TR group rats were subjected to 4 weeks of moderate intensity treadmill exercise(20 m/min).After the CUMS and tread-mill exercise,the eight-arm radial maze test(RAM)was carried out to evaluate the learning-memory functions.Subse-quently,the pathological changes of neurons in the hippocampus of rats were measured by HE staining,the protein expression of the GRP78,IRE1,XBP1,CHOP,caspase-3 and SIR in the hippocampus were measured by Western blot.Results:Compared with the control group,the time of finishing the eight-arm radial maze prolonged and the num-ber of total memory errors increased(P＜0.05)in the RAM of the CUMS group rats.The morphology and structure of neurons in the hippocampus of CUMS group rats showed obvious pathological changes,the protein expression levels of GRP78,IRE1,XBP1,CHOP and caspase-3 significantly increased and SIR protein expression decreased(P＜0.05)in hippocampus in the CUMS group compared with the control group rats.4-week treadmill exercise significantly im-prove the learning and memory ability and pathological changes of neurons in hippocampus of CUMS+TR group rats.Compared with CUMS group rats,the protein expression levels of GRP78,IRE1,XBP1,CHOP and caspase-3 signifi-cantly decreased,the SIR expression in hippocampus significantly increased in the CUMS+TR group rats(P＜0.05).Conclusion:Treadmill exercise can improve cognitive impairment in CUMS rats,and its mechanism may be related to the up-regulation of SIR expression and the inhibition of ERS-mediated apoptosis pathway in hippocampus.

Objective:To explore the effective active components of Qi Bi Anshen decoction(QAT)in the treatment of autism spectrum disorder(ASD)and its effects on ASD behaviors and related lipid metabolism abnormalities.Methods:24 adult male Sprague-Dawley rats were randomly divided into 3 groups:Control group,PPA group and PPA+QAT group.The ASD rat model was established by intracerebroventricular injection of propionic acid,and the QAT administration group was given intragastric administration for 7 days.Behavioral tests were conducted to detect the so-cial,repetitive stereotyped and anxiety-like behaviors of rats.UPLC-MS was used to analyze the differential metabolites and enriched pathways of rats.Network pharmacology was used to screen the effective active monomer components of QAT involved in regulating ASD.10 sexually mature C57BL/6J mice were randomly paired in male-female cages.The ASD mouse model was established by a single intraperitoneal injection of sodium valproate to pregnant mice.The preg-nant mice were randomly divided into 3 groups:Control group,VPA group and VPA+QUE group.The administration group was given QUE in the drinking water of pregnant mice until the end of the perinatal period.Behavioral tests were conducted to detect ASD-like behaviors in mice.Reagent kits were used to detect the contents of alkaline phosphatase(AKP),alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglycerides(TG)and total cholesterol(TC)in the liver and serum of mice.Oil red O staining was used to observe the morphology of liver cells.Results:QAT administration could improve the ASD-like behaviors induced by PPA(P＜0.05).UPLC-MS analysis showed that the differential metabolites of each group of rats were mainly enriched in lipid metabolism pathways.Network pharmacol-ogy screening identified QUE as the effective active monomer component.QUE administration could improve the ASD-like behaviors induced by VPA(P＜0.05).QUE administration could reverse the abnormal changes in AKP,TC and TG in the liver induced by VPA(P＜0.05)and reduce lipid droplet deposition in the liver.Conclusion:The active monomer component QUE in QAT has therapeutic effects on ASD behaviors and related liver lipid metabolism abnormali-ties.