Objective:To observe the effects of eye acupuncture on neurological deficits and mitophagy in neurons of ischemic cortical brain tissue in rats with cerebral ischemia reperfusion injury (CIRI), and to explore the possible mechanism of eye acupuncture in improving CIRI. Methods:Forty-eight SD rats were randomly divided into a sham operation group, a model group, an eye acupuncture group, and an inhibitor group, with 12 rats in each group. The modified middle cerebral artery occlusion method was used to establish CIRI model in the model group, eye acupuncture group, and inhibitor group. Before modeling, the rats in the sham operation group, model group, and eye acupuncture group were intraperitoneally injected with phosphate-buffered solution, while the rats in the inhibitor group were intraperitoneally injected with Compound C solution (adenosine monophosphate-activated protein kinase [AMPK] inhibitor). After successful modeling, the rats in the eye acupuncture group and inhibitor group received eye acupuncture intervention. Bilateral Shangjiao, Xiajiao, Gan (CO12) and Shen (CO10) were selected, with needles retained for 30 min without manipulation, once every 12 h, for a total of 7 interventions. Longa and Bederson scores were used to evaluate neurological deficits in rats. TTC staining was used to observe cerebral infarction. Nissl staining was used to observe the number of Nissl bodies in cortical brain tissue in the ischemic penumbra area. Transmission electron microscopy was used to observe the ultrastructure of neuronal mitochondria in cortical brain tissue in the ischemic penumbra area. Western blot was used to detect the protein expressions of AMPK, phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK), unc-51-like autophagy activating kinase 1 (ULK1), phosphorylated unc-51-like autophagy activating kinase 1 (p-ULK1), FUN14 domain containing 1 (FUNDC1), phosphorylated FUN14 domain containing 1 (p-FUNDC1), and microtubule-associated protein light chain 3βⅡ (LC3-Ⅱ). Results:Compared with the sham operation group, Longa and Bederson scores in the model group were increased (P<0.05), cerebral infarct volume was increased (P<0.05), the number of Nissl bodies in cortical brain tissue in the ischemic penumbra area was reduced (P<0.05), autophagosomes appeared, and the ratios of p-AMPK/AMPK, p-ULK1/ULK1, p-FUNDC1/FUNDC1, as well as LC3-Ⅱ protein expression in cortical brain tissue in the ischemic penumbra area were increased (P<0.05). Compared with the model group and inhibitor group, Longa and Bederson scores in the eye acupuncture group were reduced (P<0.05), cerebral infarct volume was reduced (P<0.05), the number of Nissl bodies in cortical brain tissue in the ischemic penumbra area was increased (P<0.05), a small number of autophagosomes were observed, and the ratios of p-AMPK/AMPK, p-ULK1/ULK1, p-FUNDC1/FUNDC1, as well as LC3-Ⅱ protein expression in cortical brain tissue in the ischemic penumbra area were increased (P<0.05). Conclusion:The eye acupuncture may improve CIRI by upregulating the AMPK/ULK1/FUNDC1 signaling axis, promoting mitophagy, and alleviating neuronal injury.

BackgroundElectroacupuncture (EA) demonstrates efficacy in alleviating diarrhea-predominant irritable bowel syndrome (IBS-D), yet its mechanisms concerning low-grade intestinal inflammation remain insufficiently elucidated. This study investigated whether EA ameliorates IBS-D symptoms by modulating the long non-coding RNA Taurine Upregulated Gene 1 (lncRNA TUG1)/microRNA-127 -3p (miR-127-3p)/nuclear factor-kappa B p65 (NF-κB p65) axis.MethodsAn IBS-D rat model was established using maternal separation, acetic acid enema, and chronic restraint stress. Rats were randomly allocated into control, model, EA (at ST-25 and ST-37), drug (rifaximin), and PDTC (NF-κB p65 inhibitor) groups. Behavioral assessments (body weight, loose stool rate, abdominal withdrawal reflex) were conducted. Molecular analyses included dual-luciferase reporter assays, RT-qPCR, western blot, ELISA, and immunofluorescence to evaluate the TUG1/miR-127-3p/NF-κB p65 axis, inflammatory factors (TNF-α, NLRP3, IL-6), and tight junction proteins (occludin, claudin-1, ZO-1). Intestinal ultrastructure was examined by electron microscopy.ResultsEA significantly improved general status, reduced diarrhea and visceral hypersensitivity in IBS-D rats, comparable to rifaximin and PDTC. Mechanistically, EA upregulated colonic lncRNA TUG1 expression, which sequesters miR-127-3p and partially derepresses NF-κB negative regulators (IκBα, A20), consistent with attenuated NF-κB pathway activation. This was associated with downregulated downstream pro-inflammatory mediators (NF-κB p65, TNF-α, NLRP3, IL-6) in serum and colon tissues. Furthermore, EA restored intestinal barrier integrity, as evidenced by improved mucosal ultrastructure and increased expression of tight junction proteins.ConclusionEA alleviates low-grade intestinal inflammation and visceral hypersensitivity in IBS-D rats. The therapeutic effect is mediated, at least in part through upregulation of lncRNA TUG1, which sponges miR-127-3p to inhibit the NF-κB p65 signaling pathway, thereby reducing inflammatory cytokine release and repairing the intestinal epithelial barrier. These findings suggest the TUG1/miR-127/NF-κB axis as a candidate therapeutic target for EA in IBS-D.

Objective:To evaluate the comparative effectiveness of different acupuncture therapies in stroke rehabilitation using network meta-analysis. Methods:We systematically searched major databases for RCTs (2016-2025) comparing six acupuncture interventions. Frequentist network meta-analysis was conducted in Stata 17.0, with interventions ranked by SUCRA values. Primary outcomes included NIHSS, BI, mRS, MoCA, and MMSE scores. Results:Analysis of 120 RCTs (n = 15,848) demonstrated distinct efficacy profiles: EA ranked highest for neurological recovery (NIHSS SUCRA 95.7%) and BI improvement (74.8%); WNM was optimal for disability reduction (mRS SUCRA 100%) and comprehensive cognition (MoCA 94.9%); while SA excelled in MMSE improvement (86.1%). Conclusion:Distinct acupuncture modalities exhibit unique advantages in the context of stroke rehabilitation. EA ranked first in the restoration of neurological function, whereas WNM and SA were identified as the optimal interventions for global disability and cognitive impairment, respectively. Given that the included studies originated exclusively from China, the external validity of the present findings in other populations warrants further verification. Systematic review registration:https://www.crd.york.ac.uk/PROSPERO, CRD42025112036.

Objective:To analyze the risk factors for pulmonary infection in convalescent patients with cerebral infarction under acupuncture and moxibustion intervention, and to develop a nomogram model. Methods:According to a modeling-to- verification set ratio of 7∶3, 350 patients with cerebral infarction treated in the hospital from January 2022 to January 2024 were selected as the modeling set. At the same time, 150 patients with cerebral infarction admitted from February 2024 to December 2024 were selected as the verification set. General patient information, laboratory indicators, and acupuncture intervention variables were collected. After screening variables using LASSO regression in the modeling set, multivariate logistic regression was performed. Based on the results of the multivariate regression, a nomogram prediction model was established, and the models were evaluated in the modeling and validation set, respectively. Results:The incidence of pulmonary infection in patients with cerebral infarction during convalescence was 26.20%(131/500), including 95 patients in the model group. A total of 147 pathogen strains were detected, mainly Gram- negative bacteria. Modeling concentration, LASSO regression, and multivariate stepwise logistic regression were used to identify factors influencing pulmonary infection: increased age (OR = 1.043, 95%CI: 1.010-1.077), higher Eating Assessment Tool 10 (EAT-10) score (OR = 1.249, 95%CI: 1.125-1.387), invasive procedures (OR = 2.715, 95%CI: 1.340-5.501), higher National Institutes of Health Stroke Scale (NIHSS) score (OR = 2.477, 95%CI: 1.951-3.144), lower albumin (OR = 0.856, 95%CI: 0.786-0.931), lower prealbumin (OR = 0.992, 95%CI: 0.987-0.997), needle retention time of 30 min (OR = 2.439,95%CI: 1.146-5.191), acupuncture course less than 14 d (OR = 2.463,95%CI: 1.157-5.243), and initiation of intervention acupuncture treatment more than >3 d after onset (OR = 2.660, 95%CI: 1.190-5.942). Based on these factors, a nomogram model was constructed, with areas under the curve (AUC) of 0.916 (95%CI: 0.877-0.954) and 0.923 (95%CI: 0.874-0.973). The calibration curve showed that the predicted probabilities of the model were in close agreement with the observed probabilities, and its application could yield positive net clinical benefits. Conclusion:The nomogram model constructed used Lasso-logistic regression can identify the risk of pulmonary infection in patients with cerebral infarction during convalescence after acupuncture treatment. Quantifying risk factors can guide patients in optimizing acupuncture treatment plans, achieving precise intervention, and reducing the risk of infection.

Objective To observe the effect of electroacupuncture(EA)on the trimethylamine-N-oxide(TMAO)-high mobility group box 1 protein(HMGB1)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway in cerebral ischemia(CI)rats,so as to explore its mechanism in brain-gut co-treatment.Methods Male SD rats were randomly divided into sham operation,model and EA groups,with 15 rats in each group.The CI model was established by occlusion of the middle cerebral artery(MCAO).EA was applied to"Baihui"(GV20),"Tianshu"(ST25),"Shangjuxu"(ST37),and"Dazhui"(GV14)for 20 min,once every other day for 14 d.Neurological deficit was assessed using the Zea-Longa score.The cerebral infarction volume was detected by TTC staining,the pathological changes in the ischemic cerebral cortex and colon tissues were observed by HE staining.The contents of TMAO,HMGB1,NLRP3,interleukin(IL)-1β,zonula occludens-1(ZO-1),and Occludin in serum,colon,and ischemic cerebral cortex were detected by ELISA.The protein and mRNA expression levels and positive expression of HMGB1,NLRP3,IL-1β,ZO-1 and Occludin in colon tissue and ischemic cerebral cortex were detected by Western blot,real-time fluorescent quantitative PCR,and immunofluorescence staining,respectively.Results After modeling and compared with the sham operation group,the Zea-Longa score and the percentage of cerebral infarction volume,the contents of TMAO,HMGB1,NLRP3 and IL-1β in serum,colon and ischemic cerebral cortex,the expression levels and positive expressions of HMGB1,NLRP3,IL-1β protein and mRNA in colon and ischemic cerebral cortex were significantly increased(P<0.01)in the model group,while the contents of ZO-1 and Occludin,and the expression levels and positive expressions of ZO-1,Occludin protein and mRNA were decreased(P<0.01).After 14 days'treatment,all the increased and decreased levels of the indexes mentioned above were reversed in the EA group relevant to the model group(P<0.01).HE staining showed disordered arrangement of neurons in the ischemic cerebral cortex,loosened structure,widened intercellular spaces,with inflammatory cell infiltration;as well as mucosal damage in colonic tissues,disordered glandular arrangement,reduced number of goblet cells,accompanied by inflammatory cell infiltration in the model group,which were relatively milder in the EA group.Conclusion EA mediating brain-gut co-treatment can reduce the content of intestinal flora metabolite TMAO,down-regulate the expressions of HMGB1/NLRP3 in intestinal tissue,and reduce peripheral inflammatory factors,thereby alleviating the inflammatory response in brain tissue of MCAO rats.

Objective:To observe the effect of electroacupuncture (EA) on the activation of mast cells (MCs) in the colon of rats with diarrhea-predominant irritable bowel syndrome (IBS-D) and to explore the underlying mechanisms of its protective effect on the intestinal mucosal barrier. Methods:Of 74 offsprings from 6 pregnant SD rats, 14 offsprings were randomly collected as a normal group, and the remaining 60 offsprings underwent IBS-D model prepared by maternal separation combined with acetic acid enema and chronic restraint stress. Fifty-six successfully modeled rats were randomly divided into a model group, a ketotifen group, an EA group, and an EA+ketotifen (combination) group, 14 rats in each one. In the ketotifen group, the rats received the intragastric administration of ketotifen solution (10 mL/kg). In the EA group, acupuncture was delivered at bilateral "Tianshu" (ST25) and "Shangjuxu" (ST37) with disperse-dense wave, a frequency of 2 Hz/100 Hz and a current of 0.3 mA. In the combination group, the intervention measures of the above two groups were operated in combination. In each group, the intervention was delivered once daily for 7 consecutive days. Separately, before modeling, after modeling and after intervention completion, diarrhea score and the minimum injection volume for the abdominal withdrawal reflex (AWR) in rats were evaluated in each group. After intervention, HE staining was adopted to observe the morphology of colonic mucosa; scanning electron microscopy to examine mucosal ultrastructure; Toluidine blue staining to observe MCs degranulation in the colonic mucosa; ELISA to detect serum levels of calcitonin gene-related peptide (CGRP), diamine oxidase (DAO), D-lactate (D-LA), and lipopolysaccharide (LPS); fluorescence microplate assay to detect the serum concentration of 4 kDa fluorescein isothiocyanate-dextran (FD4); Western blot, real-time quantitative PCR (RT-qPCR), and immunofluorescence to detect the expression of nerve growth factor (NGF), tyrosine kinase receptor A (TrkA), and transient receptor potential vanilloid type 1 (TRPV1) in colon tissue; and Pearson correlation coefficient to analyze the relationship between CGRP level and intestinal barrier function. Results:Compared with the normal group, the model group showed the decrease in AWR minimum injection volume (P<0.01), the increase in diarrhea score and MC degranulation rate (P<0.01), the damage of colonic mucosal structure, obvious inflammatory cell infiltration, and the increase in serum levels of CGRP, DAO, D-LA, LPS, FD4 mean fluorescence intensity (MFI), as well as the protein, mRNA, and positive expression levels of NGF, TrkA, and TRPV1 in colon tissue (P<0.01, P<0.05). Compared with the model group, the ketotifen, EA, and combination groups exhibited the increase in AWR minimum injection volume (P<0.01), the decline in diarrhea score and MC degranulation rate (P<0.01), relatively intact intestinal mucosal structure, reduced inflammatory cell infiltration, and the decrease in serum levels of CGRP, DAO, D-LA, LPS, FD4 MFI, as well as the protein, mRNA, and positive expression levels of NGF, TrkA, and TRPV1 in colon tissue (P<0.01). When compared with the ketotifen group, the EA group demonstrated lower FD4 MFI in serum and reduced protein expression of NGF, TrkA, and TRPV1 in colon (P<0.01), and no statistically significant differences were presented in the other indicators (P>0.05). In comparison with both the ketotifen and EA groups, the combination group showed the improvements in all the aforementioned indicators (P<0.01, P<0.05). Serum level of CGRP was positively correlated with the levels of DAO, D-LA, LPS, and FD4 (P<0.001). Conclusion:Electroacupuncture may ameliorate intestinal mucosal barrier function and alleviate symptoms in IBS-D rats by inhibiting MC activation, modulating the NGF/TrkA pathway, and reducing serum levels of DAO, D-LA, LPS, and FD4.

Objective Transcriptome-level insights into electroacupuncture (EA)'s mechanisms for alleviating intestinal mucosal barrier damage in diarrhea-predominant irritable bowel syndrome (IBS-D) are limited. This study aimed to construct ceRNA networks and elucidate EA's role in restoring barrier integrity via lncRNA-miRNA-mRNA regulation in IBS-D rats.Methods The IBS-D model was established by neonatal maternal separation (NMS), 4% acetic acid enema and restrain stress (RS). Rats were randomized into control, model, and EA groups. After 2-week EA treatment, colonic morphology was assessed by HE staining and TEM; intestinal barrier biomarkers were analyzed via ELISA and WB. RNA-seq identified differentially expressed RNAs (DE RNAs) to construct ceRNA networks. GO and KEGG analyzed EA-modulated DE mRNAs. RT-qPCR validated RNA-seq; WB and IF confirmed mast cell (MC) involvement in EA-regulated pathways.Results RNA-seq identified 426 up-regulated and 429 down-regulated DE mRNAs, 342 up-regulated and 362 down-regulated DE lncRNAs, and 10 up-regulated and 48 down-regulated DE miRNAs following EA. Constructed ceRNA networks included 7 DE lncRNAs-miR-139-3p-Bid and -miR-378b-Slc4a5. GO analysis linked EA to defense response, hormone regulation, and cytokine function pathways. KEGG implicated antigen processing/presentation, neuroactive ligand-receptor interaction, PPAR signaling, and glutathione metabolism. RT-qPCR validated RNA-seq results.Conclusion This RNA-seq study reveals EA mitigates IBS-D intestinal mucosal barrier damage by regulating genes and ceRNA networks, providing novel transcriptomic insights into its therapeutic mechanisms.

OBJECTIVES:To evaluate the effect of eye acupuncture on neural function and angiogenesis of ischemic cerebral tissue in rats, and explore the roles of METTL3-mediated m6A methylation and the HIF-1α/VEGF-A signal axis in mediating this effect. METHODS:Fifty SD rats were randomized into normal control group, sham-operated group, model group, eye acupuncture group and DMOG (a HIF-1α agonist) group. Rat models of cerebral ischemia/reperfusion injury (CIRI) were established using a modified thread thrombus method, and the changes in neurological deficits of the rats after interventions were evaluated. TTC and Nissl staining were used to examine the changes in infarction size and neuronal injury, and cerebral angiogenesis was detected by double-immunofluorescence staining. m6A methylation modification level in the brain tissue was detected by ELISA, and RT-qPCR and Western blotting were used to detect the mRNA and protein expressions of METTL3 and HIF-1α/VEGF-A. RESULTS:Compared with the control and sham-operated rats, the CIRI rats had significantly higher neurological deficit scores with larger cerebral infarction area, a greater number of CD31- and EDU-positive new vessels, higher expression levels of HIF-1α and VEGF-A, reduced number of Nissl bodies and m6A methylation level, and lowered METTL3 protein and mRNA expressions. All these changes were significantly improved by interventions with eye acupuncture after modeling or intraperitoneal injections of DMOG for 7 consecutive days prior to modeling, and the effects of the two interventions were similar. CONCLUSIONS:Eye acupuncture can improve neurological deficits in CIRI rat models possibly by promoting cortical angiogenesis via upregulating METTL3-mediated m6A methylation and regulating the HIF-1α/VEGF-A signal axis.

Acupuncture-moxibustion therapy has been known to ameliorate the symptoms of functional gastrointestinal disorders (FGIDs), although its mechanism remains unclear. The paper reviews the articles on acupuncture-moxibustion therapy for FGIDs in recent 5 years, and it is revealed that acupuncture-moxibustion therapy can alleviate FGIDs symptoms through regulating gastrointestinal motility, modulating visceral hypersensitivity, improving the impaired gastric-duodenal mucosal barrier and inflammation, balancing intestinal microbiota, and adjusting the gut-brain axis. Currently, the molecular mechanism of acupuncture-moxibustion therapy remains unknown for FGIDs, the specific disease target is not identified, and the interaction among various molecules is not elucidated adequately. The researches in the future should employ advanced technologies and methodologies to comprehensively and deeply explore and clarify the mechanism of acupuncture- moxibustion therapy for FGIDs.

To evaluate the effect of eye acupuncture on neural function and angiogenesis of ischemic cerebral tissue in rats, and explore the roles of METTL3-mediated m6A methylation and the HIF-1α/VEGF-A signal axis in mediating this effect. Fifty SD rats were randomized into normal control group, sham-operated group, model group, eye acupuncture group and DMOG (a HIF-1α agonist) group. Rat models of cerebral ischemia/reperfusion injury (CIRI) were established using a modified thread thrombus method, and the changes in neurological deficits of the rats after interventions were evaluated. TTC and Nissl staining were used to examine the changes in infarction size and neuronal injury, and cerebral angiogenesis was detected by double-immunofluorescence staining. m6A methylation modification level in the brain tissue was detected by ELISA, and RT-qPCR and Western blotting were used to detect the mRNA and protein expressions of METTL3 and HIF-1α/VEGF-A. Compared with the control and sham-operated rats, the CIRI rats had significantly higher neurological deficit scores with larger cerebral infarction area, a greater number of CD31- and EDU-positive new vessels, higher expression levels of HIF-1α and VEGF-A, reduced number of Nissl bodies and m6A methylation level, and lowered METTL3 protein and mRNA expressions. All these changes were significantly improved by interventions with eye acupuncture after modeling or intraperitoneal injections of DMOG for 7 consecutive days prior to modeling, and the effects of the two interventions were similar. Eye acupuncture can improve neurological deficits in CIRI rat models possibly by promoting cortical angiogenesis via upregulating METTL3-mediated m6A methylation and regulating the HIF-1α/VEGF-A signal axis.

ObjectiveThe aim of this study is to assess the impact of moxibustion on the colonic mucosal barrier and gut microbiota in a rat model of cerebral ischemic stroke (CIS).MethodThe CIS rat model was established using the modified Zea Longa suture method. Successfully modeled rats were randomly allocated into a model group and a moxibustion group, with a sham surgery group serving as the control. The moxibustion group received suspended moxibustion at Dazhui (GV 14), Baihui (GV 20), Fengfu (GV 16), and bilateral Tianshu (ST 25) and Shangjuxu (ST 37) acupoints. Neurological function was assessed using the Longa score, and brain infarct size was assessed through 2,3,5-triphenyl tetrazolium chloride staining. Gut microbiota composition was analyzed using 16S rDNA amplification sequencing. Intestinal mucosal permeability was evaluated using the FITC-Dextran tracer method. The serum ET-1 levels and the expression of Occludin and ZO-1 proteins in colonic tissues were also measured.ResultThe model group exhibited significantly higher Longa scores, larger brain infarct size, and higher serum FITC-Dextran levels and ET-1 levels when compared with the sham surgery group (p < 0.01). The model group demonstrated decreased expression of Occludin and ZO-1 in colonic tissues (p < 0.01) and changes in gut microbiota structure. When compared to the model group, the moxibustion group demonstrated significantly lower Longa scores, smaller brain infarct size, and lower serum FITC-Dextran levels and ET-1 levels (p < 0.05). Furthermore, the moxibustion group demonstrated decreased inflammatory cell infiltration in colonic tissues, increased expression of Occludin and ZO-1 proteins in colonic tissues (p < 0.05), enhanced gut microbiota structure, and a decreased Simpson index (p < 0.05).ConclusionMoxibustion can improve the neurological dysfunction in CIS model rats. The mechanism may be associated with the improvement in gut microbiota dysbiosis, reduction in colonic mucosal permeability, and restoration of intestinal mucosal barrier damage.

Objective To observe the effects of Tongdu Tiaoshen acupuncture(for unblocking the obstruction in the governor vessel and regulating the spirit)on depression-like behavior and the hippocampal Endophilin A1/reactive oxygen species(ROS)pathway in the chronic unpredictable mild stress(CUMS)model rats,and explore the mechanism of this therapy for depression.Methods Forty-eight male SD rats of SPF grade were randomly divided into a normal group(n=12)and a modeling group(n=36).In the modeling group,CUMS was performed to establish depression model.The successfully-modeled rats were randomized into a model group,a Tongdu Tiaoshen acupuncture group(referred to as the acupuncture group),and a fluoxetine group,with 12 rats in each group.In the acupuncture group,"Baihui"(GV20),"Shenting"(GV24),"Shuigou"(GV26)and"Dazhui"(GV14)were stimulated with acupuncture.This intervention measure was delivered once a day,continuously for 6 days;it was discontinued on day 7 and was completed in 28 days.In the fluoxetine group,intragastric administration was done with fluoxetine solution(2.1 mg/kg),once a day,and for 28 consecutive days.Before and after modeling,and after intervention completion,the body mass,sucrose preference rate and the total distance of movement and the boxes of horizontal crossing in the open field experiment were observed in each group.After intervention,using HE staining,the hippocampal neuron morphology was observed;using Nissl staining,the hippocampal Nissl body number was counted.The hippocampal mitochondria was observed under transmission electron microscopy.The average fluorescence intensity of ROS in hippocampal was determined using flow cytometry.With Western blot method,the protein expression of Endophilin A1,growth associated protein 43(GAP-43),and brain-derived neurotrophic factor(BDNF)in hippocampal was detected;and with RT-qPCR method,the mRNA expression of Endophilin A1,GAP-43,and BDNF was recorded.Using the immunofluorescence,the average fluorescence intensity of Endophilin A1,GAP-43,and BDNF in hippocampal tissue was determined.Results Compared with the normal group,in the model group,the body mass,sucrose preference rate,and the total distance of movement and the boxes of horizontal crossing in the open field experiment decreased(P＜0.01);the hippocampal neuronal structure was unclear,the matrix was relatively loose,and the number of Nissl body decreased(P＜0.01);mitochondrial structure was disarranged,the outer membrane was ruptured,mitochondrial cristae was irregular or missed;the average fluorescence intensity of ROS in hippocampal tissue,the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue increased(P＜0.01),while the protein and mRNA expression of GAP-43 and BDNF and its average fluorescence intensity decreased(P＜0.01).Compared with the model group,the acupuncture group and the fluoxetine group showed the increase in body mass,sucrose preference rate,the total distance of movement and the boxes of horizontal crossing in the open field experiment(P＜0.05,P＜0.01);the hippocampal neuronal structure became relatively clear,the matrix was relatively dense,and the number of Nissl body was elevated(P＜0.01);mitochondrial structure got normal and disarranged slightly,the average fluorescence intensity of ROS in hippocampal tissue,the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue were reduced(P＜0.01),while the protein and mRNA expression of GAP-43 and BDNF and the average fluorescence intensity rose(P＜0.01,P＜0.05).Compared with the fluoxetine group,the acupuncture group presented the increase in the average fluorescence intensity of ROS,the protein expression and the average fluorescence intensity of Endophilin A1,the protein expression of GAP-43 and the mRNA expression of BDNF(P＜0.01,P＜0.05),and the decrease of the protein expression and the average fluorescence intensity of BDNF,the mRNA expression of Endophilin A1,and the average fluorescence intensity of GAP-43(P＜0.01,P＜0.05).Conclusion Tongdu tiaoshen acupuncture alleviates depression-like behaviors in CUMS model rats and protects hippocampal neurons,which may be related to suppressing Endophilin A1/ROS signaling pathway and attenuating oxidative stress reactions.

To observe the efficacy of successive trigger needling combined with conventional acupuncture for cervical spondylosis of qi stagnation and blood stasis. A total of 40 patients with cervical spondylosis of qi stagnation and blood stasis were selected, and successive trigger needling at ashi points combined with conventional acupuncture at Baihui (GV20) and bilateral C3-C7 Jiaji (EX-B2), Jiquan (HT1), etc. were delivered, once a day, 5 times a week as one course, and 2 courses were required totally. Before treatment and after 1, 2 weeks of treatment, the pain visual analogue scale (VAS) score was observed, and the clinical efficacy was evaluated after treatment. After 1, 2 weeks of treatment, the VAS scores were decreased compared with that before treatment (P<0.001). After treatment, the total effective rate was 97.5% (39/40). Successive trigger needling combined with conventional acupuncture can effectively treat the cervical spondylosis of qi stagnation and blood stasis, reduce pain and improve the clinical symptoms and signs.

Objective:To systematically evaluate the efficacy and safety of acupuncture as an adjuvant therapy for osteoporosis (OP) through a comprehensive synthesis of recent randomized controlled trial (RCT) evidence. Methods:A systematic literature search was conducted across PubMed, Web of Science, CNKI, and Wanfang databases (2014 - 2024) to identify RCTs investigating acupuncture combined with conventional therapy for OP. Study quality was appraised using the Cochrane Risk of Bias tool, and meta-analyses were performed using RevMan 5.4 and Stata 15.0, with subgroup analyses stratified by intervention type, population characteristics, and treatment duration. Results:28 RCTs (n=2,758) were included. Meta-analysis revealed acupuncture significantly enhanced bone mineral density (BMD) versus controls: total (SMD = 0.47, p = 0.03), femoral neck (MD = 0.05, p = 0.01), lumbar spine (SMD = 0.40, p < 0.001), Ward's triangle (MD = 0.07, p = 0.02), and hip (SMD = 0.55, p < 0.001), with particularly marked improvements in the postmenopausal osteoporosis subgroup. Acupuncture demonstrated significant improvements in treatment efficacy, biochemical markers, pain scores, and symptom assessments, while reducing adverse events. Warm needle moxibustion outperformed controls in femoral neck (MD = 0.07, p = 0.002) and hip BMD (SMD = 0.87, p < 0.001), while electroacupuncture significantly elevated serum calcium (MD = 0.18, p = 0.02). Short-term interventions (≤ 3 months) demonstrated optimal efficacy. Conclusion:Acupuncture demonstrates efficacy and safety as an OP adjuvant therapy. Current evidence is limited by regional bias and methodological heterogeneity. Multicenter, large-sample RCTs are needed to standardize protocols and validate long-term therapeutic efficacy. Systematic review registration:https://www.crd.york.ac.uk/PROSPERO/, identifier CRD42024499354.

Objective To observe the effect of moxibustion on visceral hypersensitivity and the phosphatidylinositol-3 kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway in rats with diarrhea-predominant irritable bowel syndrome(IBS-D),so as to explore its mechanisms underlying amelioration of pain reaction of IBS-D by regulating the autophagy level of intestinal epithelial cells and protecting the intestinal mucosal barrier.Methods Forty male SD rats were randomly divided into control,model,moxibustion and moxibustion+LY294002 groups,with 10 rats in each group.The IBS-D model was established by acetic acid enema+chronic binding method.In the moxibustion group,the rats were given suspended moxibustion on bilateral"Tianshu"(ST25)and"Shangjuxu"(ST37)for 20 minutes.Rats of the moxibustion+LY294002 group received intraperitoneal injection of PI3K inhibitor LY294002(0.75 mg/kg)30 min before each moxibustion intervention.Both groups were treated once a day for 7 days.The minimum colon-rectal water injection volume(minimum water-injection volume)for achieving a score of 3 points of the abdominal withdrawal reflex(AWR)was measured to assess the rats'visceral pain reaction before and after modeling,and after the intervention.H.E.staining was used to observe inflammatory damage of the colonic mucosa.A transmission electron microscopy was employed to examine the mitochondrial structure and the number of autophagosomes in the colon mucosa.The expressions of Occludin,ZO-1,Claudin-2,p62 proteins,and the ratios of p-PI3K/PI3K,p-AKT/AKT,p-mTOR/mTOR and LC3B-Ⅱ/Ⅰ were detected by Western blot.The mRNA expression levels of PI3K,AKT,mTOR,Occludin,ZO-1 and Claudin-2 in the colon tissues were detected by qPCR.Results Compared with the control group,the minimum water injection volume,and the expression levels of p-PI3K/PI3K,p-AKT/AKT,p-mTOR/mTOR ratios,Occludin,ZO-1 and p62 proteins and the mRNA expression of PI3K,AKT,mTOR,Occludin and ZO-1 were significantly decreased(P<0.01),while the protein and mRNA expression level of Claudin-2 and the ratio of LC3B-Ⅱ/Ⅰ were strikingly increased in the model group(P<0.01).H.E.staining showed disordered arrangement of the intestinal epithelial cells,and infiltration of local inflammatory cells,and electron microscopy observation showed multiple autophagosomes in the model group.In comparison with the model group,the minimum water injection volume,the p-PI3K/PI3K,p-AKT/AKT,p-mTOR/mTOR ratios,the protein expression levels of Occludin,ZO-1 and p62 and the mRNA expressions of PI3K,AKT,mTOR,Occludin and ZO-1 in the colon tissue were considerably increased(P<0.05,P<0.01),while the protein and mRNA expression of Claudin-2 and the ratio of LC3B-Ⅱ/Ⅰ were apparently decreased in the moxibustion group(P<0.01).After administration of PI3K inhibitor,the above effects of moxibustion were eliminated(P<0.01,P<0.05).In the moxibustion group,the intestinal mucosal structure was relatively intact,inflammatory cell infiltration was decreased,and the quantity of autophagosomes under the electron microscope was reduced,while in the moxibustion+LY294002 group,the intestinal mucosal structure was relatively disordered,the inflammatory cell infiltration was increased,and the quantity of autophagosomes was increased.Conclusion Moxibustion can relieve visceral pain of IBS-D rats,which may be related to its functions in alleviating intestinal epithelial damage and protecting the intestinal mucosal barrier by activating the PI3K/AKT/mTOR signaling pathway and reducing the level of autophagy of the intestinal epithelial cells.

OBJECTIVE:To observe the efficacy of successive trigger needling combined with conventional acupuncture for cervical spondylosis of qi stagnation and blood stasis. METHODS:A total of 40 patients with cervical spondylosis of qi stagnation and blood stasis were selected, and successive trigger needling at ashi points combined with conventional acupuncture at Baihui (GV20) and bilateral C3-C7 Jiaji (EX-B2), Jiquan (HT1), etc. were delivered, once a day, 5 times a week as one course, and 2 courses were required totally. Before treatment and after 1, 2 weeks of treatment, the pain visual analogue scale (VAS) score was observed, and the clinical efficacy was evaluated after treatment. RESULTS:After 1, 2 weeks of treatment, the VAS scores were decreased compared with that before treatment (P<0.001). After treatment, the total effective rate was 97.5% (39/40). CONCLUSION:Successive trigger needling combined with conventional acupuncture can effectively treat the cervical spondylosis of qi stagnation and blood stasis, reduce pain and improve the clinical symptoms and signs.

PURPOSE:The incidence of Functional Dyspepsia (FD) is gradually increasing, yet there are currently no effective treatment methods available. This study explored the effective components, potential targets, and pathways of Shi-San-Wei-He-Zhong-Wan (SSWHZW) in the treatment of FD, aiming to provide new insights into its treatment. METHODS:First, the Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP) and GeneCards databases were utilized to identify the major active components of SSWHZW and potential therapeutic targets of FD. Subsequently, functional enrichment analyses were performed to elucidate the mechanisms of SSWHZW on FD. Molecular docking simulations were then conducted to assess the binding affinity of key targets and major active components. Next, an FD animal model was established, and the therapeutic effects of SSWHZW were validated using Hematoxylin and Eosin (HE) staining and Enzyme-linked Immunosorbent Assay (ELISA). Finally, Western blot analysis was performed to validate the involvement of key signaling pathways. RESULTS:A total of 229 active ingredients and 283 putative targets were identified from SSWHZW, of which 173 overlapped with the targets of FD and were considered potential therapeutic targets. Key ingredients, such as quercetin, kaempferol, wogonin, and baicalein, were identified as pivotal components of SSWHZW, potentially acting on the 173 overlapping targets and influencing FD through related signaling pathways. Functional enrichment analysis revealed that the PI3K-Akt signaling pathway, VEGF signaling pathway, and NF-kappa B signaling pathway may be involved in the mechanism of SSWHZW in treating FD. Molecular docking predicted that all five ingredients could firmly bind with the top-ranked target TP53 in the Protein- protein Interaction (PPI) network. Further experiments demonstrated that SSWHZW protected the intestinal tissues of FD rats from inflammatory damage by inhibiting the PI3K/AKT signaling pathway. CONCLUSION:Based on network pharmacology, this study explored the multi-component, multitarget, and multi-pathway characteristics of SSWHZW in treating FD. The findings suggest that SSWHZW exerts its anti-FD effects by inhibiting the expression of the PI3K/AKT signaling pathway, providing new insights and methods for further research on the mechanism of SSWHZW in treating FD.

The study aimed to identify genes linked to cell senescence in CIRI. DEGs were analysed by establishing a rat model of CIRI and by transcriptome analysis. The GSE 16561 dataset was obtained from the GEO database and controls for the association analysis of CS with CIRI. CS-related genes were collected from the GeneCards database. The GEO dataset with CS-related genes was analysed for the intersection of differential genes with aging-related genes using Venn diagrams, and 16 CS-related hub genes were screened. The expression correlation heatmap of these intersected genes was constructed to explore their interrelationships. The interactions between the overlapping genes were investigated by PPI network analysis. KEGG enrichment analysis and GSEA were meticulously performed on the core intersecting genes to explore their potential biological functions and intricate pathways. The KEGG enrichment analysis highlighted key metabolic pathways, while GSEA revealed significant gene sets associated. The clinical potential of core intersecting genes as markers of cerebral ischemia-reperfusion senescence was evaluated using ROC analysis, and six core genes regulating senescence in CIRI cells were identified. In conclusion, FCGR2A, FOS, MMP9, PTGS2, SPI1, and STAT3 can be used as potential therapeutic targets for CIRI, a novel diagnostic biomolecular marker for CS.

BACKGROUND:Electroacupuncture (EA) is a form of physical therapy rooted in traditional Chinese medicine, which has been widely used in clinical practice. This study aimed to explore the effect of EA treatment on synaptic plasticity in mice subjected to middle cerebral artery occlusion (MCAO) and to elucidate the associated molecular mechanisms. METHODS:After MCAO modeling, C57BL/6 mice underwent EA treatment and/or miR-670-3p mimic injection, followed by assessment of neurological deficit by modified neurological severity score (mNSS) and cerebral infarction areas were evaluated via TTC staining. The changes of synaptic ultrastructure related parameters were observed using transmission electron microscopy (TEM). The expression levels of miR-670-3p, HMGB1, TLR4/NF-κB pathway-related proteins, and synapse-associated proteins (Synapsin I, PSD95, BDNF, and GAP43) were quantified by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. The binding relationship between miR-670-3p and HMGB1 was assessed through dual-luciferase reporter assays and RNA pull-down assays. RESULTS:Mice that underwent EA treatment or miR-670-3p mimic injection exhibited increased miR-670-3p expression, reduced expression levels of HMGB1 and TLR4/NF-κB pathway-related proteins, improved neurological function, and enhanced synaptic plasticity. Furthermore, the combination of EA treatment and miR-670-3p mimic injection produced a synergistic effect, further amplifying these outcomes. Mechanistically, miR-670-3p was found to negatively regulate HMGB1. CONCLUSION:EA treatment enhances synaptic plasticity in MCAO mice by promoting miR-670-3p expression to negatively regulate the HMGB1/TLR4/NF-κB pathway.

Objective:To observe the effect of moxibustion on small intestinal mucosal immune barrier in rats with diarrhea-predominant irritable bowel syndrome (IBS-D) and explore its underlying mechanisms. Methods:Of 38 newborn rats from 4 healthy SPF pregnant rats, 12 neonatal rats were randomly selected in a normal group. IBS-D model was prepared by the combined measures for the rest rats, including neonatal maternal separation, acetic acid enema and chronic restraint stress. Twenty-four successfully-modeled rats were randomized into a model group and a moxibustion group, 12 rats in each one. In the moxibustion group, suspending moxibustion was delivered at bilateral "Tianshu" (ST25) and "Shangjuxu" (ST37), 20 min each time, once daily and for 7 consecutive days. Separately, before acetic acid enema (aged 35 days), after modeling (aged 45 days) and after intervention (aged 53 days), the body mass, loose stool rate (LSR) and and the minimum volume threshold when abdominal withdrawal reflex (AWR) scored 3 were observed in the rats of each group. After intervention (aged 53 days), using HE and PAS staining, the morphology of duodenum was observed, the length of villus and the depth of crypt were measured, the ratio of the length of villus to the depth of crypt was calculated; and the numbers of mucosal intraepithelial lymphocytes (IELs) and goblet cells were counted. With ELISA adopted, the contents of γ-interferon (IFN-γ), interleukin-4 (IL-4) and secretory immunoglobulin A (sIgA) in duodenal mucosa of rats were detected. The proportion of T cell subsets in duodenal mucosa was detected using flow cytometry. The microvilli and tight junctions of duodenal mucosal epithelial cells were observed by transmission electron microscopy, and the integrity of duodenal mucosa observed by scanning electron microscopy. Results:Compared with the normal group, for the rats in the model group, the body mass, the minimum volume threshold when AWR scored 3, the length of duodenal villus and the the ratio of the length of villus to the depth of crypt, as well as the proportion of CD8+ T subset were all reduced (P<0.01, P<0.05), the counts of goblet cells in duodenal mucosa decreased (P<0.01); LRS, the proportion of CD4+ T subset and CD4+/CD8+, as well as the contents of IFN-γ, IL-4 and sIgA in duodenal mucosa and IFN-γ/IL-4 were all elevated (P<0.01); and the numbers of IELs rose (P<0.01). The morphology of duodenal mucosa was irregular, the villi got shorter, sparse and scattered, with uneven density. The morphology of epithelial cells was destroyed and the tight junctions damaged, with larger spaces. When compared with the model group, in the moxibustion group, the body mass, the minimum volume threshold when AWR scored 3, the length of duodenal villus and the ratio of the length of villus to the depth of crypt, as well as the counts of goblet cells in duodenal mucosa increased (P<0.01); LRS, the proportion of CD4+ T subset, and CD4+/CD8+, as well as the contents of IFN-γ, IL-4 and sIgA in duodenal mucosa and IFN-γ/IL-4 were reduced (P<0.01); and the numbers of IELs was dropped (P<0.01). The morphology of duodenal mucosa was more regular, the villi were grew, got longer and arranged regularly, with even density. The morphology of epithelial cells was slightly destroyed, and the tight junctions partially damaged. Conclusion:Moxibustion at "Tianshu" (ST25) and "Shangjuxu" (ST37) can reduce visceral hypersensitivity in IBS-D rats and relieve abdominal pain, diarrhea and other symptoms. Its effect mechanism may be related to the repair of small intestinal mucosal immune barrier and the improvement in the immune function in IBS-D.