The ventrolateral periaqueductal gray , located within the midbrain, is a brain region implicated in various physiological and pathological processes, including pain modulation. This area integrates nociceptive signals from multiple brain regions such as the prefrontal cortex, amygdala, and hypothalamus, and inhibits nociceptive input through descending pathways. It is also involved in the analgesic effects of electroacupuncture. This review summarizes and analyzes the role of this area in pain modulation and electroacupuncture-induced analgesia, offering insights into the central mechanisms underlying pain and electroacupuncture analgesia, as well as providing recommendations for future basic and clinical research.

IntroductionTrigeminal neuralgia (TN) is extremely vulnerable to inducing anxiety disorders in TN patients owing to the severe intensity of pain and recurrent episodes. The efficacy of the routine management of TN with anxiety disorders is still unsatisfactory. Acupuncture has been widely utilized in the treatment of TN for many years; however, the effectiveness of Electroacupuncture (EA) for TN with anxiety disorders remains uncertain. Therefore, this randomized, controlled, single-blinded trial was designed to investigate the efficacy and safety of EA for TN with anxiety disorders.MethodsThis single-center, prospective, randomized controlled trial will recruit 60 TN patients with anxiety. All patients will be randomly assigned to the EA or sham EA group in a 1:1 ratio via a computerized central randomization system. The total trial duration is 18 weeks, including a 2-week baseline phase, a 4-week treatment phase, and a 12-week follow-up phase. Participants who meet the inclusion criteria will receive active EA or sham EA treatment for 60 min, three times a week, for 4 weeks.Planned OutcomesThe primary outcome is the change of Beck Anxiety Inventory score and Visual Analog Scale score from baseline, while the BPI-Facial scores will be regarded as the secondary outcomes. All adverse effects will also be assessed during the study period. Outcome measures will be assessed at pre-intervention, post-intervention (week 4), and at the end of follow-up (week 16).Trial RegistrationChinese Clinical Trial Registry: ChiCTR2300079048. This study was registered on December 25, 2023.

Objective To explore the mechanism of hyperalgesia priming(HP)and evaluate the intervention effect of electroacupuncture(EA)on pain transition.Methods Fifty-nine male C57 mice were used for 4 separate experiments.In the first experiment,the mice were divided into a control group,a sham sensitization group,and a sensitization group,with 5 mice per group.In the second experiment,the mice were divided into a sham sensitization group,a sensitization group,and a sensitization+EA group,with 5 mice per group.The third experiment consisted of a sensitization group and a sensitization+minocycline group,with 7 mice per group.In the fourth experiment,the mice were divided into a sensitization group,a sensitization+EA+PBS group,and a sensitization+EA+colony-stimulating factor(CSF)group,with 5 mice per group.A HP mouse model was established via consecutive intraplantar injections of carrageenan and prostaglandin E2.EA was applied to bilateral"Zusanli"(ST36)and"Kunlun"(BL60)for 30 min,once daily for 8 d.Mechanical paw withdrawal thresholds(PWTs)were measured at different time points to assess pain transition and the effect of EA.Immunofluorescence was used to detect the activation of microglia marker(Iba1)and astrocyte marker(GFAP)in the spinal cord dorsal horn.Intrathecal injection of minocycline was also administered,and its impacts on Iba1 activation and PWTs were tested.After the experiment,skeleton and Sholl analyses were performed to analyze the morphology of microglia.To further examine the role of CSF-1,the factor was administered intrathecally to assess its influence on the effects of EA.Results In the pain transition mouse model,the PWTs at various time points decreased significantly(P<0.01).After modeling,the microglia was activated(P<0.01),but the astrocyte was not.EA increased the PWTs in pain transition mice(P<0.01)and suppressed microglia activation(P<0.01).Intrathecal minocycline administration inhibited microglia activation,reduced Iba1 expression in the spinal cord dorsal horn(P<0.01),and effectively reversed the PWTs reduction(P<0.01).Subsequent intrathecal CSF-1 injection promoted microglia activation and blocked the analgesic effect of EA in the model mice(P<0.01).Conclusion Microglia activation is critical in pain transition.EA can alleviate hyperalgesia in HP mice by inhibiting microglia activation.

The spinoparabrachial tract is a major ascending pathway that transmits nociceptive signals from the spinal cord to the brain, but whether electroacupuncture (EA) relieves pain-anxiety comorbidity by modulating this pathway remains unclear. Here, we investigated the effects of EA on mechanical allodynia and anxiety-like behaviors in Complete Freund's adjuvant (CFA)-induced inflammatory pain mice using von Frey testing, the open field test, and the elevated plus maze, combined with chemogenetic manipulation of glutamatergic projections from the spinal dorsal horn (SDH) to the lateral parabrachial nucleus (LPBN). EA attenuated CFA-induced mechanical allodynia and anxiety-like behaviors and reduced c-Fos expression in both the SDH and LPBN. Chemogenetic inhibition of LPBN glutamatergic neurons mimicked the analgesic and anxiolytic effects of EA, whereas their activation abolished these effects. Activation of either ipsilateral or contralateral SDH→LPBN glutamatergic projections induced pain- and anxiety-like phenotypes that depended on intact LPBN glutamatergic output. Moreover, selective activation of spinoparabrachial glutamatergic terminals was sufficient to block the therapeutic efficacy of EA. These findings indicate that EA alleviates CFA-induced pain-anxiety comorbidity by inhibiting the SDH-LPBN glutamatergic circuit, identifying a key ascending pathway through which peripheral EA signals modulate central pain and affective processing.

We evaluated the effectiveness of cartilage tympanoplasty type 1 combined with platelet-rich plasma (PRP) compared to cartilage tympanoplasty type 1 alone in dry tympanic membrane (TM) perforation due to chronic otitis media, demonstrating the safety and efficacy of PRP in tissue repair and regeneration. Data were retrieved from the literature up to April 2025. Outcomes of interest were success rate (complete closure of the TM postoperatively), postoperative air-conduction (AC) threshold, and changes in air–bone gap (ABG) and AC threshold (preoperatively to postoperatively). Pooled effects were calculated using standardized mean differences (SMDs) and odds ratios (ORs) with 95

Yurong Jiang, Lexuan Li, Jianqiao Fang The Third School of Clinical Medicine, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People&rsquo;s Republic of ChinaCorrespondence: Jianqiao Fang, The Third School of Clinical Medicine, Zhejiang Chinese Medical University, 548 Binwen Road, Hangzhou, Zhejiang, 310053, People&rsquo;s Republic of China, Email fangjianqiao7532@163.com

OBJECTIVE:This study investigated the role of the cannabinoid receptor type 2 (CB2R) in the spinal cord dorsal horn (SCDH) using a spared nerve injury (SNI) model of neuropathic pain (NP). The aim was to clarify the analgesic effects of different electroacupuncture (EA) frequencies in SNI rats and to explore whether the higher frequency exerts analgesic effects by modulating the CB2R signaling pathway. METHODS:An SNI rat model was initially established. Mechanical allodynia was assessed using the paw withdrawal threshold (PWT), and motor function was assessed using a gait analysis system. Expression levels of CB2R, phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK), cluster of differentiation 11b (CD11b), ionized calcium-binding adapter molecule 1 (Iba1), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were quantified via immunofluorescence staining and Western blot analysis. The functional role of the CB2R/p38 MAPK pathway was further investigated through intrathecal administration of the CB2R agonist AM1241 and the p38 MAPK inhibitor SB203580. The analgesic effects of low-frequency (2 Hz) and high-frequency (100 Hz) EA were observed in SNI rats, and the modulatory effect of the superior frequency (2 Hz EA) on this pathway was explored. The CB2R antagonist AM630 was co-administered with low-frequency EA to evaluate its impact on pathway activity. RESULTS:CB2R and p-p38 MAPK expression increased after SNI and they were mainly expressed in microglia. The SNI group exhibited a substantial increase in Iba1 and CD11b staining intensity, indicating robust microglial activation, along with elevated levels of the pro-inflammatory cytokine IL-6. Intrathecal administration of either the CB2R agonist AM1241 or the p-p38 MAPK inhibitor SB203580 increased the PWT, inhibited microglial activation and decreased the release of inflammatory cytokines in the SCDH. Both low-frequency EA and high-frequency EA were effective in alleviating pain induced by SNI. Low-frequency EA was significantly more effective than high-frequency EA in improving PWT. Low-frequency EA improves SNI-induced gait disturbances. Low-frequency EA ameliorated hyperalgesia in SNI rats by enhancing CB2R expression and suppressing p-p38 MAPK activation. AM630 reversed this analgesic effect. Moreover, EA attenuated microglial activation and lowered the levels of pro-inflammatory cytokines, specifically TNF-α and IL-6, in the SCDH. It also suppressed the phosphorylation of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT). Notably, CB2R antagonism abolished all these effects. CONCLUSION:CB2R participates in SNI-induced NP by activating the p38 MAPK pathway. Low-frequency EA shows superior efficacy compared with high-frequency EA and alleviates NP through CB2R-mediated suppression of the p38 MAPK and PI3K/AKT signaling in the SCDH. Please cite this article as: Wei ZY, Zhang JJ, Lin C, Li SY, Kang YR, Liang JQ, Liu ZQ, He XF, Shao XM, Fang JQ, Jiang YL. Low-Frequency electroacupuncture alleviates neuropathic pain via cannabinoid receptor type 2 signaling in the spinal cord dorsal horn. J Integr Med. 2026; Epub ahead of print.

Introduction Postoperative pain is common after surgery, with a high incidence and risk of becoming chronic. Current multimodal analgesia has drawbacks, including limited efficacy from single agents and opioid side effects and addiction risk. These issues have led to opioid-sparing multimodal analgesia. Transcutaneous auricular vagus nerve stimulation (taVNS) is non-invasive and convenient. Studies have shown it can reduce postoperative pain, improve mood and lower adverse events. However, taVNS lacks a comprehensive evaluation and standardised protocols, so further research is needed to provide reliable evidence.Methods and analysis This study strictly adheres to the guidelines of the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols. To identify suitable randomised controlled trials (RCTs), eight credible databases will be searched, including four English databases (Web of Science, PubMed, Cochrane Central Register of Controlled Trials, EMBASE) and four Chinese databases (China National Knowledge Infrastructure, VIP Database for Chinese Technical Periodicals, Wanfang Database, Chinese Biomedical Literature Database). RevMan V.5.3 will be employed to integrate the retrieved data and conduct meta-analyses. The methodological quality of included RCTs will be evaluated using the Cochrane Risk of Bias Assessment 2.0 tool. Additionally, the Grading of Recommendations, Assessment, Development and Evaluation system will be applied to assess the strength and certainty of the evidence. We will also conduct publication bias analyses, sensitivity analyses and subgroup analyses.Ethics and dissemination No ethical review is required as no private or confidential patient data will be included. Results of this study will be disseminated through a peer-reviewed journal.PROSPERO registration number CRD420251207651.

Chemotherapy-induced nausea and vomiting (CINV) remain difficult to control after discharge despite guideline-based antiemetics. This multicenter exploratory single-arm two-stage trial evaluated patient-controlled, real-time transcutaneous electrical acupoint stimulation (TEAS) at Neiguan (PC6) as an adjunct to standard antiemetic therapy in patients with breast cancer. Forty-eight patients who had experienced CINV and were scheduled at enrollment to receive the same chemotherapy and antiemetic regimens during two consecutive cycles were analyzed. Cycle 1 served as the observation period, and cycle 2 added on-demand wrist-worn TEAS initiated by patients at nausea or vomiting onset during 0–120 h after chemotherapy. Compared with the observation period, TEAS was associated with lower nausea frequency and visual analogue scale scores on days 1–5 and higher nausea response rates on days 1–4. Vomiting frequency and severity were lower on days 1–3, but between-period differences were not significant on days 4–5. SF-36 scores increased and SAS/SDS scores decreased on day 5. Two transient local numbness events occurred, with no serious TEAS-related adverse events. These findings suggest that patient-controlled TEAS is feasible and warrants confirmation in randomized sham-controlled trials.

OBJECTIVES:To observe temporal changes of pain-related behaviors in mice with chronic postsurgical pain (CPSP) and identify its key mediators in the dorsal root ganglion (DRG). METHODS:In mouse models of CPSP induced by plantar incision (INC) followed by a dorsal foot injection of prostaglandin E2 (PGE2) and sham-operated mice, mechanical paw withdrawal thresholds (PWTs), thermal paw withdrawal latencies (PWLs), and cold withdrawal durations (WDs) were measured at different time points after modeling. Gene expression profiling of the DRG with RNA sequencing was performed on day 1 and day 8 after PGE2 injection. Bioinformatics analyses were conducted to explore the key mediators in the DRG for regulating CPSP, and the candidate genes and proteins were validated using RT-qPCR and ELISA. The effects of intrathecal injection of a CX3CL1-neuralizing antibody or JMS-17-2 (a CX3CR1 antagonist) on CPSP were observed. RESULTS:In CPSP mouse models, incision-induced pain was resolved within 14 days, and PWTs and WDs decreased progressively till day 10 and day 12 after PGE2 injection, respectively, without significant changes in PWLs. RNA-Seq identified 975 differentially expressed genes (DEGs) on day 1 and 895 on day 8 following CPSP modeling, including 524 intersecting DEGs enriched in cell membrane, plasma membrane, and CX3C chemokine receptor binding. Cx3cl1 and Cxcl14 were the top two upregulated chemokine-related DEGs in early CPSP, whose mRNA and protein expression increased significantly in the ipsilateral DRG on day 1 but declined on day 8. Intrathecal injection of the CX3CL1-neutralizing antibody before PGE2 injection prevented CPSP development, while JMS-17-2 partially reversed CPSP during the maintenance phase. CONCLUSIONS:This CPSP mouse model shows persistent mechanical and cold allodynia for at least 10 days after PGE2 injection without significant changes in heat hypersensitivity. CX3CL1 and related chemokine signaling in the DRG may contribute to the development of CPSP.

Background Chronic pain is frequently accompanied by cognitive dysfunction, while whether pain-related memory impairment depends on task difficulty remains unclear. According to the limited cognitive resource hypothesis, persistent pain may consume finite neural processing capacity and thereby impair cognitive performance mainly under high cognitive demand. Electroacupuncture (EA) alleviates neuropathic pain and may improve cognitive function, but its effect on task-dependent memory deficits associated with chronic neuropathic pain remains insufficiently defined. This study investigated whether spared nerve injury (SNI)-induced neuropathic pain selectively impairs memory under high cognitive demand and whether repeated EA reverses these deficits. Results SNI induced robust mechanical hypersensitivity in rats but did not produce detectable impairments in novel object recognition (NOR) or post-NOR novel object location (NOL) performance under low-difficulty conditions. While under high-difficulty conditions, SNI rats showed impaired long-term object recognition memory, whereas post-NOR location memory remained unaffected. When object novelty and spatial relocation were combined in a higher-demand NOR + NOL task, SNI rats exhibited deficits in both object recognition and object-location memory. Repeated 2-Hz EA at ST36 and SP6 attenuated mechanical hypersensitivity and restored impaired performance in high-demand memory tasks. Immunofluorescence analysis showed that SNI reduced the percentage of CaMKII/c-Fos double-positive neurons among CaMKII-positive neurons in the rostral anterior cingulate cortex (rACC), whereas EA increased CaMKII/c-Fos co-localization to a level comparable with that in sham-operated rats. Conclusions Chronic neuropathic pain selectively disrupts long-term memory under high cognitive demand while producing no detectable impairment under lower task demand, supporting a resource-competition model of pain-cognition interaction. Repeated EA alleviates mechanical hypersensitivity and reverses task-dependent memory deficits, and these behavioral effects are associated with increased CaMKII/c-Fos co-localization in rACC CaMKII-positive neurons. These findings suggest that EA may represent a superior efficacy for chronic pain-associated cognitive dysfunction by increasing the CaMKII-positive neuronal activation in the rACC.

Hengyu Chi,1,&ast; Qunqi Hu,1,&ast; Jingqi Liang,1,&ast; Jingjing Zhang,1 Siying Qu,1 Hanzhi Wang,2 Yurong Kang,1 Zhouqi Liu,1 Yuran Chen,1 Yongliang Jiang,1 Xiaofen He,1 Jianqiao Fang,1 Yinhang Zhong31Key Laboratory of Acupuncture and Neurology of Zhejiang Province, The Third Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, People’s Republic of China; 2Department of Gynecology, Tongde Hospital of Zhejiang Province, Hangzhou, Zhejiang, 310012, People’s Republic of China; 3Department of Traditional Chinese Pharmacy, Zhejiang Xiaoshan Hospital, Hangzhou, Zhejiang, 311200, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yinhang Zhong, Department of Traditional Chinese Pharmacy, Zhejiang Xiaoshan Hospital, Hangzhou, Zhejiang, 311200, People’s Republic of China, Email yinhzh@qq.com Jianqiao Fang, Key Laboratory of Acupuncture and Neurology of Zhejiang Province, The Third Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, People’s Republic of China, Email fangjianqiao7532@163.comPurpose: Perimenopause represents a critical neuroendocrine transition stage in the lives of women, characterized by metabolic disorders and physiological vulnerability, which also commonly manifests as depression and cognitive impairment. This study explores associations between the triglyceride-glucose-frailty index (TyGFI) and depression/cognitive impairment in perimenopausal women.Methods: Data were obtained from the 2015 China Health and Retirement Longitudinal Study (CHARLS) and 2,596 perimenopausal women were included. Logistic regression models were used to evaluate the association between TyGFI and depression/cognitive impairment. Restricted cubic spline (RCS) curves were plotted to visualize the dose–response relationship. Subgroup analyses and receiver operating characteristic (ROC) curves were used to assess interaction effects and evaluate the discriminatory ability of TyGFI. The results were expressed as odds ratios (OR) with 95% confidence intervals (CI).Results: Perimenopausal women with higher TyGFI levels exhibited a greater tendency towards depression and cognitive impairment, older age, higher blood pressure, and poorer self-reported health. In the fully adjusted model, each one-point increase in TyGFI was associated with elevated risk of depression (OR = 3.70, 95% CI: 3.18– 4.32) and cognitive impairment (OR = 1.51, 95% CI: 1.34– 1.71). RCS curves confirmed non-linear, positive associations and clear dose–response trends. Subgroup analysis revealed that the associations remained largely robust across most subgroups. ROC analysis demonstrated moderate-to-good discriminatory ability, with area under the curve (AUC) of 0.786 for depression and 0.634 for cognitive impairment.Conclusion: The TyGFI was concurrently correlated with depression and cognitive impairment in Chinese perimenopausal women, which may serve as a potential indicator for clinical risk screening.Keywords: perimenopause, triglyceride-glucose index, frailty index, depression, cognitive impairment, CHARLS

Background:Trigeminal neuralgia (TN) is a clinically intractable disorder characterized by episodes of severe pain that often significantly impairs patients' quality of life. Although previous studies suggest that acupuncture may alleviate pain, it is particularly important to conduct an updated meta-analysis to confirm the effectiveness and safety of acupuncture in light of updated clinical evidence in recent years. Methods:A systematic search was conducted across eight databases: four English-language (PubMed, Cochrane Library, Embase, and Web of Science) and four Chinese-language (Wanfang Data, China National Knowledge Infrastructure, China Science and Technology Journal Database, and China Biomedical Literature Service System). The search period for each database spans from its inception to 19 December 2025. Meta-analysis was conducted by using Stata 17.0. Results:A total of 38 randomized controlled trials involving 2836 patients with TN were included. Meta-analysis results showed that compared with the control group, the acupuncture group demonstrated significant differences in the following indicators: higher overall response rate (RR = 1.14, 95% CI 1.11-1.18), and lower visual analog scale scores (SMD = -1.19, 95% CI -1.62 to -0.76), frequency of pain episodes (SMD = -1.31, 95% CI -2.18 to -0.44), adverse events (RR 0.28, 95% CI: 0.16-0.50), and BPI-facial pain score (SMD = -0.53, 95% CI -0.96 to -0.09) were reduced. Conclusion:Acupuncture combined with medication may improve efficacy and reduce adverse events compared with monotherapy. However, the quality of existing evidence is low due to methodological limitations. Rigorous, multicenter randomized controlled trials with standardized protocols are needed to confirm the role of acupuncture for TN management.

BackgroundThe prevalence of adolescent major depressive disorder (MDD) is rising; however, diagnosis relies on subjective measures due to a lack of objective biomarkers. This study explored infrared thermography (IRT) as a non-invasive tool to quantify thermal radiation characteristics of acupoints in adolescents with MDD. The objective was to establish diagnostic models based on acupoint temperature-derived biomarkers.MethodsA prospective, multi-center observational study enrolled 108 participants (65 adolescents with MDD and 43 healthy controls [HCs]). We first examined correlations between acupoint temperatures and depression severity using Pearson analysis. Multiple linear and binary logistic regression models were developed to diagnose MDD and assess severity. The diagnostic model for MDD was visualized as a nomogram and validated using Receiver Operating Characteristic (ROC) curves, Hosmer-Lemeshow tests, calibration plots, and decision curve analysis (DCA). Internal validation was performed using the bootstrap method.ResultsAmong 27 acupoints analyzed, adolescents with MDD exhibited altered acupoint temperatures at Taiyang (EX-HN5), Quchi (LI11), Yanggu (SI5), and Waiqiu (GB36). Subsequent Pearson correlation analysis revealed negative correlations between the infrared relative temperatures of Taiyang (EX-HN5), Quchi (LI11), and Waiqiu (GB36) and depression severity (P = 0.001, r = -0.319; P = 0.022, r = -0.229; P = 0.001, r = -0.325) and a weak positive correlation between the infrared relative temperature of Yanggu (SI5) and depression severity (P = 0.043, r = 0.202). Building on these findings, two diagnostic models were developed: a linear regression model for depression severity of adolescents (Y = 52.25-9.52*TEX-HN5-13.07*TGB36) and a logistic regression model for adolescents with MDD diagnosis (P = ex/(1+ex), x = 0.22-1.14*TEX-HN5+0.45*TSI5-2.19*TGB36). The nomogram-based model demonstrated good calibration (Hosmer-Lemeshow P = 0.855), discrimination (AUC = 0.785, 95%CI: 0.693 - 0.876), and clinical utility. Internal validation using the bootstrap method produced a C-index of 0.752 (95% CI: 0.617 - 0.877), further confirming the model’s robustness.ConclusionsIn conclusion, acupoint temperature-based models show promising efficacy for the objective and non-invasive diagnosis and severity quantification of adolescents with MDD, offering valuable tools for early clinical intervention. Future studies should validate these findings across diverse populations and integrate multi-modal biomarkers to enhance diagnostic precision.Clinical Trial RegistrationClinicalTrials.gov, identifier NCT06750640.

Background: Primary insomnia (PI) is the most common clinical sleep disorder that can be effectively treated with acupuncture. However, the selection of optimal acupoints remain contentious. Objective: To investigate the pain sensitization phenomenon of specific acupoints commonly used in treating PI, we evaluated the threshold and sensitization rate of acupoint sensitization and their correlations with sleep questionnaire scores, polysomnography (PSG) and heart rate variability (HRV) to identify the optimal sensitized points. Methods: This study recruited 73 PI patients and 73 age- and sex-matched healthy controls in the outpatient acupuncture clinic of the Third Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine from January 31, 2024 to March 31, 2025. Pressure pain thresholds (PPT) were tested at 10 specific points commonly used in PI. The primary outcome measure was calculated as the acupoint hyperalgesia rate using the receiver operating characteristic (ROC) curve. Secondary outcome measures included PSG and HRV indices of some subjects. Results: Compared with the control group, PI patients showed significantly lower PPTs at specific acupoints such as RN14 and BL15, along with increased pain sensitivity rates. The pain sensitization rate in the T5-T8 nerve segments of PI patients was markedly elevated. Additionally, the degree of acupoint pain sensitization was negatively correlated with age and positively correlated with disease duration. Higher PSQI scores were associated with a greater likelihood of pain sensitization at EX-HN3, EX-HN0, and HT7. The probability of GV20 pain sensitization increased with higher SDS scores but decreased with increased high-frequency components in HRV. Conclusion: Pain sensitization was observed at specific points commonly used in PI patients. BL15 and RN14 may be the best acupoints for clinical diagnosis and treatment. Furthermore, acupoint pain sensitization was correlated with age, high-frequency, disease duration, PSQI, and SDS. Acupoint pain sensitization can be used as a screening and assessment index for acupuncture treatment in insomnia patients, providing an objective basis for individualized treatment.

Objective To observe the effect of electro-acupuncture(EA)at different acupoints on chronic inflammatory pain-anxiety comorbidity in mice,and to investigate the effect of naloxone on EA induced activation of neurons of the anterior cingulate cortex(ACC),medial prefrontal cortex(mPFC),paraventricular nucleus of the thalamus(PVT),and mediodorsal thalamus nucleus(MD),so as to explore whether the anti-anxiety effect of EA depends on its analgesic mechanism.Methods In the first part of the experiment,the efficacy of EA at different acupoints was compared.Sixty male C57BL/6 mice were randomly divided into control,model,EA Zusanli(ST36),EA Baihui(GV20),and EA Shengmen(HT7)groups(12 mice/group).In the second part,the impact of naloxone on the analgesic and anxiety effects of EA at ST36 was investigated.Forty male C57BL/6 mice were randomly divided into control,model,EA ST36,and EA ST36+naloxone groups(10 mice/group).Except for those in the control group,all the mice were subjected to a chronic inflammatory pain-anxiety comorbidity by subcutaneous injection of 20 μL emulsified complete Freund's adjuvant(CFA)into the hind paw.Mice in the control group received an equivalent volume of PBS.EA(2 Hz/100 Hz,0.2-0.4 mA)intervention was initiated on the 12th day after modeling,and applied to"Zusanli"(ST36,bilateral)or"Baihui"(GV20),or"Shengmen"(HT7,bilateral).The EA intervention was conducted for 30 min,once daily for 6 d.Naloxone was intraperitoneally injected 30 min before EA intervention.Paw withdrawal thresholds(PWTs)were measured using von Frey filaments.The anxiety-like behaviors were assessed using the open field(OF)test and elevated plus maze(EPM)test,separately.The expression of c-Fos in the ACC,mPFC,PVT,and MD was detected by immunofluorescence staining.Results Compared with the control group,the model group exhibited a significant decrease in the PWTs,number of entries into the open arms and time spent in the open arms in the EPM test,and in the number of entries into the central area,time spent in the central area,and distance traveled in the central area in the OF test(P<0.01),and a significant increase in the number of c-Fos positive neurons in the bilateral ACC,mPFC and MD(P<0.05).The total distance traveled of OF test remained unchanged.Compared with the model group,1)EA at ST36,GV20,and HT7 significantly increased the PWTs(P<0.01,P<0.05),2)EA at ST36 significantly increased the number of entries into the open arms,time spent in the open arms in the EPM test,and increased the number of entries,time spent,and distance traveled in the central area in the OF test(P<0.01 P<0.05),3)EA at GV20 significantly increased the time spent in the open arms in the EPM test(P<0.01),4)EA at HT7 obviously increased the time spent in the open arms of the EPM test and increased the time spent in the central area of the OF test(P<0.01),and 5)EA at ST36 significantly decreased the expression of c-Fos in the bilateral ACC,contralateral mPFC,bilateral PVT,and bilateral MD(P<0.05).Compared with the EA ST36 group,the EA ST36+naloxone group showed a significant decrease in the PWTs(P<0.01),number of entries into the open arms,total distance traveled and time spent in the open arms in the EPM test(P<0.05),and in the number of entries into the central area and time spent in the central area in the OF test(P<0.01).The expression levels of c-Fos in the bilateral ACC and ipsilateral MD were strikingly higher in the EA ST36+naloxone group than those in the EA ST36 group(P<0.05),suggesting a disappearance of the suppressive effect of EA after administration of naloxone.Conclusion EA at ST36 has a better analgesic and emotional relief effects on mice with comorbid pain and emotion compared to EA at GV20 and HT7.The underlying mechanism may involve the inhibition of neuronal excitability in the ACC,mPFC,MD and PVT.Additionally,the emotional relief effect of EA at ST36 is dependent on its analgesic efficacy,which may be related to the inhibition of neuronal excitability in the ACC and MD.

BACKGROUND:The analgesic effects of multiple electroacupuncture (EA) sessions and single EA sessions differ significantly in pain management. Area 24b (A24b) of the anterior cingulate cortex (ACC) is crucial in pain processing. EA relieves pain by targeting and modulating the neuronal activity within this subregion. However, whether the cumulative effect of EA antinociception is connected to A24b mechanisms has remained unclear. METHODS:In our study, we used the Complete Freund's Adjuvant (CFA) model to induce inflammatory pain and the Spared Nerve Injury (SNI) model to induce neuropathic pain, and adult male C57BL/6, FosTRAP, and FosTRAP:Ai9 mice were used as experimental subjects to investigate the cumulative effect of EA antinociception and whether multiple EA sessions and a single EA session regulate different neuronal populations in the A24b. RESULTS:We observed that EA effectively alleviated pain in mice, with three EA sessions yielding superior analgesic effects compared to a single session. Using chemical genetics combined with FosCreER technology to activate EA-TRAPed cells in the A24b, we found that pain relief was more pronounced with three EA sessions. Moreover, chemogenetic inhibition of EA-TRAPed cells in the A24b reversed the analgesic effects of a single EA session but not those of three EA sessions. Fluorescent in situ hybridization results indicated that three EA sessions significantly increased the number of GABAergic neurons in the A24b compared with a single session. Additionally, retrograde tracing revealed that the A24b circuit that monosynaptically innervates EA-TRAPed cells included projections from the central lateral nucleus (CL), lateral mediodorsal thalamic nucleus (MDL), lateral habenula (LHb), dorsal raphe nucleus (DR), caudal linear nucleus of the raphe (CLi), dorsal tuberomamillary nucleus (DTM), periventricular hypothalamic nucleus (Pe) and hippocampal fields CA1, CA2, and CA3. These findings suggest that multiple EA sessions and single EA sessions activated different neuronal populations in the A24b. The enhanced analgesic effect of multiple EA sessions may be attributed to an increase in the proportion of GABAergic neurons within the A24b. CONCLUSIONS:Multiple and single EA sessions recruit distinct neuronal populations in A24b, with the stronger analgesic effect of repeated EA linked to a higher proportion of GABAergic neurons in this region.

Introduction:Trigeminal neuralgia (TN) is a severe neuropathic pain disorder characterized by unpredictable pain paroxysms, significantly impairing patients' quality of life through associated anxiety and depression. Electroacupuncture (EA) has demonstrated clinical value in managing chronic neuropathic pain. However, its specific efficacy as a pharmacotherapy-sparing strategy for TN remains to be systematically evaluated. This study aims to evaluate the short-term effectiveness and safety of an EA regimen for TN, and to examine changes in exploratory neuro-immune biomarkers, including IL-6 and 5-HT, related to pain and affective symptoms. Methods and Analysis:This is a multicenter, randomized, double-simulation, placebo-controlled trial designed to compare the efficacy of EA plus placebo with sham EA plus carbamazepine (CBZ) for TN. A total of 126 adults with TN, maintained on a stable, low-to-moderate CBZ dose (200-400 mg/day) to control for baseline medication-masking effects, will be randomly allocated to the EA plus placebo group or the sham EA plus CBZ group at a 1:1 ratio. Participants in the EA plus placebo group will receive EA plus placebo for 2 weeks, while those in the sham EA plus CBZ group will receive sham EA plus active CBZ on the same schedule, followed by a 6-week follow-up. The primary outcome will be the proportion of participants achieving a ≥50% reduction in visual analog scale (VAS) score at week 2. Secondary outcomes will include daily pain diary metrics (attack frequency, intensity, rescue medication use), Patient Global Impression of Change, Brief Pain Inventory-Facial, Short-Form McGill Pain Questionnaire, and psychological assessments (Self-Rating Anxiety and Depression Scales). Furthermore, plasma biomarkers (interleukin-6 and serotonin) and safety profiles (hepatic and renal functions) will be evaluated at baseline and week 2. All analyses will be conducted in accordance with the intention-to-treat principle. Trial Registration:ClinicalTrials.gov Identifier: NCT06977932 (March 2, 2026).

Postoperative pain develops after surgical operation. Inadequately managed postoperative pain is oftentimes associated with deterioration in life quality and delayed rehabilitation. Clinical studies confirmed analgesic effects of acupuncture on patients with postoperative pain. We hereby explored mechanisms underlying how acupuncture alleviates postoperative pain. A mouse model of skin incision-induced pain was established to mimic postoperative pain condition. Electroacupuncture (EA) or sham EA was administered at ST36 and BL60 acupoints in model mice. A combination of RNA-Sequencing (RNA-Seq), immunostaining, biochemical assay, in vivo imaging and behavioral test were applied for mechanism investigations. 2/100 Hz EA intervention ameliorated mechanical allodynia and improved cumulative pain of incisional pain model mice. Sham EA exerted no obvious analgesic effect. Incisional pain model mice develop gait impairment, which was improved by EA intervention. RNA-Seq identified EA significantly reduced Il33 gene overexpression in the incised skin tissues of model mice. IL-33 was produced from keratinocytes upon skin incision. EA reduced IL-33 overproduction from keratinocytes, resulting in less macrophage infiltration and less ROS accumulation in the incised skin. Adoptive transfer of macrophages into the incised tissue abrogated the ameliorative effects of EA on macrophage infiltration as well as ROS accumulation in incised skin and further reversed EA-induced analgesia on incisional pain model mice. Additionally, EA intervention did not affect skin wound healing process. These results demonstrate that EA ameliorates incisional pain via suppressing IL-33 overproduction in incised skin tissue. EA-induced suppression of IL-33 overproduction results in less macrophage infiltration and ROS accumulation that contribute to analgesia. Our work helps to support EA as an alternative and green therapy for postoperative pain management.

Objective To observe the effect of electro-acupuncture (EA) at different acupoints on chronic inflammatory pain-anxiety comorbidity in mice, and to investigate the effect of naloxone on EA induced activation of neurons of the anterior cingulate cortex (ACC), medial prefrontal cortex (mPFC), paraventricular nucleus of the thalamus (PVT), and mediodorsal thalamus nucleus (MD), so as to explore whether the anti-anxiety effect of EA depends on its analgesic mechanism. Methods In the first part of the experiment, the efficacy of EA at different acupoints was compared. Sixty male C57BL/6 mice were randomly divided into control, model, EA Zusanli (ST36), EA Baihui (GV20), and EA Shengmen (HT7) groups (12 mice/group). In the second part, the impact of naloxone on the analgesic and anxiety effects of EA at ST36 was investigated. Forty male C57 BL/6 mice were randomly divided into control, model, EA ST36, and EA ST36 + naloxone groups (10 mice/group). Except for those in the control group, all the mice were subjected to a chronic inflammatory pain-anxiety comorbidity by subcutaneous injection of 20 u03BCL emulsified complete Freundu2019s adjuvant (CFA) into the hind paw. Mice in the control group received an equivalent volume of PBS. EA (2 Hz/100 Hz, 0.2u20140.4 mA) intervention was initiated on the 12th day after modeling, and applied to u201CZusanliu201D (ST36, bilateral) or u201CBaihuiu201D (GV20), or u201CShengmenu201D (HT7, bilateral). The EA intervention was conducted for 30 min, once daily for 6 d. Naloxone was intraperitoneally injected 30 min before EA intervention. Paw withdrawal thresholds (PWTs) were measured using von Frey filaments. The anxiety-like behaviors were assessed using the open field (OF) test and elevated plus maze (EPM) test, separately. The expression of c-Fos in the ACC, mPFC, PVT, and MD was detected by immunofluorescence staining. Results Compared with the control group, the model group exhibited a significant decrease in the PWTs, number of entries into the open arms and time spent in the open arms in the EPM test, and in the number of entries into the central area, time spent in the central area, and distance traveled in the central area in the OF test (Pu0026lt;0.01), and a significant increase in the number of c-Fos positive neurons in the bilateral ACC, mPFC and MD (Pu0026lt;0.05). The total distance traveled of OF test remained unchanged. Compared with the model group, 1) EA at ST36, GV20, and HT7 significantly increased the PWTs (Pu0026lt;0.01, Pu0026lt;0.05), 2) EA at ST36 significantly increased the number of entries into the open arms, time spent in the open arms in the EPM test, and increased the number of entries, time spent, and distance traveled in the central area in the OF test (Pu0026lt;0.01 Pu0026lt;0.05), 3) EA at GV20 significantly increased the time spent in the open arms in the EPM test (Pu0026lt;0.01), 4) EA at HT7 obviously increased the time spent in the open arms of the EPM test and increased the time spent in the central area of the OF test (Pu0026lt;0.01), and 5) EA at ST36 significantly decreased the expression of c-Fos in the bilateral ACC, contralateral mPFC, bilateral PVT, and bilateral MD (Pu0026lt;0.05). Compared with the EA ST36 group, the EA ST36 + naloxone group showed a significant decrease in the PWTs (Pu0026lt;0.01), number of entries into the open arms, total distance traveled and time spent in the open arms in the EPM test (Pu0026lt;0.05), and in the number of entries into the central area and time spent in the central area in the OF test (Pu0026lt;0.01). The expression levels of c-Fos in the bilateral ACC and ipsilateral MD were strikingly higher in the EA ST36 + naloxone group than those in the EA ST36 group (Pu0026lt;0.05), suggesting a disappearance of the suppressive effect of EA after administration of naloxone. Conclusion EA at ST36 has a better analgesic and emotional relief effects on mice with comorbid pain and emotion compared to EA at GV20 and HT7. The underlying mechanism may involve the inhibition of neuronal excitability in the ACC, mPFC, MD and PVT. Additionally, the emotional relief effect of EA at ST36 is dependent on its analgesic efficacy, which may be related to the inhibition of neuronal excitability in the ACC and MD.