ObjectiveMigraine is a leading neurological disorder and the fourth most common cause of years lived with disability worldwide, affecting nearly 116 million individuals. Although pharmacological treatments are available, their efficacy is often limited by side effects and variable response rates. Repetitive transcranial magnetic stimulation (rTMS) over the dorsolateral prefrontal cortex (DLPFC) offers a safe, non-invasive alternative for migraine management. However, the neurophysiological mechanisms, particularly how rTMS modulates local cortical excitability and distributed pain-related circuits, remain poorly understood. Elucidating these mechanisms is essential for optimizing treatment protocols and improving clinical outcomes. MethodsThis study employed concurrent transcranial magnetic stimulation and electroencephalography (TMS-EEG) to investigate neuroplastic and neurocircuitry mechanisms of DLPFC-rTMS in migraine. Study 1 compared 30 migraineurs and 28 healthy controls to identify abnormalities in TMS-evoked potentials (TEPs) and significant current density (SCD) within sensory-discriminative regions including the primary somatosensory cortex (S1) and posterior insula (pINS), cognitive-affective regions including the anterior insula (aINS) and midcingulate cortex (MCC), and a descending modulatory region, the periaqueductal gray (PAG). Study 2 used a single-blind, crossover, sham-controlled design in 34 healthy participants. Each participant received both active (10 Hz, 80% RMT, 1 500 pulses) and sham DLPFC-rTMS in counterbalanced order. TMS-EEG and cold pain tolerance were assessed before and after each session. ResultsIn Study 1, migraineurs showed a significantly less negative N120 amplitude compared to healthy controls (P=0.027, Cohen’s d=0.60), indicating local intracortical disinhibition. No group differences were observed for N40, P60, or P180 components. At the source level, migraineurs exhibited significantly higher SCD in the S1, pINS, aINS, and MCC (allQ<0.05), but not in the ventroposterior thalamus (vpTHAL), mediodorsal thalamus (mdTHAL), or PAG. In Study 2, active rTMS significantly reduced SCD from pre- to post-stimulation in the S1, aINS, and MCC (all Q<0.05). Sham stimulation also reduced SCD in the S1 (Q<0.05) but not in the aINS or MCC. Although no significant group-level analgesic effect was observed between active and sham conditions (P=0.107), correlation analyses revealed that greater SCD reductions in the S1 and MCC were significantly associated with higher post-rTMS pain tolerance (R=-0.487 and -0.495, both Q<0.01) and larger improvements in pain tolerance(R=-0.487 and -0.451, both Q<0.05). No such correlations were found following sham stimulation, suggesting that the behavioural relevance of neural changes is specific to active rTMS. ConclusionThis study provides novel evidence that migraineurs exhibit both local neuroplastic abnormalities (reduced N120 amplitude) and hyperactivity in key pain-processing regions (S1, pINS, aINS, MCC). A single session of DLPFC-rTMS reduced hyperactivity in the aINS, MCC, and S1. Notably, greater reductions in the S1 and MCC were associated with improved pain tolerance. These findings identify distinct cortical circuitries, particularly within the cognitive-affective pain network, that may serve as potential biomarkers for optimizing rTMS treatment in migraine and other chronic pain conditions. Future studies should validate these results in patient populations experiencing spontaneous migraine attacks and explore multi-session or accelerated rTMS protocols.

This study was aimed at establishing a sensitive and specific sandwich ELISA detection method for Brucella.We screened monoclonal capture antibodies and detection antibodies for Brucella detection,and optimized and determined the opti-mal antibody coating time and concentration,as well as the optimal blocking solution,blocking time,and yin-yang critical val-ue.The specificity of this method was verified by examination of other bacteria prone to cross-reacting with Brucella.The sen-sitivity of the method was verified by detection of a gradient dilution of inactivated Brucella.Moreover,the sandwich ELISA detection results were compared with test tube agglutination and qPCR results.The selected capture antibody was 4A12,and the selected detection antibody was 6C12.Experimental analysis indicated that the optimal coating concentration for the 4A12 capture antibody was 5 μg/mL,and the optimal dilution ratio for the 6C12 detection antibody was 1∶2000.The optimal coating conditions were overnight at 4℃,and blocking with 5％skim milk powder for 2 hours.The established double antibody sand-wich ELISA method reacted with only Brucella but not other bacteria,thus demonstrating the method's good specificity.Inac-tivated Brucella solution was still detectable after dilution to 1 × 105 CFU/mL,thus demonstrating the method's good sensitiv-ity.The intra-and inter batch coefficients of variation were both below 10％,thus indicating the method's good repeatability.Thus,this study successfully established a dual antibody sandwich ELISA method for Brucella detection,which has good spe-cificity and sensitivity,and might provide an effective approach for the precise diagnosis and effective prevention and control of brucellosis.

Blood from a case of group B epidemic cerebrospinal meningitis identified in February 2024 in Ganzhou City,Jiangxi Province,and throat swabs from close contacts were collected for isolation and culture.The isolates were subjected to serogrouping,drug sensitivity testing,and whole genome sequencing and analysis,to provide a basis for epidemiological inves-tigation and clinical drug use.One strain of Neisseria meningitidis was isolated from the blood of the case and denoted group B.The MLST type was ST-7962,with no clonal group attribution.The phylogenetic tree showed that it was genetically close to the 1977 Shanghai carrier isolate(id-52231).Drug sensitivity results indicated that the strain was sensitive to 8 drugs:azithro-mycin,cefotaxime,minocycline,ceftriaxone,chloramphenicol,meropenem,rifampicin,and benzylpenicillin;resistant to cot-rimoxazole,levofloxacin,and ciprofloxacin;and showed an intermediate response to penicillin.This report describes the first case of ST-7962 group B meningoencephalitis found in Jiangxi Province.Monitoring of Neisseria meningitidis carriage,drug re-sistance,and molecular characteristics of strains in the healthy population in this region should be strengthened,to provide la-boratory support for the clinical use of medications,traceability,and control of the pathogen underlying meningoencephalitis infection.

Icaritin(ICT)is an 8-isopentenylflavonoid,which is the main effective component of the tra-ditional Chinese medicine Epimedium.Previously,we found that Icaritin inhibits the growth of glioblasto-ma(GBM)cells.Herein we aim to study the in vivo anti-GBM effectiveness of Icaritin and explore its mechanism.The results of MTT assay,flow cytometry,comet assay and cellular immunofluorescence as-say in vitro showed that ICT inhibited the proliferation of four kinds of GBM cells,U87,U251,U118 and A172,induced early apoptosis(P＜0.001)and late apoptosis(P＜0.05)in U87 cells,induced DNA damage in U87 cells,and blocked the growth of U87 cells at the G0/G1 phase(P＜0.0001)in a concen-tration-time-dependent manner.In vivo subcutaneous tumor transplantation tumor experiments showed that feeding 200 mg/kg(P＜0.01)and 400 mg/kg(P＜0.001)ICT had a significant inhibitory effect on the growth of GBM subcutaneous tumors,and had no significant toxic effects on heart,liver,spleen,lung and kidney tissues.The results of network pharmacological analysis,molecular docking and cellular thermodynamic experiments showed that there were 26 possible target proteins between ICT and GBM,a-mong which the expression of p53 in GBM tissues was significantly(P＜0.001)higher than in normal tis-sues,and the binding energy of ICT and p53 was lower;cellular thermodynamic experiments verified that ICT significantly enriched the level of p53 in the living cells of GBM,which indicated that ICT could tar-get p53.The expression of key proteins in the DNA damage repair and apoptosis-associated FOXO signa-ling pathway was detected by ICT.The results showed that the expression of ATR(P＜0.01),P53(P＜0.001),P21(P＜0.05)and γ-H2AX(P＜0.05)was up-regulated,whereas the expression of Cyc-lin E1(P＜0.01),E2F1(P＜0.05),CDK2(P＜0.01),Rb(P＜0.001),p-Rb(P＜0.0001)and WRN(P＜0.0001)expression were down-regulated.There was no significant change in the expres-sion of FOXO 1 in the FOXO pathway or a significant down-regulation of its phosphorylation level.This study demonstrated that ICT could effectively inhibit the growth of GBM cells in vivo.It targets p53 to regulate the DNA damage repair pathway and FOXO signaling pathway to induce GBM cell cycle arrest and apoptosis.

Objective To evaluate the status of latent tuberculosis infection(LTBI)with Mycobacterium tubercu-losis and the risk of LTBI activation after glucocorticoid therapy in patients with anti-neutrophil cytoplasmic antibody(ANCA)-associated vasculitis(AAV).Methods Clinical data of AAV patients who visited Xiangya Hospital of Central South University from May 2018 to May 2023 were retrospectively analyzed.According to the results of in-terferon-γ release assay(IGRA),they were divided into a LTBI group and a non-LTBI group.The LTBI group was subdivided into an active TB group(ATB group)and a non-ATB group according to the presence of ATB.General conditions and biochemical indicator characteristics of the two groups were compared.The COX regression model was used to analyze the risk factors for LTBI activation in AAV patients.Results A total of 302 AAV patients were included in this study,with an age of 64.0(53.0,71.0)years old,169 male cases(55.96％),and an IGRA positive rate of 24.17％(n=73).The proportion of patients with renal insufficiency in the LTBI group was higher than that in the non-LTBI group(79.45％vs 60.70％),with statistically significant difference(P＜0.05).In the LTBI group,the age of patients was 62.0(53.5,72.0)years old,44 cases were male(60.27％),out of which 11 cases(15.07％)had ATB.In the non-LTBI group,4 male cases(1.75％)had ATB.In the LTBI group,there were statistically significant differences between the ATB group and the non-ATB group in terms of daily average glucocorticoid levels,age,renal dysfunction,neutrophils,and blood creatinine(all P＜0.05).COX univariate re-gression analysis showed that statistically significant differences existed among the groups in terms of daily average glucocorticoid levels,daily average glucocorticoid levels＞11.75 mg/day,age,age＞65 years old,and blood crea-tinine levels ≥150 μmol/L(all P＜0.05).COX multivariate regression analysis showed that daily average glucocor-ticoid levels＞11.75 mg/day(OR=0.14,95％CI:1.30-79.01)and age＞65 years(OR=0.08,95％CI:0.01-0.85)were independent influencing factors for LTBI activation.Conclusion AAV patients have a high rate of LT-BI,and daily average glucocorticoid levels＞11.75 mg/day is an independent risk factor for LTBI activation in AAV patients.Age＞65 years may benefit from receiving lower doses of immunosuppressive therapy,but it is necessary to expand sample size for further study.

Objective:According to the"Technology-Organization-Environment"framework,it aims to analyze barriers of meaningful use of digital technologies in primary care facilities and propose corresponding optimization strategies.Methods:Nvivo 11.0 Software is used to conduct thematic framework analysis on interview data of primary healthcare and administrative staff in a district of Jiangsu Province.Results:The barriers are as follows:(1)technology level:infrastructure needs improvement,unified technology standards for data,low level of technology usability and usefulness,and limited economic benefits;(2)organizational level:insufficient digital literacy of medical staff,low organizational innovation ability,lack of training,inertia of healthcare staff,and limited size of primary care facilities;(3)environmental level:insufficient government support,high government requirements of performance evaluation,low digital literacy of residents,not covered by medical insurance schemes,constraints on interests among stakeholders,and mismatched digital scenarios.The main reason for insufficient effective use of digital technologies includes insufficient adaptability between digital technologies and the practical needs of grassroots health services,lack of organizational change and proactive personnel behavior,low level of external environment optimization and institutional support.Conclusion:Policy makers and implementers should be demand-oriented,continuously improve digital technologies maturity in primary healthcare system,integrate organizational resources,achieve changes in primary healthcare models,optimize the external environment,and enhance collaborative governance levels.

With the rapid development of generative artificial intelligence(GAI)technologies,their widespread application in academic research and writing is continuously expanding the boundaries of sci-entific inquiry.However,this trend has also raised a series of ethical and regulatory challenges,inclu-ding issues related to authorship,content authenticity,citation accuracy,and accountability.In light of the growing involvement of AI in generating academic content,establishing an open,controllable,and trustworthy ethical governance framework has become a key task for safeguarding research integrity and maintaining trust within the academic community.This expert consensus outlines ethical requirements across key stages of AI-assisted academic writing-including topic selection,data management,citation practices,and authorship attribution.It aims to clarify the boundaries and ethical obligations surrounding AI use in academic writing,ensuring that technological tools enhance efficiency without compromising in-tegrity.The goal is to provide guidance and institutional support for building a responsible and sustainable research ecosystem.

This study was aimed at establishing a sensitive and specific sandwich ELISA detection method for Brucella.We screened monoclonal capture antibodies and detection antibodies for Brucella detection,and optimized and determined the opti-mal antibody coating time and concentration,as well as the optimal blocking solution,blocking time,and yin-yang critical val-ue.The specificity of this method was verified by examination of other bacteria prone to cross-reacting with Brucella.The sen-sitivity of the method was verified by detection of a gradient dilution of inactivated Brucella.Moreover,the sandwich ELISA detection results were compared with test tube agglutination and qPCR results.The selected capture antibody was 4A12,and the selected detection antibody was 6C12.Experimental analysis indicated that the optimal coating concentration for the 4A12 capture antibody was 5 μg/mL,and the optimal dilution ratio for the 6C12 detection antibody was 1∶2000.The optimal coating conditions were overnight at 4℃,and blocking with 5％skim milk powder for 2 hours.The established double antibody sand-wich ELISA method reacted with only Brucella but not other bacteria,thus demonstrating the method's good specificity.Inac-tivated Brucella solution was still detectable after dilution to 1 × 105 CFU/mL,thus demonstrating the method's good sensitiv-ity.The intra-and inter batch coefficients of variation were both below 10％,thus indicating the method's good repeatability.Thus,this study successfully established a dual antibody sandwich ELISA method for Brucella detection,which has good spe-cificity and sensitivity,and might provide an effective approach for the precise diagnosis and effective prevention and control of brucellosis.

Objective To evaluate the status of latent tuberculosis infection(LTBI)with Mycobacterium tubercu-losis and the risk of LTBI activation after glucocorticoid therapy in patients with anti-neutrophil cytoplasmic antibody(ANCA)-associated vasculitis(AAV).Methods Clinical data of AAV patients who visited Xiangya Hospital of Central South University from May 2018 to May 2023 were retrospectively analyzed.According to the results of in-terferon-γ release assay(IGRA),they were divided into a LTBI group and a non-LTBI group.The LTBI group was subdivided into an active TB group(ATB group)and a non-ATB group according to the presence of ATB.General conditions and biochemical indicator characteristics of the two groups were compared.The COX regression model was used to analyze the risk factors for LTBI activation in AAV patients.Results A total of 302 AAV patients were included in this study,with an age of 64.0(53.0,71.0)years old,169 male cases(55.96％),and an IGRA positive rate of 24.17％(n=73).The proportion of patients with renal insufficiency in the LTBI group was higher than that in the non-LTBI group(79.45％vs 60.70％),with statistically significant difference(P＜0.05).In the LTBI group,the age of patients was 62.0(53.5,72.0)years old,44 cases were male(60.27％),out of which 11 cases(15.07％)had ATB.In the non-LTBI group,4 male cases(1.75％)had ATB.In the LTBI group,there were statistically significant differences between the ATB group and the non-ATB group in terms of daily average glucocorticoid levels,age,renal dysfunction,neutrophils,and blood creatinine(all P＜0.05).COX univariate re-gression analysis showed that statistically significant differences existed among the groups in terms of daily average glucocorticoid levels,daily average glucocorticoid levels＞11.75 mg/day,age,age＞65 years old,and blood crea-tinine levels ≥150 μmol/L(all P＜0.05).COX multivariate regression analysis showed that daily average glucocor-ticoid levels＞11.75 mg/day(OR=0.14,95％CI:1.30-79.01)and age＞65 years(OR=0.08,95％CI:0.01-0.85)were independent influencing factors for LTBI activation.Conclusion AAV patients have a high rate of LT-BI,and daily average glucocorticoid levels＞11.75 mg/day is an independent risk factor for LTBI activation in AAV patients.Age＞65 years may benefit from receiving lower doses of immunosuppressive therapy,but it is necessary to expand sample size for further study.

Blood from a case of group B epidemic cerebrospinal meningitis identified in February 2024 in Ganzhou City,Jiangxi Province,and throat swabs from close contacts were collected for isolation and culture.The isolates were subjected to serogrouping,drug sensitivity testing,and whole genome sequencing and analysis,to provide a basis for epidemiological inves-tigation and clinical drug use.One strain of Neisseria meningitidis was isolated from the blood of the case and denoted group B.The MLST type was ST-7962,with no clonal group attribution.The phylogenetic tree showed that it was genetically close to the 1977 Shanghai carrier isolate(id-52231).Drug sensitivity results indicated that the strain was sensitive to 8 drugs:azithro-mycin,cefotaxime,minocycline,ceftriaxone,chloramphenicol,meropenem,rifampicin,and benzylpenicillin;resistant to cot-rimoxazole,levofloxacin,and ciprofloxacin;and showed an intermediate response to penicillin.This report describes the first case of ST-7962 group B meningoencephalitis found in Jiangxi Province.Monitoring of Neisseria meningitidis carriage,drug re-sistance,and molecular characteristics of strains in the healthy population in this region should be strengthened,to provide la-boratory support for the clinical use of medications,traceability,and control of the pathogen underlying meningoencephalitis infection.