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.

Objective·To investigate the effects and molecular mechanisms of phosphatidylethanolamine(PE)on macrophage senescence and its senescence-associated secretory phenotype(SASP),as well as its pathophysiological role in liver injury.Methods·A macrophage senescence model was established using doxorubicin(DOX),followed by PE treatment.A mouse liver injury model was generated via intraperitoneal co-administration of PE and lipopolysaccharide(LPS)to investigate the effects of PE on liver injury.Senescence markers and SASP factors,including senescence-associated β-galactosidase(SA-β-gal),cell cycle inhibitor p21,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6),were evaluated using SA-β-gal staining,quantitative real-time PCR,and Western blotting.RNA sequencing(RNA-seq)was performed,followed by Gene Ontology(GO)cellular component enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,Gene Set Variation Analysis(GSVA),and Gene Set Enrichment Analysis(GSEA),to explore the molecular mechanisms and signaling pathways by which PE promotes macrophage senescence.The expression of endoplasmic reticulum(ER)stress-related proteins,including inositol-requiring enzyme 1 α(IRE1α),spliced X-box binding protein 1(XBP1s),activating transcription factor 6(ATF6),ATF4,and C/EBP homologous protein(CHOP),was analyzed through in vivo and in vitro experiments.Results·PE significantly promoted the expression of senescence markers SA-β-gal,p21,p16 and SASP factors.RNA-seq analysis revealed that ER stress was involved in PE-induced promotion of SASP.Further experiments demonstrated that PE activated the ER stress signaling pathway,promoting macrophage senescence and the expression of SASP factors.In vivo experiments further confirmed that PE exacerbated LPS-induced liver injury in mice through ER stress.Conclusion·PE promotes macrophage senescence and the expression of SASP factors by activating ER stress signaling pathway,thereby aggravating LPS-induced liver injury.

Objective To analyze the association between lifestyle and the risk of type 2 diabetes(T2D)among adult residents.Methods The data was sourced from the Shanghai Suburban Adult Cohort and Biobank.A total of 42 096 adult residents who had not developed T2D were recruited from four districts of Shanghai(Songjiang,Jiading,Minhang,and Xuhui)between 2016 and 2019.The follow-up ended on Feb 28,2023.A structured questionnaire was used to collect information on six lifestyle-related items,including smoking,alcohol consumption,BMI,waist circumference(WC),physical activity,and diet.The unhealthy lifestyle scores(UHLS)were calculated by counting the number of all the unhealthy lifestyle items,with a range of 0-6.New-onset T2D events diagnosed by physicians were obtained through the medical information system.Cox proportional hazards regression model and restricted cubic spline model were utilized to evaluate the association between unhealthy lifestyles and the risk of T2D incidence.Results About 28.1%of the participants led 4-6 unhealthy lifestyles.A total of 1 752 new T2D cases were identified during 218 513.4 person-years of follow-up.Analysis of single unhealthy lifestyle showed that abnormal WC(HR=1.5,95%CI:1.4-1.7)and abnormal BMI(HR=1.3,95%CI:1.2-1.5)were associated with an increased risk of T2D.Compared with individuals with a UHLS of 0-1,those with a UHLS of 3 and 4-6 had 30%(95%CI:1.1-1.6)and 50%(95%CI:1.2-1.8)higher risks of T2D,respectively.Each additional unhealthy lifestyle was associated with a 10%increase in T2D incidence risk(HR=1.1,95%CI:1.1-1.2).Conclusion The risk of T2D in adult residents increases with the cumulative number of unhealthy lifestyles.Adult residents with abnormal WC or BMI,or have three or more unhealthy lifestyles accumulated,will increase the risk of new-onset T2D.

The scientific research and innovation capabilities of medical students are intrinsically linked to the sustained and high-quality development of national healthcare initiatives.Cultivating outstanding medi-cal students with independent scientific capabilities and innovative consciousness is a critical component in the education and training of high-level medical professionals.Our investigation revealed that within the imperfections of the cultivating model,some faculty and students at medical schools have an insufficient understanding of scientific research and innovation and lack motivation for engaging in such activities,which hinder the progression of scientific research activities.Consequently,we initiated a teaching practice and exploratory study on the"tutorial system"aimed at fostering medical students'scientific research and innovation abilities.Based on the principle of"research informing teaching,teaching and research advan-cing together,"this study implements a"tutorial system"coordinated by tutors,supplemented by graduate and undergraduate student mentors,to cultivate innovative thinking,stimulate interest in scientific re-search,and enhance practical and research skills among medical students.Through collaborative efforts within"scientific research innovation teams,"various educational methods—including preliminary re-search,in-class and extracurricular activities,intra-group and inter-group interactions,and theoretical and practical applications—are employed to improve and strengthen the cultivation of medical students'scientif-ic research and innovation abilities.This study aims to provide valuable references for optimizing medical education management systems and enhancing the quality of medical student training.

This study was aimed at investigating the effects and mechanisms of Toxoplasma gondii type Ⅰ(RH strain)ROP16 protein on proliferation and the cell cycle in mouse alveolar macrophage MH-S cells.We constructed a Toxoplasma gondii type Ⅰ(RH)ROP16 overexpression lentivirus,transduced MH-S cells,and then screened cells with puromycin to obtain a cell line stably overexpressing ROP16Ⅰ.RT-qPCR and western blotting were used to verify expression effects,CCK-8 assays were used to detect cell proliferation activity,and flow cytometry was used to detect cell cycle changes.Western blotting and RT-qPCR were used to detect the expression levels of p53,p21,CDK6,Cyclin D1,STAT3,p-STAT3(Y705),and JAK1 proteins or genes,and immunofluorescence was used to detect the expression levels of ROP16Ⅰ and p-STAT3(Y705)and their subcellular co-localization in MH-S cells.ROP16Ⅰ protein and gene expression were detected in MH-S cells transduced with lentivirus for ROP16Ⅰ overexpression.CCK-8 assays revealed that ROP16Ⅰ promoted the proliferation of MH-S cells(P＜0.01)and enhanced cell viability.Flow cytometry revealed that ROP16Ⅰ overexpression decreased the G0/G1 phase and elevated the G2 and S phases of the cell cycle in MH-S cells(P＜0.01 or P＜0.05).Compared with the MH-S cell group and MH-S-empty vector group,the MH-S-ROP16 cell group showed lower expression of p53 and p21 proteins;higher expression of CDK6,Cyclin D1,p-STAT3(Y705),and JAK1 proteins;lower expression of p53 and p21 mRNAs;and higher expression of CDK6 and Cyclin D1 mRNAs(all P＜0.01).Immunofluorescence revealed that ROP16Ⅰ co-localized with p-STAT3(Y705)in the nucleus and surrounding cytoplasm.Therefore,Toxoplasma gondii type Ⅰ(RH)ROP16Ⅰ protein activates the JAK-STAT3 pathway;shortens the G0/G1 phase and lengthens the G2/S phase of the cell cycle;and promotes cell proliferation.These findings provide a theoretical basis for revealing the mechanism of immune evasion of Toxoplasma gondii,and lay a foundation for research on the prevention and treatment of Toxoplasma gondii pneumonia.

Objectives:To investigate the safety and efficacy of Cuffstent in the treatment of immediate type Ia endoleak during endovascular aneurysm repair(EVAR)of abdominal aortic aneurysm.Methods:The clinical data of 24 patients with immediate type Ia endoleak during EVAR treated with Cuffstent at the Second Xiangya Hospital of Central South University from January 2019 to December 2022 were retrospectively analyzed.There were 18 males and 6 females with a mean age of(70.3±7.1)years.Results:Of the 24 patients,22 underwent conventional proximal Cuffstent implantation,and 2 underwent fenestrated Cuffstent to preserve unilateral renal artery.Emergency EVAR was performed in 7 patients.The procedure was technically successful in all patients.Immediate disappearance of type Ia endoleak was achieved in 23 patients,whereas 1 patient exhibited mild persistent type Ia endoleak,which was managed conservatively with watchful follow-up.All the 24 patients finished an average of(39.0±15.1)months follow-up,and all patients were alive during the follow-up period.Two patients received aortic reintervention,and one patient received thoracic aortic stent graft implantation due to thoracic aortic penetrating ulcer,and one patient received abdominal aortic aneurysm resection and iliac artery stent implantation due to type II endoleak and iliac artery rupture,respectively.There were no other aorta-related complications and reintervention.Conclusions:Proximal addition of Cuffstent for the management of immediate type Ia endoleak during EVAR is safe and effective.

This study was aimed at investigating the effects and mechanisms of Toxoplasma gondii type Ⅰ(RH strain)ROP16 protein on proliferation and the cell cycle in mouse alveolar macrophage MH-S cells.We constructed a Toxoplasma gondii type Ⅰ(RH)ROP16 overexpression lentivirus,transduced MH-S cells,and then screened cells with puromycin to obtain a cell line stably overexpressing ROP16Ⅰ.RT-qPCR and western blotting were used to verify expression effects,CCK-8 assays were used to detect cell proliferation activity,and flow cytometry was used to detect cell cycle changes.Western blotting and RT-qPCR were used to detect the expression levels of p53,p21,CDK6,Cyclin D1,STAT3,p-STAT3(Y705),and JAK1 proteins or genes,and immunofluorescence was used to detect the expression levels of ROP16Ⅰ and p-STAT3(Y705)and their subcellular co-localization in MH-S cells.ROP16Ⅰ protein and gene expression were detected in MH-S cells transduced with lentivirus for ROP16Ⅰ overexpression.CCK-8 assays revealed that ROP16Ⅰ promoted the proliferation of MH-S cells(P＜0.01)and enhanced cell viability.Flow cytometry revealed that ROP16Ⅰ overexpression decreased the G0/G1 phase and elevated the G2 and S phases of the cell cycle in MH-S cells(P＜0.01 or P＜0.05).Compared with the MH-S cell group and MH-S-empty vector group,the MH-S-ROP16 cell group showed lower expression of p53 and p21 proteins;higher expression of CDK6,Cyclin D1,p-STAT3(Y705),and JAK1 proteins;lower expression of p53 and p21 mRNAs;and higher expression of CDK6 and Cyclin D1 mRNAs(all P＜0.01).Immunofluorescence revealed that ROP16Ⅰ co-localized with p-STAT3(Y705)in the nucleus and surrounding cytoplasm.Therefore,Toxoplasma gondii type Ⅰ(RH)ROP16Ⅰ protein activates the JAK-STAT3 pathway;shortens the G0/G1 phase and lengthens the G2/S phase of the cell cycle;and promotes cell proliferation.These findings provide a theoretical basis for revealing the mechanism of immune evasion of Toxoplasma gondii,and lay a foundation for research on the prevention and treatment of Toxoplasma gondii pneumonia.

Objectives:To investigate the safety and efficacy of Cuffstent in the treatment of immediate type Ia endoleak during endovascular aneurysm repair(EVAR)of abdominal aortic aneurysm.Methods:The clinical data of 24 patients with immediate type Ia endoleak during EVAR treated with Cuffstent at the Second Xiangya Hospital of Central South University from January 2019 to December 2022 were retrospectively analyzed.There were 18 males and 6 females with a mean age of(70.3±7.1)years.Results:Of the 24 patients,22 underwent conventional proximal Cuffstent implantation,and 2 underwent fenestrated Cuffstent to preserve unilateral renal artery.Emergency EVAR was performed in 7 patients.The procedure was technically successful in all patients.Immediate disappearance of type Ia endoleak was achieved in 23 patients,whereas 1 patient exhibited mild persistent type Ia endoleak,which was managed conservatively with watchful follow-up.All the 24 patients finished an average of(39.0±15.1)months follow-up,and all patients were alive during the follow-up period.Two patients received aortic reintervention,and one patient received thoracic aortic stent graft implantation due to thoracic aortic penetrating ulcer,and one patient received abdominal aortic aneurysm resection and iliac artery stent implantation due to type II endoleak and iliac artery rupture,respectively.There were no other aorta-related complications and reintervention.Conclusions:Proximal addition of Cuffstent for the management of immediate type Ia endoleak during EVAR is safe and effective.

Aim To investigate the regulatory effects and underlying mechanisms of 4-methoxybenzyl alcohol(4-MBA),an active ingredient of Gastrodia elata,on hepatic cholesterol metabolism.Methods Acute hy-perlipidemia mouse models were established via egg yolk emulsion induction,and hyperlipidemia rat models were constructed using a high-fat diet.Serum and he-patic total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),and high-den-sity lipoprotein cholesterol(HDL-C)levels were quan-tified via enzymatic assays.Hepatic histopathological changes were evaluated through hematoxylin-eosin(HE)and Oil Red O staining.Interactions between 4-MBA and key cholesterol metabolism targets were sim-ulated using molecular docking.mRNA and protein ex-pression levels of LDL receptor(LDLR),proprotein convertase subtilisin/kexin type 9(PCSK9),liver X receptor α(LXRα),peroxisome proliferator-activated receptor γ(PPARγ),ATP-binding cassette transporter G1(ABCG1),and cholesterol 7α-hydroxylase(CYP7A1)were assessed using quantitative polymer-ase chain reaction(qPCR)and immunohistochemis-try.Results In acute hyperlipidemic mice,4-MBA administration significantly reduced serum TG and LDL-C levels while elevating HDL-C(P＜0.05).Hy-perlipidemic rats exhibited decreased serum TG and LDL-C,increased HDL-C(P＜0.01),reduced hepatic LDL-C(P＜0.01),and elevated hepatic HDL-C(P＜0.01).Although TC levels showed a downward trend,the difference lacked statistical significance.He-patic lipid accumulation and steatosis were alleviated.Upregulated mRNA and protein expression of LDLR,PPARγ,LXRα,and ABCG1(P＜0.01),alongside downregulated PCSK9(P＜0.05),were observed.Conclusion 4-MBA modulates cholesterol metabolism primarily via the LDLR/PCSK9 pathway to enhance cholesterol uptake and the PPARγ-LXRα-CYP7A1/ABCA1 axis to promote cholesterol utilization and ef-flux.

Aim To study the effect of evodiamine(EVO)regulating forkhead box protein Ml(FOXM1)on the proliferation and apoptosis of colorectal cancer-resistant cells HCT8/5-FU.Methods CCK-8 assay and EdU assay were used to detect the effect of EVO on cell proliferation ability.Clone formation assay was employed to detect the effect of EVO on the clone for-mation ability of cells.Flow cytometric counting was applied to detect apoptosis.Western blot was utilized to detect the expression of cellular Bcl-2,Bax,FOXM1,β-catenin,c-MYC,and CyclinD1;Molecular docking was used to explore the EVO-FOXM1 interac-tion.Nude mouse transplant tumor model was estab-lished to validate the effect of EVO on HCT8/5-FU cells in vivo.Results CCK-8 assay showed that EVO inhibited the proliferation of HCT8/5-FU cells in a time-and concentration-dependent manner.EdU assay found that the newly proliferated cells in the EVO-trea-ted group were significantly reduced.The results of the clone formation assay showed that EVO inhibited the clone-forming ability of HCT8/5-FU cells.Flow cyto-metric counting found that apoptosis rate of the cells in the EVO group significantly increased.Western blot showed that FOXM1 and β-catenin were significantly highly expressed in HCT8/5-FU cells,and EVO down-regulated the expression of FOXM1,β-cateniin,c-MYC,CyclinD1,and Bcl-2,and up-regulated the ex-pression of Bax.Molecular docking revealed strong in-teractions between EVO and FOXM1.The in vivo ex-perimental results demonstrated that EVO exerted a substantial inhibitory effect on the growth of subcutane-ously implanted HCT8/5-FU xenograft tumors and regulated the expression of related proteins.HE stai-ning revealed significant nuclear consolidation and fragmentation of tumor cells in the EVO group.Con-clusions The findings suggest that EVO could sup-press the activation of the Wnt signaling pathway through a mechanism involving the downregulation of FOXM1 protein expression,thus inhibiting the prolifer-ation of HCT8/5-FU cells and induce their apoptosis.