Objective Early-life developmental factors may influence the risk of cerebrovascular disease in adulthood,but the causal relationship between birth weight and different subtypes of ischemic stroke remains unclear.This study used Mendelian randomization(MR)to investigate the causal association between birth weight and the risk of ischemic stroke(IS)and its major clinical subtypes in adulthood.Methods A two-stage analytical design combining univariable and multivariable MR(UVMR and MVMR)was adopted.Birth weight data were obtained from the Early Growth Genetics(EGG)Consortium,with a sample size of 143 677.Stroke outcome data were obtained from the GIGASTROKE consortium,with a total sample size of approximately 1.29 million,including overall IS and 3 major clinical subtypes:cardioembolism(CE),large artery atherosclerosis(LAA),and small vessel disease(SVD).To ensure sufficient instrumental variables in the multivariable analysis,the screening threshold for birth weight instruments was set at P＜1×10-5.Adult body mass index(BMI)and systolic blood pressure(SBP)were included as covariates in the MVMR model to adjust for adult body size and blood pressure.Causal inference was mainly performed using the random-effects inverse variance weighted(IVW)method.F-statistics,MR-Egger intercept test,and MR-Plelotropy RESidual Sum and Outlier(MR-PRESSO)test were used to assess instrument strength,horizontal pleiotropy,and potential outlier loci.The significance threshold was set at P＜0.012 5 after Bonferroni correction.Fetal-only genetic effect estimates derived from structural equation modeling were further used in a supplementary sensitivity analysis to preliminarily assess the potential influence of maternal genetic effects on the results.Results In the univariable analysis,no significant causal association was observed between birth weight and IS or any of its subtypes.The multivariable analysis included 19 to 21 independent core single-nucleotide polymorphisms(SNPs).The univariable F-statistics were all＞10,whereas the conditional F-statistic for birth weight was＜10,suggesting a potential risk of weak instruments in the conditional model.After adjusting for adult BMI and SBP,MVMR analysis showed that genetically predicted higher birth weight was associated with a reduced risk of LAA(OR=0.736,95％CI:0.587 to 0.922,P=0.007 6).No significant associations were observed with IS(OR=0.943,95％CI:0.869 to 1.023,P=0.156 6),CE(OR=1.018,95％CI:0.857 to 1.210,P=0.836 9),or SVD(OR=0.850,95％CI:0.688 to 1.050,P=0.132 3).Supplementary sensitivity analysis using fetal-only genetic effect estimates within the multivariable framework showed a direction consistent with the main analysis for the association between birth weight and reduced LAA risk.However,because only a limited number of core SNPs were available and the association reached nominal significance(P=0.033),this result was used only to support the directional consistency of the main analysis.Conclusion After adjusting for adult BMI and SBP,genetically predicted higher birth weight is associated with a reduced risk of LAA in adulthood,whereas no significant causal associations are observed with the risk of overall IS,CE,or SVD in adulthood.
Objective On the land battlefield,non-compressible torso hemorrhage is the leading cause of preventable death from combat injuries,yet frontline military physicians generally lack effective endovascular hemostatic skills.This study aims to develop a standardized training program for resuscitative endovascular balloon occlusion of the aorta(REBOA)by combining intelligent sensor-equipped simulation manikins with animal models,and to verify its efficacy in improving REBOA proficiency among frontline medical physicians with no prior interventional experience,with a goal of addressing the bottleneck in extending advanced combat casualty care to the frontline.Methods A self-controlled before-and-after study design was adopted.Twenty on-duty primary-level military physicians from the Army(representing 18 brigade-and regiment-level medical units)were enrolled in June 2023.The participants first completed 10 full-procedure practice sessions on an intelligent simulator to establish skill acquisition curves;subsequently underwent stepwise reinforcement training in femoral artery puncture and catheterization using 10 experimental pigs(weighing 30 to 40 kg);and finally,were assessed using 10 porcine hemorrhagic models(weighing 30 to 40 kg).Digital subtraction angiography(DSA)was applied to observe the accuracy of balloon position.Training effectiveness was evaluated by comparative analysis of trainees'procedural time at different stages of simulator practice,puncture time and procedural success rate during REBOA training on the porcine model,as well as puncture time,balloon positioning accuracy,procedural success rate,and complications during assessment on the porcine hemorrhagic model.Results All the 20 trainees successfully completed the training.① Skill acquisition efficiency:During the simulator training phase,the single-operator procedural time was decreased from 10.8±1.2 min at the first attempt to 5.3±0.4 min at the tenth attempt(P＜0.001);The goodness-of-fit of the single-phase decay model was R2=0.98,indicating that skill plateau was reached after the 7th practice session.② Real-world transfer capability:The trainees completed stepwise skill intensive practice on live experimental pigs,with a mean femoral artery puncture duration of 4.8±1.2 min and a success rate of 95％(19/20);Under DSA fluoroscopy guidance,the mean time for balloon positioning at aortic Zone Ⅰ(timed from balloon entry into the arterial sheath)was 3.8±0.9 min,with a success rate of 95％(19/20).In the assessment on the live porcine hemorrhagic model,the average total procedure time was 7.8±1.4 min(95％CI:7.2 to 8.4);19 out of 20 trainees(95％)achieved visual confirmation of successful hemostasis of the contralateral femoral artery.③ Safety:The complication rate during the assessment phase was 10％(2/20),and the overall pass rate in the final comprehensive evaluation reached 90％(18/20).Conclusion Junior military physicians with no prior interventional experience can acquire preliminary proficiency in REBOA technique through the training program of intelligent sensor-equipped simulator combined with animal model;This program provides a replicable training pathway for disseminating REBOA technology to grassroots units.
Objective To address the dilemma of insufficient recognition of the minority class caused by class imbalance in predicting depression tendency among college students,and systematically compare the predictive performance of 7 oversampling,undersampling,and hybrid sampling methods combined with 12 machine learning models on a class-imbalanced dataset of music APP listening habits and depression tendency in them,providing a reference for method selection in handling class-imbalanced data.Methods This study adopted a cross-sectional study design.Data were collected from questionnaire surveys of college students from 29 provinces,autonomous regions,and municipalities in China in 2023,with college students primarily being undergraduate and postgraduate students aged 18 to 24 years.Univariate analysis was used to screen 10 features significantly associated with depression tendency in the participants.Three oversampling methods including synthetic minority over-sampling technique(SMOTE),adaptive synthetic sampling approach(ADASYN),and synthetic minority over-sampling technique for regression with Gaussian noise(SMOGN),2 undersampling methods including edited nearest neighbors(ENN)and Tomek Links,and 2 hybrid sampling methods including SMOTEENN and ADASYN-Tomek were applied to process the raw data.The raw data and processed data were respectively used to construct prediction models with 12 classification algorithms,including logistic regression(LR),support vector machine(SVM),random forest(RF),decision tree(DT),Light Gradient Boosting Machine(LightGBM),K-Nearest Neighbors(KNN),eXtreme Gradient Boosting(XGBoost),Lasso logistic regression(LASSO),ridge regression(Ridge),elastic net(ENet),multilayer perceptron(MLP),and stacking ensemble model(Stacking).Area under the receiver operating characteristic curve(AUC),Recall,F1-score,and balanced accuracy were selected as evaluation metrics to assess model performance.The optimal model was screened by comparing various combinations of class-imbalance handling techniques and machine learning strategies.SHapley Additive exPlanations(SHAP)analysis was applied for interpretability analysis for the optimal model.Results Univariate analysis identified 10 features significantly associated with depression tendency in college students:gender,grade,major category,preference for Children's songs,approximate length since music listening habits,preference for Chinese style songs,usual music styles,daily listening duration,period of listening during a day,and frequency of comments on music APP.The comparison of 84 combined models with the 12 models based on raw data showed that hybrid sampling outperformed single sampling methods among the 7 resampling strategies;among the 12 classification algorithms,the stacking ensemble model demonstrated better comprehensive performance than individual classifiers.The combination of SMOTEENN hybrid sampling with Stacking achieved the best performance(AUC=0.95,F1=0.89,Recall=0.88,balanced accuracy=0.89).SHAP analysis showed that"usual listening style"contributed the highest in predicting depression tendency among college students.Conclusion Based on the comparison of 84 combination models,the SMOTEENN+stacking ensemble model can accurately predict depression tendency in college students.The comparative analysis provides a methodological basis and practical reference for method selection in class-imbalanced data scenarios and for early identification of mental health risk based on music listening behavior data.
Objective Medical undergraduates are burdened with academic and career pressures,and their mental health deserves close attention.This study aims to analyze the current status of mental health among these students and examine how academic-career experience,interpersonal relationships and family finance influence their mental health,as well as the underlying pathways of action.Methods A multicenter cross-sectional survey was performed from April to September 2025.A total of 1 853 undergraduates were recruited from one medical university each in Southwest,Northwest and East China by convenience sampling.Chinese College Students Mental Health Scale(CCSMHS)was used to assess mental health status.Demographic data were collected,including gender,grade,major category,only-child status,and family residence.Academic-career experiences encompassed major satisfaction,course difficulty,academic pressure,attitude toward employment prospects,and career planning clarity.Interpersonal relationships involved relationships with classmates,typical responses to interpersonal conflicts,and psychological feelings brought by romantic relationships.Family financial factors included family atmosphere,family members' usual pratices when disagreement arise,monthly family income,monthly living expenses,and the impact of family economic status on study and life.The scores of each dimension of the scale for medical undergraduate were compared with the norms for ordinary college students.Intergroup difference tests were performed to analyze disparities in psychological scores among students with different demographic characteristics.Spearman correlation analysis was employed to explore correlations between mental health status and its influencing factors.SPSS Amos 28 was applied to construct a structural equation model for mediating effect analysis,and the Bootstrap method with 5 000 resamples was adopted to test the significance of mediating effect.Results A total of 13.00％of the participants were at high risk for mental health problems.Compared with the norms for ordinary college students,medical students had significantly higher scores in the inferiority and paranoia dimensions(P＜0.001),lower scores in 9 dimensions including anxiety,depression,sexual dysfunction,obsession,dependence,impulsivity,psychotic tendency,social withdrawal,and social aggression(P＜0.05),but somatization showed no significant intergroup difference(P=0.824).Female students,freshmen and non-clinically majors presented significantly higher overall scores of psychological symptoms(P＜0.05).There were no statistically significant differences in scores between only-child and non-only-child students or between urban and rural students.Spearman correlation analysis revealed that academic-career experiences,interpersonal relationships,and family financial factors were all significantly correlated with inferiority and paranoia(P＜0.001).The structural equation model showed good fit(x2/df=2.880,CFI=0.980,TLI=0.972,RMSEA=0.032).Academic-career experiences positively predicted mental health(β=1.501,95％CI:0.828 to 2.178,P＜0.001),and also positively predicted interpersonal relationships(β=0.211,95％CI:0.161 to 0.263,P＜0.001).Interpersonal relationships positively predicted mental health(β=6.027,95％CI:3.875 to 8.499,P＜0.001).The indirect effect of academic-career experiences on mental health through interpersonal relationships was significant(β=1.274,95％CI:0.829 to 1.855,P＜0.001),accounting for 34.68％of the total effect.Conclusion Most medical undergraduates maintain good mental health,with stratified differences by gender,grade and major.Academic-career experiences,interpersonal relationships and family finances are all influencing factors for their mental health;interpersonal relationships exert a mediating role between academic-career experiences and mental well-being.Positive academic-career experiences can directly improve their mental health,and can also indirectly reduce psychological distress through good interpersonal relationships.Accordingly,medical universities should improve interpersonal support systems and implement targeted tiered psychological interventions.
Objective Oxidative stress-induced cardiomyocyte apoptosis is a central event in myocardial ischemia-reperfusion(MIR)injury.MicroRNA-27b(miR-27b)is aberrantly expressed in this process,yet its role and regulatory mechanism in MIR injury remain to be elucidated.This study aims to clarify the molecular mechanism by which miR-27b regulates H2O2-induced cardiomyocyte apoptosis via the PPAR-γ/Bcl-2 signaling pathway,thereby providing a novel target for intervention in MIR injury.Methods ① Healthy male C57BL/6J mice(10 to 12 weeks old,weighing 24 to 30 g)were randomly divided into a MIR group and a sham operation group,with 6 animals in each group.The MIR group underwent ligation of the left anterior descending coronary artery for 45 min followed by reperfusion to induce myocardial injury,while the Sham group underwent the identical surgical procedure without ligation.The heart was harvested 24 h postoperatively,and the entire heart was sectioned and stained with TTC for 30 min to assess infarct size.The ischemic myocardial tissue from the anterior wall of the left ventricle was used for total RNA extraction.② Primary neonatal mouse ventricular cells(PNMVCs)were isolated from 1-to 3-day-old mice and transfected for 24 h with miR-27b mimics or anti-miR-27b oligonucleotides(AMO-miR-27b).The cells were subsequently treated with 100 pumol/L hydrogen peroxide(H2O2)for 24 h to induce cardiomyocyte apoptosis.Transfected cardiomyocytes were treated with the PPAR-γ antagonist T0070907(10.0 nmol/L),and apoptosis was assessed 48 h later.Cell experiments were divided into normal control group,H2O2 model group,miR-27b mimic transfection+H2O2 group,AMO-miR-27b transfection+H2O2 group,AMO-miR-27b+H2O2+T0070907 group,and negative control(NC)+H2O2 group.RT-qPCR was performed to measure the expression levels of miR-27b and PPAR-γ.Cell viability was assessed using MTT assay.Cell apoptosis was quantified by TUNEL assay combined with DAPI staining.Western blotting was employed to detect the expression of PPAR-γ,Bcl-2,Bax,and Cyt-c,and other related proteins.Results ① Compared with the Sham group,the MIR group exhibited significantly larger infarct size in TTC-stained heart sections,and upregulated miR-27b expression in the ischemic myocardial tissue of the left ventricular anterior wall(P＜0.05).② Compared to the normal control group,miR-27b expression levels were significantly upregulated in H2O2-treated PNMVCs(P＜0.01).Overexpression of miR-27b in PNMVCs induced cardiomyocyte damage,as evidenced by decreased cell viability[(48.75±5.55)％vs(87.20±3.52)％,P＜0.01]and increased cell apoptosis[(41.08±2.84)％vs(1.75±0.29)％,P＜0.01],accompanied by reduced expression of PPAR-γ at both mRNA(P＜0.01)and protein levels(P＜0.01).In contrast,inhibition of miR-27b expression enhanced PPAR-γ expression at both mRNA(P＜0.05)and protein levels(P＜0.01).MTT assay showed that AMO-miR-27b attenuated H2O2-induced decrease in cell viability[(79.46±6.41)％vs(60.29±4.83)％,P＜0.01];TUNEL assay further demonstrated that after AMO-miR-27b transfection,H2O2-induced cardiomyocyte apoptosis was significantly reduced[(19.60±2.95)％vs(42.96±5.80)％,P＜0.01].H2O2 treatment caused significantly decreased Bcl-2 levels(P＜0.01)and elevated Bax levels(P＜0.01)in cardiomyocytes,which were reversed by AMO-miR-27b.Furthermore,cytoplasmic Cyt-c protein levels were significantly increased in cardiomyocytes after H2O2 exposure(P＜0.01),and the downregulatory effect of AMO-miR-27b on cytoplasmic Cyt-c(P＜0.05)and its protective effect against cardiomyocyte apoptosis(P＜0.01)were abolished by the PPAR-γ antagonist T0070907.Conclusion MiR-27b is highly expressed in myocardial tissues and cardiomyocytes after oxidative stress injury,which exacerbates tissue damage and cell apoptosis.Suppression of the PPAR-γ/Bcl-2 signaling pathway may be a primary mechanism underlying this effect,suggesting that targeted inhibition of miR-27b may be a potential novel strategy for MIR injury intervention.
Objective Sepsis-associated acute kidney injury(SA-AKI)is a life-threatening condition with high mortality.This study aims to develop and validate an interpretable machine-learning model for assessing mortality risk in SA-AKI patients,and facilitate early identification of high-risk patients to support clinical decision-making.Methods A retrospective cohort study was conducted on the clinical data derived from the Medical Information Mart for Intensive Care Ⅳ(MIMIC-Ⅳ)and the eICU Collaborative Research Database(eICU-CRD).A total of 24 487 patients from MIMIC-Ⅳ and 13 757 ones from eICU-CRD were included for analysis.Predictive variables were selected through univariate analysis and least absolute shrinkage and selection operator(LASSO)regression.The MIMIC-ⅣV dataset was randomly divided into training and internal testing sets in a ratio of 8∶2.Ten machine-learning algorithms were used to construct mortality risk prediction models,including logistic regression(LR),random forest(RF),light gradient boosting machine(LightGBM),K-nearest neighbors(KNN),eXtreme gradient boosting(XGBoost),ridge regression(Ridge),elastic net(ENet),decision tree(DT),stacking ensemble model(Stacking),and multilayer perceptron(MLP).Model performance was evaluated using the area under the receiver operating characteristic curve(AUC),accuracy,sensitivity,specificity,and with calibration curve and decision curve analyses.SHapley Additive exPlanations(SHAP)were adopted to analyze the contribution and directional effect of each variable on prediction.External validation was performed using SA-AKI patients meeting inclusion criteria from the eICU-CRD database.Results A total of 31 predictive variables were selected through univariate analysis combined with LASSO regression,and 10 machine-learning models for mortality risk prediction were constructed.The LightGBM model achieved an AUC value of 0.817(95％CI:0.803 to 0.830)in the internal testing set and 0.735(95％CI:0.725 to 0.745)in the external validation set based on the eICU-CRD database,outperforming other models.After comprehensive comparison of discrimination,calibration,and decision curve analyses across all models,the LightGBM model demonstrated optimal performance,suggesting favorable predictive efficiency and clinical applicability in mortality risk prediction for SA-AKI patients.SHAP analysis revealed that the most important variables for 28-day mortality risk included sequential organ failure assessment(SOFA)score,Charlson comorbidity index,Glasgow coma scale(GCS)score,mean body temperature,mean blood urea nitrogen,mean serum sodium,mean respiratory rate,mean creatinine,Oxford acute severity of illness score(OASIS),and red blood cell distribution width(RDW).Conclusion For SA-AKI patients,the mortality risk prediction model based on the LightGBM algorithm demonstrates optimal performance.The SOFA score,Charlson comorbidity index,and other variables are important indicators for predicting 28-day mortality in these patients,providing a reference for early identification of high-risk patients.
Coagulation disorders constitute a core pathological event in sepsis, and the progression of sepsis-induced coagulopathy (SIC) to disseminated intravascular coagulation (DIC) is a major driver of multi-organ failure and death in septic patients. With deepening understanding of disease pathogenesis, the diagnosis and treatment model of SIC has gradually shifted from controlling bleeding risk-oriented supportive therapy for primary diseases towards a new precision anticoagulation model centering on excessive coagulation activation and emphasizing individualized intervention. Current routine coagulation indicators and scoring systems widely used in clinical practice are mostly static and outdated assessment tools, which fail to reflect the dynamic progression and pathological heterogeneity of the disease. These tools also present prominent limitations such as insufficient early identification and limited accuracy in SIC classification. Novel biomarkers, point-of-care testing for functional coagulation, and artificial intelligence-powered early warning models provide new modalities for early warning, dynamic monitoring and precise classification of SIC. In terms of treatment, the limitations of conventional anticoagulation have become increasingly prominent, and targeted intervention strategies focusing on stratified anticoagulation, endothelial protection and immunothrombosis regulation are progressively emerging as research hotspots, with a variety of novel therapeutic modalities demonstrating potential clinical value. Future efforts should further improve the molecular phenotyping and precise diagnosis and treatment system of SIC. Early identification, dynamic evaluation and individualized intervention are expested to improve the clinical outcomes of SIC patients.
Objective Sepsis-induced myocardial contractile dysfunction is one of the major causes of high mortality in septic patients,yet effective targeted therapeutic agents remain unavailable.This study aims to investigate the ameliorative effect and underlying mechanism of ginsenoside Rg5 on myocardial contractile dysfunction in sepsis.Methods ① A mouse sepsis model was established using cecal ligation and puncture(CLP).Ninety-six male C57BL/6 mice(10 weeks old,weighing 20 to 30 g)were randomly divided into Sham group,sepsis group,and Rg5 treatment group.Cardiac function was evaluated by echocardiography.Serum cardiac troponin I(cTn I)level was detected by ELISA,and the 72-h survival rate of mice was observed.Cardiomyocytes were isolated using a Langendorff perfusion system,and their contractile function and calcium transients were subsequently detected using a cell microtensiometer.② Network pharmacology was employed to screen the potential targets of Rg5 for the treatment of sepsis and to perform functional enrichment analysis.A protein-protein interaction(PPI)network analysis was further conducted using STRING,and hub targets were identified with Cytoscape.Western blotting was used to detect the expression of the hub target protein epidermal growth factor receptor(EGFR)across groups,and molecular docking was carried out to investigate the interaction between Rg5 and EGFR.Results ① Compared with the sham group,the sepsis group showed significantly reduced left ventricular ejection fraction(LVEF)and fractional shortening(LVFS)(P＜0.05),and markedly elevated levels of the myocardial injury marker cTn I(P＜0.05),indicating obvious myocardial dysfunction after sepsis.Rg5 treatment significantly restored myocardial function in septic mice,as evidenced by increased LVEF and LVSF values,reduced serum cTn I levels,and an improved the 72-h survival rate from 25％to 50％(P＜0.05).Compared with the sham group,the cardiomyocytes in the sepsis group exhibited a significantly reduced calcium transient amplitude and calcium sensitivity,along with markedly diminished contractile amplitude and slower maximal contraction and relaxation velocities(P＜0.05).The treatment significantly restored calcium transient capacity in septic cardiomyocytes,ameliorated myofilament calcium desensitization,and markedly increased contractile amplitude as well as maximal contraction and relaxation velocities(P＜0.05).② Network pharmacology identified 53 potential targets of Rg5 for the treatment of sepsis.GO enrichment analysis showed that these targets were mainly involved in biological processes such as muscle cell proliferation,regulation of blood pressure,and the adenylate cyclase-activating G protein-coupled receptor signaling pathway.KEGG enrichment analysis indicated that these targets were significantly enriched in pathways related to myocardial contraction,including the calcium,Rap1,and Ras signaling pathways.PPI analysis and Cytoscape screening identified the top 10 candidate targets,including EGFR,STAT3,MMP9,IL2,FGF2,MAPK1,BCL2L1,MMP2,PTPN11,and MDM2,among which EGFR was the hub target.Western blotting results showed that EGFR protein level in myocardial tissue was significantly decreased after sepsis(P＜0.05),whereas Rg5 treatment markedly upregulated its expression(P＜0.05).Molecular docking predicted a strong interaction between Rg5 and EGFR,with a binding energy of-8.0 kcal/mol.Conclusion Ginsenoside Rg5 effectively ameliorates sepsis-induced myocardial contractile dysfunction,attenuates myocardial injury,and improves survival in septic mice.The underlying mechanism may be associated with its interaction with EGFR and upregulation of EGFR expression.
Objective Ovarian endometriotic cyst(OEC)is a common chronic gynecological disease in women of reproductive age,which is closely associated with chronic pelvic pain and infertility.Previous single-modality treatment,such as medications,surgical enucleation,and puncture sclerotherapy are at risks for incomplete lesion removal or functional impairment of the ovarian.Therefore,postoperative combined medical therapy has become a research hotspot in clinical practice.This study aims to retrospectively analyze the clinical efficacy of ultrasound-guided sclerotherapy with lauromacrogol combined with different maintenance medications for OEC,and to identify the optimal postoperative medication regimen.Methods A retrospective cohort study was conducted.A total of 185 patients diagnosed with OEC in the Day Ward of our department from September 2021 to December 2025 were enrolled initially.After excluding 17 cases lost to follow-up and 15 due to other reasons,153 patients were finally included.According to the type of postoperative maintenance medication regimens,they were divided into a dienogest group(n=55),a gonadotropin-releasing hormone agonist(GnRH-a)group(n=30),a dydrogesterone group(n=33),and a control group without pharmacological treatment(n=35).All of them were followed up for 6 months.Transvaginal color Doppler ultrasound examination was performed to measure the maximum diameter and volume of OEC before,and at 1,3,and 6 months after operation,and the serum carbohydrate antigen 125(CA125)levels were detected by chemiluminescence immunoassay at the same time points.Baseline characteristics of the patients were compared across the 4 groups.The efficacy and short-term complications in each group after lauromacrogol sclerotherapy were summarized.A mixed-effects model for repeated measures(MMRM)was adopted to analyze differences in the observed indicators at each follow-up time point.Results ① There were no significant differences in baseline data among the 4 groups except for OEC cyst fluid viscosity(mild:31 cases,20.26％;moderate:69 cases,45.10％;severe:53 cases,34.64％;P=0.008).(2)Puncture sclerotherapy was ineffective in 11 cases(7.19％),including 3 cases in the control group,5 in the dienogest group,1 in the GnRH-a group,and 2 in the dydrogesterone group.One patient in the dydrogesterone group still failed to respond to the drug after 6 months of medication.The main short-term postoperative complications were fever[32.03％(49/153)],abdominal pain[15.03％(23/153)]and puncture site bleeding[5.88％(9/153)].③ The maximum diameter and volume of OEC in all groups were decreased gradually after operation,with no significant intergroup difference at 1 month postoperatively.At 3 months after operation,the diameter(95％CI:3.45 to 15.65,P=0.015 1)and volume(95％CI:5.04 to 23.34,P=0.016 7)were significantly lower in the dienogest group than the control group.The GnRH-a group showed statistically significant differences in the maximum diameter(95％CI:5.21 to 18.62,P=0.003 5)and the cyst volume(95％CI:4.83 to 25.87,P=0.029 2),with even more pronounced differences at 6 months postoperatively[dienogest group(95％CI:3.66 to 15.22,P=0.009 9;95％CI:5.89 to 19.17,P=0.001 9);GnRH-a group(95％CI:7.44 to 21.47,P=0.000 5;95％CI:5.99 to 21.23,P=0.003 8)].No significant differences were found among the 3 medication groups,and either between the dydrogesterone group and other groups.④ Preoperative serum CA125 levels were mostly elevated,but no significant differences were observed across groups.Postoperative CA125 levels were decreased in all groups.The CA125 levels were significantly decreased at 1 month after operation in the GnRH-a group(95％CI:39.99 to 87.81,P＜0.000 1)and dydrogesterone group(95％CI:18.00 to 53.40,P=0.000 4),with no significant intergroup difference.After continuous maintenance medication,the dienogest group(95％CI:12.68 to 37.72,P=0.000 4),the GnRH-a group(95％CI:32.67 to 58.93,P＜0.000 1),and the dydrogesterone group(95％CI:23.10 to 42.50,P＜0.000 1)all showed significant differences compared with their own 1-month postoperative controls,but no significant intergroup differences were observed.Conclusions Maintenance therapy with dienogest or GnRH-a following ultrasound-guided lauromacrogol sclerotherapy demonstrates equally significant efficacy for OEC.OEC volume may serve as a superior imaging indicator for therapeutic response assessment.
Objective Intestinal ischemia-reperfusion(IR)injury is a common and critical pathological process in clinical practice.Ferroptosis,a form of cell death characterized by iron-dependent accumulation of uncontrolled lipid peroxidation,has been confirmed to be involved in intestinal IR injury.Nuclear factor erythroid 2-related factor 2(Nrf2)is a key molecule in regulating ferroptosis,and its activation can inhibit this form of cell death,thereby alleviating tissue injury.Sulforaphane(SFN),a classical Nrf2 activator,exerts antioxidant protective effects.This study aims to investigate whether SFN alleviates intestinal IR injury by activating the Nrf2 pathway to inhibit ferroptosis.Methods ① SFN-related target genes,ferroptosis-related genes,and intestinal IR injury-related genes were retrieved and integrated from online databases,including TCMSP,GeneCards and OMIM,and the intersection genes were identified and presented in a Venn diagram.Then the intersection genes were imported into STRING to construct a protein-protein interaction(PPI)network,and topological analysis was performed using Cytoscape(v3.9.1)to screen core targets.Molecular docking was employed to predict the binding affinity between SFN and the key target Nrf2.② Twenty-four SPF-grade male C57BL/6 mice(6 to 8 weeks old,weighing 22±2 g)were randomly divided into 4 groups(n=6):Sham group,IR group,IR+SFN group,and IR+SFN+ML385(a specific Nrf2 inhibitor)group.Except for the Sham group,the other groups were inflicted with clamping the superior mesenteric artery for 45 min followed by reperfusion for 30 min to establish an IR injury model.The IR+SFN and IR+SFN+ML385 groups were given an intraperitoneal injection of SFN(5 mg/kg)1 h before clamping,while the IR+SFN+ML385 group additionally received intraperitoneal injection of ML385(30 mg/kg)2 h before clamping.After 30 min of reperfusion,small intestinal tissues were harvested.Histopathological changes were observed by light microscopy after HE staining,and the severity of intestinal injury was evaluated using Chiu's score.Levels of malondialdehyde(MDA),superoxide dismutase(SOD),and reduced glutathione(GSH)were measured using biochemical reagent kits.Western blotting was performed to detect the protein expression of acyl-CoA synthetase long-chain family member 4(ACSL4),glutathione peroxidase 4(GPX4),Nrf2,and heme oxygenase-1(HO-1).Results ① Network pharmacology analysis identified 162 SFN target genes,2 772 ferroptosis-related genes,and 2 100 intestinal IR injury-related genes,with 43 intersection genes.PPI network topological analysis revealed 36 core genes,including Nrf2.Molecular docking showed that SFN could form a binding conformation with Nrf2,with a binding energy of-3.3 kcal/mol.② In vivo results demonstrated that compared with the Sham group,the IR group exhibited significantly aggravated intestinal mucosal injury,as evidenced by an increased Chiu's score(P＜0.05),elevated MDA level,and decreased SOD and GSH activities(P＜0.05).Meanwhile,the expression of GPX4 was downregulated while that of ACSL4 was upregulated(P＜0.05),suggesting the occurrence of ferroptosis in intestinal IR.SFN pretreatment ameliorated the above pathological and biochemical changes,upregulated Nrf2 and HO-1 expression(P＜0.05),restored GPX4 expression and downregulated ACSL4 expression(P＜0.05).The protective effects of SFN and its regulatory effects on the Nrf2/HO-1 axis and ferroptosis-related proteins were partially reversed by the addition of ML385(P＜0.05).Conclusion SFN can alleviate intestinal IR injury,and its effect may be related to enhancing the antioxidant response associated with the Nrf2/HO-1 signaling axis and ameliorating ferroptosis-related molecular changes.
Objective Transmembrane member 16A(TMEM16A)is involved in intestinal epithelial ion transport and secretory homeostasis,but its effects on enteroendocrine cell(EEC)function and gut-brain axis-related behaviors remain unclear.This study aims to investigate the effects of intestinal epithelial TMEM16A deficiency on the differentiation phenotype of chromogranin A(CgA)-positive EECs,related gut-brain signaling molecules,hippocampal inflammatory signals and anxiety-like behavior,and to analyze the role of the vagal afferent pathway in this process.Methods Male intestinal epithelial TMEM16A conditional knockout mice(KO)and littermate control mice(wild type,WT),aged 8 to 10 weeks and weighing 24 to 26 g,were subjected.Terminal ileal mucosal tissues from WT mice were treated with DMSO,T16Ainh-A01 or CaCCinh-A01,and Ca2+-activated Cl-secretion were measured using short-circuit current recording.Intestinal epithelial TMEM16A expression between the WT and KO mice was compared with Western blotting and immunofluorescence staining.Behavioral parameters were compared between the WT and KO mice using open field,elevated plus maze and light-dark box tests.The WT and KO mice were treated with cholecystokinin-saporin conjugate(CCK-SAP)or blank saporin to establish WT+Blnk-SAP,WT+CCK-SAP,KO+Blnk-SAP and KO+CCK-SAP groups(n=10),and to evaluate the involvement of CCK-sensitive vagal afferent pathways.The differentiation phenotype of CgA+EEC in intestinal tissues between the 2 types of mice was compared by immunohistochemistry and qPCR.WT-and KO-derived organoids were divided into untreated and bone morphogenetic protein 4(BMP4)-treated groups to detect organoid differentiation phenotypes and levels of 5-hydroxytryptamine(5-HT),tryptophan hydroxylase 1(TPH1),glucagon-like peptide-1(GLP-1)and peptide YY(PYY).Immunohistochemical staining and Western blotting were performed to detect the expression of intestinal epithelial renewal-related molecules and those in inflammatory signals in the hippocampal CA3 region.Results Compared with the DMSO group,TMEM16A inhibitors reduced Ca2+-activated Cl secretion in intestinal epithelium(P＜0.000 1).Compared with the WT group,the KO group exhibited significantly decreased intestinal epithelial TMEM16A expression(P＜0.000 1),and weakened TMEM16A immunofluorescence signals in intestinal villus and crypt epithelium;decreased total distance traveled,reduced time in the center,and lower center entry frequency in the open field test(P＜0.000 1);decreased total distance traveled and open-arm time(P=0.017 1,P＜0.000 1)and increased closed-arm time(P=0.046 0)in the elevated plus maze test;and decreased light-box entries and shortened time spent in the light box in the light-dark box(P＜0.000 1),but no significant change was observed in nose-poke frequency.Compared with the KO+Blnk-SAP group,the KO+CCK-SAP group showed an increase in open-arm time from 1.75％to 13.16％(P＜0.000 1),decreased closed-arm time(P=0.004 5),increased total distance traveled and center-zone time in the open field test(P=0.000 2,P=0.023 8),and increased light-box time and entries(P=0.002 3,P=0.005 5).Compared with the WT mice,the KO mice showed decreased numbers of intestinal CgA+EECs and CgA mRNA expression(P＜0.000 1),reduced proportion of mature budding organoids from 77.80％in the WT group to 23.60％in the KO group(P＜0.000 1),and decreased 5-HT and TPH1 levels(P＜0.000 1).BMP4 treatment increased 5-HT,TPH1,GLP-1 and PYY levels in WT organoids(P＜0.000 1,P＜0.000 1,P＜0.000 1,P=0.006 0),whereas no significant changes were observed in the KO+BMP4 group.In the KO group,intestinal CgA,Lgr5 and Asc12 expression levels were decreased(P＜0.000 1,P＜0.000 1,P=0.001 6),whereas Ki67,Wnt3a,β-catenin,Dll1 and EGF expression levels were increased(P＜0.000 1),and NLRP3 and IL-1β expression levels in the hippocampal CA3 region were enhanced(P＜0.000 1).Conclusion Intestinal epithelial TMEM16A deficiency attenuates the differentiation phenotype of CgA+EECs,reduces 5-HT,TPH1,GLP-1 and PYY levels,and is accompanied by enhanced NLRP3/IL-1β inflammatory signals in the hippocampal CA3 region and anxiety-like behavior.The CCK-sensitive vagal afferent pathway is involved in TMEM16A deficiency-related anxiety-like behavior.
Objective High-grade clear cell renal cell carcinoma(ccRCC)is characterized by rapid progression and high recurrence risk,and the optimal surgical approach for such patients remains controversial.This study aims to compare the prognostic impact of partial nephrectomy(PN)and radical nephrectomy(RN)in such patients and provide evidence-based guidance for clinical decision-making.Methods This study was a multi-hospital retrospective cohort study.A total of 187 patients with high-grade(Fuhrman grade 3 to 4)ccRCC who underwent PN or RN at the urological departments of Daping Hospital,General Hospital of Central Theater Command,and Xiangyang First People's Hospital between June 2010 and May 2023 were retrospectively enrolled.Meanwhile,11 580 patients were extracted from the Surveillance,Epidemiology,and End Results(SEER)database(2010-2015)for external cohort validation.Patients were divided into the PN group and RN group according to the surgical approach,with overall survival(OS)as the primary endpoint.A 1∶1 propensity score matching(PSM)was performed to balance the influence of baseline confounders.Kaplan-Meier analysis was used to plot survival curves and compare OS,and Cox proportional hazards regression analysis was employed to identify independent prognostic factors.Results In the multi-hospital retrospective cohort,36 patients underwent PN and 151 underwent RN before PSM.The PN group exhibited significantly smaller tumors(4.00 vs 6.00 cm,P＜0.000 1)and lower T stage(P＜0.000 1).Kaplan-Meier analysis showed that the PN group had a survival benefit compared with the RN group(P=0.046),but neither group obtained the median survival.After PSM(34 patients in each group),there were no significant differences between the 2 groups in terms of age,preoperative creatinine,sex,smoking history,diabetes,hypertension,N stage,tumor size,or tumor laterality(all P＞0.05);Kaplan-Meier analysis showed no statistical difference in OS between the 2 groups(P=0.26),with the median survival not reached in either group.In the SEER cohort,the patients from the RN group were older and had greater tumor burden before PSM(both P＜0.000 1).After PSM(2 807 pairs),baseline characteristics were well balanced;Kaplan-Meier analysis showed that significantly longer OS was observed in the PN group than the RN group(P＜0.000 1),but the median survival was still not reached in either group.Multivariable Cox regression analysis indicated that PN was an independent protective factor,whereas RN was an independent risk factor.RN was associated with a 33％increase in mortality risk compared with PN(HR=1.33,95％CI:1.21 to 1.46,P＜0.001).Conclusion Based on the results of our multi-hospital retrospective cohort and the large-sample SEER database cohort analysis,for patients with high-grade ccRCC,PN demonstrates an OS benefit compared with RN,suggesting the potential value of PN and supporting the possible adoption of PN under strict adherence to surgical indications.
Objective Helicobacter pylori(H.pylori)infection is a major cause of chronic gastritis.This study aims to determine whether H.pylori infection induces the expression of G protein-coupled receptor class C group 5 member C(GPRC5C)in gastric epithelial cells,to verify the regulatory role of the upstream NF-κB signaling pathway,and to elucidate the biological function of GPRC5C in H.pylori-associated gastritis,thereby providing a potential new therapeutic target for the prevention and treatment of this disease.Methods A total of 190 SPF-grade female C57BL/6J mice(6 to 8 weeks old,weighing 18 to 22 g)were randomly divided into an uninfected control group(gavaged with PBS)and an H.pylori-infected group(gavaged with H.pylori),with 95 mice per group.The mice were gavaged 5 times consecutively,twice a day.To establish a chronic in vivo infection model,5 mice from each group were randomly euthanized weekly from week 1 to week 19 post-infection,for 19 weeks.The expression of GPRC5C in mouse gastric mucosal tissues was examined by quantitative real-time PCR,Western blotting,and immunohistochemical staining.Human gastric tissue specimens were obtained from normal gastric tissues at least 5 cm from the tumor margin in gastric cancer patients hospitalized in Department of General Surgery of First Affiliated Hospital of Army Medical University.Immunohistochemical staining was employed to evaluate the protein expression of GPRC5C in human gastric tissues.Mouse primary gastric epithelial cells were isolated from gastric tissues of uninfected mice by enzymatic digestion and mechanical dissociation,followed by filtration to prepare single-cell suspensions,which were then purified by CD326 based magnetic-activated cell sorting(MACS)system.An H.pylori infection model was established in both the GES-1 cell line and mouse primary gastric epithelial cells.The expression of GPRC5C in GES-1 cells and mouse primary gastric epithelial cells was examined by qRT-PCR and Western blotting.Signaling pathway blocking experiments were performed to screen and verify the intracellular signaling pathway activated during H.pylori-induced GPRC5C expression in gastric epithelial cells.Dual-luciferase reporter gene assay was used to analyze the GPRC5C promoter activity of the full-length and truncated sequences of GPRC5C promoter region after H.pylori infection,and to determine the effect of signaling pathway inhibitors on GPRC5C promoter activity.Furthermore,another female C57BL/6J WT mice and Gprc5c-/-mice(n=5,6 to 8 weeks old,weighing 18 to 22 g)were used to establish the H.pylori infection model.Gastric tissue inflammation was observed after HE staining to preliminarily investigate the effector function of GPRC5C in H.pylori infection-associated gastritis.Results Compared with the uninfected group,the mRNA levels of GPRC5C in mouse gastric mucosal tissues were significantly upregulated at week 14 post-H.pylori infection(P＜0.000 1),and the protein levels were also elevated in both mouse and human gastric mucosal tissues.In vitro experiments showed that,compared with the corresponding uninfected group,H.pylori infection upregulated the expression of GPRC5C at both mRNA and protein levels in GES-1 cells and mouse primary gastric epithelial cells,with mRNA expression levels increased 4.1-fold and 3.1-fold,respectively(P＜0.01).This upregulation was dependent on the bacterial dose and infection duration(P＜0.05).In terms of the underlying mechanism,pretreatment with an NF-κB signaling pathway inhibitor significantly suppressed H.pylori-induced GPRC5C upregulation in gastric epithelial cells,with an inhibitory rate of 64.0％(P＜0.01).Dual-luciferase reporter assay revealed that H.pylori infection significantly increased GPRC5C promoter activity to 4.1 times of that in the uninfected group(P＜0.001).When the NF-κB signaling pathway was blocked,this promoter activity was significantly suppressed,with an inhibitory rate of 66.1％(P=0.000 2);In functional validation,compared with H.pylori-infected WT mice,Gprc5c-/-mice exhibited significantly reduced gastric inflammation after infection(P＜0.001).Conclusion H.pylori infection induces GPRC5C expression in gastric epithelial cells through activation of the NF-κB signaling pathway,thereby promoting the progression of H.pylori-associated gastritis.
Sepsis-induced coagulopathy(SIC)is a critical intermediate stage in the progression from sepsis to disseminated intravascular coagulation(DIC),characterized by an imbalance between coagulation activation and fibrinolysis inhibition,with widespread formation of microthrombi as a core feature.The incidence of SIC exceeds 60％,and its mortality rate is as high as 30％.In this article,we systematically review 4 major issues in current research on the diagnosis and treatment of SIC:① For assessment and intervention of platelet function,introducing various platelet function tests,along with controversies in antiplatelet and pro-platelet therapies;② In mechanisms and evaluation of altered fibrinolysis,elaborating the fibrinolysis inhibition caused by plasminogen activator inhibitor-1(PAI-1)overexpression and the associated detection dilemmas;③ About issues and optimization of SIC diagnostic criteria,comparing the International Society on Thrombosis and Haemostasis(ISTH)-SIC criteria,modified Chinese SIC criteria,and the 2025 ISTH-DIC criteria,pointing out shortcomings such as insufficient specificity,exclusion of D-dimer,and susceptibility of prothrombin time-international normalized ratio(PT-INR)to interference,and proposing optimization strategies including dynamic scoring,individualized thresholds,novel molecular biomarkers,and machine learning;④ Concerning precision anticoagulation strategies,emphasizing the importance of anticoagulation timing,drug selection,and dynamic monitoring[e.g.,anti-activated factor Ⅹ(Ⅹa)activity,thromboelastography],and analyzing the application of heparin,low molecular weight heparin,nafamostat,and other agents.SIC patients frequently face dual risks of bleeding and thrombosis,which can rapidly shift.Clinical management should move beyond conventional static indicators,integrate immunothrombosis mechanisms,novel molecular biomarkers[e.g.,thrombin-antithrombin complex(TAT),plasmin-α2-plasmin inhibitor complex(PIC),soluble thrombomodulin(sTM)]and dynamic monitoring technologies to seize the"therapeutic window"for anticoagulation,and implement stratified management and precision diagnosis and treatment based on individualized,multidimensional assessment.
Objective Oral and maxillofacial venous malformations are common congenital vascular disorders in children that seriously impair their physical and mental health.Sclerotherapy is currently the preferred treatment,yet clinical practice has shown that bleomycin alone may not be effective in some cases.This study aims to compare the clinical efficacy and safety of 1％polidocanol foam combined with bleomycin versus bleomycin monotherapy for sclerotherapy of oral and maxillofacial venous malformations in children under the guidance of digital subtraction angiography(DSA),aiming to explore the clinical value of the combination regimen.Methods A prospective cohort study design was adopted.A total of 38 children with oral and maxillofacial venous malformations undergoing polidocanol foam combined with bleomycin sclerotherapy in our department from May 2024 to December 2025 were consecutively enrolled and served as the observation group.Another 38 children who received bleomycin sclerotherapy alone at our hospital during the same period were selected as the control group.All patients were regularly followed up for 4 to 12 months.Clinical baseline data,treatment response rate,lesion area,number of treatment sessions,cosmetic satisfaction,recurrence rate,and adverse events were observed and compared between groups.Binary logistic regression analysis was used to analyze clinical factors influencing the efficacy for oral and maxillofacial venous malformations in children.Results ① No statistically significant differences were observed between the 2 groups in terms of age,sex,lesion location,or Puig classification.② The overall treatment response rate was significantly higher in the observation group than in the control group(94.74％vs 78.95％,P=0.042),yet no significant difference was found in the distribution of treatment efficacy grades(Z=1.949,P=0.051).There was no significant difference in pre-treatment lesion area between the 2 groups(P=0.725).After treatment,the area was notably reduced in both groups compared with their corresponding baseline values(P<0.001).The observation group had a smaller lesion area than the control group at the corresponding post-treatment time point[(1.48±0.42)cm2 vs(2.32±0.61)cm2,P<0.001],and the rate of lesion area reduction was also markedly higher in the observation group[(51.24±10.47)％vs(37.82±5.43)％,P<0.001].Additionally,the number of treatment sessions required in the observation group was notably lower than that in the control group(2.05±0.26 vs 2.90±0.21,P<0.001).③ Compared with the control group,the observation group showed no statistical difference in overall aesthetic satisfaction[38(100.00％)vs 34(89.47％),P=0.115],yet,the distribution of satisfaction grades was superior(Z=2.394,P=0.017).While,no statistically significant difference was observed in the recurrence rate between the 2 groups[1(2.78％)vs 4(13.33％),P=0.109].④ There existed no statistical differences in individual adverse events or total events between the observation group and the control group[5 cases(13.16％)vs 3 cases(7.89％),P=0.711].⑤ Binary logistic regression analysis revealed that combination therapy was an independent protective factor for overall treatment efficacy(OR=5.13,95％CI:3.08 to 8.62,P=0.014).Conclusion Compared with bleomycin sclerotherapy alone,the application of polidocanol foam combined with bleomycin sclerotherapy for oral and maxillofacial venous malformations in children can significantly improve treatment response rate,reduce lesion area,and decrease the number of treatment sessions,without increasing the risk of complications.
Objective Identifying the stressors of postgraduate students is essential for understanding the causes of mental health problems and implementing targeted interventions.Based on the stress process theory,this study aims to develop a stressor scale applicable to contemporary postgraduate students and test its reliability and validity.The scale is further applied to medical postgraduates to explore their stress levels and the impacts of stressors on anxiety and depression.Methods This study established the scale dimensions and developed the initial scale through literature review and semi-structured interviews with 20 postgraduate students.Following revision based on a pilot survey of 200 postgraduate students,exploratory factor analysis and reliability analysis were conducted on 267 postgraduate students to finalize the formal scale.A cross-sectional study was conducted using stratified cluster sampling.A total of 666 medical postgraduate students from two medical colleges in Chongqing were surveyed using the newly developed Stressor Scale,Patient Health Questionnaire-9 items(PHQ-9),and Generalized Anxiety Disorder-7(GAD-7).Confirmatory factor analysis was performed to assess the levels of stressors,anxiety,and depressive mood.Hierarchical regression analysis was employed to examine the influence of stressors on anxiety and depressive mood.Results Exploratory factor analysis of the postgraduate students identified 5 distinct dimensions:academic stressor(5 items),employment and career stressor(7 items),financial stressor(4 items),supervisory stressor(8 items),family and romantic stressor(5 items).Reliability analysis showed strong internal consistency,with Cronbach's ɑ coefficients ranging from 0.85 to 0.94 and split-half coefficients of 0.79 to 0.93 for both the total scale and individual dimensions.Confirmatory factor analysis conducted on medical postgraduate students demonstrated a good fit,with χ2/df=3.88,comparative fit index(CFI)=0.93,Tucker-Lewis Index(TLI)=0.92,root mean square error of approximation(RMSEA)=0.06,and standardized root mean squared residual(SRMR)=0.07.The convergent validity was good,with average variance extracted(AVE)=0.50 to 0.72 and construct reliability(CR)=0.83 to 0.93.The discriminant validity was satisfactory,with the square roots of AVE for all dimensions ranging from 0.71 to 0.85,all of which were greater than the maximum correlation coefficients between the corresponding factor and other factors.The survey showed that academic stress(2.17±0.74),employment and future stress(1.67±0.68),and financial stress(1.59±0.76)were the 3 major stressors for medical postgraduates.Hierarchical regression analysis indicated that after adjusting for demographic variables and negative emotions(depression and anxiety),employment and future stressor(β=0.19,P<0.001)and family and romantic stressor(β=0.07,P<0.05)independently and positively predicted anxiety;academic stress(β=0.07,P<0.05),employment and future stress(β=0.12,P<0.01),and financial stress(β=0.08,P<0.05)independently positively predicted depression in medical postgraduates.Conclusion The newly developed Five-Factor Postgraduate Stressor Scale demonstrates good reliability and validity,and can be used to assess the stressor levels in postgraduate students.Academic stress is the predominant stressor for medical postgraduates.Stressors can independently and positively predict their anxiety and depression.Different stressors exert differential impacts on various negative emotions(anxiety and depression).
Objective Oral squamous cell carcinoma(OSCC)presents with insidious early symptoms,and most patients have developed lymph node metastasis at the time of initial diagnosis,resulting in poor prognosis and low survival rates.Therefore,exploring novel biomarkers for early diagnosis and targeted therapy strategies is crucial in this field.Clathrin heavy chain(CLTC)is a key regulatory protein in the endocytic process;however,its expression level,clinical significance,and role in invasion and migration in OSCC remain to be elucidated.Revealing these aspects may provide novel targets for OSCC-targeted therapy.Methods Transcriptomic data and corresponding clinical information from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)databases were integrated to analyze the expression level of CLTC in OSCC patients.Kaplan-Meier survival analysis was performed to evaluate the impact of CLTC expression on the prognosis of OSCC patients.A nomogram prognostic prediction model was constructed by integrating CLTC expression levels and clinicopathological parameters including patient age,tumor stage,tumor size,and lymph node metastasis status to individually evaluate prognosis in OSCC patients.Gene Set Enrichment Analysis(GSEA)was applied based on Kyoto Encyclopedia of Genes and Genomes(KEGG)database to explore the signaling pathways associated with CLTC.Immunohistochemical assay and Western blotting were employed to detect the protein level of CLTC in OSCC tissues,and analyze its relationship with the clinicopathological features in OSCC patients.Western blotting and RT-qPCR were utilized to measure CLTC expression in HOK,HGE,HSC3,SCC9,CAL27 and SCC25 cell lines.Three small interfering RNAs(siRNAs)were designed according to the CLTC gene sequence.Transfection efficiency was subsequently validated by Western blotting and RT-qPCR.Scratch wound healing assay and Transwell assay were utilized to evaluate the migration and invasion of OSCC cells,respectively.Western blotting was conducted to detect the expression of ERK signaling pathway-related proteins(ERK and pERK).Results Bioinformatics analysis showed that CLTC was highly expressed in various cancers,including oral cancer,hepatocellular carcinoma and lung adenocarcinoma,and its high expression was significantly correlated with poor prognosis in patients(P=0.033,HR=1.45,95％CI:1.03 to 2.03).The nomogram prediction model for the TCGA cohort showed the AUC values for 1-,3-,and 5-year survival predictions were 0.660,0.753,and 0.793,respectively,with calibration curves indicating good consistency between predicted and actual survival probabilities.The nomogram prediction model in the GEO cohort showed the 1-,3-,and 5-year survival prediction AUC values of 0.722,0.762,and 0.760,respectively;and calibration curves similarly demonstrated favorable consistency.GSEA results indicated that the differential gene set for CLTC was significantly enriched in multiple cancer-related KEGG pathways.Notably,cancer pathways,Wnt,TGF-β,and MAPK signaling pathways showed significant enrichment effects with high statistical credibility.Clinical specimen detection demonstrated that CLTC expression level in the OSCC tissues was significantly higher than that in paired adjacent non-cancerous tissues(P<0.001),and its expression level was significantly correlated with tumor differentiation degree and lymph node metastasis(P<0.05).CLTC knockdown resulted in markedly inhibited migration and invasion abilities of SCC25 cells(P<0.001)and significantly downregulated levels of ERK signaling pathway-related proteins(P<0.001).Rescue experiment results demonstrated that addition of the ERK signaling pathway activator LM22B-10 partially restored the decreased cell invasion and migration abilities(P<0.01).Conclusion CLTC is aberrantly highly expressed in OSCC tissues and cells,and its expression level is significantly correlated with tumor differentiation degree and lymph node metastasis status.CLTC regulates the invasion and migration of OSCC cells,and this effect may be associated with the inhibition of ERK signaling pathway activation.
Objective Preoperative risk assessment of cesarean scar pregnancy(CSP)in early pregnancy mainly relies on two-dimensional ultrasound classification;however,it fails to fully reflect blood supply and myometrial invasion,thereby limiting surgical decision-making.This study aims to investigate whether a novel classification based on contrast-enhanced ultrasound(CEUS)can more accurately identify high-risk cases and reduce unnecessary invasive surgical procedures.Methods A prospective cohort study design was adopted.A total of 128 patients diagnosed with CSP according to the 2016 Expert Consensus were enrolled from Department of Obstetrics and Gynecology of Yingshang People's Hospital during August 2021 and January 2026.The 2016 consensus classification categorized CSP into Types Ⅰ to Ⅲ mainly based on the relationship between the gestational sac and cesarean scar location as well as the residual myometrial thickness between the sac and the bladder.Our CEUS-based classification was developed based on the positional relationship between the peak"early and hyper-enhanced"trophoblastic perfusion area within 2 min after contrast injection and the scar myometrium,as well as the morphology of gestational tissues.Thus,the CSP cases were classified into 7 subtypes:Ⅰa,Ⅰb,Ⅰc,Ⅱa,Ⅱb,Ⅲa,and Ⅲb,and planned surgical approaches were formulated accordingly.Weighted Kappa test was used to evaluate the consistency of recommended surgical approaches between the 2 systems,and Pearson's χ² test was employed to compare the distribution of recommendations.The rate and direction(downgrading vs upgrading)of CEUS-induced changes in classification and surgical strategy were calculated with 95％confidence intervals(95％CIs).Results The 2016 Expert Consensus identified 60 cases(46.9％)as Type Ⅰ,43(33.6％)as Type Ⅱ,and 25(19.5％)as Type Ⅲ.The CEUS classification identified 35 Type Ia cases(27.3％),26 Type Ib(20.3％),10 Type Ⅰ c(7.8％),35 Type Ⅱ a(27.3％),11 Type Ⅱ b(8.6％),2 Type Ⅲ a(1.6％),and 9 Type Ⅲ b(7.0％).CEUS reclassification changed the classification in 55 cases(43.0％)and the final surgical approach in 56 cases(43.7％,95％CI:35.4％to 52.0％),including 47 downgraded(36.7％,95％CI:28.5％to 44.9％)and 9 upgraded cases(7.0％,95％CI:3.1％to 10.9％).Compared with the 2016 Consensus,CEUS-based classification reduced the planned proportion of invasive scar repair from 53.1％(68/128)to 17.2％(22/128)and increased the ratio of minimally invasive dilation and curettage(D&C)/hysteroscopic strategies from 46.9％(60/128)to 82.8％(106/128).The weighted Kappa was 0.582(95％CI:0.441 to 0.723),indicating moderate consistency,and Pearson's χ² was 23.44(df=1,P<0.001),indicating a significant difference in recommendation distribution.Conclusion Our new CEUS pattern-based classification improves preoperative risk stratification beyond conventional morphologic assessment,markedly reduces the planned proportion of invasive scar repair,and retains the ability to identify high-risk cases.This system may provide imaging evidence for individualized and minimally invasive management of early CSP.
Objective The etiology of recurrent spontaneous abortion(RSA)is complex,with embryonic chromosomal abnormalities representing a major contributing factor.This study aims to characterize the embryonic chromosomal abnormalities products of conception(POC)from RSA patients,compare the composition of abnormalities across various etiological subtypes,and analyze the association between maternal clinical factors and different types of chromosomal abnormalities.Methods A retrospective cohort study was conducted with predefined inclusion and exclusion criteria,a unified etiological evaluation protocol,standardized copy number variation sequencing(CNV-seq)and abnormality interpretation criteria,and systematic clinical variable extraction and statistical analysis to ensure quality control.A total of 534 RSA patients involving 629 pregnancy events who underwent POC CNV-seq in our hospital between January 2021 and December 2024 were recruited.According to systematic etiological evaluation,the patients were classified into a maternal/parental factor-explained recurrent spontaneous abortion(MPF-eRSA)group and a maternal/parental factor-unexplained recurrent spontaneous abortion(MPF-uRSA)group.The distribution of embryonic chromosomal abnormalities was compared between the 2 groups.Mixed-effects logistic regression models were used to evaluate the associations of maternal age,gestational age,number of previous miscarriages,mode of conception,and maternal risk factors with different types of chromosomal abnormalities.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were further performed on genes covered by RSA-related pathogenic CNVs(PCNVs),and the selected candidate genes were preliminarily validated by qRT-PCR.Results Among the 629 POC samples,chromosomal abnormalities were detected in 321 cases,with an overall detection rate of 51.0％.Numerical abnormalities were the predominant type,accounting for 75.1％of abnormal cases.Univariate analysis showed that the MPF-uRSA group had higher proportions of aneuploidy and single trisomy than the MPF-eRSA group(P<0.001),whereas mosaicism was more frequent in the MPF-eRSA group(P=0.015).Mixed-effects logistic regression analysis revealed a lower risk of overall chromosomal abnormalities in the MPF-eRSA group than the MPF-uRSA group(OR=0.41,95％CI:0.21 to 0.82,P=0.011).Maternal age was positively associated with the risk of overall chromosomal abnormalities(OR=1.48,95％CI:1.22 to 1.80,P<0.001),aneuploidy(OR=1.44,95％CI:1.17 to 1.77,P<0.001),and single trisomy(OR=1.55,95％CI:1.20 to 2.00,P<0.001).The risks of aneuploidy(OR=0.24,95％CI:0.11 to 0.51,P<0.001)and single trisomy(OR=0.14,95％CI:0.05 to 0.36,P<0.001)were significantly lower in the MPF-eRSA group than the MPF-uRSA group.Maternal endocrine factors were associated with an increased risk of single trisomy(OR=2.87,95％CI:1.19 to 6.90,P=0.018),while maternal genetic factors were associated with increased risks of single trisomy(OR=3.75,95％CI:1.20 to 11.69,P=0.023)and PCNVs(OR=9.91,95％CI:2.87 to 34.21,P<0.001).GO/KEGG enrichment analyses showed that the genes covered by RSA-related PCNVs were mainly involved in DNA damage repair,extracellular matrix homeostasis,and inflammation-related processes.qRT-PCR showed that RAD21,LIG4,and COL4A1 were downregulated,whereas CTSB and GSDMD were upregulated in the RSA group compared with the control group(P<0.05).Conclusion Embryonic chromosomal abnormalities in POC from RSA patients are predominantly numerical abnormalities,and their distribution differs across etiological subtypes.Aneuploidy is more common in MPF-uRSA patients.Maternal age is an important influencing factor for embryonic chromosomal abnormalities,mainly associated with the risk of aneuploidy.POC genetic testing combined with etiological classification may help improve etiological evaluation,risk stratification,and clinical management of RSA.
Objective Previous studies have suggested that vitamin D deficiency is common in patients with recurrent spontaneous abortion(RSA)and may be associated with immune-inflammatory imbalance and a prothrombotic state.In light of this,this study aims to investigate the relationship between serum 25-hydroxyvitamin D[25(OH)D]levels and inflammatory response as well as prothrombotic state in RSA patients.Methods A retrospective cohort study was conducted on 220 RSA patients who visited our reproductive medicine center between January 2022 and August 2023.Clinical data were analyzed,and serum levels of 25(OH)D,inflammatory markers(IL-6,TNF-α,IL-10,hsCRP),prothrombotic state indicators[total homocysteine(tHCY),D-dimer],glucolipid metabolism parameters,thyroid function,immune cell subsets and ovarian reserve function were assessed.According to 25(OH)D levels,these RSA patients were stratified into a severe deficiency group(<10 ng/mL,n=14),a deficiency group(10 to<20 ng/mL,n=140),an insufficiency group(20 to<30 ng/mL,n=53),and a sufficiency group(≥30 ng/mL,n=13).The above-mentioned indicators were compared among the groups.Mann-Whitney U test was used for comparisons between 2 groups.For comparisons among more than 2 groups,Kruskal-Wallis H test was performed followed by Dunn's post hoc test.Spearman and partial correlation analyses were applied to evaluate associations between 25(OH)D and related indicators.Results Vitamin D deficiency or insufficiency was observed in 94.09％of the cohort.Compared with the patients with 25(OH)D≥20 ng/mL,those with levels<20 ng/mL had significantly higher tHCY levels[8.35(7.23 to 9.60)µmol/L vs 7.60(6.35 to 8.95)µmol/L,P=0.007].No statistical differences were observed in the intergroup comparisons in glucolipid metabolism,thyroid function,inflammatory markers,immune cell subsets or ovarian reserve function.Spearman correlation analysis showed that serum 25(OH)D levels were negatively correlated with tHCY(r=-0.222,P<0.01),and this association remained significant after adjusting for age and body mass index(r=-0.188,P=0.019).In the patients with 25(OH)D<30 ng/mL,a weak negative correlation was observed between 25(OH)D and IL-6(r=-0.159,P<0.05),but this difference was not statistically significant after adjustment.Conclusion Vitamin D deficiency is quite common in RSA patients.Lower 25(OH)D levels are associated with an altered prothrombotic profile,particularly elevated tHCY,whereas their association with inflammatory markers appears limited.These findings suggest that vitamin D status may be involved in the regulation of thrombotic risk associated with RSA.
