ObjectiveTo explore the role of cucurbitacin B （CuB） in inducing ferroptosis in 4T1 cells and its mechanism. MethodsThe effects of CuB（0.2， 0.4， 0.8 μmol·L^-1）on the proliferation ability of 4T1 cells in vitro were detected using the methyl thiazolyl tetrazolium （MTT） assay. The clonogenic ability of 4T1 cells was detected by the plate cloning assay， and the levels of lactate dehydrogenase （LDH） in 4T1 cells were detected by the use of a kit. The mitochondrial membrane potential and reactive oxygen species （ROS） levels in 4T1 cells were detected by flow cytometry， and the mitochondrial ultrastructure of 4T1 cells was observed by transmission electron microscopy. The western blot was used to detect the expression of ferroptosis-related protein p53 in 4T1 cells， solute carrier family 7 member 11 （SCL7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， transferrin receptor protein 1 （TFR1）， and ferritin heavy chain 1 （FTH1）. ResultsCompared with that in the blank group， the survival rate of 4T1 cells in CuB groups was significantly decreased （P<0.05）， and the number of cell clones in CuB groups was significantly reduced （P<0.01）. In addition， compared with that in the blank group， the leakage of LDH in cells in CuB groups was significantly increased （P<0.01）， and the mitochondrial membrane potential of cells in CuB groups decreased significantly （P<0.01）. Cellular ROS levels were significantly elevated in CuB groups （P<0.01）. The mitochondria of cells in CuB groups were obviously wrinkled， and the mitochondrial cristae were reduced or even disappeared. Compared with that in the blank group， the protein expression of p53， ACSL4， and TFR1 were significantly up-regulated in CuB groups （P<0.05）， and that of SLC7A11， GPX4， and FTH1 were significantly down-regulated （P<0.05）. ConclusionCuB may inhibit SLC7A11 and GPX4 expression by up-regulating the expression of p53， which in turn regulates the p53/SLC7A11/GPX4 signaling pathway axis and accelerates the generation of lipid peroxidation substrate by up-regulating the expression of ACSL4. It up-regulates TFR1 expression to promote cellular uptake of Fe³⁺ and down-regulates the expression of FTH1 to reduce the ability of iron storage， resulting in an elevated free Fe²⁺ level. It catalyzes the Fenton reaction， generates excess ROS， imbalances the antioxidant system and iron metabolism， and then induces ferroptosis in 4T1 cells.

ObjectiveTo explore the role of cucurbitacin B （CuB） in inducing ferroptosis in 4T1 cells and its mechanism. MethodsThe effects of CuB（0.2， 0.4， 0.8 μmol·L^-1）on the proliferation ability of 4T1 cells in vitro were detected using the methyl thiazolyl tetrazolium （MTT） assay. The clonogenic ability of 4T1 cells was detected by the plate cloning assay， and the levels of lactate dehydrogenase （LDH） in 4T1 cells were detected by the use of a kit. The mitochondrial membrane potential and reactive oxygen species （ROS） levels in 4T1 cells were detected by flow cytometry， and the mitochondrial ultrastructure of 4T1 cells was observed by transmission electron microscopy. The western blot was used to detect the expression of ferroptosis-related protein p53 in 4T1 cells， solute carrier family 7 member 11 （SCL7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， transferrin receptor protein 1 （TFR1）， and ferritin heavy chain 1 （FTH1）. ResultsCompared with that in the blank group， the survival rate of 4T1 cells in CuB groups was significantly decreased （P<0.05）， and the number of cell clones in CuB groups was significantly reduced （P<0.01）. In addition， compared with that in the blank group， the leakage of LDH in cells in CuB groups was significantly increased （P<0.01）， and the mitochondrial membrane potential of cells in CuB groups decreased significantly （P<0.01）. Cellular ROS levels were significantly elevated in CuB groups （P<0.01）. The mitochondria of cells in CuB groups were obviously wrinkled， and the mitochondrial cristae were reduced or even disappeared. Compared with that in the blank group， the protein expression of p53， ACSL4， and TFR1 were significantly up-regulated in CuB groups （P<0.05）， and that of SLC7A11， GPX4， and FTH1 were significantly down-regulated （P<0.05）. ConclusionCuB may inhibit SLC7A11 and GPX4 expression by up-regulating the expression of p53， which in turn regulates the p53/SLC7A11/GPX4 signaling pathway axis and accelerates the generation of lipid peroxidation substrate by up-regulating the expression of ACSL4. It up-regulates TFR1 expression to promote cellular uptake of Fe³⁺ and down-regulates the expression of FTH1 to reduce the ability of iron storage， resulting in an elevated free Fe²⁺ level. It catalyzes the Fenton reaction， generates excess ROS， imbalances the antioxidant system and iron metabolism， and then induces ferroptosis in 4T1 cells.

Objective To develop and compare the predictive performance of five machine learning models for adverse postoperative outcomes in cardiac surgery patients, and to identify key decision factors through SHapley Additive exPlanations (SHAP) interpretability analysis. Methods A retrospective collection of perioperative data (including demographic information, preoperative, intraoperative, and postoperative indicators) with 88 variables was conducted from adult cardiac surgery patients at the First Affiliated Hospital of Xinjiang Medical University in 2023. Adverse postoperative outcomes were defined as the occurrence of acute kidney injury and/or in-hospital mortality during the postoperative hospitalization period following cardiac surgery. Patients were divided into an adverse outcome group and a favorable outcome group based on the presence of adverse postoperative outcomes. After screening feature variables using the least absolute shrinkage and selection operator (LASSO) regression method, five machine learning models were constructed: eXtreme gradient boosting (XGBoost), random forest (RF), gradient boosting machine (GBM), light gradient boosting machine (LightGBM), and generalized linear model (GLM). The dataset was randomly divided into a training set and a test set at a 7 : 3 ratio using stratified sampling, with postoperative outcome as the stratification factor. Model performance was evaluated using receiver operating characteristic curves, decision curve analysis, and F1 Score. The SHAP method was applied to analyze feature contribution. Results A total of 639 patients were included, comprising 395 males and 244 females, with a median age of 62 (55, 69) years. The adverse outcome group consisted of 191 patients, while the favorable outcome group included 448 patients, resulting in an adverse postoperative outcome incidence of 29.9%. Univariate analysis showed no significant differences between the two groups for any variables (P>0.05). Using LASSO regression, 16 feature variables were selected (including cardiopulmonary bypass support time, blood glucose on postoperative day 3, creatine kinase-MB isoenzyme, systemic inflammatory response index, etc.), and five machine learning models (GLM, RF, GBM, LightGBM, XGBoost) were constructed. Evaluation results demonstrated that the XGBoost model exhibited the best predictive performance on both the training set (n=447) and test set (n=192), with area under the curve values of 0.761 [95%CI (0.719, 0.800) ] and 0.759 [95%CI (0.692, 0.818) ], respectively. It also significantly outperformed other models in positive predictive value, and balanced accuracy in the test set. Decision curve analysis further confirmed its clinical utility across various risk thresholds. SHAP analysis indicated that variables such as cardiopulmonary bypass support time, blood glucose on postoperative day 3, creatine kinase-MB isoenzyme, and inflammatory markers (SIRI, NLR, CAR) had high contributions to the prediction. Conclusion The XGBoost model effectively predicts adverse postoperative outcomes in cardiac surgery patients. Clinically, attention should be focused on cardiopulmonary bypass support time, postoperative blood glucose control, and monitoring of inflammatory levels to improve patient prognosis.

Objective:To investigate the efficacy and safety of endovascular treatment (EVT) in young patients with symptomatic intracranial atherosclerotic stenosis (sICAS).Methods:Young patients with sICAS admitted to the Department of Neurology, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University, Medical School from January 2020 to July 2024 were included retrospectively. According to the therapeutic modalities, they were divided into a best medical treatment (BMT) group and an EVT group. The efficacy outcome was any stroke recurrence or death within 30 days and 1 year. The safety outcome was symptomatic intracranial hemorrhage (sICH) within 30 days and restenosis within 1 year.Results:A total of 113 patients were enrolled, including 85 males (75.2%), with a median age of 43 (interquartile range, 37-48) years; 44 patients (38.9%) received EVT, and 69 (61.1%) received BMT. Among the 44 patients who underwent EVT, 8 (18.2%) underwent balloon angioplasty and 36 (81.8%) underwent stenting. There was no significant difference in the incidence of stroke recurrence or death within 30 days (2.9% vs. 2.3%) and sICH incidence (0% vs. 2.3%) between the BMT group and the EVT group. However, the 1-year stroke recurrence or death rate in the EVT group was significantly lower than that in the BMT group (18.8% vs. 4.5%; P=0.029). Cox proportional hazards regression analysis showed that EVT was independently associated with a lower incidence of stroke recurrence or death within 1 year (hazard ratio 0.225, 95% confidence interval 0.051-0.996; P<0.05). The median age of the balloon angioplasty group was significantly lower than that of the stenting group (33.5 years vs. 46 years; P=0.007), while there were no significant differences in other demographic and baseline data. There was no significant difference in all efficacy and safety outcome between the balloon angioplasty group and the stenting group. Conclusions:For young patients with sICAS who have an unsatisfactory response to drug treatment, EVT can reduce the risk of stroke recurrence or death within 1 year without increasing the risk of sICH. The safety and efficacy of balloon angioplasty and stenting are similar.

Objective To investigate the effects of body weight and length of SD rats on the carotid artery balloon injury-induced vascular stenosis model in order to provide a reference for replicating an ideal vascular stenosis mod-el.Methods Male rats were divided into three groups based on body weight and length.The CONQUEROR? SC PTCA balloon catheter was employed,with a fixed balloon inflation volume of 0.2 mL to induce injury in the left common carotid artery,while the right side served as a control.As soon as surgery operation,one rat from each group was selected for Evans Blue dye verification.Fourteen days later,the injured and contra lateral common ca-rotid arteries from remaining rats were harvested for HE staining to check the extent of stenosis.Based on these find-ings,six rats within the optimal range of body weight and length were selected for further validation.Results Rats with body weights ranging from 280 to 380 g(corresponding body lengths of 21.0-26.5 cm)underwent balloon catheter injury,resulting in endothelial detachment and varying degrees of stenosis in the common carotid artery.In rats weighing 280-300 g(body lengths of 21.0-22.5 cm)had severe stenosis or occlusion of the common carotid artery with thrombosis.In rats weighing 320-340 g(body lengths of 23.0-24.5 cm),the internal and external elastic plates of the common carotid artery were ruptured and the vascular morphology was abnormal.Conversely,rats weighing 360-380 g(body lengths of 25.0-26.5 cm)did not show any ruptured elastic laminae or thrombus formation in the common carotid artery,and the extent of vascular stenos in rats with a body weight of 360 g was moderate and uniform.The results of the repeated validation experiments were consistent.Conclusions Rats with a body weight range of 360 g(corresponding body length of 25.0-26.5 cm)are suitable for development of an ideal vascular stenosis model.

Objective To investigate the detection rate and related influencing factors of gastrointestinal diseases in grass-roots personnel.Methods A total of 481 grass-roots personnel were enrolled and examined by magnetic-controlled capsule endoscopy(MCCE).Multivariate logistic regression were used to analyze the influencing factors of gastrointestinal diseases detected by MCCE.Results All personnel completed MCCE,and gastrointestinal diseases were detected in 154(32.0%)cases,including 106 cases of erosive gastritis,25 cases of chronic atrophic gastritis,17 cases of digestive tract polyp,16 cases of gastric ulcer,5 cases of reflux esophagitis,4 cases of cardia,1 case of duodenitis,and 1 case of enteritis.Gastrointestinal diseases was correlated with special operation posts,long-term tasks within recent 6 months,abdominal distension,belching,nausea and vomiting,diarrhea and other symptoms(all P＜0.05).Multivariate logistic regression analysis showed that the risk factors of gastrointestinal diseases were working in special operation posts,performing long tasks within 6 months,belching,nausea and vomiting symptoms(all P＜0.05).Conclusion The overall detection rate of gastrointestinal diseases(mainly acid related diseases)is relatively high,and its incidence is closely related to working in special operation posts and performing long-term tasks within recent 6 months.Personnel working in special operation posts should be more alert to gastrointestinal diseases.Gastrointestinal symptoms have reference value for the prediction of lesions,but more attention should be paid to the identification of functional gastrointestinal diseases with endoscopy.

Objective:To estimate the economic burden of 14 chronic diseases among middle-aged and elderly people in China.Methods:This cross-sectional study was based on longitudinal data from five waves (2011 to 2020) of the China Health and Retirement Longitudinal Study (CHARLS). Participants aged 45 and over who had been diagnosed with exactly one target chronic disease were included in the study. The 14 chronic diseases included: hypertension, dyslipidemia, diabetes or elevated blood glucose, malignant tumors, chronic pulmonary diseases, liver diseases, heart diseases, stroke, kidney diseases, digestive system diseases, emotional and mental diseases, memory-related diseases, arthritis or rheumatic diseases, and asthma. The economic burden of disease was measured in terms of both direct and indirect economic burden, with the results adjusted using a healthcare-specific Consumer Price Index (CPI). The direct economic burden included direct medical and non-medical burden. The human capital method was employed to calculate the indirect economic burden. A generalized linear mixed model (GLMM) was conducted to compare the differences in the economic burden between urban and rural areas, with provinces and prefecture-level cities set as random effects and residence (urban or rural areas) as fixed effects to control for the effects of geographic hierarchical structure. Gender, age and educational attainment were also included as covariates to control for confounding factors. The model′s robustness was assessed by comparing the significance of urban-rural differences before and after adding the covariates.Results:The median annual economic burden of the 14 chronic diseases among the middle-aged and elderly population in China ranged from 7 565 to 17 174 CNY, of which the direct economic burden ranged from 6 909 to 16 565 CNY, and the indirect burden ranged from 284 to 1 276 CNY. The direct economic burden was primarily driven by direct medical burden (83.67% to 95.01% of direct economic burden). Out-of-pocket expenses for outpatient medical burden ranged from 50% to 100%, while those for inpatient ranged from 36.30% to 61.29%. GLMM analysis revealed no statistically significant difference in the overall economic burden between urban and rural areas across diseases. However, the burden of inpatient medical burden for arthritis or rheumatism was higher in urban areas than in rural areas (5 338 vs. 3 898 CNY; LR=6.04, P=0.014). Similarly, the burden of self-treatment for hypertension was also higher in urban areas than in rural areas (324 vs. 238 CNY; LR=8.30, P=0.004). The outpatient non-medical burden for diabetes or elevated blood glucose (59 vs. 149 CNY; LR=5.99, P=0.014), stroke (0 vs. 307 CNY; LR=4.55, P=0.033), and digestive system diseases (45 vs. 107 CNY; LR=9.58, P=0.002) was lower in urban areas than rural. Conclusions:Chronic diseases cause heavy economic burden on middle-aged and elderly people in both urban and rural areas of China, with direct economic burden accounting for the majority of expenditure. The outpatient medical burden accounts for a higher proportion of out-of-pocket expenses than the inpatient.

Objective:To screen and analyze the carcinogenesis-related differential expression profile of long non-coding RNAs(LncRNAs)in oral lichen planus(OLP)mucosa tissue,and preliminarily analyze their functions,to explore their possible role in the development of OLP.Methods:High-throughput sequencing technology was used to construct differential expression profile from 5 cases of erosive OLP lesions and 5 of normal oral mucosa.LncRNAs that are closely related to the carcinogenesis of OLP were ob-tained by bioinformatics analysis.Results:400 LncRNAs associated with OLP were screened,of which 250 were up-regulated and 150 were down-regulated,and 5 LncRNAs were obtained with differential expression associated with OLP carcinogenesis:LncRNA 54055,100128560,399717,378825 and 100130231.Conclusion:400 LncRNAs are differentially expressed in the mucosa of erosive OLP lesions,and 5 of them are related to the incidence and carcinogenesis tendency of OLP.

Objective:To construct a perioperative pain nursing protocol for thoracoscopic surgery patients, providing a reference for clinical pain nursing practice.Methods:An evidence-based approach was used to search relevant guidelines and extract the best evidence. The initial draft was created through discussions among the research team, followed by two rounds of Delphi expert consultations. Based on the experts' suggestions, the protocol was revised and the best plan was finalized.Results:A total of 10 guidelines were included, and 22 experts participated in two rounds of consultations. The response rate for the consultation questionnaires was 100.00% (22/22) , with expert authority coefficients of 0.94 and 0.95 for the two rounds, respectively. The coefficient of variation for all indicators in the second round ranged from 0.04 to 0.24. The final pain nursing protocol included four primary indicators: personnel preparation, pain assessment, pain education, and pain intervention, with 10 secondary indicators and 27 tertiary indicators.Conclusions:The constructed perioperative pain nursing protocol for thoracoscopic surgery patients is significant, scientific, comprehensive, and targeted. It provides theoretical support and practical guidance for pain management, helping to reduce postoperative pain in patients.

Objective:To screen and analyze the carcinogenesis-related differential expression profile of long non-coding RNAs(LncRNAs)in oral lichen planus(OLP)mucosa tissue,and preliminarily analyze their functions,to explore their possible role in the development of OLP.Methods:High-throughput sequencing technology was used to construct differential expression profile from 5 cases of erosive OLP lesions and 5 of normal oral mucosa.LncRNAs that are closely related to the carcinogenesis of OLP were ob-tained by bioinformatics analysis.Results:400 LncRNAs associated with OLP were screened,of which 250 were up-regulated and 150 were down-regulated,and 5 LncRNAs were obtained with differential expression associated with OLP carcinogenesis:LncRNA 54055,100128560,399717,378825 and 100130231.Conclusion:400 LncRNAs are differentially expressed in the mucosa of erosive OLP lesions,and 5 of them are related to the incidence and carcinogenesis tendency of OLP.