Objective To investigate the effects of scribble planar cell polarity protein(SCRIB)on proliferation,apoptosis,and autophagy of glioblastoma(GBM)and elucidate its potential underlying mechanisms.Methods The expression level of SCRIB in GBM tissue was queried through the Biomarker Exploration of Solid Tumors(BEST)database.Lentivirus-mediated shRNA interference was employed to downregulate SCRIB expression in human glioblastoma cell lines U87 and U251,which were divided into negative control group(mock)and SCRIB shRNA interference groups(kd1 and kd2).SCRIB expression levels were detected using Western blotting(WB)and quantitative polymerase chain reaction(qPCR).EdU incorporation and cell apoptosis rates were detected by flow cytometry(FCM).CCK-8 assay was used to detect the proliferation vitality of U87 and U251 cells,and WB was used to detect the expression of proliferation-related proteins.Immunofluorescence(IF)staining was conducted to detect the expression of autophagy-related proteins LC3 and p62,followed by quantitative analysis across multiple fields.WB was also used to detect the expression levels of LC3,p62,and proteins in the JAK-STAT3 signaling pathway.Results Compared with that of normal tissues,SCRIB mRNA expression level was significantly upregulated in GBM tissues(P＜0.05).FCM results showed that EdU incorporation rates were significantly reduced(P＜0.001)while cell apoptosis rates were markedly increased(P＜0.001)in U87 and U251 cells with SCRIB knockdown.CCK-8 results indicated that compared with the mock group,the proliferation vitality of U87 and U251 cells in the SCRIB knockdown group was significantly downregulated(P＜0.001).IF staining showed that LC3 fluorescence aggregation was significantly enhanced(P＜0.001),while p62 fluorescence aggregation was significantly reduced(P＜0.001)in the SCRIB knockdown group.WB results showed that compared with the mock group,the protein expression levels of p27,LC3,p-JAK2 and p-STAT3 were upregulated,while C-Myc,Cyclin D1,MCM,PCNA and p62 were downregulated,with statistically significant differences(P＜0.05).Conclusion Downregulation of SCRIB may induce autophagy and apoptosis in glioblastoma cells by inhibiting the JAK-STAT3 signaling pathway,thereby suppressing cell proliferation.

Objectives:QT interval prolongation during treadmill test exercise is one of the clinical feature of patients with long QT syndrome(LQTS).This study aimed to explore the feasibility and efficacy of treadmill exercise testing as an auxiliary diagnosis tool for LQTS in clinical practice.Methods:We enrolled normal healthy individuals,common cardiovascular disease patients,and clinically diagnosed or suspected LQTS patients,who underwent treadmill exercise test from July 2023 to July 2024 at Fuwai Hospital.A special treadmill exercise testing procedure was designed to record the QT interval correction(QTc)intervals of the twelve lead electrocardiogram at 6 time points when performing the exercise tablet,including supine,sitting,standing,peak exercise,and recovery at 1-minute and 4-minute.The differences in QTc intervals among healthy group,cardiovascular diseases group,and suspected LQTS group were compared.Results:A total of 80 cases were consecutively enrolled,including 37 normal healthy controls,25 patients with common cardiovascular disease,and 18 patients with suspected LQTS.The QTc intervals at 6 points did not differ significantly between normal healthy controls and patients with cardiovascular disease,with QTc intervals less than 480 ms at all measurement.For patients with suspected LQTS,67.7%(12/18)of these patients presented a QTc interval≥480 ms at the 4-minute during recovery period.Among them,5 cases were confirmed to have pathogenic gene mutations of LQTS by genetic testing(including 1 case with a lying electrocardiogram QTc interval of 489 ms diagnosed with LQTS 1 type and a QTc interval of 636 ms during the 4-minute recovery period after exercise);5 clinically diagnosed patients(negative or undetectable in genetic testing)with a Schwartz score≥4,and the remaining 2 patients had a Schwartz score of 3.The remaining 5/18 patients,include 2 patients with clinical Schwartz scores≥4 and 3 patients with clinical suspicion(Schwartz scores 2-3)had a 4 min QTc interval of 445-480 ms during exercise recovery.Another patient with clinical suspicion(Schwartz score 3)had a 4 min QTc interval of<445 ms during exercise recovery and a negative genetic test at a later stage.Receiver operating characteristic curve analysis showed a sensitivity of 83.3%and specificity of 98.4%for QTc interval≥482 ms during the 4-minute recovery period of exercise as the LQTS diagnostic cutoff.Conclusions:This study results suggest that this special treadmill exercise testing protocol is effective in identifying LQTS and has strong feasibility and generalizability for clinical practice.

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.

Outpatient services reflect the overall management level of hospitals and is a key link affecting patients′ healthcare experiences. To address the challenges faced by traditional outpatient services and meet patients′ diverse medical needs, a tertiary hospital implemented personalized outpatient services in March 2024. By utilizing information technologies such as big data and artificial intelligence, patient profile and a personalized service knowledge base was constructed, and a personalized outpatient service platform was built (including various functional modules such as intelligent guidance, pre consultation, abnormal indicator management, and full course management, as well as intelligent recommendation engines). This platform could provide personalized health guidance, customer service, and diagnosis and treatment services for outpatient throughout the entire process from pre-diagnosis to post diagnosis.After the launch of the personalized outpatient service platform, patient satisfaction in the oncology and pediatric departments increased from 90% and 86% in May 2024 to 94% and 91% in May 2025, respectively. From May 2024 to May 2025, the intelligent customer service addressed 740 000 patient inquiries, with a resolution rate of 90%. The online conversion rate (the proportion of patients switching from offline visits to online follow-ups relative to the total number of offline visits during the same period) rose from 37% in May 2023 to 43% in May 2025. The platform had enhanced the quality and efficiency of outpatient services, effectively improved patients′ healthcare experiences, and could serve as a reference for other hospitals seeking to upgrade their outpatient services.

Objective:To analyze the expression of tumor-associated transmembrane protein 61 (TMEM61) in cholangiocarcinoma tissues and its influence on prognosis and immune infiltration, as well as the effect on the proliferation of cholangiocarcinoma cells.Methods:In the cholangiocarcinoma gene chip dataset (TCGA-CHOL), differentially expressed genes between cholangiocarcinoma tissues and normal bile duct tissues were screened, and the upregulated TMEM61 gene was selected for further analysis. Based on the TMEM61 expression, cholangiocarcinoma patients higher than the median value were classified as the high-expression group ( n=17), and those lower than the median value were classified as the low-expression group ( n=18). The Kaplan-Meier survival curve was plotted. Functional and pathway enrichment analyses were conducted on differentially expressed genes related to TMEM61, and the correlations between TMEM61 expression and immune cells and immune molecules were respectively analyzed. The expression level of TMEM61 in cholangiocarcinoma tissues was analyzed by immunohistochemistry; The effect of TMEM61 expression on the proliferation of cholangiocarcinoma cells was detected by Western blotting, CCK-8, clone formation assay, etc. Results:Compared with normal tissues, the expression of TMEM61 mRNA in cholangiocarcinoma tissues was significantly upregulated ( t=18.31, P<0.001). The overall survival rate of patients in the high-expression group of TMEM61 was significantly lower than that in the low-expression group, and the difference was statistically significant ( χ2=7.23, P=0.007). The differentially expressed genes related to TMEM61 were involved in cell proliferation, cell cycle and DNA replication, etc. Compared with normal tissues, regulatory T cells ( t=10.21, P<0.001) and M0-type macrophages ( t=5.89, P=0.008) were significantly increased in cholangiocarcinoma tissues. Plasma cells ( t=7.34, P=0.002), γδT cells ( t=9.87, P<0.001), and M2-type macrophages ( t=11.53, P<0.001) were significantly decreased in cholangiocarcinoma tissues. The expression of TMEM61 was correlated with neurociliary protein 1, tumor necrosis factor ligand superfamily member 15 and B7 homologous protein 3 (all P<0.05). The proportion of positive staining area of TMEM61 protein in normal tissues was (10.15±2.27) %, and that in cholangiocarcinoma tissues was (69.43±11.66) %. The difference was statistically significant ( t=14.97, P<0.001). Inhibition of TMEM61 expression led to a decrease in the number of cholangiocarcinoma cell clones and proliferation activity, and the differences were statistically significant (both P<0.01). Conclusion:The expression of TMEM61 is elevated in cholangiocarcinoma tissues and is associated with poor prognosis. The abnormally high expression of TMEM61 affects the infiltration of immune cells and promotes the proliferation of cholangiocarcinoma cells. TMEM61 is expected to become a potential biomarker for the prognosis assessment of cholangiocarcinoma.

Objectives:QT interval prolongation during treadmill test exercise is one of the clinical feature of patients with long QT syndrome(LQTS).This study aimed to explore the feasibility and efficacy of treadmill exercise testing as an auxiliary diagnosis tool for LQTS in clinical practice.Methods:We enrolled normal healthy individuals,common cardiovascular disease patients,and clinically diagnosed or suspected LQTS patients,who underwent treadmill exercise test from July 2023 to July 2024 at Fuwai Hospital.A special treadmill exercise testing procedure was designed to record the QT interval correction(QTc)intervals of the twelve lead electrocardiogram at 6 time points when performing the exercise tablet,including supine,sitting,standing,peak exercise,and recovery at 1-minute and 4-minute.The differences in QTc intervals among healthy group,cardiovascular diseases group,and suspected LQTS group were compared.Results:A total of 80 cases were consecutively enrolled,including 37 normal healthy controls,25 patients with common cardiovascular disease,and 18 patients with suspected LQTS.The QTc intervals at 6 points did not differ significantly between normal healthy controls and patients with cardiovascular disease,with QTc intervals less than 480 ms at all measurement.For patients with suspected LQTS,67.7%(12/18)of these patients presented a QTc interval≥480 ms at the 4-minute during recovery period.Among them,5 cases were confirmed to have pathogenic gene mutations of LQTS by genetic testing(including 1 case with a lying electrocardiogram QTc interval of 489 ms diagnosed with LQTS 1 type and a QTc interval of 636 ms during the 4-minute recovery period after exercise);5 clinically diagnosed patients(negative or undetectable in genetic testing)with a Schwartz score≥4,and the remaining 2 patients had a Schwartz score of 3.The remaining 5/18 patients,include 2 patients with clinical Schwartz scores≥4 and 3 patients with clinical suspicion(Schwartz scores 2-3)had a 4 min QTc interval of 445-480 ms during exercise recovery.Another patient with clinical suspicion(Schwartz score 3)had a 4 min QTc interval of<445 ms during exercise recovery and a negative genetic test at a later stage.Receiver operating characteristic curve analysis showed a sensitivity of 83.3%and specificity of 98.4%for QTc interval≥482 ms during the 4-minute recovery period of exercise as the LQTS diagnostic cutoff.Conclusions:This study results suggest that this special treadmill exercise testing protocol is effective in identifying LQTS and has strong feasibility and generalizability for clinical practice.

Objective To investigate the effects of scribble planar cell polarity protein(SCRIB)on proliferation,apoptosis,and autophagy of glioblastoma(GBM)and elucidate its potential underlying mechanisms.Methods The expression level of SCRIB in GBM tissue was queried through the Biomarker Exploration of Solid Tumors(BEST)database.Lentivirus-mediated shRNA interference was employed to downregulate SCRIB expression in human glioblastoma cell lines U87 and U251,which were divided into negative control group(mock)and SCRIB shRNA interference groups(kd1 and kd2).SCRIB expression levels were detected using Western blotting(WB)and quantitative polymerase chain reaction(qPCR).EdU incorporation and cell apoptosis rates were detected by flow cytometry(FCM).CCK-8 assay was used to detect the proliferation vitality of U87 and U251 cells,and WB was used to detect the expression of proliferation-related proteins.Immunofluorescence(IF)staining was conducted to detect the expression of autophagy-related proteins LC3 and p62,followed by quantitative analysis across multiple fields.WB was also used to detect the expression levels of LC3,p62,and proteins in the JAK-STAT3 signaling pathway.Results Compared with that of normal tissues,SCRIB mRNA expression level was significantly upregulated in GBM tissues(P＜0.05).FCM results showed that EdU incorporation rates were significantly reduced(P＜0.001)while cell apoptosis rates were markedly increased(P＜0.001)in U87 and U251 cells with SCRIB knockdown.CCK-8 results indicated that compared with the mock group,the proliferation vitality of U87 and U251 cells in the SCRIB knockdown group was significantly downregulated(P＜0.001).IF staining showed that LC3 fluorescence aggregation was significantly enhanced(P＜0.001),while p62 fluorescence aggregation was significantly reduced(P＜0.001)in the SCRIB knockdown group.WB results showed that compared with the mock group,the protein expression levels of p27,LC3,p-JAK2 and p-STAT3 were upregulated,while C-Myc,Cyclin D1,MCM,PCNA and p62 were downregulated,with statistically significant differences(P＜0.05).Conclusion Downregulation of SCRIB may induce autophagy and apoptosis in glioblastoma cells by inhibiting the JAK-STAT3 signaling pathway,thereby suppressing cell proliferation.

Objective:To analyze the expression of tumor-associated transmembrane protein 61 (TMEM61) in cholangiocarcinoma tissues and its influence on prognosis and immune infiltration, as well as the effect on the proliferation of cholangiocarcinoma cells.Methods:In the cholangiocarcinoma gene chip dataset (TCGA-CHOL), differentially expressed genes between cholangiocarcinoma tissues and normal bile duct tissues were screened, and the upregulated TMEM61 gene was selected for further analysis. Based on the TMEM61 expression, cholangiocarcinoma patients higher than the median value were classified as the high-expression group ( n=17), and those lower than the median value were classified as the low-expression group ( n=18). The Kaplan-Meier survival curve was plotted. Functional and pathway enrichment analyses were conducted on differentially expressed genes related to TMEM61, and the correlations between TMEM61 expression and immune cells and immune molecules were respectively analyzed. The expression level of TMEM61 in cholangiocarcinoma tissues was analyzed by immunohistochemistry; The effect of TMEM61 expression on the proliferation of cholangiocarcinoma cells was detected by Western blotting, CCK-8, clone formation assay, etc. Results:Compared with normal tissues, the expression of TMEM61 mRNA in cholangiocarcinoma tissues was significantly upregulated ( t=18.31, P<0.001). The overall survival rate of patients in the high-expression group of TMEM61 was significantly lower than that in the low-expression group, and the difference was statistically significant ( χ2=7.23, P=0.007). The differentially expressed genes related to TMEM61 were involved in cell proliferation, cell cycle and DNA replication, etc. Compared with normal tissues, regulatory T cells ( t=10.21, P<0.001) and M0-type macrophages ( t=5.89, P=0.008) were significantly increased in cholangiocarcinoma tissues. Plasma cells ( t=7.34, P=0.002), γδT cells ( t=9.87, P<0.001), and M2-type macrophages ( t=11.53, P<0.001) were significantly decreased in cholangiocarcinoma tissues. The expression of TMEM61 was correlated with neurociliary protein 1, tumor necrosis factor ligand superfamily member 15 and B7 homologous protein 3 (all P<0.05). The proportion of positive staining area of TMEM61 protein in normal tissues was (10.15±2.27) %, and that in cholangiocarcinoma tissues was (69.43±11.66) %. The difference was statistically significant ( t=14.97, P<0.001). Inhibition of TMEM61 expression led to a decrease in the number of cholangiocarcinoma cell clones and proliferation activity, and the differences were statistically significant (both P<0.01). Conclusion:The expression of TMEM61 is elevated in cholangiocarcinoma tissues and is associated with poor prognosis. The abnormally high expression of TMEM61 affects the infiltration of immune cells and promotes the proliferation of cholangiocarcinoma cells. TMEM61 is expected to become a potential biomarker for the prognosis assessment of cholangiocarcinoma.

Outpatient services reflect the overall management level of hospitals and is a key link affecting patients′ healthcare experiences. To address the challenges faced by traditional outpatient services and meet patients′ diverse medical needs, a tertiary hospital implemented personalized outpatient services in March 2024. By utilizing information technologies such as big data and artificial intelligence, patient profile and a personalized service knowledge base was constructed, and a personalized outpatient service platform was built (including various functional modules such as intelligent guidance, pre consultation, abnormal indicator management, and full course management, as well as intelligent recommendation engines). This platform could provide personalized health guidance, customer service, and diagnosis and treatment services for outpatient throughout the entire process from pre-diagnosis to post diagnosis.After the launch of the personalized outpatient service platform, patient satisfaction in the oncology and pediatric departments increased from 90% and 86% in May 2024 to 94% and 91% in May 2025, respectively. From May 2024 to May 2025, the intelligent customer service addressed 740 000 patient inquiries, with a resolution rate of 90%. The online conversion rate (the proportion of patients switching from offline visits to online follow-ups relative to the total number of offline visits during the same period) rose from 37% in May 2023 to 43% in May 2025. The platform had enhanced the quality and efficiency of outpatient services, effectively improved patients′ healthcare experiences, and could serve as a reference for other hospitals seeking to upgrade their outpatient services.