Background and Aims:Pancreatic ductal adenocarcinoma(PDAC)is a highly malignant digestive system tumor with an inferior prognosis,and its early diagnosis and treatment remain significant challenges.In recent years,RNA methylation modifications(such as m6A and m5C)have attracted considerable attention for their roles in tumor development;however,their regulatory mechanisms and clinical significance in PDAC remain unclear.This study was conducted to identify prognosis-related long noncoding RNAs(lncRNAs)associated with m6A and m5C in PDAC,construct a reliable prognostic prediction model,and explore their relationship with the tumor immune microenvironment.Methods:Based on RNA-seq data from the TCGA-PDAC cohort,differentially expressed lncRNAs(DElncRNAs)related to m6A and m5C were identified through differential expression analysis and Pearson correlation analysis.The samples were randomly divided into a training set(n=89)and a validation set(n=89).Key DElncRNAs were selected using LASSO-Cox regression to construct a prognostic model,and patients were categorized into high-and low-risk groups based on risk scores.Kaplan-Meier survival analysis,ROC curves,and multivariate Cox regression were used to evaluate the model's predictive performance.Furthermore,CIBERSORT and ESTIMATE scores were used to analyze immune cell infiltration characteristics and tumor microenvironment(TME)differences between the high-and low-risk groups.Results:To construct the prognostic model,four m6A-and m5C-related DElncRNAs(LINC00857,LINC02038,TSPOAP1-AS1,and TRPC7-AS1)were identified.Patients in the high-risk group had significantly lower overall survival than those in the low-risk group(P<0.05),and the risk score was an independent prognostic factor for PDAC(HR=1.551,95%CI=1.297-1.854,P<0.001).ROC curve analysis showed that the risk score model exhibited high predictive efficiency in both the training and validation sets(AUC values for 1,3,and 5 years:0.766,0.875,0.879;0.685,0.711,0.792,respectively).Immune analysis revealed increased infiltration of M0 macrophages with lower TME scores in the high-risk group(all P<0.05),suggesting an immunosuppressive microenvironment.Conclusion:This study successfully established a PDAC prognostic model based on m6A-and m5C-related DElncRNAs and confirmed its independent predictive value.High-risk patients exhibited M0 macrophage enrichment and immunosuppressive microenvironment characteristics,possibly contributing to poor prognosis.

Objectives:To investigate the predictive value of epicardial fat volume(EFV)and atrioventricular groove fat thickness(AVGT)—morphological biomarkers of epicardial adipose tissue—for major adverse cardiovascular events(MACE)in patients with dilated cardiomyopathy(DCM).Methods:This study enrolled 216 DCM patients.EFV and AVGT were obtained from cardiac magnetic resonance imaging(CMR).Patients were divided into event-free group(n=142)and event group(n=74)based on MACE occurrence during follow-up.Receiver operating characteristic(ROC)curve analysis was used to determine optimal cutoff values.Survival differences were assessed using Kaplan-Meier analysis,Cox proportional hazards regression analysis was used to identify independent risk factors,and restricted cubic spline(RCS)models were used to evaluate dose-response relationships.Results:AVGT and EFV were significantly higher in the event group than in event-free group(both P<0.05).ROC analysis identified optimal MACE-predicting cutoffs as follows:AVGT≥7.74 mm(area under the curve[AUC]=0.57)and EFV≥78.6 ml(AUC=0.62).Kaplan-Meier analysis revealed significantly lower MACE-free survival rates in patients with AVGT≥7.74 mm and EFV≥78.6 ml(both P<0.05).Cox regression analysis confirmed that AVGT(HR=2.18,95%CI:1.34-3.54)and EFV(HR=1.81,95%CI:1.11-2.96)were independent MACE risk factors(both P<0.05)in this patient cohort.RCS models demonstrated the significant linear associations between EFV/AVGT and MACE risk(bothoverall P<0.05).Conclusions:EFV and AVGT,the non-invasive imaging biomarkers quantifying and characterizing fat distribution,are independently correlated with elevated MACE risk in DCM patients.These metrics serve as potential prognostic indicators,enriching risk stratification indicators for early identification of high-risk patients and guiding personalized medication strategies.

Objectives:To investigate the predictive value of epicardial fat volume(EFV)and atrioventricular groove fat thickness(AVGT)—morphological biomarkers of epicardial adipose tissue—for major adverse cardiovascular events(MACE)in patients with dilated cardiomyopathy(DCM).Methods:This study enrolled 216 DCM patients.EFV and AVGT were obtained from cardiac magnetic resonance imaging(CMR).Patients were divided into event-free group(n=142)and event group(n=74)based on MACE occurrence during follow-up.Receiver operating characteristic(ROC)curve analysis was used to determine optimal cutoff values.Survival differences were assessed using Kaplan-Meier analysis,Cox proportional hazards regression analysis was used to identify independent risk factors,and restricted cubic spline(RCS)models were used to evaluate dose-response relationships.Results:AVGT and EFV were significantly higher in the event group than in event-free group(both P<0.05).ROC analysis identified optimal MACE-predicting cutoffs as follows:AVGT≥7.74 mm(area under the curve[AUC]=0.57)and EFV≥78.6 ml(AUC=0.62).Kaplan-Meier analysis revealed significantly lower MACE-free survival rates in patients with AVGT≥7.74 mm and EFV≥78.6 ml(both P<0.05).Cox regression analysis confirmed that AVGT(HR=2.18,95%CI:1.34-3.54)and EFV(HR=1.81,95%CI:1.11-2.96)were independent MACE risk factors(both P<0.05)in this patient cohort.RCS models demonstrated the significant linear associations between EFV/AVGT and MACE risk(bothoverall P<0.05).Conclusions:EFV and AVGT,the non-invasive imaging biomarkers quantifying and characterizing fat distribution,are independently correlated with elevated MACE risk in DCM patients.These metrics serve as potential prognostic indicators,enriching risk stratification indicators for early identification of high-risk patients and guiding personalized medication strategies.

Background and Aims:Pancreatic ductal adenocarcinoma(PDAC)is a highly malignant digestive system tumor with an inferior prognosis,and its early diagnosis and treatment remain significant challenges.In recent years,RNA methylation modifications(such as m6A and m5C)have attracted considerable attention for their roles in tumor development;however,their regulatory mechanisms and clinical significance in PDAC remain unclear.This study was conducted to identify prognosis-related long noncoding RNAs(lncRNAs)associated with m6A and m5C in PDAC,construct a reliable prognostic prediction model,and explore their relationship with the tumor immune microenvironment.Methods:Based on RNA-seq data from the TCGA-PDAC cohort,differentially expressed lncRNAs(DElncRNAs)related to m6A and m5C were identified through differential expression analysis and Pearson correlation analysis.The samples were randomly divided into a training set(n=89)and a validation set(n=89).Key DElncRNAs were selected using LASSO-Cox regression to construct a prognostic model,and patients were categorized into high-and low-risk groups based on risk scores.Kaplan-Meier survival analysis,ROC curves,and multivariate Cox regression were used to evaluate the model's predictive performance.Furthermore,CIBERSORT and ESTIMATE scores were used to analyze immune cell infiltration characteristics and tumor microenvironment(TME)differences between the high-and low-risk groups.Results:To construct the prognostic model,four m6A-and m5C-related DElncRNAs(LINC00857,LINC02038,TSPOAP1-AS1,and TRPC7-AS1)were identified.Patients in the high-risk group had significantly lower overall survival than those in the low-risk group(P<0.05),and the risk score was an independent prognostic factor for PDAC(HR=1.551,95%CI=1.297-1.854,P<0.001).ROC curve analysis showed that the risk score model exhibited high predictive efficiency in both the training and validation sets(AUC values for 1,3,and 5 years:0.766,0.875,0.879;0.685,0.711,0.792,respectively).Immune analysis revealed increased infiltration of M0 macrophages with lower TME scores in the high-risk group(all P<0.05),suggesting an immunosuppressive microenvironment.Conclusion:This study successfully established a PDAC prognostic model based on m6A-and m5C-related DElncRNAs and confirmed its independent predictive value.High-risk patients exhibited M0 macrophage enrichment and immunosuppressive microenvironment characteristics,possibly contributing to poor prognosis.

Background Multiple studies have shown a close relationship between changes in gut microbiota composition and obesity, and research results are influenced by factors such as race and geographical location, but there are few studies on children. Objective To analyze the diversity of gut microbiota related to obesity in a population of 2-6 years old, observe the distribution characteristics and species differences of gut microbiota between obese/overweight and normal weight groups, and explore the association betweenobese/overweight and gut microbiota diversity. Methods Fecal samples were collected from 74 children aged 2-6 years in Shanghai, including 18 obese/overweight individuals, 6 males and 12 females (male to female ratio of 1∶2), and 56 normal weight individuals, 18 males and 38 females (male to female ratio is nearly 1∶2). The 16S rDNA was extracted from bacteria in fecal samples, followed by PCR amplification, cDNA construction, and high-throughput sequencing. Naive Bayes algorithm was used to perform taxonomic analysis (phylum, class, order, family, genus, species) and community diversity analysis (Sobs index, Shannon index, Shannoneven index, Coverage index, PD index, and principal co-ordinates analysis) on representative sequences and abundance of amplicon sequence variants (ASV). Wilcoxon rank sum test, P-value multiple test correction, and analysis of similarities were used to test differences between the two groups to obtain information on the distribution characteristics and species differences of intestinal microbiota in children. Results Seventy-four fecal samples were sequenced, and the sequencing results were subjected to quality control and filtering. A total of 4905306 optimized sequences were obtained, resulting in 1860 ASVs. The diversity data analysis of ASVs generated 889 species annotation results at 8 taxonomic levels. The alpha diversity analysis showed that the richness (Sobs index), diversity (Shannon index), evenness (Shannoneven index), and phylogenetic diversity (PD index) of fecal community of the obese/overweight children were increased compared to those of the normal weight children, but there were no statistical differences between the two groups (P>0.05). The beta diversity analysis showed that there was little difference in the composition of microbial species between the two groups, and no significant clustering separation was observed. The results of species composition analysis at phylum, order, family, and genus levels of 74 samples showed a consistent core microbiota structure in the two groups of gut microbiota, but there were differences in microbiota composition. The differences in microbial community composition between the two groups were manifested at the taxonomic levels of order, family, and genus, among which phylum Firmicutes, order Erysipelotrichales, family Erysipelatocyclostridiaceae, genus Erysipelotrichaceae_ UCG-003 and genus Catenibacterium were significantly enriched in the obese/overweight group and contributed significantly to the phenotypic difference of obese/overweight [linear discriminant analysis (LDA)=3.72, P<0.01; LDA=3.29, P<0.05). Phylum Proteobacteria, order Enterobacterales, family Enterobacteriaceae, genus unclassified was significantly enriched in the normal weight group and contributed significantly to the phenotypic difference of normal body weight (LDA=3.93, P<0.05). Conclusion The richness and diversity of gut microbiota in obese/overweight children aged 2-6 years in Shanghai are increased, but there is no difference compared to normal weight children. There is a difference in the composition of gut microbiota between the obese/overweight group and the normal weight group.

Intermittent theta burst stimulation (iTBS), a time-saving and cost-effective repetitive transcranial magnetic stimulation regime, has been shown to improve cognition in patients with Alzheimer's disease (AD). However, the specific mechanism underlying iTBS-induced cognitive enhancement remains unknown. Previous studies suggested that mitochondrial functions are modulated by magnetic stimulation. Here, we showed that iTBS upregulates the expression of iron-sulfur cluster assembly 1 (ISCA1, an essential regulatory factor for mitochondrial respiration) in the brain of APP/PS1 mice. In vivo and in vitro studies revealed that iTBS modulates mitochondrial iron-sulfur cluster assembly to facilitate mitochondrial respiration and function, which is required for ISCA1. Moreover, iTBS rescues cognitive decline and attenuates AD-type pathologies in APP/PS1 mice. The present study uncovers a novel mechanism by which iTBS modulates mitochondrial respiration and function via ISCA1-mediated iron-sulfur cluster assembly to alleviate cognitive impairments and pathologies in AD. We provide the mechanistic target of iTBS that warrants its therapeutic potential for AD patients.
Humans ; Mice ; Animals ; Transcranial Magnetic Stimulation ; Alzheimer Disease/therapy* ; Cognitive Dysfunction/therapy* ; Cognition ; Sulfur ; Iron ; Iron-Sulfur Proteins ; Mitochondrial Proteins

Aim To explore the effect of total flavonoid extract(TFE)of Ampelopsis megalophylla on the pro-liferation and apoptosis of human breast cancer cells and its mechanism in autophagy inhibition.Methods For human cervical cancer cell Hela,human lung cancer cell A549,human liver cancer cell SMMC-7721,human breast cancer cell MCF-7,MDA-MB-231 and human normal liver cell L-02,MTT method was used to select sensitive cell lines.The inhibitory effect of TFE combined with autophagy inhibitor 3-methylade-nine(3-MA)on sensitive cell proliferation was detec-ted using MTT assay.The morphological changes of cells were observed using transmission electron micros-copy and Hoechst 33258 single staining method.The changes in cell apoptosis rate were detected using An-nexin V-FITC/PI dual staining method.The expression levels of apoptosis related proteins and pathway pro-teins(death receptor pathway,mitochondrial pathway,endoplasmic reticulum stress pathway)were detected uisng Western blot.The expression of the key protein Cyt-c in mitochondrial pathway was determiend by im-munofluorescence,and the autophagy agonist rapamy-cin was selected for reverse validation.Results TFE could inhibit the proliferation of human breast cancer cells in a concentration-dependent manner,and MCF-7 cells were sensitive cell lines.Compared with the TFE group,the TFE+3-MA group significantly increased the inhibition rate of MCF-7 cells at 24,48,and 72 h(P＜0.01).The number of cells decreased,the gap increased,the number of apoptotic bodies increased,and the apoptosis rate increased(P＜0.01).The ex-pression levels of Bax/Bcl-2(P＜0.01),cleaved-caspase3(P＜0.01),Cyt-c(P＜0.05),FADD,and cleaved-caspase-12 all increased,and the expres-sion of apoptotic protein Cyt-c in nucleus increased.The fluorescence of the TFE+RA group decreased,re-versing the mitochondrial pathway apoptosis induced by TFE.Conclusions TFE can significantly inhibit the proliferation of human breast cancer cells.When inhib-iting autophagy,it may promote the apoptosis of MCF-7 cells through the mitochondrial pathway,and activa-ting autophagy can reverse apoptosis.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in healthcare facilities in major regions of China in 2023.Methods Clinical isolates collected from 73 hospitals across China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2023 Clinical & Laboratory Standards Institute (CLSI) breakpoints.Results A total of 445199 clinical isolates were collected in 2023,of which 29.0％ were gram-positive and 71.0％ were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species (excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi) (MRSA,MRSE and MRCNS) was 29.6％,81.9％ and 78.5％,respectively.Methicillin-resistant strains showed significantly higher resistance rates to most antimicrobial agents than methicillin-susceptible strains (MSSA,MSSE and MSCNS).Overall,92.9％ of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 91.4％ of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis had significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 93.1％ in the isolates from children and and 95.9％ in the isolates from adults.The resistance rate to carbapenems was lower than 15.0％ for most Enterobacterales species except for Klebsiella,22.5％ and 23.6％ of which were resistant to imipenem and meropenem,respectively .Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.6％ to 10.0％.The resistance rate to imipenem and meropenem was 21.9％ and 17.4％ for Pseudomonas aeruginosa,respectively,and 67.5％ and 68.1％ for Acinetobacter baumannii,respectively.Conclusions Increasing resistance to the commonly used antimicrobial agents is still observed in clinical bacterial isolates.However,the prevalence of important crabapenem-resistant organisms such as crabapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a slightly decreasing trend.This finding suggests that strengthening bacterial resistance surveillance and multidisciplinary linkage are important for preventing the occurrence and development of bacterial resistance.

AIM To investigate the effects of ampelopsin-mediated autophagy and apoptosis of human cervical cancer SiHa cells.METHODS After 24 h corresponding administration and culture among the control group,the 3-MA (5 mmol/L) group,the Z-VAD-FMK (50μmol/L) group,the ampelopsin (80μmol/L) group,the 3-MA+ampelopsin group and the Z-VAD-FMK+ampelopsin group,the cells had their cell proliferation inhibition rate detected by MTT method.After 24 h corresponding administration and culture among the control group,the 3-MA (5 mmol/L) group,the ampelopsin (80 μmol/L) group and the 3-MA+ampelopsin group,the cells had their morphological changes observed under electron microscope and their apoptosis detected by Hoechst33258 and AnnexinV-FITC/PI staining.After 24 h corresponding administration and culture among the control group,the Z-VAD-FMK (50μmol/L) group,the ampelopsin (80μmol/L) group and the Z-VAD-FMK+ampelopsin group,the cells had their autophagy and ultrastructure observed by MDC method and transmission electron microscopy.After 12 h corresponding administration and culture among the control group,the 3-MA (5 mmol/L) group,the ampelopsin (80 μmol/L) group,the 3-MA+ampelopsin group or control group,the Z-VAD-FMK (50 μmol/L) group,the ampelopsin (80 μmol/L) group,and the Z-VAD-FMK+ampelopsin group,the cells had their protein expressions of cleaved-PARP,cleaved-Caspase3,Bax,Bcl-2,Atg13,Beclin-1,LC3,and P62 detected by Western blot.RESULTS Compared with the control group,the ampelopsin group displayed enhanced proliferation inhibition of SiHa and C-33A cells (P<0.01).Compared with the ampelopsin group,the groups intervened with 3-MA+ampelopsin and Z-VAD-FMK+ampelopsin showed more significantly inhibited proliferation of the two cell lines (P<0.01),and decreased number of living cells.Compared with the ampelopsin group,the 3-MA+ampelopsin group showed increased bright blue fluorescence and apoptosis rate of SiHa cells (P<0.05),increased cleaved PARP,Bax,and P62 protein expressions (P<0.01),and decreased LC3Ⅱ/LC3Ⅰ ratio and Bcl-2 protein expression (P<0.01).Compared with the ampelopsin group,the Z-VAD-FMK+ampelopsin group demonstrated increased green dot fluorescence and number of autophagosomes and autopolysosomes,increased LC3Ⅱ/LC3Ⅰ ratio,Atg13 and Beclin-1 protein expression (P<0.05,P<0.01);and decreased protein expressions of P62,cleaved-PARP,cleaved-Caspase3 (P<0.05,P<0.01).CONCLUSION Being an antagonist of human cervical carcinoma SiHa cells,ampelopsin can induce autophagy and apoptosis of the cells through its key target on Beclin-1/Bcl-2.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in healthcare facilities in major regions of China in 2023.Methods Clinical isolates collected from 73 hospitals across China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2023 Clinical & Laboratory Standards Institute (CLSI) breakpoints.Results A total of 445199 clinical isolates were collected in 2023,of which 29.0％ were gram-positive and 71.0％ were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species (excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi) (MRSA,MRSE and MRCNS) was 29.6％,81.9％ and 78.5％,respectively.Methicillin-resistant strains showed significantly higher resistance rates to most antimicrobial agents than methicillin-susceptible strains (MSSA,MSSE and MSCNS).Overall,92.9％ of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 91.4％ of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis had significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 93.1％ in the isolates from children and and 95.9％ in the isolates from adults.The resistance rate to carbapenems was lower than 15.0％ for most Enterobacterales species except for Klebsiella,22.5％ and 23.6％ of which were resistant to imipenem and meropenem,respectively .Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.6％ to 10.0％.The resistance rate to imipenem and meropenem was 21.9％ and 17.4％ for Pseudomonas aeruginosa,respectively,and 67.5％ and 68.1％ for Acinetobacter baumannii,respectively.Conclusions Increasing resistance to the commonly used antimicrobial agents is still observed in clinical bacterial isolates.However,the prevalence of important crabapenem-resistant organisms such as crabapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a slightly decreasing trend.This finding suggests that strengthening bacterial resistance surveillance and multidisciplinary linkage are important for preventing the occurrence and development of bacterial resistance.