ObjectiveTo investigate the pathogenic spectrum and epidemiological characteristics of diarrheal disease in Fengxian District of Shanghai, and to provide scientific basis for the prevention and control of diarrheal diseases. MethodsBasic information of the initial adult cases visited diarrheal disease surveillance sentinel hospital in Fengxian District, Shanghai, was collected from August 2013 to 2023, and fecal samples were collected at 1∶5 sampling intervals to isolate and identify 5 kinds of diarrheagenic Escherichia coli (DEC), Salmonella (SAL), Vibrio parahaemolyticus, Campylobacter, Vibrio cholerae, Shigella and Yersinia enterocolitica (YE). Simultaneously, nucleic acid detection was performed for 3 kinds of rotavirus, 2 kinds of norovirus, intestinal adenovirus, astrovirus and sapovirus. ResultsA total of 1 861 cases of newly diagnosed diarrheal disease were reported, with the peak in July to August. Additionally, 704 surveillance samples were detected, with a total positive detection rate of 50.57%. The detection rates of bacterial, viral and mixed infection were 25.14%, 21.02% and 4.40%, respectively. Among the pathogens detected, DEC accounted for the highest (17.61%, 124/704), followed by norovirus (16.48%, 116/704), rotavirus (6.39%, 45/704), SAL (5.97%, 42/704) and Campylobacter (3.84%, 27/704). DEC detected were mainly enteroaggregative Escherichia coli and enterotoxigenic Escherichia coli, with no detection of Vibrio cholerae, Shigella and YE. The highest total pathogen detection rate was observed from June to September, and the detection peaks of norovirus were from March to June and from October to December, whereas that of DEC was from June to October. The detection rate of rotavirus peaked from January to February, but which was not detected between 2020‒2023. The SAL positive rate peak was in September, whereas that of Campylobacter was from July to September. ConclusionThe main pathogens detected in Fengxian District from 2013‒2019 are DEC, norovirus, rotavirus, SAL and Campylobacter. Different pathogens have different detection peaks, with bacteria predominating in summer and viruses in winter and spring. Prevention and control measures should be carried out according to the epidemiological characteristics of different seasons.

Objective　To analyze the epidemiological characteristics and infection pathways and virulence factors of one cholera outbreak in Fengxian District, Shanghai, China, in 2024, and to provide scientific basis for epidemic control and prevention. Methods　Epidemiologic data of cases of one cholera outbreak in Fengxian District in 2024 were collected using on-site epidemiologic survey methods; RT-PCR nucleic acid testing and bacterial culture were applied to carry out pathogenicity testing of cases, close contacts, environment, and food samples; and the genome sequences of the strains were obtained using second-generation gene sequencing. Results　The case was a 62-year-old woman, who presented to the doctor with diarrhea for 4 consecutive days, 4-5 times a day, with watery stools, which was not effectively relieved by self-administered medication. There was no history of traveling away from Shanghai for 5 days before the onset of the disease, and she was engaged in the preparation and delivery of food for rural banquets during the period. Vibrio cholerae O139 was detected in the anal swab sample of the case and the septic tank of the workplace on the 4th day after the onset of the disease; samples of turtle and links in the store selling turtle were cultured for Vibrio cholerae O139. The isolate carried several virulence-related genes such as ctxA, ctxB, HlyA, zot, rtxA, hapA, nanH, tdh, and T3SS. Comparison of the isolate with the O139 strain of cholera cluster within our country through the National Pathogenic Bacteria Recognition Network (NPBN) in recent years suggests that the closest environmental or aquatic animal isolate to the sequence of this strain is the turtle isolate uploaded at a place in Guangdong. Conclusion　This outbreak was a disseminated outbreak caused by the case's contact with turtle contaminated with Vibrio cholerae O139, and early detection of enteric infectious diseases such as cholera can be achieved by relying on the outpatient enteric cholera surveillance network.

Given the increasing concern regarding antibacterial resistance, the antimicrobial properties of naphthoquinones have recently attracted significant attention. While 1,4-naphthoquinone and its derivatives have been extensively studied, the antibacterial properties of 5,8-dihydroxy-1,4-naphthoquinone derivatives remain relatively unexplored. This study presents a comprehensive in vitro and in vivo analysis of the antibacterial activity of 35 naturally sourced and chemically synthesized derivatives of 5,8-dihydroxy-1,4-naphthoquinone. Kirby-Bauer antibiotic testing identified three compounds with activity against methicillin-resistant Staphylococcus aureus (MRSA), with one compound (PNP-02) demonstrating activity comparable to vancomycin in minimum inhibitory concentration, minimum bactericidal concentration (MBC), and time-kill assays. Microscopic and biochemical analyses revealed that PNP-02 adversely affects the cell wall and cell membrane of MRSA. Mechanistic investigations, including proteomic sequencing analyses, Western blotting, and RT-qPCR assays, indicated that PNP-02 compromises cell membrane integrity by inhibiting arginine biosynthesis and pyrimidine metabolism pathways, thereby increasing membrane permeability and inducing bacterial death. In an in vivo mouse model of skin wound healing, PNP-02 exhibited antibacterial efficacy similar to vancomycin. The compound demonstrated low toxicity to cultured human cells and in hemolysis assays and remained stable during serum incubation. These findings suggest that PNP-02 possesses promising bioactivity against MRSA and represents a potential novel antibacterial agent.
Methicillin-Resistant Staphylococcus aureus/genetics* ; Anti-Bacterial Agents/chemistry* ; Naphthoquinones/administration & dosage* ; Animals ; Microbial Sensitivity Tests ; Mice ; Humans ; Staphylococcal Infections/microbiology* ; Molecular Structure

Objective: To analyze the effects of 12week highintensity fitness exercise on body composition,lipid metabolism and gut microbiota in obese adolescents, so as to provide references for improving the health levels of obese adolescents. Methods: From January to June 2023, 20 obese adolescents from Huaifeng Vocational and Technical School in Huaian City were recruited for the study. Participants were assigned to an exercise group (n=10) and a control group (n=10) for a 12week exercise intervention by random number table method, and both groups had the same diet during the intervention period. The exercise group engaged in three exercises every week, mainly consisting of moderate to highintensity aerobic exercise combined with highintensity intervals. In the first week, there was a 30 minutes of aerobic exercise, followed by 10 minutes of highintensity interval training in the total intervention time each week, and the rest of the time was aerobic exercise with a total intervention time of 60 minutes to maintain; the control group did not receive specific interventions. Body composition was measured using bioelectrical impedance analysis, and lipid levels were determined using an automatic biochemical analyzer. The expression levels of serum inflammatory factors were measured at baseline and after 12 weeks of intervention, and gut microbiota was analyzed using 16S rRNA gene sequencing. Statistical analysis was performed using t test and Chisquare test. Results: After 12 weeks of intervention, the levels of triglycerides (TG), and lowdensity lipoprotein cholesterol (LDL-C) in obese adolescents in the exercise group decreased from (1.7±0.6, 3.5±0.8) mmol/L to (0.9±0.3, 2.6±0.4) mmol/L, while highdensity lipoprotein cholesterol (HDL-C) increased from (1.1±0.2) mmol/L to (1.4±0.2) mmol/L; and serum interleukin-1 receptor antagonist(IL-1Rn) decreased from (8.4±1.6) to (4.5±0.4) ng/mL in the exercise group (t=7.34,2.49,-3.05,2.56, P<0.05). The α-diversity results showed that the Chao index (268.00±22.67) and Ace index (243.98±38.64) in the exercise group were higher than those in the control group (184.52±19.28, 171.43±23.33), and the differences were statistically significant (t=2.48, 2.53, P<0.05). The Shannon index (5.36±1.41) in the exercise group was higher than that in the control group (4.73±1.12), and the Simpson index (0.78±0.10) was lower than that in the control group (0.89±0.10), but the differences were not statistically significant (t=1.83, -2.10, P>0.05). The β-diversity results showed that the intergroup differences in gut microbiota structure between the exercise group and the control group were greater than the intragroup differences, and the differences in gut microbiota structure between the exercise group and the control group were statistically significant (R2=0.083，P<0.05). After intervention, there were significant differences in the relative abundances at the levels of phylum, class, genus, and species in gut microbiota among obese adolescents between the exercise group and the control group (P<0.05). Conclusion The 12week highintensity fitness exercise can alleviate obesity symptoms in obese adolescents through the gut microbiota-lipid metabolism pathway and improve mild chronic inflammatory status.

Objective As important components in the microenvironment of hepatocellular carcinoma(HCC),hepatic stellate cells(HSCs)and hydrogen sulfide(H2S)participate in various biological processes that regulate the development and progression of HCC.Through the co-culture of HSCs and HCC cells,this article aims to investigate the role and mechanism of HSCs in regulating the apoptosis of HCC cells by secreting H2S.Methods The HSC cell line(LX-2)and HCC cell lines(HepG2 and PLC/PRF/5)were used for experiment.RT-qPCR and Western Blot(WB)were used to measure the mRNA and protein expression levels of cystathionine γ-lyase(CSE),a key synthase for H2S;ELISA was used to measure the concentration of H2S in supernatant;next-generation sequencing,cell immunofluorescence assay,chromatin immunoprecipitation(ChIP),and WB were used to measure the JNK/JunB-TNFSF14 signaling pathway genes,binding sites,and related proteins after HepG2 cells were treated by H2S.LX-2 cells were co-cultured with HepG2 or PLC/PRF/5 cells in a Transwell chamber;CCK-8 assay and flow cytometry were used to measure the viability and apoptosis of HCC cells,and WB was used to measure the H2S-TNFSF14 signaling pathway-related proteins.All cell experiments were repeated three times.The independent-samples t test was used for comparison of continuous data between two groups;a one-way analysis of variance or the analysis of variance with repeated measures was used for comparison between multiple groups,and the Dunnett-t test was used for further comparison between two groups.Results LX-2 cells synthesized H2S mainly through CSE,and the concentration of H2S in supernatant of LX-2 cells gradually increased over time(22.89±0.08 pg/mL vs 28.29±0.15 pg/mL vs 36.19±1.90 pg/mL,F=79.63,P<0.05).In LX-2 cells,the mRNA expression level of CSE was significantly higher than that of CBS and MPST(1.008±0.13 vs 0.320±0.014 vs 0.05±0.02,F=80.84,P<0.05).When CSE was inhibited by PPG,the concentration of H2S decreased with the increase in the concentration of PPG(P<0.05).LX-2 cells were co-cultured with HepG2 or PLC/PRF/5 cells,and over the time of culture,there were significant reductions in the viability of HepG2 cells(87.48%±0.82%vs 70.48%±0.641%vs 52.89%±0.57%vs 45.20%±0.69%,F=1 517.13,P<0.001)and PLC/PRF/5 cells(92.41%±0.48%vs 74.10%±0.73%vs 53.70%±0.60%vs 44.00%±0.27%,F=2626.21,P<0.001)and significant increases in the apoptosis of HepG2 cells(12.88%±0.64%vs 15.5%±0.16%vs 18.43%±0.37%vs 13.01%±0.58%,F=142.15,P<0.001)and PLC/PRF/5 cells(8.51±0.05 vs 12.80±0.33 vs 15.59±0.21 vs 10.72±0.30,F=676.40,P<0.001),with the most significant changes on day 3.Next-generation sequencing showed that endogenous H2S and NaHS(endogenous H2S donor)were involved in regulating the expression of various genes in HepG2 cells.By releasing H2S,NaHS and LX2 activated the JNK/JunB signaling pathway and upregulated the expression of the apoptosis gene TNFSF14 in HCC cells,with increased binding between p-JunB and the transcriptional regulatory regions of the TNFSF14 gene.Conclusion In the microenvironment of HCC,HSCs activate the JNK/JunB signaling pathway in HCC cells through the signal molecules CSE/H2S,and there is an increase in the expression of TNFSF14,thereby promoting the apoptosis of HCC cells.

Objective:To search, evaluate and summarize the best evidence of intervention and management of eating behavior in children with autism, so as to provide evidence for medical staff and caregivers of children with autism to manage eating behavior problems.Methods:According to the evidence pyramid "6S" model, the clinical decisions, recommended practices, guidelines, evidence summaries, expert consensus, systematic reviews and randomized controlled trials on the intervention and management of eating behavior in children with ASD were systematically searched from domestic and foreign databases, related guide websites and professional association websites. The search period was from the establishment of the database to March 31th, 2023. The quality of the guide was independently evaluated by 4 researchers, and the remaining articles were evaluated by 2 researchers. Content extraction, evidence integration and grading of the included articles were carried out, and the level of evidence recommendation was formed through the expert meeting.Results:A total of 19 articles were included, including 3 guidelines, 1 expert consensus, 13 systematic reviews and 2 randomized controlled trials. This paper summarized 25 best evidence from 5 aspects, including the common eating behavior problems of children with autism, the assessment of eating behavior problems, causes, consequences and intervention methods.Conclusions:The best evidence summarized in this study provides a basis for the management of eating behavior problems in children with autism. It is suggested that clinical staff should fully consider the clinical context when applying the evidence, and formulate more detailed and feasible eating behavior intervention programs combined with the children and their families.

Objective:To review and summarize the best evidence of prevention and control measures of carbapenem-resistant Enterobacteriaceae (CRE) hospital infection, and provide evidence-based reference for health care workers to prevent and control CRE infection. Methods:A literature search strategy was developed to systematically search BMJ Best Clinical Practice, UpToDate, the World Health Organization, the Centers for Disease Control and Prevention website, the European Centers for Disease Control and Prevention website, the International Guidelines Collaboration website, the National Practice Technical Guidelines Library of the United States, the National Institute of Health and Clinical Optimization in the United Kingdom, the Ontario Medical Association of Canada, Maimai Tong, Cochrane Library, Australian Joanna Briggs Institute Evidence-Based Health Care Center database, PubMed, Web of Science, China National Knowledge Network, Wanfang database, VIP database, Chinese Biomedical Literature database of evidence related to the prevention and control of CRE hospital infection. The search period was from January 1, 2018 to October 1, 2023. The literature quality was evaluated independently by two research members, and evidence extraction and synthesis were carried out for the included literature.Results:A total of 10 articles were included, including 6 guidelines, 3 expert consensus articles, and 1 systematic review. 29 best evidences were formed from 6 aspects: preventive monitoring, active screening, contact isolation, environmental cleaning and disinfection, bacterial colonization, and hand hygiene measures.Conclusions:This study summarized the best evidence on the prevention and control measures of CRE infection in hospitals. Clinical staff and management departments should take active and effective prevention and control measures based on the local CRE epidemiological situation and resource requirements to reduce the risk of CRE infection.

Objective To explore the method for in vitro culture of organoid model of lung cancer,and use the organoid model for preliminary detection of personalized drug sensitivity.Methods Lung cancer patient tumor tissues were obtained by techniques such as surgery,fiberoptic bronchoscopy and CT-guided lung puncture biopsy.The lesion tissues were washed,cut,digested and filtered to ob-tain lung cancer cells.The lung cancer cells were resuspended in matrigel and inoculated in culture dishes for 3D organoid culture.ki-67 and CD133 immunohistochemistry were used to compare the correlation between organoids and parent tumor tissues.Chemotherapeutic drugs(vinorelbine,cisplatin,paclitaxel,gefitinib and pemetrexed)were used to analyze drug sensitivity results.Results Lung cancer organoids showed obvious nuclear atypia,consistent with parent lung cancer tissues.Lung cancer organoids showed strong positive CD133 immunohistochemistry,suggesting that lung cancer organoids were enriched with tumor stem cells.The proportion of ki-67 positive cells exceeded 50％,indicating that the organoids had certain proliferation ability.After treatment with chemotherapeutic drugs,lung cancer organoids were highly sensitive to cisplatin,gefitinib and paclitaxel,but less sensitive to vinorelbine and pemetrexed.Conclusion This study successfully established an in vitro culture system of lung cancer organoids,which can be preliminarily used for in vitro drug sensi-tivity test.

Objective To monitor the antifungal resistance of clinical isolates of Candida spp.in the East China region.Methods MALDI-TOF MS or molecular methods were used to re-identify the strains collected from January 2018 to December 2022.Antifungal susceptibility testing was performed using the broth microdilution method.The susceptibility test results were interpreted according to the breakpoints of 2022 Clinical and Laboratory Standards Institute(CLSI)documents M27 M44s-Ed3 and M57s-Ed4.Results A total of 3 026 strains of Candida were collected,65.33％of which were isolated from sterile body sites,mainly from blood(38.86％)and pleural effusion/ascites(10.21％).The predominant species of Candida were Candida albicans(44.51％),followed by Candida parapsilosis complex(19.46％),Candida tropicalis(13.98％),Candida glabrata(10.34％),and other Candida species(0.79％).Candida albicans showed overall high susceptibility rates to the 10 antifungal drugs tested(the lowest rate being 93.62％).Only 2.97％of the strains showed dose-dependent susceptibility(SDD)to fluconazole.Candida parapsilosis complex had a SDD rate of 2.61％and a resistance rate of 9.42％to fluconazole,and susceptibility rates above 90％to other drugs.Candida glabrata had a SDD rate of 92.01％and a resistance rate of 7.99％to fluconazole,resistance rates of 32.27％and 48.24％to posaconazole and voriconazole non-wild-type strains(NWT),respectively,and susceptibility rates above 90％to other drugs.Candida tropicalis had resistance rates of 29.55％and 26.24％to fluconazole and voriconazole,respectively,resistance rates of 76.60％and 21.99％to posaconazole and echinocandins non-wild-type strains(NWT),and a resistance rate of 2.36％to echinocandins.Conclusions The prevalence and species distribution of Candida spp.in the East China region are consistent with previous domestic and international reports.Candida glabrata exhibits certain degree of resistance to fluconazole,while Candida tropicalis demonstrates higher resistance to triazole drugs.Additionally,echinocandins resistance has emerged in Candida albicans,Candida glabrata,Candida tropicalis,and Candida parapsilosis.