Purpose: Erectile dysfunction (ED) is a common male sexual dysfunction. Gut microbiota plays an important role in various diseases. To investigate the effects and mechanisms of intestinal flora dysregulation induced by high-fat diet (HFD) on erectile function. Materials and Methods: Male Sprague–Dawley rats aged 8 weeks were randomly divided into the normal diet (ND) and HFD groups. After 24 weeks, a measurement of erectile function was performed. We performed 16S rRNA sequencing of stool samples. Then, we established fecal microbiota transplantation (FMT) rat models by transplanting fecal microbiota from rats of ND group and HFD group to two new groups of rats respectively. After 24 weeks, erectile function of the rats was evaluated and 16S rRNA sequencing was performed, and serum samples were collected for the untargeted metabolomics detection. Results: The erectile function of rats and the species diversity of intestinal microbiota in the HFD group was significantly lower, and the characteristics of the intestinal microbiota community structure were also significantly different between the two groups. The erectile function of rats in the HFD-FMT group was significantly lower than that of rats in the ND-FMT group. The characteristics of the intestinal microbiota community structure were significantly different. In the HFD-FMT group, 27 metabolites were significantly different and they were mainly involved in the several inflammation-related pathways. Conclusions Intestinal microbiota disorders induced by HFD can damage the intestinal barrier of rats, change the serum metabolic profile, induce low-grade inflammation and apoptosis in the corpus cavernosum of the penis, and lead to ED.

Purpose: Erectile dysfunction (ED) is a common male sexual dysfunction. Gut microbiota plays an important role in various diseases. To investigate the effects and mechanisms of intestinal flora dysregulation induced by high-fat diet (HFD) on erectile function. Materials and Methods: Male Sprague–Dawley rats aged 8 weeks were randomly divided into the normal diet (ND) and HFD groups. After 24 weeks, a measurement of erectile function was performed. We performed 16S rRNA sequencing of stool samples. Then, we established fecal microbiota transplantation (FMT) rat models by transplanting fecal microbiota from rats of ND group and HFD group to two new groups of rats respectively. After 24 weeks, erectile function of the rats was evaluated and 16S rRNA sequencing was performed, and serum samples were collected for the untargeted metabolomics detection. Results: The erectile function of rats and the species diversity of intestinal microbiota in the HFD group was significantly lower, and the characteristics of the intestinal microbiota community structure were also significantly different between the two groups. The erectile function of rats in the HFD-FMT group was significantly lower than that of rats in the ND-FMT group. The characteristics of the intestinal microbiota community structure were significantly different. In the HFD-FMT group, 27 metabolites were significantly different and they were mainly involved in the several inflammation-related pathways. Conclusions Intestinal microbiota disorders induced by HFD can damage the intestinal barrier of rats, change the serum metabolic profile, induce low-grade inflammation and apoptosis in the corpus cavernosum of the penis, and lead to ED.

Objective This study aimed to investigate the antimicrobial resistance profiles of bacterial strains isolated from pediatric intensive care units(PICU)in China for better antimicrobial therapy.Methods Clinical isolates were collected from 17 institutions,including tertiary care children's hospitals and pediatric department of tertiary general hospitals in China from January 1,2020 to December 31,2022.Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems.Results were interpreted according to the breakpoints released by the Clinical and Laboratory Standards Institute(CLSI)in 2020.Results A total of 10 688 isolates were collected,including gram-positive organisms(39.2％)and gram-negative organisms(60.8％).The top three organisms were S.aureus(13.6％,1 453/10 688),A.baumannii(10.0％,1 067/10 688),and coagulase-negative Staphylococcus(9.9％,1 058/10 688).Multi-drug resistant organisms(MDROs)were very common in children.The prevalence of methicillin-resistant Staphylococcus aureus(MRSA),carbapenem-resistant Enterobacterales(CRE),carbapenem-resistant E.coli,carbapenem-resistant K.pneumoniae(CRKP),carbapenem-resistant A.baumannii(CRAB),and carbapenem-resistant P.aeruginosa(CRPA)was 41.1％,19.4％,8.8％,30.9％,67.4％,and 28.8％,respectively.Overall,more than 50％of Enterobacteriales isolates were resistant to cephalosporins,while nearly 25％of Enterobacteriales isolates were resistant to carbapenems.MDROs were highly resistant to commonly used antibiotics.More than 80％of CRE and CRAB strains were resistant to all beta-lactam antibiotics.CRE and CRAB showed low resistance rates to tigecycline and polymyxin.CRPA showed lower resistance rates to piperacillin,beta-lactamase inhibitor combinations than the resistance rates to third and fourth generation cephalosporins.All of the Staphylococcus and Enterococcus isolates were susceptible to vancomycin and tigecycline.None of PRSP strains isolated from meningitis and nonmeningitis samples were resistant to rifampicin,vancomycin,or linezolid.The prevalence of β-lactamase-negative ampicillin-resistant(BLNAR)strains was 43.3％in Haemophilus influenzae.Conclusions MDROs were prevalent in PICU.It is necessary to establish an effective multidisciplinary team(MDT)to control the antimicrobial resistance.

Recent rapid developments in molecular biological techniques have allowed the use of gene editing,as a means of genome modification,for the establishment of experimental animal models,with high efficiency,accuracy,and flexibility.This article mainly summarizes the construction and application of the latest gene-editing techniques in animal models,including pigs,non-human primates and dogs.It provides a theoretical reference for the application and in-depth study of gene-editing techniques in large experimental animals,which may better simulate human diseases,and for further studies of the potential pathogenesis of biomedical and human complex diseases.

Objective:To explore the effectiveness of positive psychological intervention based on the positive emotion, engagement, relationship, meaning, accomplishment (PERMA) model in patients with chronic heart failure (CHF) .Methods:From January 2022 to December 2023, 230 patients with CHF in the Department of Cardiovascular Medicine of the Second Affiliated Hospital of Wenzhou Medical University were selected by convenience sampling method. The patients were divided into an experimental group ( n=115) and a control group ( n=115) based on the randomized numerical table method. Control group was given conventional intervention and experimental group was treated with positive psychological intervention based on the PERMA model on the basis of control group, both for four weeks. The Self-Perceived Burden Scale (SPBS) , Self-Management Scale of Heart Failure Patients, Warwick-Edinburgh Mental Well-being Scale (WEM-WBS) , Six-Minute Walk Test, and Newcastle Satisfaction with Nursing Scale (NSNS) were used to compare the effects of the intervention. Results:After the intervention, the SPBS score of experimental group was lower than that of control group, the Self-Management Scale of Heart Failure Patients score and WEM-WBS score were higher than those of control group, the Six-Minute Walk Test distance was longer than that of control group, and the satisfaction with nursing services was higher than that of control group, and the differences were all statistically significant ( P<0.05) . Conclusions:Positive psychological interventions based on the PERMA model can reduce the self-perceived burden of CHF patients and improve their self-management, positive psychological state, and satisfaction.

OBJECTIVE To explore the effect and potential mechanisms of berberine on formation of biofilms of clinical methicillin-resistant Staphylococcus aureus(MRSA)isolates.METHODS Totally 95 clinical MRSA iso-lates were collected from Shanghai Eighth People's Hospital from Jan.2023 to Dec.2023.The 14 biofilm forma-tion-related genes in the strains were detected by polymerase chain reaction(PCR)and multiplex PCR,the mini-mum inhibitory concentration(MIC)of berberine was determined by microbroth dilution method,the effect of berberine on resistance of biofilm formation was evaluated by crustal violet staining,fluorescence microscope,Congo red agar plate and extracelluar DNA(eDNA).The transcription levels of 9 biofilm formation-related genes were detected by real-time fluorescent quantitative reverse transcription PCR.RESULTS All of the 95 strains of MRSA carried eno,clfA,clfB and icaA genes,the most widespread gene profile was bbp-eno-ebpS-fnbA-fib-clfA-clfB-icaA-sasG,and 29 strains had the phenotypes with strong capability of biofilm formation.The MIC score of berberine ranged between 64 and 1 024 μg/ml.Berberine with the concentration of 1/2 MIC could inhibit the biofilm formation of MRSA(P＜0.001),and the inhibiting rate of biofilm formation of the MIC ≥512 μg/ml group was higher than that of the MIC≤128 μg/ml group and the MIC 256 μg/ml group(all P＜0.05).The re-sult under the fluorescence microscope showed that the fluorescence intensity of biofilms decreased with the rise of berberine concentration.Berberine could reduce the formation of amyloid fibrils and the release of eDNA,down-regulating the transcription levels of ica A,sasG,ebpS,fib,eno,clfA,clfB,bbp and fnbA genes(P＜0.05).CONCLUSION Berberine may inhibit the biofilm formation of MRSA by downregulating expression levels of related genes,interfering the formation of amyloid fibrils and blocking the release of eDNA,which may provide experimental bases for development of drugs resisting to MRSA biofilms.

Objective:To analyze the biological characteristics and genomic features of a Streptococcus pyogenes strain isolated from the pus of a child with an atypical brain abscess. Methods:The Streptococcus pyogenes strain, named 21SPY7071 ( emm22 type), was isolated from the brain abscess specimen of a child undergoing surgery for brain abscess. The biological characteristics of this strain were analyzed using blood agar culture, Gram staining, growth curve measurement, and hemolytic analysis. After whole-genome sequencing, core genome multilocus sequence typing (cgMLST) was performed to analyze the cgST type of the strain, and the distribution of virulence genes and drug resistance genes were analyzed. qRT-PCR was used to detect the differences in the expression of virulence genes at mRNA level between the strain 21SPY7071 and two Streptococcus pyogene strains of ATCC 19615 ( emm80 type) and GAS029 ( emm22 type, clinical isolate). Results:The strain 21SPY7071 was a Gram-positive Streptococcus with atypical weak β-hemolysis. Whole-genome sequencing and cgMLST revealed that this isolate belonged to the cgST type of 65937, containing 187 virulence genes and 97 drug resistance genes, with high sequence similarity (95.16%~100.00%) to the main virulence factor-encoding genes in the Virulence Factor Database. Compared with the strains of ATCC 19615 and GAS029, the strain 21SPY7071 showed reduced expression of genes encoding streptolysin S ( sagABCDEFGHI), streptolysin O ( slo), and hyaluronate lyase ( endA/ sdaB). Besides, compared with the strain ATCC 19615, the strain 21SPY7071 also showed decreased expression of virulence genes including speG, ideS/ mac, hylA, smeZ, lmb, and scpA/ scpB. Conclusion:This Streptococcus pyogenes strain isolated from the pus of a child with atypical brain abscess exhibits weak hemolysis and lower expression of virulence genes, which may be related to the mild clinical symptoms observed in the child.

Objective To obtain the recombinant Argonaute protein(TtAgo)from Thermus thermophilus using gene cloning and pro-tein purification techniques and detect its endonuclease activity against the KRAS 12D mutation in vitro,thereby to provide experimental evidence for its subsequent application in the detection of gene mutations.Methods The TtAgo gene sequence was amplified from the genome of Thermus thermophilus by the PCR technology.Then,the amplified sequence was inserted into the prokaryotic expression vec-tor pET-28a to construct the recombinant vector pET-28a-TtAgo.After the recombinant TtAgo protein was induced to be expressed,it was purified sequentially by the nickel ion affinity chromatography(Ni-NTA),heparin affinity chromatography,and size-exclusion chromatography.The high-specificity nuclease activity of the recombinant TtAgo protein mediated by a short DNA guide strand(gDNA)was verified in vitro.Results The prokaryotic expression vector pET-28a-TtAgo was successfully constructed by the gene cloning tech-nique,and the soluble expression of the recombinant TtAgo protein was achieved.With the help of protein purification technology,the high-purity recombinant TtAgo protein was obtained.Under the mediation of gDNA,the recombinant TtAgo protein could specifically cleave wild-type single-stranded DNA(ssDNA),double-stranded DNA(dsDNA),and RNA molecules,while had no cleavage activi-ty for tumor KRAS 12D mutant molecules.Conclusion The high-purity recombinant TtAgo protein with nuclease activity is successful-ly obtained,and its targeted cleavage activity towards the KRAS 12D mutation site is detected in vitro.

Objective:To investigate the effects of lactulose and oral rehydration salt powder (Ⅲ) on bowel preparation for colonoscopy and their impact on the detection rate of lesions.Methods:A retrospective study was conducted with 120 patients who underwent colonoscopy at The Second Affiliated Hospital of Wenzhou Medical University from January 2023 to April 2024. The patients were randomly divided into a control group and an experimental group, with 60 patients in each group. The control group received Hengkang Zhengqing and simethicone for bowel preparation, while the experimental group received lactulose, oral rehydration salt powder (Ⅲ), and simethicone for bowel preparation. The Boston Bowel Preparation Scale (referred to as the Boston scale) was used to assess bowel cleanliness, the Bubble scale was used to assess the presence of gas in the bowel, and a self-designed questionnaire was used to evaluate patient satisfaction after colonoscopy. The Boston scale scores, Bubble scale scores, patient satisfaction, lesion detection rates, and the incidence of adverse reactions were compared between the two groups. Results:The total score on the Boston scale in the experimental group was significantly higher than that of the control group [(7.23 ± 1.30) vs. (5.44 ± 1.17), t = 7.92, P < 0.001]. The Bubble scale score in the experimental group was significantly higher than that of the control group [(2.60 ± 0.15) vs. (2.21 ± 0.10), t = 16.75, P < 0.001]. The rates of acceptable taste and tolerance in the experimental group were 96.6% (58/60) and 95.0% (57/60), respectively, both of which were higher than those in the control group at 86.6% (52/60) and 81.6% (49/60) ( χ2 = 3.92, 5.17, P = 0.048, 0.023). The detection rate of colonic polyps in the experimental group was significantly higher than that in the control group [31.67% (19/60) vs. 15.00% (9/60), χ2 = 4.65, P = 0.031]. The incidence of adverse reactions in the experimental group was significantly lower than that in the control group [3.33% (2/60) vs. 13.33% (8/60), χ2 = 3.92, P = 0.048]. The patient satisfaction rate after examination in the experimental group was significantly higher than that in the control group [96.67% (58/60) vs. 85.00% (52/60), χ2 = 4.90, P = 0.027]. Conclusions:The intestinal preparation for colonoscopy using the lactulose and oral rehydration salt powder (Ⅲ) regimen can significantly improve the quality of bowel preparation and the detection rate of lesions. Additionally, it offers better taste, higher patient tolerance, and greater satisfaction, along with fewer adverse reactions compared with the Hengkang Zhengqing and simethicone regimen.

Turning to critical illness is a common stage of various diseases and injuries before death. Patients usually have complex health conditions, while the treatment process involves a wide range of content, along with high requirements for doctor′s professionalism and multi-specialty teamwork, as well as a great demand for time-sensitive treatments. However, this is not matched with critical care professionals and the current state of medical care in China. Telemedicine, which shortens the distance of medical professionals and the gap of disease diagnosis and treatments in various regions through electronic information, can effectively solve the current problem. Therefore, there is an urgent need to develop a standardized, high-quality visualization telemedicine round system .Therefore, experts have been organized to search domestic and foreign literature on telemedicine round for critically ill patients and to form this consensus based on clinical experiences so as to further improve the level of critical care treatments in regions.