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 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.

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.

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 construct a risk prediction model for post-stroke dysphagia(PSD)based on clinical and laboratory indicators of elderly stroke patients with explainable machine learning.Methods A retrospective analysis was conducted on 3994 stroke patients hospitalized in Depart-ment of Neurology of First Medical Center of Chinese PLA General Hospital from October 2010 to December 2021.Among them,the 1390 cases admitted during January 2019 and December 2021 were assigned into an external validation set,and the 2604 cases admitted during October 2010 to January 2019 were into a training group.Those from the training group were further divided into a training set(1823 cases)and an internal validation set(781 cases)in a 7∶3 ratio,and also grouped into a PSD subgroup(773 cases)and a non-PSD group(1831 cases).With occurrence of swallowing difficulties as an endpoint,risk prediction models were constructed using random for-est(RF),eXtreme Gradient Boosting(XGBoost),Support Vector Machine(SVM),and logistic regression.ROC curve analysis was employed to evaluate the performance of our models.After the optimal model was selected,SHAP was employed to interpret feature contributions.Results There were significant differences in muscle strength,right/left-sided stroke,and area of brain in-jury between the PSD and the non-PSD groups(P＜0.01).The PSD group had obviously larger proportions of hypertension,diabetes,and drinking history,increased neutrophil counts,and de-creased levels of potassium and albumin when compared with the non-PSD group(P＜0.05,P＜0.01).Multivariate logistic regression analysis showed that age,drinking history,diabetes,hyper-tension,muscle strength grade,area of brain injury,hemispheric stroke,neutrophil count,and al-bumin and potassium levels were risk factors for PSD(P＜0.05,P＜0.01).The external validation results showed that the area under curve value of the RF model,XGBoost model,SVM model,and our logistic model was 0.883,0.902,0.877,and 0.868,respectively.The distribution of SHAP value showed that drinking history,hypertension and diabetes were positively correlated with PSD risk;Muscle strength was negatively correlated with the risk;Age growth was positively correlated with the risk;Subtentorial lesions showed stronger predictive efficacy than supratentorial lesions and entire lesions;The bilateral and right-sided stroke had higher risk for PSD than the left-sided stroke.Conclusion The model based on the XGBoost model shows best performance in predicting the risk for swallowing disorders in elderly patients after stroke.

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.

Objective To construct a risk prediction model for post-stroke dysphagia(PSD)based on clinical and laboratory indicators of elderly stroke patients with explainable machine learning.Methods A retrospective analysis was conducted on 3994 stroke patients hospitalized in Depart-ment of Neurology of First Medical Center of Chinese PLA General Hospital from October 2010 to December 2021.Among them,the 1390 cases admitted during January 2019 and December 2021 were assigned into an external validation set,and the 2604 cases admitted during October 2010 to January 2019 were into a training group.Those from the training group were further divided into a training set(1823 cases)and an internal validation set(781 cases)in a 7∶3 ratio,and also grouped into a PSD subgroup(773 cases)and a non-PSD group(1831 cases).With occurrence of swallowing difficulties as an endpoint,risk prediction models were constructed using random for-est(RF),eXtreme Gradient Boosting(XGBoost),Support Vector Machine(SVM),and logistic regression.ROC curve analysis was employed to evaluate the performance of our models.After the optimal model was selected,SHAP was employed to interpret feature contributions.Results There were significant differences in muscle strength,right/left-sided stroke,and area of brain in-jury between the PSD and the non-PSD groups(P＜0.01).The PSD group had obviously larger proportions of hypertension,diabetes,and drinking history,increased neutrophil counts,and de-creased levels of potassium and albumin when compared with the non-PSD group(P＜0.05,P＜0.01).Multivariate logistic regression analysis showed that age,drinking history,diabetes,hyper-tension,muscle strength grade,area of brain injury,hemispheric stroke,neutrophil count,and al-bumin and potassium levels were risk factors for PSD(P＜0.05,P＜0.01).The external validation results showed that the area under curve value of the RF model,XGBoost model,SVM model,and our logistic model was 0.883,0.902,0.877,and 0.868,respectively.The distribution of SHAP value showed that drinking history,hypertension and diabetes were positively correlated with PSD risk;Muscle strength was negatively correlated with the risk;Age growth was positively correlated with the risk;Subtentorial lesions showed stronger predictive efficacy than supratentorial lesions and entire lesions;The bilateral and right-sided stroke had higher risk for PSD than the left-sided stroke.Conclusion The model based on the XGBoost model shows best performance in predicting the risk for swallowing disorders in elderly patients after stroke.

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.