Mannheimia varigena(M.varigena)is a significant pathogen causing bovine respiratory disease.Transferrin binding protein B(TbpB)is a lipoprotein directly exposed to the outer mem-brane of the cell,which is not only involved in the bacterial iron metabolism pathway,but also an important virulence factor.This study aims to lay the groundwork for developing novel subunit vaccines by conducting site-directed mutagenesis on M.varigena TbpB binding-related residues and evaluating their immunogenicity.Based on whole-genome sequencing of a bovine M.varigena i-solate,its iron uptake pathway was predicted.Key amino acid residues of M.varigena TbpB that play a role for binding to bovine transferrin(bTf)was identified by bioinformatics.We constructed two M.varigena TbpB mutants(Y205A and Y258A)and assessed their bTf binding activity and immunogenicity through dot blot assays and mouse immunization studies.Dot blot assays result showed that the Y258A mutation caused TbpB to lose its ability to bind to bTf.Mouse immuniza-tion studies showed that,compared to wild-type TbpB,the mutants of TbpB induced higher levels of specific antibodies.In challenge experiments,mice immunized with mutant TbpB exhibited high-er survival rates.These results demonstrate that site-directed mutagenesis can enhance the immu-nogenicity of TbpB.This study provides a novel approach for developing new subunit vaccines a-gainst M.varigena.

AIM:To investigate the effects of puerarin(PU)on fecal metabolomics in atherosclerosis(AS)model mice.METHODS:Apolipoprotein E gene knockout(ApoE-/-)mice were fed a high-fat,high-cholesterol diet to es-tablish an AS model.The mice were randomly divided into control,model,and PU treatment groups,with five mice in each group.Body weight changes were recorded weekly.After 12 weeks,plasma inflammatory cytokines,lipid profiles,and aortic plaque area were measured,and fecal metabolomic analysis was performed using untargeted metabolomics.RE-SULTS:(1)The high-fat,high-cholesterol diet successfully induced the AS model,with significant increases in body weight(P<0.01),plasma levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1β(P<0.01),as well as higher levels of low-density lipoprotein cholesterol(LDL-C),total cholesterol(TC),and triglycerides(TG),and reduced levels of high-density lipoprotein cholesterol(HDL-C)(P<0.01).The plaque area in both the whole aorta and the aortic sinus was significantly increased(P<0.01).(2)PU treatment significantly reduced body weight(P<0.05)and inflammatory cytokine levels(P<0.01),and improved lipid metabolism disorders.Specifically,TC,TG,and LDL-C lev-els significantly decreased(P<0.05 or P<0.01),while HDL-C levels significantly increased(P<0.01).In addition,the plaque area in both the whole aorta and the aortic sinus was significantly reduced in the PU-treated group(P<0.01).(3)Metabolomic analysis showed that PU regulated the levels of L-valine,L-phenylalanine,and L-histidine,restoring abnor-mal amino acid metabolism pathways.Additionally,PU downregulated abnormal levels of phosphatidylcholine and ce-ramide,promoting lipid metabolism balance.PU also modulated bile acid metabolism and oxidative stress mechanisms,improving purine and pyrimidine metabolism and related energy metabolism pathways.CONCLUSION:PU significantly ameliorates metabolic abnormalities in AS mouse models by regulating lipid metabolism,amino acid metabolism,bile acid metabolism,and gut microbiota-related metabolic pathways.

Low-level laser therapy(LLLT),a noninvasive photobiomodulation technique,employs red or near-infrared(NIR)light(600-1 000 nm)with power outputs ranging from 5 to 500 mW.It exerts therapeutic effects through molecular mechanisms,specifically the activation of cytochrome C oxidase(CCO)and the modulation of intracellular signaling pathways.By enhancing mitochondrial adenosine triphosphate(ATP)synthesis,LLLT mitigates oxidative stress,regulates the reactive oxygen species(ROS)/glutathione peroxidase(GSH-Px)/superoxide dismutase(SOD)axis,and activates key path-ways,including phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)and mitogen-activated pro-tein kinase/extracellular signal-regulated kinase(MAPK/ERK).These mechanisms confer antioxidant,anti-inflammatory,and pro-regenerative properties to LLLT,making it a viable intervention for dermato-logical conditions,oncological therapies,and musculoskeletal disorders.Recent preclinical studies un-derscore LLLT's potential in male reproductive health.Specifically,it ameliorates cavernosal fibrosis and endothelial dysfunction in erectile dysfunction(ED)models by upregulating the PI3K/Akt and MAPK/ERK pathways.In the context of sperm biology,LLLT enhances motility and acrosomal integrity in both fresh and cryopreserved spermatozoa.This is achieved through mitochondrial metabolic reprogramming,such as CCO-mediated electron transport chain activation,redox homeostasis restoration,and epigenetic modulation involving DNA methylation and histone acetylation.Additionally,LLLT alleviates scrotal heat-induced oligospermia by promoting seminiferous epithelial differentiation,elevating serum testoster-one levels,and suppressing lipid peroxidation.These findings highlight the translational potential of LLLT in regenerative medicine,particularly for male sexual and reproductive disorders.Future research efforts should focus on interdisciplinary collaborations spanning life sciences,engineering,and physics.The goal is to optimize laser parameters,including wavelength,irradiance,and treatment duration,and establish standardized protocols.Rigorous preclinical and clinical investigations are paramount to validate the safety,efficacy,and long-term outcomes of LLLT,ultimately paving the way for its integration into precision medicine frameworks for urological and reproductive therapies.

AIM:To investigate the effects of puerarin(PU)on fecal metabolomics in atherosclerosis(AS)model mice.METHODS:Apolipoprotein E gene knockout(ApoE-/-)mice were fed a high-fat,high-cholesterol diet to es-tablish an AS model.The mice were randomly divided into control,model,and PU treatment groups,with five mice in each group.Body weight changes were recorded weekly.After 12 weeks,plasma inflammatory cytokines,lipid profiles,and aortic plaque area were measured,and fecal metabolomic analysis was performed using untargeted metabolomics.RE-SULTS:(1)The high-fat,high-cholesterol diet successfully induced the AS model,with significant increases in body weight(P<0.01),plasma levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1β(P<0.01),as well as higher levels of low-density lipoprotein cholesterol(LDL-C),total cholesterol(TC),and triglycerides(TG),and reduced levels of high-density lipoprotein cholesterol(HDL-C)(P<0.01).The plaque area in both the whole aorta and the aortic sinus was significantly increased(P<0.01).(2)PU treatment significantly reduced body weight(P<0.05)and inflammatory cytokine levels(P<0.01),and improved lipid metabolism disorders.Specifically,TC,TG,and LDL-C lev-els significantly decreased(P<0.05 or P<0.01),while HDL-C levels significantly increased(P<0.01).In addition,the plaque area in both the whole aorta and the aortic sinus was significantly reduced in the PU-treated group(P<0.01).(3)Metabolomic analysis showed that PU regulated the levels of L-valine,L-phenylalanine,and L-histidine,restoring abnor-mal amino acid metabolism pathways.Additionally,PU downregulated abnormal levels of phosphatidylcholine and ce-ramide,promoting lipid metabolism balance.PU also modulated bile acid metabolism and oxidative stress mechanisms,improving purine and pyrimidine metabolism and related energy metabolism pathways.CONCLUSION:PU significantly ameliorates metabolic abnormalities in AS mouse models by regulating lipid metabolism,amino acid metabolism,bile acid metabolism,and gut microbiota-related metabolic pathways.

In September 2023， the Measures for Scientific and Technological Ethics Review （Trial Implementation） was issued， revising the provisions related to the simplified procedure for ethical review in Chapter 3， Section 3. This revision of these provisions provides systematic guarantees for further optimizing ethical review work， ensuring that ethical review procedure is well-regulated， and improving scientific research efficiency. The “simplified procedure” does not mean reducing the quality and requirements of the review. Instead， based on always following internationally recognized ethical standards and emphasizing not violating national laws and regulations， improving the efficiency of ethical review and subsequent research work， and promoting the development of life sciences and medical research involving humans. In practical work， it introduces numerous new opportunities and challenges for the improvement of ethics review ability， such as new tests on the judgment and decision-making power of ethics committees， how to ensure the reliability and controllability of the conditions related to the simplified review procedure， and how to determine the basic conditions for adopting the simplified review procedure for review. Therefore， to actively respond to the challenges and possible risks brought by the simplified procedure review， efforts should be made to achieve three “unifications”， including the unification of researchers’ moral autonomy and the heteronomy of supervision implemented by relevant departments； the unification of the standard formulation of the simplified procedure review and the review work in practice； and the unification of ethical responsibility and legal responsibility.

Mannheimia varigena(M.varigena)is a significant pathogen causing bovine respiratory disease.Transferrin binding protein B(TbpB)is a lipoprotein directly exposed to the outer mem-brane of the cell,which is not only involved in the bacterial iron metabolism pathway,but also an important virulence factor.This study aims to lay the groundwork for developing novel subunit vaccines by conducting site-directed mutagenesis on M.varigena TbpB binding-related residues and evaluating their immunogenicity.Based on whole-genome sequencing of a bovine M.varigena i-solate,its iron uptake pathway was predicted.Key amino acid residues of M.varigena TbpB that play a role for binding to bovine transferrin(bTf)was identified by bioinformatics.We constructed two M.varigena TbpB mutants(Y205A and Y258A)and assessed their bTf binding activity and immunogenicity through dot blot assays and mouse immunization studies.Dot blot assays result showed that the Y258A mutation caused TbpB to lose its ability to bind to bTf.Mouse immuniza-tion studies showed that,compared to wild-type TbpB,the mutants of TbpB induced higher levels of specific antibodies.In challenge experiments,mice immunized with mutant TbpB exhibited high-er survival rates.These results demonstrate that site-directed mutagenesis can enhance the immu-nogenicity of TbpB.This study provides a novel approach for developing new subunit vaccines a-gainst M.varigena.

Antimicrobial resistance(AMR)has been a difficult problem of globally public health that can threaten human health.If its spread could not be effectively curbed,it will lead to increasing limitation in the options of clinical treatment.In the increasingly complex environment of diagnosis and treatment of infectious diseases,it is particularly urgent to build a rapid,accurate and widely popularized diagnosis system for pathogen,which is the key prerequisite of realizing accurate assessment for susceptibility of antimicrobial drug.This article comprehensively reviewed the main rapid diagnostic techniques currently used in AMR monitoring and detecting for antimicrobial susceptibility on the basis of the above conditions.It not only provided a detailed analysis of molecular typing methods represented by gene sequencing and phenotype identification methods based on conventional culture,but also focused on a new joint diagnostic strategy that combined genomic information with functional verification.Through comparative study for various technical principles,operating procedures and clinical applicability,the purpose of this paper was to provide scientific basis for optimizing the rapid detection scheme of drug susceptibility in clinical microbiology laboratories,and promote a comprehensive improvement of the level of diagnosis and treatment for infectious diseases,and ultimately improve the multi-layered network system of prevention and control for nosocomial infection.

Objective:To analyze the death cases in Beijing municipal hospitals, identify possible medical quality issues in unexpected death cases, so as to provide a basis for medical quality management in medical institutions.Methods:A total of 432 death cases from Beijing municipal hospitals were included, and their acute physiology and chronic health status were evaluated. Based on the evaluation results and the death risk level determined by the diagnosis-related groups, an average of 10 cases were selected from each hospital. A total of 186 cases from 20 hospitals were selected for clinical diagnosis and treatment process evaluation, those cases with problems in the diagnosis and treatment process were defined as unexpected deaths.Results:According to expert judgment, 19.89% (37/186) of the examined deaths had medical quality issues. The main medical quality issues included delayed diagnosis and treatment of patient condition changes (15 cases), inadequate preoperative evaluation (12 cases), failure to consult as required(11 cases), and inadequate prevention of venous thromboembolism (7 cases).Conclusions:Medical institutions should attach importance to the analysis of death cases, address the medical quality issues in the diagnosis and treatment of unexpected death cases, continuously improve medical quality management.

Objective To draw the genome-wide distribution and remodeling characteristics of H3K27me3 silencers in signet-ring cell carcinoma of the stomach(SRCC)through epigenetic sequencing technology,and to investigate their roles in transcriptional regulation in order to elucidate the regulatory mechanism of SRCC malignant progression.Methods The study was conducted on 35 gastric samples obtained by gastroendoscopic biopsy(15 normal and 20 SRCC tissues)from Department of Gastroenterology of Army Medical Center of PLA between January 2021 and December 2023.Multi-omics analyses,including assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq),cleavage under targets and tagmentation(CUT&Tag)and transcriptome sequencing(RNA-seq),were performed to identify chromatin accessibility,H3K27me3 silencer regions,and transcriptional changes,with aid of Illumina NovaSeq 6000.H3K27me3 related differentially expressed genes(|Log2FC|>1,FDR<0.05)were screened using DESeq2.Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were employed to analyze the enrichment function,and Homer was employed to identify transcription factor motifs.A regulatory network was constructed using Cytoscape,and then validated using immunohistochemistry to explore its regulatory mechanism.Results H3K27me3 silencers were primarily located in distal intergenic regions(37.06%)in SRCC.Compared with the normal tissues,SRCC showed a significant reduction in H3K27me3 silencer signals(95%CI:1.34～2.30,P=0.007)with 6 257 lost sites(FDR<0.01).Integrating CUT&Tag and RNA-seq revealed 380 up-regulated immune-related genes,particularly in T cell receptor signaling(OR=4.2,95%CI:2.8～6.3,P=0.002).Immunohistochemistry confirmed elevated expression of transcription factor EHF(P<0.05).Conclusion There is the remodeling of H3K27me3 silencers in SRCC,and EHF may potentially play a crucial role in the SRCC malignant progression.

Objective:To analyze the death cases in Beijing municipal hospitals, identify possible medical quality issues in unexpected death cases, so as to provide a basis for medical quality management in medical institutions.Methods:A total of 432 death cases from Beijing municipal hospitals were included, and their acute physiology and chronic health status were evaluated. Based on the evaluation results and the death risk level determined by the diagnosis-related groups, an average of 10 cases were selected from each hospital. A total of 186 cases from 20 hospitals were selected for clinical diagnosis and treatment process evaluation, those cases with problems in the diagnosis and treatment process were defined as unexpected deaths.Results:According to expert judgment, 19.89% (37/186) of the examined deaths had medical quality issues. The main medical quality issues included delayed diagnosis and treatment of patient condition changes (15 cases), inadequate preoperative evaluation (12 cases), failure to consult as required(11 cases), and inadequate prevention of venous thromboembolism (7 cases).Conclusions:Medical institutions should attach importance to the analysis of death cases, address the medical quality issues in the diagnosis and treatment of unexpected death cases, continuously improve medical quality management.