Animal experiments are an essential component of life sciences and medical research. However, the external validity and reliability of individual animal studies are frequently challenged by inherent limitations such as small sample sizes, high design heterogeneity, and poor reproducibility, which impede the effective translation of research findings into clinical practice. Systematic reviews and meta-analysis represent a key methodology for integrating existing evidence and enhancing the robustness of conclusions. Currently, however, the application of systematic reviews and meta-analysis in the field of animal experiments lacks standardized guidelines for their conduct and reporting, resulting in inconsistent quality and, to some extent, diminishing their evidence value. To address this issue, this paper aims to systematically delineate the reporting process for systematic reviews and meta-analysis of animal experiments and to propose a set of standardized recommendations that are both scientific and practical. The article's scope encompasses the entire process, from the preliminary preparatory phase [including formulating the population， intervention， comparison and outcome （PICO） question, assessing feasibility, and protocol pre-registration] to the key writing points for each section of the main report. In the core methods section, the paper elaborates on how to implement literature searches, establish eligibility criteria, perform data extraction, and assess the risk of bias, based on the Preferred Reporting Items for Systematic Reviews and Meta-analysis （PRISMA) statement, in conjunction with relevant guidelines and tools such as Animal Research： Reporting of in Vivo Experiments （ARRIVE) and a risk of bias assessment tool developed by the Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE). For the presentation of results, strategies are proposed for clear and transparent display using flow diagrams and tables of characteristics. The discussion section places particular emphasis on how to scientifically interpret pooled effects, thoroughly analyze sources of heterogeneity, evaluate the impact of publication bias, and cautiously discuss the validity and limitations of extrapolating findings from animal studies to clinical settings. Furthermore, this paper recommends adopting the Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology to comprehensively grade the quality of evidence. Through a modular analysis of the entire reporting process, this paper aims to provide researchers in the field with a clear and practical guide, thereby promoting the standardized development of systematic reviews and meta-analysis of animal experiments and enhancing their application value in scientific decision-making and translational medicine.

Background Grassroots medical and health personnel are an important component of China's public health system, and guaranteeing their physical and mental health will have a profound impact on the development of China's health service. Objective To identify potential influencing factors of occupational stress, anxiety, depression, and insomnia as well as their interactions. Methods In August 2021, a cross-sectional survey was conducted among all the staff (2675 medical and health personnel) at 7 secondary public hospitals and 8 health institutions in Xiong’an New Area of Hebei Province by cluster sampling. Occupational stress, anxiety, depression, and insomnia were evaluated by the Effort-Reward Imbalance Questionnaire (ERI), the Generalized Anxiety Disorder-7 (GAD-7), the Patient Health Questionnaire-9 (PHQ-9), and the Athens Insomnia Scale (AIS). The bnlearn and gmodels packages of R4.2.1 software were used to construct Bayesian networks on work and personal factors-occupational stress- health effects. The Bayesian network model was visualized by Netica32.0. Results Among the 3018 questionnaires distributed, a total of 2675 valid questionnaires were recovered, with an effective recovery rate of 88.63%. The positive rates of occupational stress, anxiety, depression, and insomnia were 51.48%, 62.13%, 62.50%, and 56.37%, respectively in the grassroots medical and health personnel. The positive rate of occupational stress among the medical and health workers varied by age, educational level, and organization type (P<0.05); the positive rate of anxiety varied by age, marital status, and daily working hours (P<0.05); and the positive rates of depression and insomnia varied by gender, educational level, and overcommitment (P<0.05). The final Bayesian network contained 14 nodes and 18 directed edges, and its prediction accuracy was 85.4%. .Job title, daily working hours, overcommitment and exercise directly associated with occupational stress, and other work and personal factors associated with occupational stress indirectly. Occupational stress associated with insomnia directly or indirectly via anxiety. Anxiety and insomnia associated with reporting depression. Conclusion Grassroots medical and health personnel of Xiong'an New Area, Hebei Province report high levels of occupational stress, anxiety, depression, and insomnia. Occupational stress directly or indirectly associates with anxiety, depression, and insomnia. Work and personal factors associate with occupational stress, and associate with anxiety, depression, and insomnia via occupational stress. Occupational stress may be an early warning factor of general work and personal factors affecting anxiety, depression, insomnia, and other mental health problems. Attention should be paid to the assessment and intervention of occupational stress in medical and health personnel to prevent mental health problems.

Objective:To analyze the genetic characteristics of the complete genome of a strain of human respiratory syncytial virus (HRSV) in Qinghai province in 2024.Methods:A total of 300 samples were collected during 2024 influenza surveillance in Qinghai province sentinel hospitals from patients with fever accompanied by severe respiratory infection symptoms. We used real-time fluorescent quantitative reverse transcription polymerase chain reaction RT-PCR) method to screen out HRSV subtype B (HRSVB) positive specimens, whole genome sequencing was performed on positivespecimens meeting the requirements for the sequencing. After downloading the global representative HRSVB genotypes at GenBank database, sequence alignment was performed, related evolutionary tree was built and the calculation and analyses of genetic distance were done, analyses of HRSVB sequencing of sequence homology of nucleotides, amino acids and amino acid mutation were performed.Results:The first strain in Qinghai, China/qinghai/2024-03 had a complete sequence of 15 140 bp nucleotides, with HRSV′s all structural characteristics, and subtype HRSVA prototype strain Long strains of nucleotide the lowest homology was 80.0%, and subtype HRSVB prototype strain nucleotide homology was above 94.7%. The result indicated that the first strain in Qinghai belonged to HRSVB subtype. Genetic evolution shows China/qinghai/2024-03 and USA/WA-S23450/2021 (OR326803.1) and Germany/2021 (OR795235.1) all belong to a branch, they have the closest relationship. Phylogenetic analysis of G gene showed that the strain belonged to BA9 genotype of HRSVB subtype, and the hypervariable regions of the genome were SH and G genes.Conclusions:In this study, the complete genome sequence of HRSV China/qinghai/2024-03 was obtained for the first time, and the basic molecular structural characteristics were elucidated, which filled the gaps in the gene and amino acid data of HRSV in our province, and also provided a basis for HRSV epidemiology.

Metabolic syndrome is a complex group of metabolic disorders with an increasing global incidence rate,posing a serious threat to human health.Sodium-glucose linked transporter 2(SGLT2)inhibitors are a new type of oral hypoglycemic drug.SGLT2 inhibitors not only lower blood glucose level in a non-insulin-dependent manner by inhibiting glucose reabsorption by renal proximal convoluted tubular epithelial cell to promote urinary glucose excretion,but also by improving islet β cell function,reducing inflammatory responses,and inhibiting oxidative stress.In addition,SGLT2 inhibitors can reduce body weight through osmotic diuresis and increase fat metabolism;reduce blood pressure by inhibiting excessive activation of sympathetic nervous system and by improving vascular function.They can also improve blood lipids by increasing degradation of triacylglycerol;reduce blood uric acid by promoting uric acid excretion in kidney and intestine,and by reducing uric acid synthesis.This article reviews the effects and mechanisms of SGLT2 inhibitors on multiple metabolic disorders in metabolic syndrome and explores their potential application in metabolic syndrome treatment.

Single-cell transcriptome sequencing (scRNA-seq) can resolve the expression characteristics of cells in tissues with single-cell precision, enabling researchers to quantify cellular heterogeneity within populations with higher resolution, revealing potentially heterogeneous cell populations and the dynamics of complex tissues. However, the presence of a large number of technical zeros in scRNA-seq data will have an impact on downstream analysis of cell clustering, differential genes, cell annotation, and pseudotime, hindering the discovery of meaningful biological signals. The main idea to solve this problem is to make use of the potential correlation between cells and genes, and to impute the technical zeros through the observed data. Based on this, this paper reviewed the basic methods of imputing technical zeros in the scRNA-seq data and discussed the advantages and disadvantages of the existing methods. Finally, recommendations and perspectives on the use and development of the method were provided.
Cluster Analysis ; Transcriptome

Background Grassroots center for disease control and prevention (CDC) staff undertake intensive work of disease prevention and control, and may be susceptible to occupational stress, anxiety, depression, and other health problems. Objective To understand the current situation of occupational stress, anxiety, and depression among grassroots CDC staff, and to identify potential risk factor configurations for occupational stress, anxiety, and depression using fuzzy set qualitative comparative analysis (fsQCA), so as to provide a basis for effective intervention. Methods The staff working in county/district-level CDCs in Hebei Province were the target population of the current study. Stratified random cluster sampling method was used to select 1860 staff members of the target population. A questionnaire of general situation, Job Content Scale, 7-item Generalized Anxiety Disorder Scale, and Patient Health Questionnaire-9 were used. Risk factor configurations associated with health outcomes of interest were identified by fsQCA3.0 software. Results The positive rates of occupational stress, anxiety, and depression were 42.69%, 44.25%, and 47.96%, respectively. Marital status was a necessary condition for occupational stress, anxiety, and depression in the grassroots CDC staff (the necessity values were 0.911, 0.939, and 0.933, respectively). There were two types of risk factor configurations for occupational stress: "self-improvement" and "disease burden"; the risk factor configurations for anxiety were "disease burden" and "economic-disease burden"; while the risk factor configurations for depression were "disease burden", "economic-disease burden", and "self-improvement". The overall consistency scores of occupational stress, anxiety, and depression were 0.941, 0.820, and 0.774, respectively. Regarding outstanding components, "self-improvement" included pressure of job requirements and promotion, "disease burden" included impact of chronic illness on psychological state, and "economic-disease burden" included not only impact of chronic illness but also financial support for CDC staff. Conclusion All positive rates of occupational stress, anxiety, and depression are high among grassroots CDC staff in Hebei Province. Occupational stress, anxiety, and depression of grassroots CDC staff are the results of multiple influencing factors, so targeted intervention measures should be formulated.

Background Since the outbreak of the COVID-19 epidemic, staff of the centers for disease control and prevention (CDC) have been burdened with heavy epidemic prevention control, and excessive occupational stress can cause depression and other psychological problems. Objective To explore the status of occupational stress, resilience, and depression of CDC staff and potential relationships between them. Methods From December 2020 to April 2021, a survey was conducted at provincial and municipal levels, and the stratified cluster sampling method was used at county (district) level to select a total of 3514 samples. Their occupational stress, resilience, and depression status were evaluated using the Chinese Effort-Reward Imbalance Scale (ERI), the Chinese Connor-Davidson Resilience Scale (CD-RISC), and the Patient Health Questionnaire (PHQ-9). Independent sample t test, analysis of variance, χ² test, Pearson correlation analysis, and mediation test (structural equation model) were conducted. Results The positive rate of occupational stress was 34.29% in the CDC staff, the resilience score was 66.28±15.32, and the positive rate of depression was 48.58%. Significant differences were found in the positive rates of occupational stress among different groups of gender, age, education background, marital status, administrative duty, weekly exercise frequency, chronic disease prevalence, and participation in epidemic control (P<0.05); in the resilience scores among different groups of gender, age, administrative duty, weekly exercise frequency, chronic disease prevalence, and participation in epidemic control (P<0.05); in the positive rates of depression among different groups of gender, age, educational background, personal monthly income, weekly exercise frequency, chronic disease prevalence, and participation in epidemic control (P<0.05). Occupational stress was negatively correlated with resilience (r=−0.165, P<0.01). Resilience was negatively correlated with depression (r=−0.383, P<0.01). Occupational stress was positively correlated with depression (r=0.343, P<0.01). The structural equation embracing a partial mediating effect of resilience on the relationship occupational stress and depression was established, and the partial mediating effect was 0.039, accounting for 10.46% of the total effect. Conclusion High positive rates of occupational stress, reduced resilience, and depression are shown among CDC staff in the context of the COVID-19 epidemic, and resilience partially mediates the effect of occupational stress on depression. The study findings suggest that improving resilience may reduce occupational stress and depression in CDC staff.

Objective:To examine the influencing factors of endothelial injury using the structural equation model (SEM).Methods:A total of 6 861 asymptomatic individuals free of cardiovascular disease underwent health examinations at the health management center of the third Xiangya hospital, Central South University from May 2015 to August 2020. And collected their questionnaire and checkup data. Spearman′s rank correlation coefficient was used to analyze metabolic factors and brachial artery flow-mediated dilatation (FMD). Exploratory factor analysis (3 430/6 861) and confirmatory factor analysis (3 431/6 861) were conducted on the diet items. An SEM was constructed using the diet pattern data, cardiovascular risk factors and FMD, and using multi-path regression analysis to determine the correlation between the indicators.Results:Based on the factor analysis, diet items were divided into three patterns: healthy food, meat, and supplementary food. The SEM indicated that age ( β=0.27) and blood pressure ( β=0.12) had obvious effects on low FMD. Triglyceride ( β=0.03), fasting blood glucose ( β=0.04), and body mass index ( β=0.08) were positively correlated with low FMD. On the upstream, healthy food was negatively correlated with blood pressure ( β=-0.04) and body mass index ( β=-0.04), meat was positively correlated with triglyceride ( β=0.33), blood pressure ( β=0.06), fasting blood glucose ( β=0.20), and body mass index ( β=0.16), and supplementary food was negatively correlated with fasting blood glucose ( β=-0.30). This was the only pattern that was directly correlated with FMD ( β=0.05). Conclusions:SEM is an effective method to analyze the influence of various risk factors on the population and the relationship between individual indicators. This study revealed direct and indirect correlations between age, diet pattern, cardiovascular-metabolic risk, and FMD impairment. Comprehensive control of dietary patterns and metabolic indicators could prevent and improve early cardiovascular injury.

Objective:To understand the prevalence and genotype characteristics of enterovirus and Coxsackie virus in hand, foot and mouth disease (HFMD) patients in Qinghai province from 2015 to 2018.Methods:The throat swabs of HFMD patients were collected for virus isolation and RT-PCR in Qinghai province. The nucleotide sequence of the amplified products was determined and analyzed, and the gene evolution tree was constructed with reference to the sequence of some strains of NCBI.Results:From 2015 to 2018, 1 738 samples of clinical diagnosis positive cases were collected, including 326 EV-A71 cases, accounting for 18.76%, 237 CV-A16 cases, accounting for 13.64%, 628 CV-A6 cases, accounting for 36.13%. There were statistically significant differences among different genotypes in 4 years (EV-A71, χ2=245.315, P<0.001; CV-A16, χ2=27.680, P<0.001; CV-A6, χ2=702.713, P<0.001). A total of 317 cell culture isolates were obtained after isolation with RD cells. After sequence, DNA sequences of typical genotypes were selected for sequence analysis, from 2017 to 2018. The homology of all genotypes of VP1 was between 59% and 100%, the homology was 93% to 100% in 19 strains of EV-A71, the homology was 92% to 100% in 20 strains of CV-A16, the homology was 97% to 99% in 6 strains of CV-A6. According to the evolutionary tree, the sequences of EV-A71 strains are all on the C4a branch of the evolutionary tree in Qinghai province; 18 strains of CV-A16 were B1b and 2 stains were 1D; 5 strains of CV-A6 were D3 and 1 was B1. Conclusions:From 2015 to 2018, the prevalent genotypes of HFMD shifted from EV-A71 to CV-A16 and CV-A6 in Qinghai province, The EV-A71 genotype in Qinghai province has always been C4a genotype. There are different genotypes of CV-A16 and CV-A6. The nucleotide sequence differences between different genotypes are large, and the sequence variation among the same gene is small.

Objective:To analyze the epidemiological characteristics of rubella and the genetic characteristics of rubella virus (RV) circulated in Qinghai province in 2020, so as to provide scientific basis for optimization and improvement of local rubella prevention and control strategy.Methods:The rubella epidemiological characteristics were analyzed by summarizing the data on the rubella incidence in Qinghai province in 2020 from the National Notifiable Disease Reporting System. Through Qinghai provincial measles and rubella laboratory network, throat swab samples from susceptible rubella outbreak and sporadic cases in 2020 were collected and identified. RV strains were obtained after three passages of virus isolation from positive samples. After extracting the viral RNA, the 739 nucleotide fragments within the E1 gene were amplified and determined to identify the genotype and sub-genotype of the Qinghai strains in 2020 and further analyzed the molecular differences between Qinghai strains and the RV strains circulated in China.Results:In 2020, the rubella incidence in Qinghai province had shown an obvious upward trend, and the age of onset had shifted to adolescents in the 10-19 years of age group (accounting for 94.9%). Totally 29 RV strains were isolated from four high incidence areas of rubella in Qinghai province. All RV strains were identified as sub-genotype 2B-L2c, which is also the dominant subtype of RV circulated in China. In addition, virological surveillance data showed that there were different transmission chains of sub-genotype 2B-L2c in Qinghai province in 2020, and an outbreak might be caused by different transmission chain viruses.Conclusions:The accumulation of rubella susceptible population aged 10-19 years and the transmission of new imported 2B-L2c virus had led to the rubella reemergence and outbreaks in several cities in Qinghai province in 2020.