Background Frailty serves as a significant precursor to falls, disability, and mortality. Epidemiological evidence examining the impact of working hours on frailty remains scarce. Objective To explore the prevalence of frailty and the relationship between frailty and working hours among full-time middle-aged and older workers in China. Methods Data were derived from the 2020 China Health and Retirement Longitudinal Study (CHARLS). The study included full-time workers aged 45 years and above with a weekly working duration exceeding 35 h. Frailty was assessed using the Frailty Index (FI). First, the dose-response relationship between working hours and FI was explored using a generalized additive model (GAM). Second, univariate analyses were performed using t-tests, χ² tests, and the Jonckheere–Terpstra trend test. Restricted cubic splines (RCS) were introduced for modeling. Based on ordinal logistic regression models, covariates were adjusted sequentially to examine whether differences in frailty prevalence existed across different weekly working hour categories. Finally, subgroup analyses were performed. Results Among the 5504 included participants, the mean age was (56.41±7.30) years, with 2304 being female (41.86%). A total of 807 participants (14.66%) were identified as frail, and 2 686 participants (48.80%) were pre-frail. After standardization for gender and age according to the seventh national census data, the standardized prevalence of frailty and prefrailty were 18.89% and 47.25%, respectively. Significant higher proportions of frail individuals were found among those in non-managerial (73.73%), agricultural (64.06%), and non-employed (65.68%) occupations. Long working hours were prevalent: 2596 (47.17%) workers worked 41–60 h per week, 1649 (29.96%) worked >60 h per week, and only 1259 (22.87%) worked 35–40 h per week. GAM analysis showed a non-linear relationship between weekly working hours and FI. Restricted cubic spline analysis showed a statistically significant association between weekly working hours and frailty status (P=0.022). Logistic regression analysis revealed that individuals working more than 60 h per week had a 23% higher odds of frailty compared to those working 41–60 h (OR=1.23, 95%CI: 1.09, 1.38, P<0.01), while those working 35–40 h per week showed a 15% higher odds of frailty (OR=1.15, 95%CI: 1.01, 1.31, P=0.034). The subgroup analysis revealed that the positive association between working more than 60 h per week and frailty remained generally consistent across sex, age, urban/rural residence, occupational attributes, industry, or employment type, although the association did not reach statistical significance among non-rural residents and individuals aged 60 years and older. Conclusion The prevalence of frailty among full-time middle-aged and older labor force in China is relatively high, and the odds of frailty increases significantly among those who work more than 60 h per week. It is recommended to implement frailty prevention and management strategies targeting long-working-hour workers and to prioritize the impact of working duration on frailty.

Objective:To identify strain-specific features of Klebsiella pneumoniae by analyzing metagenomics next generation sequencing (mNGS) data, thereby expanding the downstream applications of mNGS. Methods:The sequences of K.pneumoniae strains were organized from both the self-built database of the long-term multi-center research cohort in China established by the Peking University People′s Hospital from 2009 to 2020 (with 2 345 sequences) and the public databases (with 19 648 sequences). The existing large-scale databases were compressed, and a set of strains representative of clonal groups were screened. A strain genome information library was constructed based on k-mer features, and the most matching representative sequences in the database were searched for the raw mNGS data. The search results of the self-built library and public library were merged and optimized to update the prediction of antimicrobial-resistance characteristics and avoid the impact of uneven data distribution on the results. A total of 314 clinical samples from patients with K.pneumoniae detected by mNGS in the Clinical Microbiology Laboratory of Peking University People′s Hospital from 2022 to 2024 were retrospectively collected, and 101 samples with positive clinical culture results were selected to validate the prediction results. The antimicrobial-resistance phenotypes were verified by clinical antimicrobial susceptibility test results. Whole-genome sequencing was performed on the culture strains of 14 samples randomly selected using random numbers to verify the genotypes. Single nucleotide polymorphism distance analysis was used to verify the occurrence of outbreak events. The χ2 test and Mann-Whitney U test were used for statistical analysis. Results:A representative strain sequence k-mer feature library containing self-built and public sub-libraries was constructed. The library construction required only about 1 hour with <3 GB storage, with a high compression ratio and low update cost. Using k-mer-based analysis, mNGS data achieved precise strain characterization within 4 minutes and and <5 GB memory occupation. There was a significant difference in the antimicrobial-resistance rates to more than half of the antibiotics between the self-built database (90.8%, 2 130/2 345) and the public database (22.7%, 4 457/19 648) ( χ2=4 634.1, P<0.001). After optimizing the search results, the mean category agreement, sensitivity, and specificity of the prediction for eight antibiotics reached 84.8% (323/381), 78.9% (131/166), and 91.2% (196/215), respectively. The target genotypes were successfully detected in 10 out of 12 samples, and two outbreak events (2 samples per event) were successfully identified. Conclusions:An independent analysis process adapted to the needs of identifying the features of K. pneumoniae strains in mNGS data was developed. This process requires minimal computational resources and processing time and can directly achieve the simultaneous analysis of the antimicrobial-resistance phenotypes of K. pneumoniae at the strain level and their corresponding genomic characteristic profiles based on the raw mNGS reads.

BACKGROUND: The main cause of restenosis after percutaneous transluminal angioplasty (PTA) is the excessive proliferation and migration of vascular smooth muscle cells (VSMCs). Lin28a has been reported to play critical regulatory roles in this process. However, whether CCAAT/enhancer-binding proteins β (C/EBPβ) binds to the Lin28a promoter and drives the progression of restenosis has not been clarified. Therefore, in the present study, we aim to clarify the role of C/EBPβ-Lin28a axis in restenosis. METHODS: Restenosis and atherosclerosis rat models of type 2 diabetes ( n   =  20, for each group) were established by subjecting to PTA. Subsequently, the difference in DNA methylation status and expression of C/EBPβ between the two groups were assessed. EdU, Transwell, and rescue assays were performed to assess the effect of C/EBPβ on the proliferation and migration of VSMCs. DNA methylation status was further assessed using Methyltarget sequencing. The interaction between Lin28a and ten-eleven translocation 1 (TET1) was analysed using co-immunoprecipitation (Co-IP) assay. Student's t -test and one-way analysis of variance were used for statistical analysis. RESULTS: C/EBPβ expression was upregulated and accompanied by hypomethylation of its promoter in restenosis when compared with atherosclerosis. In vitroC/EBPβ overexpression facilitated the proliferation and migration of VSMCs and was associated with increased Lin28a expression. Conversely, C/EBPβ knockdown resulted in the opposite effects. Chromatin immunoprecipitation assays further demonstrated that C/EBPβ could directly bind to Lin28a promoter. Increased C/EBPβ expression and enhanced proliferation and migration of VSMCs were observed after decitabine treatment. Further, mechanical stretch promoted C/EBPβ and Lin28a expression accompanied by C/EBPβ hypomethylation. Additionally, Lin28a overexpression reduced C/EBPβ methylation via recruiting TET1 and enhanced C/EBPβ-mediated proliferation and migration of VSMCs. The opposite was noted in Lin28a knockdown cells. CONCLUSION Our findings suggest that the C/EBPβ-Lin28a axis is a driver of restenosis progression, and presents a promising therapeutic target for restenosis.
Animals ; Cell Proliferation/genetics* ; Cell Movement/genetics* ; Muscle, Smooth, Vascular/metabolism* ; Rats ; DNA Methylation/physiology* ; CCAAT-Enhancer-Binding Protein-beta/genetics* ; Male ; Myocytes, Smooth Muscle/cytology* ; Rats, Sprague-Dawley ; RNA-Binding Proteins/genetics* ; Cells, Cultured ; Coronary Restenosis/metabolism*

BACKGROUND: Valvular heart disease (VHD) has become increasingly common with the aging in China. This study aimed to evaluate regional differences in the clinical features, management strategies, and outcomes of patients with VHD across different regions in China. METHODS: Data were collected from the China-VHD Study. From April 2018 to June 2018, 12,347 patients who presented with moderate or severe native VHD with a median of 2 years of follow-up from 46 centers at certified tertiary hospitals across 31 provinces, autonomous regions, and municipalities in Chinese mainland were included in this study. According to the locations of the research centers, patients were divided into five regional groups: eastern, southern, western, northern, and central China. The clinical features of VHD patients were compared among the five geographical regions. The primary outcome was all-cause mortality or rehospitalization for heart failure. Kaplan-Meier survival analysis was used to compare the cumulative incidence rate. RESULTS: Among the enrolled patients (mean age, 61.96 years; 6877 [55.70%] male), multiple VHD was the most frequent type (4042, 32.74%), which was mainly found in eastern China, followed by isolated mitral regurgitation (3044, 24.65%), which was mainly found in northern China. The etiology of VHD varied significantly across different regions of China. The overall rate of valve interventions was 32.67% (4008/12,268), with the highest rate in southern China at 48.46% (205/423). In terms of procedure, the proportion of transcatheter valve intervention was relatively low compared to that of surgical treatment. Patients with VHD in western China had the highest incidence of all-cause mortality or rehospitalization for heart failure. Valve intervention significantly improved the outcome of patients with VHD in all five regions (all P  <0.05). CONCLUSIONS: This study revealed that patients with VHD in China are characterized by significant geographic disparities in clinical features, treatment, and clinical outcomes. Targeted efforts are needed to improve the management and prognosis of patients with VHD in China according to differences in geographical characteristics. REGISTRATION ClinicalTrials.gov , NCT03484806.
Aged ; Female ; Humans ; Male ; Middle Aged ; China/epidemiology* ; Heart Valve Diseases/therapy* ; Kaplan-Meier Estimate ; Tertiary Care Centers ; Treatment Outcome

Distinct from the complementary inhibition mechanism through binding to the target with three-dimensional conformation of small molecule inhibitors, targeted protein degradation technology takes tremendous advantage of endogenous protein degradation pathway inside cells to degrade plenty of “undruggable” target proteins, which provides a novel route for the treatment of many serious diseases, mainly including proteolysis-targeting chimeras, lysosome-targeting chimeras, autophagy-targeting chimeras, antibody-based proteolysis-targeting chimeras, etc. Unlike proteolysis-targeting chimeras first found in 2001, which rely on ubiquitin-proteasome system to mainly degrade intracellular proteins of interest, lysosome-targeting chimeras identified in 2020, which was act as the fastly developing technology, utilize cellular lysosomal pathway through endocytosis mediated by lysosome-targeting receptor to degrade both extracellular and membrane proteins. As an emerging biomedical technology, nucleic acid-driven lysosome-targeting chimeras utilize nucleic acids as certain components of chimera molecule to replace with ligand to lysosome-targeting receptor or protein of interest, exhibiting broad application prospects and potential clinical value in disease treatment and drug development. This review mainly introduced present progress of nucleic acid-driven lysosome-targeting chimeras technology, including its basic composition, its advantages compared with antibody or glycopeptide-based lysosome-targeting chimeras, and focused on its chief application, in terms of the type of lysosome-targeting receptors. Most research about the development of nucleic acid-driven lysosome-targeting chimeras focused on those which utilized cation-independent mannose-6-phosphonate receptor as the lysosome-targeting receptor. Both mannose-6-phosphonate-modified glycopeptide and nucleic aptamer targeting cation-independent mannose-6-phosphonate receptor, even double-stranded DNA molecule moiety can be taken advantage as the ligand to lysosome-targeting receptor. The same as classical lysosome-targeting chimeras, asialoglycoprotein receptor can also be used for advance of nucleic acid-driven lysosome-targeting chimeras. Another new-found lysosome-targeting receptor, scavenger receptor, can bind dendritic DNA molecules to mediate cellular internalization of complex and lysosomal degradation of target protein, suggesting the successful application of scavenger receptor-mediated nucleic acid-driven lysosome-targeting chimeras. In addition, this review briefly overviewed the history of lysosome-targeting chimeras, including first-generation and second-generation lysosome-targeting chimeras through cation-independent mannose-6-phosphonate receptor-mediated and asialoglycoprotein receptor-mediated endocytosis respectively, so that a clear timeline can be presented for the advance of chimera technique. Meantime, current deficiency and challenge of lysosome-targeting chimeras was also mentioned to give some direction for deep progress of lysosome-targeting chimeras. Finally, according to faulty lysosomal degradation efficiency, more cellular mechanism where lysosome-targeting chimeras perform degradation of protein of interest need to be deeply explored. In view of current progress and direction of nucleic acid-driven lysosome-targeting chimeras, we discussed its current challenges and development direction in the future. Stability of natural nucleic acid molecule and optimized chimera construction have a great influence on the biological function of lysosome-targeting chimeras. Discovery of novel lysosome-targeting receptors and nucleic aptamer with higher affinity to the target will greatly facilitate profound advance of chimera technique. In summary, nucleic acid-driven lysosome-targeting chimeras have many superiorities, such as lower immunogenicity, expedient synthesis of chimera molecules and so on, in contrast to classical lysosome-targeting chimeras, making it more valuable. Also, the chimera technology provides new ideas and methods for biomedical research, drug development and clinical treatment, and can be used more widely through further research and optimization.

To explore the interaction between ASFV capsid protein M1249L and host from the host cellular perspective,M1249L was selected for constructing the bait plasmid(pGBKT7-M1249L)to screen the bone marrow-derived macrophages(BMDMs)cDNA library.After again co-transform and sequence alignment,20 candidate interacting host proteins were screened,such as IL-1β,CTSB and DNAJA3.And then,co-immunoprecipitation assay was performed to verify the interaction be-tween M1249L and host proteins.GO ontology(GO)and KEGG pathway enrichment analyses re-vealed that biological regulation,cellular communication and response to stimulus and others were enriched in biological processes.And these host proteins could share some pathways,including toll-like receptor signaling pathway and Nod-like receptor signaling pathway.Therefore,the results provides the theoretical basis for further research on the mechanism of ASFV M1249L in viral in-fection and immune regulation.

Objective To evaluate the value of risk score model based on clinical and multimodal ultrasound features for differentiating benign and malignant breast imaging reporting and data system(BI-RADS)4A lesions in breast.Methods Totally 177 patients with BI-RADS 4A lesions detected by multimodal breast ultrasound were prospectively enrolled,and the patients were divided into training set(n=123)and test set(n=54)at the ratio of 7∶3.Univariate and multivariate logistic regression analyses were used to analyze patients'clinical,gray-scale ultrasound,CDFI and elastography ultrasound parameters of lesions,and the independent risk factors for differentiating benign and malignant BI-RADS 4A lesions in breast were screened,so as a risk score model was constructed.Based on the results of sungical pathology or follow-up,the cut-off value of the model for differentiating benign and malignant BI-RADS 4A lesions in breast was obtained by receiver operating characteristic(ROC)curve,and the patients were divided into high-risk and low-risk subgroups according to the cut-off value,and the efficacy of the model was evaluated.Results Among 177 cases,39 were with benign lesions and 138 were with malignant lesions.Patients'age＞58 years,diameter of lesion＞15.1 mm,lesions with irregular shape,lesions with grade 1 or 2 of blood flow and standard deviation of sound touch elastography of a 2 mm circular area around the lesion(shell)(shell-STESD)＞16.33 kPa were all independent risk factors of malignant BI-RADS 4A lesions in breast.The cut-off value of risk score model for differential diagnosis was 6.5 points,and its sensitivity was 84.69%(83/98)and specificity was 88.00%(22/25)in distinguishing high-risk and low-risk subgroups in training set,while its sensitivity was 77.50%(31/40)and specificity was 71.43%(10/14)in test set.Conclusion Risk score model based on clinical and multimodal ultrasound features could effectively differentiate benign and malignant BI-RADS 4A lesions in breast.

With the increasing use of nirmatrelvir-ritonavir in older COVID-19 patients, adverse drug reactions due to drug-drug interactions have become more frequent. This report describes a patient who experienced severe bradycardia and hypotension following the concurrent use of heart rate-control medications and nirmatrelvir-ritonavir during an active COVID-19 infection. This suggests that clinicians should be cautious when dealing with nirmatrelvir-ritonavir and try their best to avoid serious adverse reactions.

OBJECTIVE To establish skin contamination models with sulfur mustard analogue 1-chloro-2-ethylsulfanyl ethane(CEES),and evaluate the therapeutic efficacy of reactive skin decontamination lotion kit(RSDL).METHODS ①Kunming,BALB/c,BALB/c-nu,and C57BL/6N were contaminted with CEES 75 μL·kg-1 using exposed method and covered method for 10 min on the dorsal skin.Wound healing times and areas were assessed to determine the stock of mice and exposure method.② BALB/c mice were exposed to a gradient of CEES at the doses of 75,150,250,350 and 500 μL·kg-1(10 min)using the covered method.Wound healing times,areas and Kaplan-Meier survival curves were used to determine the optimal dose.③ BALB/c mice were exposed to CEES 150 μL·kg-1 for varying durations(5,10 and 20 min)using the covered method.Wound healing times,areas and Kaplan-Meier survival curves were analyzed to determine the optimal exposure time.④ BALB/c mice were divided into four groups:control(exposed with the covered method at 150 μL·kg-1,10 min),treatment(20 μL RSDL treatment after exposure,as the control group),5 min treatment(20 μL RSDL treatment after 5 min of exposure,followed by 5 min of coverage)and immediate-treatment(exposed with the exposed method at 150 μL·kg-1,immediately treated with 20 μL RSDL,followed by 10 min of coverage).The therapeutic efficacy was evaluated based on the wound area,subcutaneous microvesicle count,epidermal thick-ness,and inflammatory cytokine(IL-6 and TNF-α)expressions.RESULTS ① The covered method caused more severe and prolonged wounds than the exposed method.BALB/c mice exhibited a high sensitivity to CEES with delayed wound healing and were therefore selected as the model animal.② Survival rates in BALB/c mice dropped below 50%at doses of 250,350 and 500 μL·kg-1,whereas an 83.3%survival rate was observed at 150 μL·kg-1.③ The mice exposed to CEES(150 μL·kg-1,20 min)died within 3 days.The wound area was consistently smaller in the 5 min covered group than in the 10 min covered group.④CEES-induced skin injury led to epidermal nuclear pyknosis,follicular disrup-tion,inflammatory infiltration,and microvesicle formation in both treatment and poisoned control groups.As more immediate treatment,the wound area significantly decreased.While the IL-6 expres-sion showed no significant intergroup difference,the TNF-α expression was significantly higher in the treatment group.CONCLUSION A CEES-induced skin contamination model has been established in BALB/c mice using the covered method(150 μL·kg-1,10 min covered).However,RSDL should be ad-ministered in 10 min post-contamination.

Objective To analyze the relationship between the visceral adiposity index(VAI)and nocturia in the US adult population.Methods A cross-sectional study was performed.Data from subjects aged≥20 years in the National Health and Nutrition Examination Survey(NHANES)database from 2007 to 2020 were collected,including waist circumference,triglyceride,body mass index(BMI),high-density lipoprotein,age,gender,race,poverty income ratio,education level,marital status,smoking,alcohol consumption,sleep disorders,depression,occupation,hypertension,diabetes,congestive heart failure,cancer,and nocturnal urination frequency.Weighted analysis,multivariate logistic regression,generalized additive model(GAM),and curve fitting were employed to evaluate the association between VAI and nocturia,adjusting for age,gender,race,poverty income ratio,education level,marital status,smoking,alcohol consumption,sleep disorders,depression,occupation,hypertension,diabetes,congestive heart failure,and cancer.Subgroup analyses were conducted based on age,gender,race,hypertension and diabetes to further evaluate the relationship between VAI and the risk of nocturia.Results A total of 29,196 American adults were included.All subjects were divided into 4 groups based on VAI quartiles:Q1 group(0.32≤VAI<1.01),Q2 group(1.01≤VAI<1.70),Q3 group(1.70≤VAI<2.95),and Q4 group(2.95≤VAI<13.59),with nocturia prevalence rates of 28.5%,31.4%,33.3%,and 34.9%,respectively.In subgroup analyses,the risk of nocturia significantly increased with higher VAI in the 20-40 age group,females and other Hispanics(OR=1.04,95%CI 1.01-1.08,P=0.006;OR=1.02,95%CI 1.00-1.04,P=0.035;OR=1.05,95%CI 1.01-1.09,P=0.026).GAM analysis results showed a nonlinear relationship between VAI and nocturia.Conclusion VAI is positively associated with the risk of nocturia,and may be an effective indicator for predicting the risk of nocturia occurrence.