The circadian clock plays a critical role in regulating various physiological processes, including gene expression, metabolic regulation, immune response, and the sleep-wake cycle in living organisms. Post-translational modifications (PTMs) are crucial regulatory mechanisms to maintain the precise oscillation of the circadian clock. By modulating the stability, activity, cell localization and protein-protein interactions of core clock proteins, PTMs enable these proteins to respond dynamically to environmental and intracellular changes, thereby sustaining the periodic oscillations of the circadian clock. Different types of PTMs exert their effects through distincting molecular mechanisms, collectively ensuring the proper function of the circadian system. This review systematically summarized several major types of PTMs, including phosphorylation, acetylation, ubiquitination, SUMOylation and oxidative modification, and overviewed their roles in regulating the core clock proteins and the associated pathways, with the goals of providing a theoretical foundation for the deeper understanding of clock mechanisms and the treatment of diseases associated with circadian disruption.
Protein Processing, Post-Translational/physiology* ; Circadian Rhythm/physiology* ; Humans ; Animals ; CLOCK Proteins/physiology* ; Circadian Clocks/physiology* ; Phosphorylation ; Acetylation ; Ubiquitination ; Sumoylation

Researchers commonly use cyclization recombination enzyme/locus of X-over P1 (Cre/loxP) technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis. However, the shortcomings of these genetic tools include high costs, lengthy experimental periods, and limited accessibility for researchers. Therefore, exploring alternative gene silencing techniques is of great practical value. In this study, we employed adeno-associated virus (AAV) as a vector for gene silencing in Sertoli and Leydig cells. Our findings demonstrated that AAV serotypes 1, 8, and 9 exhibited high infection efficiency in both types of testis cells. Importantly, we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection. We achieved cell-specific knockouts of the steroidogenic acute regulatory ( Star ) and luteinizing hormone/human chorionic gonadotropin receptor (Lhcgr) genes in Leydig cells, but not in Sertoli cells, using AAV9-single guide RNA (sgRNA)-mediated gene editing in Rosa26-LSL-Cas9 mice. Knockdown of androgen receptor ( Ar ) gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpin RNA (shRNA)-mediated targeting. Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.
Animals ; Male ; Leydig Cells/metabolism* ; Mice ; Dependovirus/genetics* ; Sertoli Cells/metabolism* ; Gene Silencing ; Genetic Vectors ; Testis/cytology*

BACKGROUND: Mild cognitive impairment (MCI) is common in atrial fibrillation (AF) patients and may develop earlier in those with multiple cardiovascular comorbidities, potentially impairing self-management and treatment adherence. This study aimed to characterize the prevalence and profile of MCI in AF patients, examine its associations with cardiovascular comorbidities, and assess how these comorbidities influence specific cognitive domains. METHODS: This cross-sectional study analyzed data from AF patients who underwent cognitive assessment between 2017 and 2021. Cognitive status was categorized as MCI or non-MCI based on the Montreal Cognitive Assessment. Associations between comorbidities and MCI were assessed by logistic regression, and cognitive domains were compared using the Mann-Whitney U test. RESULTS: Of 4136 AF patients (mean age: 64.7 ± 9.4 years, 64.7% male), 33.5% of patients had MCI. Among the AF patients, 31.2% of patients had coronary artery disease, 20.1% of patients had heart failure, and 18.1% of patients had hypertension. 88.7% of patients had left atrial enlargement, and 11.0% of patients had reduced left ventricular ejection fraction. Independent factors associated with higher MCI prevalence included older age (OR = 1.04, 95% CI: 1.03-1.05, P < 0.001), lower education level (OR = 1.51, 95% CI: 1.31-1.73, P < 0.001), hypertension (OR = 1.28, 95% CI: 1.07-1.52, P = 0.001), heart failure (OR = 1.24, 95% CI: 1.04-1.48, P = 0.020), and lower left ventricular ejection fraction (OR = 1.43, 95% CI: 1.04-1.98, P = 0.028). A higher CHA₂DS₂-VASc score (OR = 1.27, 95% CI: 1.22-1.33, P < 0.001; ≥ 2 points vs. < 2 points), and greater atherosclerotic cardiovascular disease burden (OR = 1.45, 95% CI: 1.02-2.08, P = 0.040; 2 types vs. 0 type) were linked to increased MCI risk. These above factors influenced various cognitive domains. CONCLUSIONS MCI is common in AF and closely associated with cardiovascular multimorbidity. Patients with multiple comorbidities are at higher risk, highlighting the importance of routine cognitive assessment to support self-management and integrated care.

Acute-on-chronic liver failure (ACLF) is an acute deterioration of liver function occurring on the basis of chronic liver disease, accompanied by failure of the liver and extrahepatic organs, and is associated with a high short-term mortality rate. Liver transplantation is the only curative treatment for patients with ACLF. However, the shortage of donor livers and limitations of the organ allocation system mean that only a minority of patients can receive transplants. The current organ allocation system based on the model for end-stage liver disease (MELD) score may underestimate the urgency of liver transplantation for ACLF patients. Therefore, it is urgent to develop better assessment tools to determine which ACLF patients are most likely to benefit from liver transplantation. This article reviews the current mainstream definitions of ACLF, the selection of candidates for liver transplantation in ACLF, and the prognostic scoring systems for liver transplantation in ACLF, both domestically and internationally, in order to provide a reference for the prognostic assessment of liver transplantation in ACLF patients.

Objective:To investigate the predictive value of serum signal lymphocyte activation molecule family member 8(SLAMF8)levels for post-stroke cognitive impairment(PSCI).Methods:The GSE122063 dataset was selected from the Gene Expression Omnibus(GEO), and key genes associated with vascular dementia were identified using STRING network and Cytoscape software.This prospective cohort study involved 123 patients with acute cerebral infarction(ACI)who were admitted to the Department of Neurology at Jiangsu University Affiliated Hospital from January to December 2023.Patients were followed up for six months and categorized into PSCI and post-stroke non-cognitive impairment(PSNCI)groups based on the occurrence of PSCI.General data from both groups at baseline, as well as the Mini-Mental State Examination(MMSE)and Montreal Cognitive Assessment Scale(MoCA)scores at the six-month follow-up, were collected.Baseline serum levels of SLAMF8 and stabilin-1(STAB1), along with serum levels of interleukin-1β(IL-1β)and interleukin-6(IL-6)at the six-month follow-up, were measured.Pearson correlation analysis was used to assess the correlation between variables, while logistic regression analysis was employed to determine baseline factors influencing the occurrence of PSCI.Receiver Operating Characteristic(ROC)curves were plotted to analyze the predictive value of variables for PSCI occurrence.Results:The Cytoscape software identified SLAMF8 and STAB1 as key genes associated with vascular dementia, utilizing maximum neighborhood component density(DNMC)and eccentricity algorithms on the GSE122063 dataset.In the cohort study, patients in the PSCI group exhibited higher baseline NIHSS scores and serum SLAMF8 levels compared to the PSNCI group( t=3.033, 5.422; P<0.01). Additionally, they demonstrated significantly lower MMSE and MoCA scores( t=16.340, 18.634; P<0.001)and higher serum levels of IL-1β and IL-6( t=2.633, 2.632; P<0.05)at the 6-month follow-up.No significant difference was observed in baseline STAB1 levels between the two groups( t=1.280, P>0.05). In the PSCI group, there was no significant correlation between baseline serum SLAMF8 levels and admission NIHSS scores( r=0.257, P=0.082); however, SLAMF8 showed a negative correlation with both MMSE scores( r=-0.711, P<0.001)and MoCA scores( r=-0.686, P<0.001)at the 6-month follow-up.Logistic regression analysis indicated that baseline serum SLAMF8 levels( OR=1.142, P=0.001)and NIHSS scores( OR=1.094, P=0.007)were risk factors for the development of PSCI in patients with acute cerebral infarction(ACI). ROC curve analysis revealed that the area under the ROC curve(AUC)for baseline serum SLAMF8 levels in predicting PSCI occurrence in ACI patients was 0.776, while the AUC for the combined prediction using both SLAMF8 and NIHSS scores was 0.796.Furthermore, baseline serum SLAMF8 levels were positively correlated with serum IL-1β levels( r=0.652, P<0.001)and IL-6 levels( r=0.710, P<0.001)at the 6-month follow-up. Conclusions:The serum SLAMF8 level, exhibiting early high expression in ACI patients, may serve as a potential biological marker for predicting the occurrence of PSCI.

Objective To determine the knee joint cartilage thickness using different methods and explore the feasibility of mathematical statistical models of dataset for the prediction of cartilage thickness.Methods A total of 304 patients diagnosed as knee osteoarthritis(OA)combined with varus deformity and undergoing unilateral total knee arthroplasty at the Second Affiliated Hospital of Army Medical University from March 2023 to March 2024 were selected for the study.All patients had complete preoperative and postoperative clinical data.The healthy cartilage at four anatomical sites of patients,including the distal femur lateral condyle,lateral tibial plateau,posterior medial femoral condyle,and posterior lateral femoral condyle were selected,and the knee joint cartilage thickness was determined based on preoperative MRI analysis,robotic navigation system tracing,tissue section of surgical specimen and digital vernier caliper.The baseline indicators of demographics,disease and imaging ffor patients were collected to construct a dataset,and four models of linear regression analysis,principal component analysis,Least Absolute Shrinkage and Selection Operator(LASSO)regression analysis,and K-nearest neighbors(KNN)analysis were established for predicting the accuracy,determination coefficient(R2)and root mean square error(RMSE),and the regression equation for predicting cartilage thickness was established.Results The knee joint cartilage thicknesses determined by preoperative MRI analysis,robotic navigation system tracing,tissue section of surgical specimen had no statistically significant difference with that by digital vernier caliper(P>0.05).The predictive efficiencies of models of linear regression analysis,principal component analysis,and LASSO regression analysis for the knee joint cartilage thickness all failed to meet the expectations(R2<0.3,RMSE>0.03).The predictive effect of KNN model on the cartilage thickness of the distal femur lateral condyle and lateral tibial plateau was not ideal(R2=0.23,RMSE=0.29),while it had potential predictive value(accuracy=0.21,accuracy=0.15).Conclusion The prediction model of knee joint cartilage thickness based on individual parameters has certain scientificity,and the feasibility of KNN model is relatively high.However,due to insufficient sample size and unclear individual parameter weight,the efficiencies of the four established prediction models are not ideal,which fails to provide definite prediction equations.Therefore,the construction scheme of the prediction model still needs to be further optimized.

The Guidelines for Occupational Hazard Assessment and Control of Active Pharmaceutical Ingredient in the Pharmaceutical Industry (T/WSJD 60—2024) is the first guiding standard in the field of health in China that focuses on occupational health protection for active pharmaceutical ingredient (API). It covers the general principles, work procedures, assessment methods, and control strategies for API occupational hazard assessment, providing practical guidance and recommendations for pharmaceutical enterprises to eliminate or reduce occupational health risks associated with API, improve working environment, and enhance refined management practices. This article interpreted and analyzed the background of standard establishment, formulation process, fundamental basis, and main content, to provide scientific and comprehensive technical support for occupational health managers in the pharmaceutical industry to better apply this standard.

Objective To design a digital intelligence-driven critical treatment platform to implement integrated treatment procedure inside and outside the hospital and intelligent whole-course managment from pre-hospital emergency care to discharge for critically ill patients.Methods The platform was designed with 5G,IoT,big data and aritificial intelligence techniques and developed with Java language,which adopted Oracle database-based data management and front-end and back-end separation mode,with the front end realized by Vue.js framework and the back end by microservice architecture.There were five functional modules for pre-hospital emergency care,multidisciplinary joint diagnosis and treatment,critical care,quality control management and post-discharge follow-up involved in the platform.Results The platform developed simplified the treatment procedure,enhanced the timeliness of emergency care,decreased the workload of nursing staffs and improved medical service efficiency and working efficiency effectively.Conclusion The platform increases first aid quality and treatment efficiency and provides critically ill patients with high-quality medical experience.[Chinese Medical Equipment Journal,2025,46(1):38-43]

AIM:This study aimed to confirm the glycolytic inhibitory activity of 3-bromopyruvic acid(3BP)and to assess whether this inhibition could ameliorate hypoxia-induced pulmonary hypertension in rats.METHODS:PAH model rats were generated from normal SD rats via exposure to normal pressure and hypoxia.Intervention groups I and II(6 rats per group)were then intraperitoneally injected with 3BP(15 mg/kg),and the normal and hypoxia groups(6 rats per group)were given the same amount of normal saline for a total of 21 d.The average pulmonary artery pressure of the rats in each group was measured via right heart catheterisation,and hilar tissue measurements.The right ventricle(RV),left ventricle,and interventricular septum(LV+S)were weighed,and the ratio of RV/(LV+S)was calculated as an index of right ventricular hypertrophy.Right lower lung tissues were fixed in 4%paraformaldehyde-PBS buffer,sec-tioned in conventional paraffin(5 μm thick),stained with HE and Masson,photographed under a microscope.Then the thickness ratio of the tunica media and the area ratio of collagen fibres were calculated.The expression of pyruvate kinase isozyme type M2(PKM2),nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),and pyruvate de-hydrogenase(PDH)proteins in the hilar tissues of each group were detected by western blot,whereas interleukin-1β(IL-1β)and IL-18 contents were detected using ELISA,and lactic acid content was detected using a lactic acid kit.RE-SULTS:The results showed that 3-brpa effectively inhibited glycolysis and significantly improved hypoxia-induced pulmo-nary hypertension in rats.Compared with the hypoxia group,in intervention group II,PKM2 expression was decreased(P<0.05),PDH expression increased significantly(P<0.01),and NLRP3 expression was decreased(P<0.05).The IL-18 and IL-1β contents decreased(P<0.05 or P<0.01,respectively).Pulmonary hemodynamic indexes showed that the pro-portion of the right ventricle and the mean pressure of the pulmonary artery decreased(P<0.05 or P<0.01,respectively).The HE and Masson staining results showed that the thickness ratio of the tunica media and the area ratio of collagen fibres decreased significantly(P<0.01).Lactic acid content was significantly decreased(P<0.01).CONCLUSION:This study showed that 3BP can inhibit glycolysis and alleviate hypoxia-induced pulmonary hypertension in rats.

Objective:To investigate the pollution status of legionella pneumophila of industrial circulating cooling towers, and explore the factors affecting the positive rate and quantity of legionella pneumophila, in order to provide data support for the risk assessment and hierarchical management of legionella pneumophila in workplaces.Methods:In February, May, August and October 2023, the cooling water samples were collected from a total of 20 industrial circulating cooling towers of 9 enterprises in an industrial park in Shanghai. The water temperature, free residual chlorine, total residual chlorine, turbidity, chroma, pH, conductivity, chemical oxygen consumption, iron content and total plate count in the water samples were detected. Meanwhile, the legionella pneumophila in the water samples were qualitatively and quantitatively analyzed, and the influence of water quality indexes on the positive rate and number of legionella pneumophila detected was analyzed by logistic regression and linear regression respectively.Results:The positive rates of legionella pneumophila in the first to fourth quarters of 20 industrial circulating cooling towers of 9 chemical enterprises were 35% (7/20), 45% (9/20), 45% (9/20), and 85% (17/20). The serotypes of legionella pneumophila detected in the cooling tower were mainly legionella pneumophila type 1 (95.2%, 40/42), and the number of legionella pneumophila ranged from 0.2 to 316.8 cfu/ml. Circulating cooling water temperature ( OR=1.36, 95% CI: 1.12-1.64, P=0.001) and conductivity ( OR=1.02, 95% CI: 1.00-1.03, P=0.026) were independent risk factors affecting the detection of legionella pneumophila. There was a positive correlation between chemical oxygen consumption and the detected amount of legionella pneumophila ( β=8.08, P=0.002) . Conclusion:The positive rate of legionella pneumophila in industrial circulating cooling water system is high, and the detected quantity fluctuates greatly, which may have health risks, suggesting that it is necessary to pay attention to the water quality management such as water temperature, conductivity and chemical oxygen consumption of industrial circulating cooling water and the monitoring and control of legionella pneumophila.