ObjectiveTo construct a stable monoclonal human embryonic kidney 293 （HEK293） cell line expressing recombinant human coagulation factor Ⅶ （rhFⅦ） and evaluate the expression level and procoagulant bioactivity of rhFⅦ. MethodsThe plasmid pCDNA3.1-EGFP-FⅦ was transfected into HEK293 cells to verify the effectiveness of the transfection system. The plasmid pCDNA3.1-FⅦ was transfected into HEK293 cells， and monoclonal stable transfected cell lines were selected using geneticin （G418）. The transcription of the FⅦ gene was identified by reverse transcription polymerase chain reaction （RT-PCR）. The expression level of rhFⅦ in the supernatant of the monoclonal stable transfected cell line was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. The concentration of rhFⅦ was determined by enzyme-linked immunosorbent assay （ELISA）， and the procoagulant activity of rhFⅦ was measured by human coagulation factor Ⅶ potency assay. ResultsHEK293 cells transfected with pcDNA3.1-EGFP-FⅦ showed green fluorescence， indicating that rhFⅦ was successfully expressed in the supernatant of HEK293 cells after transient transfection with pcDNA3.1-FⅦ. The monoclonal stable transfected cell line was obtained by G418 screening. RT-PCR identified that the FⅦ gene was integrated into the genome of the monoclonal stable transfected cell line. The cell viability was good as detected by Cell Counting Kit-8， and a single band of rhFⅦ was obtained by purification of the cell supernatant. The highest rhFⅦ expression was （1.27±0.09） mg/L， and the highest procoagulant activity was （380.29±13.80）%. ConclusionThe monoclonal HEK293 cell lines which can express rhFⅦ protein efficiently and stably with excellent procoagulant bioactivity is successfully screened.

Bile acids (BAs) serve not only as key facilitators of lipid absorption but also as crucial signaling molecules regulating glucose and lipid metabolism, inflammation, and overall energy homeostasis. Aging profoundly alters BA metabolism, characterized by shifts in biosynthetic pathways, compositional changes, disrupted receptor-mediated signaling, and alterations in gut microbiota interactions. These age-related changes contribute to the onset and progression of metabolic conditions, including type 2 diabetes, obesity, metabolic dysfunction-associated fatty liver disease, and neurodegenerative disorders. An increased abundance of hydrophobic and cytotoxic BAs has been associated with systemic inflammation, metabolic rigidity (disruption of metabolic flexibility), and organ dysfunction. Targeting BA signaling—through pharmacological modulation of farnesoid X receptor and Takeda G protein-coupled receptor 5 or microbiota-directed therapies—offers promising strategies to mitigate aging-related metabolic decline. A deeper understanding of how BA metabolism evolves over the lifespan may unveil novel interventions to promote healthy aging and prevent age-related disease.

Objective To analyze the core elements of standardized diagnosis of occupational carpal tunnel syndrome (OCTS). Methods Data of first reported OCTS case in Guangdong Province were retrospectively analyzed, including occupational exposure history, clinical manifestations, auxiliary examinations, and on-site occupational health investigation results. Furthermore, standardized diagnostic approaches for such occupational diseases were explored. Results The patient worked in a garment manufacturing enterprise, engaged in fabric cutting. The patient had a occupational history of at least three consecutive years of repetitive right wrist work, holding a 15.2 kg electric cutting tool for a cumulative 6.6 hours per day, with awkward wrist movements repeated 13 times per minute. Clinical symptoms included numbness and reduced muscle strength in the areas of median nerve distribution of the right hand, which were aggravated during work and relieved after rest. Physical examination showed inability of right hand to perform fine motor opposition movements of the thumb with the index, middle, and ring fingers; decreased tactile, pain, vibration, graphesthesia, and kinesthetic sensations below the right wrist; and positive Tinel′s sign over the median nerve at the wrist crease level, Phalen′s test, and reverse Phalen′s test of right wrist. Both wrist high-frequency color Doppler ultrasonography and nerve conduction studies and electromyography indicated damage to the right median nerve. However, only the findings from high-frequency color Doppler ultrasonography of the wrist meet the diagnostic criteria specified in GBZ 336-2025 Diagnostic Standard for Occupational Carpal Tunnel Syndrome. On-site occupational health investigation confirmed that the work involved typical repetitive wrist work, and three co-workers in similar positions exhibited comparable symptoms. The case was ultimately diagnosed as OCTS. Conclusion The definitive diagnosis of OCTS requires comprehensive evaluation of four core elements: a clear occupational exposure history, typical clinical symptoms and signs, characteristic objective examination findings, and quantitative on-site occupational health investigation data. Only when these four elements corroborate each other and non-occupational causative factors are excluded can a definitive diagnosis be made.

Skeletal muscle atrophy refers to a pathological condition characterized by muscle fiber atrophy as well as decreased muscle mass and muscle strength induced by various predisposing factors. It falls under the category of “flaccidity syndrome” in traditional Chinese medicine， and its pathogenesis is mainly associated with disturbed protein homeostasis， mitochondrial dysfunction and oxidative stress injury. Active ingredients of traditional Chinese medicine exerts synergistic regulatory effects via multiple pathways， thus possessing unique advantages in the clinical prevention and treatment of skeletal muscle atrophy. This paper reviews the mechanisms by which active ingredients of traditional Chinese medicine alleviate skeletal muscle atrophy. Existing studies have demonstrated that such active ingredients can ameliorate skeletal muscle atrophy through multiple approaches： regulating protein metabolic balance （geniposide， matrine， schisandrin A， etc.） and oxidative stress （astragaloside Ⅳ， Gastrodia elata polysaccharide， Angelica sinensis polysaccharide， etc.）， improving mitochondrial function （parthenolide， berberine， curcumin， etc.）， facilitating myogenesis and myogenic differentiation （cucurbitacin Ⅱb， saikosaponin A， saikosaponin D， etc.）， suppressing inflammatory response （ursolic acid， triptolide， emodin， etc.）， ameliorating insulin resistance （akebiasaponin D， etc.）. In the future， active ingredients of traditional Chinese medicine are expected to achieve breakthroughs in clinical treatment for skeletal muscle atrophy， providing novel ideas and strategies for the prevention and treatment of degenerative disorders.

Abstract To clarify the service targets, service contents, and safeguard mechanisms of the basic public service program for student vision health management, the study grounded in the current status of child and adolescent myopia prevention and control in China and integrated with the practical experience and innovative practices of Wuhan National Demonstration Area for Student Vision Health Management, takes education as the main thread and schools as the base, consolidating resources from multiple stakeholders including education administrative departments, schools, families, and professional technical service institutions. Centering on four core components—vision health education, monitoring and record keeping, risk prevention and control, and dynamic management, the study constructs a multi tiered basic public service system for vision health management of primary and secondary school students. It further proposes the use of an intelligent management platform and a "three tier color coded early warning" mechanism (red, yellow, blue) to prevent and control student myopia at the source, providing schools with replicable and operable implementation pathways to enhance the level of student vision health management.

Background: Alzheimer’s disease (AD) is the most prevalent neurodegenerative disorder. Cellular drug screening represents a key approach in the preclinical discovery of AD therapeutics. Moreover, selecting an appropriate cellular model is essential for exploring the mechanisms underlying AD pathogenesis. Objective: This study aimed to establish a progressive strategy integrating cell model construction, monomer evaluation, and effective component group-based drug discovery for the development of novel therapeutics targeting AD. Methods: A novel AD12 cell line stably expressing the double-transgenic genes amyloid precursor protein (APP)695/presenilin 1(PS1)-ΔE9 was first constructed and subjected to lipopolysaccharide (LPS) stimulation. The resulting LPS-induced neuroinflammatory response was characterized using metabolomic analysis. Subsequently, 16 alkaloids derived from traditional Chinese medicine(TCM) were screened with the LPS-AD12 cell model. Molecular docking was then performed to assess the binding affinity of benzoylaconitine (BAC) with APP and PS1 proteins. A targeted metabolomics approach focusing on amino acids and the tricarboxylic acid (TCA) cycle was applied to BAC-treated LPS-AD12 cells. Finally, aconite extract was administered to five-familial-AD-mutations(5xFAD) transgenic mice, and therapeutic efficacy was evaluated through phenotypic and metabolomic analyses. Results: LPS stimulation induced neuroinflammation in AD12 cells, characterized by elevated levels of interleukin-6, amyloid-β (Aβ ), and PS1. The primary metabolic pathways affected in AD12 cells included alanine, aspartate, and glutamate metabolism, as well as arginine biosynthesis. Notably, BAC significantly reduced the levels of Aβ and interleukin-6 in LPS-AD12 cells. Furthermore, molecular docking analyses revealed a strong binding affinity of BAC with APP and PS1 proteins. Targeted metabolomics analysis indicated that the pharmacological activity of BAC was primarily associated with its anti-inflammatory effects, modulation of the cholinergic signaling pathway, and regulation of energy metabolism. Behavioral experiments in 5xFAD mice, together with immunofluorescence assessments of brain tissues (amyloid-β, glial fibrillary acidic protein, and ionized calcium-binding adapter molecule 1), demonstrated that aconite extract treatment significantly alleviated AD-related symptoms. In addition, metabolomics-based mechanistic analyses revealed that serine, acetylcholine, ornithine, and gamma-aminobutyric acid exhibited consistent trends in both brain tissue and serum samples from treated mice. Conclusion: This study established a novel AD12 cell model and an effective, reliable LPS-induced cell-based screening system. Additionally, we proposed a new drug discovery strategy integrating cell model construction, monomer evaluation, and effective component group-based drug discovery. This screening system has the potential to accelerate the identification of anti-AD agents and provides a feasible framework for subsequent in-depth investigations into new drug development from TCM, thereby contributing to the modernization of TCM-derived therapeutics.

Objective: To evaluate the effects of a home-school collaborative intervention on improving sugar sweetened beverages (SSBs) consumption behaviors among primary school students in grades 3 to 5 in Chengdu, so as to provide evidence for SSBs prevention and control strategies among primary school students. Methods: Using a stratified cluster random sampling method, a total of 2 006 students in grades 3 to 5 from 4 primary schools in Chengdu were enrolled from October to December 2024, including an intervention group ( n =769) and a control group ( n =1 237). The control group received routine psychological counseling, while the intervention group received a comprehensive home-school collaborative intervention integrating time perspective cultivation and family functioning improvement. A two level linear mixed effects model was used to analyze the net intervention effect, and the EBICglasso algorithm was used to construct variable association networks to identify core intervention targets. Results: After adjusting for confounders and clustering effects, the intervention significantly reduced the weekly frequency of SSBs consumption (decreased from baseline by 1.29 in the intervention group, 95% CI =-1.84 to -0.73; by 0.84 in the control group, 95% CI =-1.21 to -0.46; interaction P =0.01, Cohen s d=-0.16), improved SSBs consumption behavioral intention (decreased from baseline by 0.62 in the intervention group, 95% CI =-1.26 to -0.02; by 0.14 in the control group, 95% CI =-0.57 to 0.29; interaction P =0.02, Cohen s d =-0.15) and increased family functioning level (increased from baseline by 0.34 in the intervention group, 95% CI =-0.16 to 0.85; decreased by 0.08 in the control group, 95% CI =-0.42 to 0.25; interaction P =0.01, Cohen s d =0.17). A permutation test with 1 000 iterations showed no statistically significant difference in global network strength between the two groups ( P =0.19), whereas the differences in overall network structure and maximum edge weight were both statistically significant (all P <0.05), with present hedonistic time perspective, SSBs consumption attitude and behavioral intention identified as the core intervention targets. Conclusion The comprehensive home-school collaborative intervention effectively reduces SSB consumption frequency among primary school students and improves behavioral intention and family functioning.

Objective:To observe the efficacy and safety of combined lienal polypeptide injection therapy in the treatment of Mycoplasma pneumoniae pneumonia（MPP）in children aged 3 to 14 years old in multiple clinical centers.Methods:A randomized，controlled，multi-center clinical study design was adopted.A total of 240 hospitalized children aged 3 to 14 years old with MPP from 7 hospitals from September 1，2023 to January 31，2024 were included.According to the severity of pneumonia，they were divided into the mild MPP group with 80 cases and the severe MPP/refractory MPP（SMPP/RMPP）group with 160 cases，and then randomly divided into the control group and the experimental group at a ratio of 1 ∶1，using the random number table method.After screening，subjects entered a treatment period of 5 to 7 days.The control group was treated with azithromycin，while the experimental group was treated with azithromycin plus lienal polypeptide injection .The recovery of lung CT，length of hospital stay，duration of fever，cough score，whether mild cases developed into severe or refractory cases，duration of hormone use，use of intravenous immunoglobulin（IVIG），bronchoscopy treatment，and immune function were observed between the two groups to evaluate the efficacy of lienal polypeptide injection.Adverse events after medication，vital signs，blood routine，urine routine，liver function，myocardial enzymes，renal function，and electrocardiogram were observed to evaluate the safety. Results:A total of 231 subjects have completed the trial in the 7 hospitals，including 118 cases in the experimental group and 113 cases in the control group.Main observation index：the rate of lung CT aggravation in the experimental group was lower than that in the control group（2.6% vs 15.3%， P<0.01），and the difference was statistically significant.Secondary indexes：there were no statistically significant differences in the length of hospital stay，duration of fever，cough score，duration of hormone use，whether IVIG treatment was used，the number of bronchoscopy treatment cases，and immunoglobulin between the two groups（all P>0.05）.However，the rate of cases of plastic bronchitis（PB）found under bronchoscopy in the experimental group was lower than that in the control group（0 vs 18.8%， P=0.03），and the difference was statistically significant.Among the mild MPP（72 cases），there were no statistically significant differences in the length of hospital stay，duration of fever，cough score，duration of hormone use，whether IVIG treatment was used，the number of bronchoscopy treatment cases，and the improvement rate of lung CT between the two groups（all P>0.05）.However，compared with the control group，the rate of cases developing into SMPP/RMPP in the experimental group was less（24.3% vs 48.6%， P=0.03），and the difference in IgG before and after treatment was small［0.53（-0.04，1.18）g/L vs 1.33（0.48，2.25）g/L， P=0.01］.Among the SMPP/RMPP cases（159 cases），the rate of cases of PB found under bronchoscopy in the experimental group was less than that in the control group（0 vs 20%， P=0.04），and the rate of cases with aggravated lung CT in the experimental group was less than that in the control group（1.3% vs 19.5%， P<0.01），and the improvement rate of lung CT in the experimental group was higher than that in the control group（88.8% vs 75.3%， P=0.03），with statistically significant differences.There were no statistically significant differences in the length of hospital stay，duration of fever，cough score，duration of hormone use，whether IVIG treatment was used，the number of bronchoscopy treatment cases，and immunoglobulin between the two groups（all P>0.05）.Two cases in the experimental group developed rashes，which improved after the drug was discontinued.There were no serious adverse reactions such as abnormal vital signs like dyspnea and cyanosis due to the use of lienal polypeptide injection.There were no obvious changes in blood routine，liver function，myocardial enzymes，renal function，electrocardiogram，and urine routine values before and after medication compared with the baseline. Conclusion:The combined use of lienal polypeptide injection in the treatment of MPP in children can reduce the probability of the transformation from mild cases to SMPP/RMPP，reduce the rate of aggravation of the image findings，promote the absorption of lung inflammation，reduce the rate of PB found under bronchoscopy，and has good safety.

Objective:To investigate the extent and characteristics of parenting stress experienced by fathers of children aged 0-6 in three provinces of Eastern China, and to compare the quality of their marital relationships within each latent class, in order to provide reference for developing intervention strategies for different categories of fathers.Methods:Using cross-sectional survey methods, fathers with children aged 0-6 years were recruited using convenience sampling from December 2020 to March 2021 from Anhui province, Zhejiang province and Jiangsu province in Eastern China. Data were collected through a general information questionnaire, the Parenting Stress Index-Short Form, and the Locke-Wallace Marital Adjustment Scale.Latent profile analysis was used to determine the classification, Kruskal-Wallis H test was used to compare the difference of marital quality. Results:A total of 657 questionnaires were distributed and 604 valid questionnaires were recovered. The age of 604 fathers was (32.50 ± 6.26) years, the score of parenting stress was (89.70 ± 17.87) and the score of marriage quality was (108.36 ± 22.28). The characteristics of parenting stress among fathers of young children were categorized into three latent profile types: "high stress-negative parenting group" (88 people accounting for 14.6%), " medium stress- poor interaction group" (329 people accounting for 54.5%), and " low stress-positive parenting group" (187 people accounting for 30.9%). The scores of marriage quality were (99.94 ± 23.81), (106.52 ± 19.84), (115.56 ± 23.70) points among these three latent classes with significant differences ( H=35.89, P<0.05). Conclusions:The study identifies three latent classes of parenting stress among fathers of children aged 0-6, each associated with varying levels of marriage quality. Therefore, targeted interventions aimed at improving fathers′ parenting stress and marriage quality, and helping create family environments where children can grow up healthy.

Objective To investigate the correlation between hemoglobin A1c(HbA1c)levels and excessive daytime sleepiness(EDS)in patients with type 2 diabetes mellitus(T2DM).Methods A total of 132 T2DM patients and 40 healthy people(NC group)who were treated in the outpatient department of Nanjing Meishan Hospital from December 2020 to December 2022 were selected.General clinical data of the subjects were collected,and their Epworth sleepiness scale(ESS)scores,Pittsburgh sleep quality index(PSQI),and apnea-hypopnea index(AHI)were measured.Based on ESS scores,T2DM patients were divided into simple T2DM group(ESS score<9,n=99)and EDS group(ESS score≥9,n=33)according to ESS score.The baseline data were compared for each group.Spearman correlation analysis and multivariate logistic regression model were used to evaluate the correlation between HbA1c levels and EDS.The receiver operating characteristic(ROC)curve was used to calculate the area under the curve(AUC)to evaluate the predictive value of HbA1c levels for EDS.Results HbA1c,fasting plasma glucose,AHI index and PSQI score in EDS group were higher than those in T2DM and NC group(P<0.05).Spearman correlation analysis showed that ESS score was positively correlated with HbA1c in T2DM patients(P<0.05).Multivariate logistic regression analysis revealed that elevated HbA1c levels emerged as a significant and independent risk factor for the onset of EDS.The ROC curve indicated that the AUC of HbA1c for predicting EDS was 0.736.Conclusions There is an independent positive correlation between HbA1c levels and EDS in T2DM patients,which provides clues for early identification and treatment of EDS in clinical practice.