ObjectiveTo investigate the mechanism of Zuogui Wan （ZGW） in improving ovarian inflammatory microenvironment and stemness of ovarian germline stem cells （OSCs） for treating ovarian aging via the cyclic guanosine monophosphate/adenosine monophosphate synthase （cGAS）/stimulator of interferon genes （STING） signaling pathway. MethodsForty C57BL/6 female mice were randomly divided into a blank group， a model group， a low-dose ZGW group （2.7 g·kg^-1）， a high-dose ZGW group （5.4 g·kg^-1）， and an estradiol valerate group （0.15 mg·kg^-1）， with 8 mice in each group. Except the blank group， all other groups received a single intraperitoneal injection of cyclophosphamide at 120 mg·kg^-1 to establish an ovarian aging mouse model. After successful modeling， each group was continuously administered for 4 weeks， once daily. The physiological status of the mice was observed， and the ovarian index was calculated. The estrus cycle of the mice was monitored. Hematoxylin-eosin （HE） staining was used to observe pathological changes in ovarian tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum sex hormone levels. Serum inflammatory factors interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and mouse interleukin-6 （IL-6） levels were detected using kits. Western blot was used to detect the protein expression of ovarian cGAS， STING， p-STING， TANK-binding kinase 1 （TBK1）， p-TBK1， interferon-induced transmembrane protein 3 （Fragilis）， and Vasa homolog protein （MVH）. Quantitative real-time polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of inflammatory factors in ovarian tissue. Immunofluorescence double labeling was performed to locate OSCs in ovarian tissues， and fluorescence intensities of OSCs markers MVH and octamer binding transcription factor 4 （Oct4） were calculated. ResultsCompared with the blank group， the model group showed reduced body weight， ovarian wet weight， and ovarian index （P<0.01） and a disordered estrus cycle （P<0.01）. In addition， the levels of serum follicle-stimulating hormone （FSH）， TNF-α， IL-6， and IL-1β were increased （P<0.01）， while anti-Müllerian hormone （AMH） and estradiol （E₂） levels were decreased （P<0.01）. The protein expression of cGAS， p-STING/STING， and p-TBK1/TBK1 in ovarian tissue was increased （P<0.05， P<0.01）， while that of OSCs stemness factors MVH and Fragilis was reduced （P<0.01）. Immunofluorescence indicated a reduction in MVH and Oct4 expression in OSCs （P<0.01）. The mRNA expression of inflammatory factors TNF-α， IL-6， and IL-1β in ovarian tissue was increased （P<0.05， P<0.01）. Compared with the model group， the treatment groups exhibited improved body weight， ovarian wet weight， and ovarian index （P<0.05） and a reduced rate of estrus cycle disorder （P<0.05， P<0.01）. The levels of serum FSH， TNF-α， IL-6， and IL-1β were decreased （P<0.05， P<0.01）， while AMH and E₂ levels were increased （P<0.01）. The protein expression levels of cGAS， p-STING/STING， and p-TBK1/TBK1 in ovarian tissue were decreased （P<0.05）， while the protein expression of MVH and Fragilis was increased （P<0.05）， and the fluorescence intensities of MVH and Oct4 were increased （P<0.05， P<0.01）. The mRNA expression of inflammatory factors in ovarian tissue was decreased （P<0.05）. ConclusionZGW alleviate ovarian inflammatory response， regulate ovarian microenvironment homeostasis， and maintain stemness of OSCs in ovarian aging mice probably by modulating the cGAS-STING signaling pathway， thereby improving ovarian function and delaying ovarian aging.

Objective To evaluate the safety and efficacy of a self-developed micro-normothermic machine perfusion (NMP) system (micro-perfusion device) for preserving isolated porcine limbs. Methods Five healthy Landrace pigs were selected, and their left and right forelimbs were randomly divided into the NMP group and static cold storage (SCS) group. The NMP group was perfused with the self-developed micro-perfusion device and polymerized hemoglobin perfusate for 32 hours at normothermia, while the SCS group was preserved at 4 ℃. Hemodynamic parameters such as perfusion pressure and flow were monitored. The pH value, partial pressure of oxygen (PO₂), lactic acid (Lac), creatine kinase (CK) and lactate dehydrogenase (LDH) in the perfusate were measured. Hematoxylin-eosin staining was used to assess the muscle tissue structure, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was employed to evaluate muscle cell apoptosis, and immunohistochemistry staining was applied to detect the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6. A mixed-effects model was used to analyze the effects of time and treatment methods on tissue structure, cell apoptosis and inflammatory factors. Results The device could stably maintain a perfusion pressure of (69±15) mmHg and a flow rate of (117±42) mL/min. The pH value and electrolytes of the perfusate were generally stable, with PO₂ maintained at a high level. Lac was maintained at 5.38(3.81, 6.45) mmol/L, while CK and LDH increased over time. After 32 hours of perfusion in the NMP group, both the myocyte spacing and apoptosis rate were better than those in the SCS group. Mixed-effects model analysis showed that there were statistically significant differences in the effects of NMP treatment and SCS treatment on myocyte spacing and apoptosis rate per unit time (both P < 0.05). There were no statistically significant differences in TNF-α and IL-6 between the two groups, and mixed-effects model analysis showed no statistically significant differences in the effects of NMP treatment and SCS treatment on TNF-α and IL-6 per unit time (both P > 0.05). Conclusions The micro-perfusion device used in this study may achieve 32-hour normothermic preservation in a porcine limb amputation model, maintain basic metabolism and ionic homeostasis, reduce muscle structural damage and cell apoptosis without inducing additional inflammatory responses. This technology is expected to significantly extend the time window for replantation of amputated limbs in disaster rescue and long-distance transportation, providing an important technical basis for clinical translation and subsequent replantation research.

BACKGROUND:Total flavonoids from Sophora flavescens have a variety of pharmacological effects,including anti-inflammatory,immunomodulatory,antioxidant,and anti-hepatic injury,but the therapeutic effects and mechanisms in non-alcoholic fatty liver disease are not clear.OBJECTIVE:To reveal the mechanism of total flavonoids from Sophora flavescens in the treatment of non-alcoholic fatty liver disease using bioinformatics,network pharmacology and zebrafish experimental validation.METHODS:A zebrafish model of non-alcoholic fatty liver disease was constructed to observe lipid accumulation,pathomorphologic changes,and expression of inflammatory genes in the liver of zebrafish after treatment with total flavonoids from Sophora flavescens.The active ingredients of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease-related targets were obtained from TCMSP,Swiss Target Prediction,and Bat-man databases.STRING was used to perform protein-protein interaction network analysis,GO functional enrichment and KEGG pathway enrichment analysis.Based on the GSE33814 dataset,the differentially expressed genes of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease intersection targets were screened out.Correlation analysis and receiver operating characteristic curve were performed using R4.3.2 software.Core genes were verified by the validation set GSE89632.RT-qPCR and western blot assays were performed to verify the expression of core pathway-related genes and proteins.RESULTS AND CONCLUSION:(1)Total flavonoids from Sophora flavescens could improve lipid accumulation in the liver of zebrafish with non-alcoholic fatty liver disease,significantly inhibited the elevation of lipid and aminotransferase levels in zebrafish(P<0.05),and regulated the expression of genes related to inflammation and lipid metabolism.(2)A total of 168 common targets were obtained using the network pharmacology,and top 10 core genes,identified by Cytoscape topology analysis,were HSP90AA1,STAT3,PIK3R1,MAPK1,AKT1,RXRA,PIK3CA,EGFR,JAK2,and ESR1.GO and KEGG analysis pathways mainly included insulin resistance,lipids,and atherosclerosis.There were a total of 59 differentially expressed genes after intersection of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease targets.The receiver operating characteristic curve and validation set analyses yielded six core targets that were significantly different between healthy individuals and patients with non-alcoholic fatty liver disease(P<0.01).(3)RT-PCR and western blot results verified that total flavonoids from Sophora flavescens inhibited the activation of the JAK2/STAT3 signaling pathway in zebrafish.To conclude,total flavonoids from Sophora flavescens may alleviate the inflammatory response through the JAK2/STAT3 signaling pathway,thus inhibiting lipid accumulation and improving non-alcoholic fatty liver disease.

Based on the concept of"two-stage,four-phase,and three-phase treatments"of in vitro fertilization and embryo transfer(IVF-ET)assisted the whole-course management of TCM by national TCM master You Zhaoling,this article analyzed the treatment strategy assisted by acupuncture in five aspects:principle,method,prescription,acupoint,and technique.Principle:syndrome differentiation and treatment follow different cases;method establish guiding methods based on principles,while following stage diagnoses and treatments;prescription:determine the prescriptions based on therapeutic principles within the context of overall regulation;acupoint:precise acupoint taking and rational acupoint matching;technique:the primary method is acupuncture,supported by various therapeutic modalities,providing a comprehensive framework and evidence-based support for the clinical application of acupuncture and moxibustion in IVF-ET.

Objective:To investigate the clinical characteristics and risk factors for disease progression after treatment in patients with post-tuberculosis chronic pulmonary aspergillosis (post-TB CPA).Methods:A retrospective cohort study was conducted on post-TB CPA patients admitted to Hangzhou Red Cross Hospital between January 2020 and December 2023. The demographic manifestation, clinical manifestation, laboratory indicators, imaging findings, and treatment strategies were collected. Patients were divided into progression group and non-progression group based on treatment outcomes, and the clinical data of the two groups were compared. Chi-square test was used for univariate analysis, and multivariate logistic regression were used to identify independent risk factors for disease progression after treatment.Results:A total of 109 post-TB CPA patients were included, and 33.9%(37/109) were in the progression group and 66.1%(72/109) in the non-progression group. Multivariate logistic regression revealed that subacute invasive aspergillosis (SAIA) (odds ratio ( OR)=14.356, 95% confidence interval ( CI) 2.923 to 70.504, P=0.001), elevated erythrocyte sedimentation rate (ESR) ( OR=5.276, 95% CI 1.505 to 18.491, P=0.009), and pulmonary fibrosis ( OR=5.030, 95% CI 1.437 to 17.612, P=0.012) were independent risk factors for disease progression. Antifungal treatment for ≥3 months was associated with a lower risk of disease progression ( OR=0.038, 95% CI 0.003 to 0.431, P=0.008). The proportion of non-progression group receiving surgical treatment was higher than that of progression group with statistical significance (31.9%(23/72) vs 5.4% (2/37), χ2=8.30, P=0.004), but the protective effect of surgery on disease progression was not confirmed by multivariate analysis ( OR=0.735, 95% CI 0.132 to 4.080, P=0.724). Conclusions:Disease progression in patients with post-TB CPA is strongly associated with SAIA, elevated ESR, and pulmonary fibrosis. Standardized anti-fungal treatment for ≥3 months significantly improves the prognosis.

ObjectiveTo explore the therapeutic mechanism of Buchong Tiaojing prescription for rats with diminished ovarian reserve （DOR） from the perspectives of nucleotide-binding oligomerization domain （NOD）-like receptor protein 3 （NLRP3） inflammasome and ferroptosis. MethodsA total of 48 female SD rats were randomly divided into a normal group， a model group， low， medium， and high dose groups of Buchong Tiaojing prescription， and an MCC950 group， with eight rats in each group. Except the normal group， all the other groups were injected subcutaneously on the back of the neck with D-galactose to prepare the DOR rat model. From the 15th day of modeling， the rats in the low， medium， and high dose groups of Buchong Tiaojing prescription were subjected to gavage daily at doses of 14.4， 28.8， 57.6 g·kg^-1， respectively. Rats in the MCC950 group were injected intraperitoneally with MCC950 at a dose of 10 mg·kg^-1， once every other day. The interventions of all the groups lasted for 4 weeks. The estrous cycle of the rats was observed with vaginal exfoliated cell smear. Hematoxylin-eosin （HE） staining was performed to observe the development of follicles and corpus luteum in the ovary. Enzyme-linked immunosorbent assay （ELISA） was performed to detect the levels of serum sex hormones and interleukin-1β （IL-1β）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot assay were performed to detect the mRNA and protein expression of NLRP3 inflammasome， acyl-CoA synthetase long-chain family member 4 （ACSL4）， transferrin receptor 1 （TFR1）， and glutathione peroxidase 4 （GPX4）， and oxidative stress kits were used to detect ovarian superoxide dismutase （SOD） and malondialdehyde （MDA） levels. ResultsDuring the experiment， one rat died in the high dose group of Buchong Tiaojing prescription， and a total of 47 rats were finally included in the index tests and statistics. Compared with those in the normal group， rats in the model group had significantly disturbed estrous cycles， increased number of atretic follicles， and significant disorder of serum sex hormones. The mRNA and protein expression of NLRP3 inflammasome， ACSL4， and TFR1 in ovarian tissue was up-regulated （P<0.01）， while that of GPX4 was significantly down-regulated （P<0.01）. The SOD content in the ovary was decreased significantly， while the MDA level was increased （P<0.01）. After drug intervention， the estrous cycle of rats was basically resumed， and the follicles at all levels were more structurally intact and significantly increased in number. Additionally， the levels of serum sex hormones and IL-1β were significantly improved. The mRNA and protein expression of NLRP3 inflammasome， ACSL4， and TFR1 were down-regulated， while that of GPX4 was significantly up-regulated， and the ovarian oxidative stress was alleviated （P<0.05， P<0.01）， especially in the high dose group of Buchong Tiaojing prescription and the MCC950 group. ConclusionInflammatory injury and ferroptosis occur in the ovaries of DOR rats， and the Buchong Tiaojing prescription is able to inhibit ovarian NLRP3 inflammasome， alleviate the degree of ovarian ferroptosis， and improve ovarian reserve.

Objective: To analyze the mediating effects of self-efficacy and self-management behaviors on the relationship between depression symptoms and glycemic control among elderly patients with type 2 diabetes mellitus (T2DM), so as to provide references for optimizing health management of elderly T2DM patients. Methods: T2DM patients aged ≥60 years from 8 community health service centers in Nanjing City were selected using a convenience sampling method. Basic information such as gender and age was collected through questionnaires. Depressive symptoms, self-efficacy, and self-management behaviors were assessed using the Patient Health Questionnaire, the Diabetes Self-Efficacy Scale, and the Diabetes Self-Management Behavior Scale, respectively. Glycated hemoglobin (HbA1c) was measured to evaluate glycemic control. A mediating effect model was constructed to analyze the mediating effects of self-efficacy and self-management behaviors on the relationship between depressive symptoms and glycemic control. Results: A total of 567 elderly T2DM patients were included, with a median age of 70.00 (interquartile range, 7.50) years. There were 248 males (43.74%) and 319 females (56.26%). The median scores of self-efficacy, self-management behaviors, depressive symptoms, and HbA1c were 3.89 (interquartile range, 0.78), 4.45 (interquartile range, 1.55), 4.00 (interquartile range, 6.00), and 6.80% (interquartile range, 1.40%), respectively. The mediating effect analysis showed that depressive symptoms indirectly affected glycemic control among elderly T2DM patients through the independent mediating effects of self-efficacy (β=0.028, 95%CI: 0.016-0.043) and self-management behaviors (β=0.009, 95%CI: 0.003-0.016), as well as the chain mediating effect of both (β=0.025, 95%CI: 0.017-0.035). The mediating effects of self-efficacy and self-management behaviors accounted for 36.66% and 11.35% of the total effect, respectively, while the chain mediating effect accounted for 32.15% of the total effect. Conclusion Self-efficacy and self-management behaviors play mediating roles in the relationship between depressive symptoms and glycemic control among elderly T2DM patients.

Objective:To analyze the factors influencing the lung function of coal miners, identify the optimal combination of indicators for evaluating lung function, develop a risk assessment model using machine learning, and offer personalized risk assessment for workers.Methods:In June 2023, through cluster sampling, male underground workers who participated in occupational health examinations at a coal mine in North China from July to August 2018 were selected as the research subjects. Their health examination results and occupational environmental data were collected. A total of 3, 320 coal miners were included. Randomly divide the research subjects into a training set (2324 people) and a validation set (996 people) in a ratio of 7∶3, and the balance of the two sets was tested. Perform LASSO regression analysis using R 4.2.2 software to select relevant important variables, and determine the model's input variables by combining them with relevant literature. Utilize Python 3.8 to construct logistic regression, random forest, support vector machine, and XG Boost models, assess the models' discriminative ability using metrics like accuracy, sensitivity, specificity, F1 score, ROC curve, and AUC, evaluate the models' calibration using Brier score, Log loss score, and calibration curve, and further analyze the clinical performance of the developed models through DCA decision curve analysis.Results:Among the 3 320 coal miners, 856 had abnormal lung function (25.78%). The XG Boost model was identified as the optimal model, achieving a training set accuracy of 87.39%, sensitivity of 86.60%, specificity of 87.67%, F1 score of 0.779, AUC of 0.945, Brier score of 0.071, Log loss of 0.267 and demonstrated good calibration curve consistency.Conclusion:The XG Boost model exhibits superior predictive performance compared to other models, and the model has high application value. The Shapley Additive Explanation (SHAP) method is employed for interpretation, making it a reliable basis for preventing abnormal lung function in coal miners.

Objective:To investigate the occurrence of work-related musculoskeletal disorders (WMSDs) among underground coal mine workers, identify the risk factors for WMSDs, and provide a scientific evidence for the prevention and treatment of WMSDs.Methods:In March 2024, through cluster sampling, the on-the-job workers who underwent questionnaire surveys and health examinations at a certain coal mine from July to August 2018 were selected as the research subjects. Basic information of employees, ergonomics-related characteristics, and the occurrence status of WMSDs in each part were collected, and multivariate logistic regression was used for analysis.Results:The incidence rate of WMSDs in at least one site among underground coal mine workers within the past year was 62.22% (219/352). The top three sites in sequence were the lower back (44.32%, 156/352), neck (26.14%, 92/352), and knee (26.14%, 92/352). Multivariate logistic regression analysis showed that frequently exerting great force with arms or hands during work ( OR=2.223, 95% CI: 1.022-4.836), prolonged static forward bending ( OR=1.544, 95% CI: 1.305-1.972), and frequently exerting great effort to operate tools or machines ( OR=2.206, 95% CI: 1.011-4.813), absence of external support systems ( OR=1.589, 95% CI: 1.349-1.996), and repetitive full-body twisting ( OR=1.523, 95% CI: 1.298-1.916) were all risk factors for the occurrence of WMSDs in the lower back ( P<0.05). Both night shift work ( OR=1.564, 95% CI: 1.339-1.939) and frequent forward neck flexion ( OR=1.532, 95% CI: 1.312-1.907) were all risk factors for the occurrence of WMSDs in the neck ( P<0.05). Lifting heavy objects above the shoulder ( OR=1.333, 95% CI: 1.142-1.782), uncomfortable posture and inability to exert force ( OR=1.873, 95% CI: 1.104-2.712), the use of vibration tools ( OR=2.958, 95% CI: 1.255-6.972), and length of service >10 years ( OR=1.525, 95% CI: 1.105-1.967) were all risk factors for the occurrence of WMSDs in the knee ( P<0.05) . Conclusion:The incidence of WMSDs among underground coal miners is relatively high, mainly concentrated in the lower back, neck and knee, and is related to factors such as poor working postures, and work organization. Coal mining enterprises should strengthen work organization, provide appropriate working equipment, and ensure reasonable distribution of workloads.

Objective:To identify risk factors influencing the incidence of hypertension among steelworkers (Homo sapiens) and establish an effective and easily implementable hypertension prediction model.Methods:In September 2023, 2214 steelworkers (Homo sapiens) were selected as study subjects. Basic demographic information, lifestyle, and occupational exposure data were collected, along with physiological measurements such as height, weight, and blood pressure. Multivariate unconditional logistic regression analysis was employed based on relevant literature to determine influencing factors for hypertension among steelworkers (Homo sapiens). Python 3.9 software was used to construct and compare logistic regression, support vector machine (SVM), random forest, extreme gradient boosting tree (XGBoost), and LGBM models. Model performance was evaluated using metrics such as receiver operating characteristic (ROC) curves, accuracy, calibration curves, and F1 scores. The Shapley Additive Explanations (SHAP) model was introduced for feature importance analysis to enhance the interpretability of the prediction model.Results:A total of 432 cases of hypertension were detected among 2214 study subjects, with a detection rate of 19.51%. Age, smoking status, salt intake, use of cooling equipment, carbon monoxide exposure, family history of hypertension, fasting blood glucose, triglycerides, and hemoglobin were identified as independent risk factors for hypertension ( P<0.05). A comparison of the five models revealed the following performance metrics: logistic regression achieved an accuracy of 0.853, F1 score of 0.680, Brier score of 0.108, and AUC of 0.907; SVM demonstrated an accuracy of 0.863, F1 score of 0.687, Brier score of 0.081, and AUC of 0.910; random forest showed an accuracy of 0.857, F1 score of 0.603, Brier score of 0.105, and AUC of 0.861; XGBoost yielded an accuracy of 0.850, F1 score of 0.684, Brier score of 0.117, and AUC of 0.899; and the LGBM model exhibited an accuracy of 0.838, F1 score of 0.625, Brier score of 0.112, and AUC of 0.870. Conclusion:The SVM model demonstrated strong predictive performance, effectively assessing the risk of hypertension among steelworkers (Homo sapiens) and facilitating targeted health management interventions.