Background: Adrenocortical hormones, particularly 11-oxygenated androgens, are pivotal in female reproductive health and fertility. Standardized detection kits and population-specific reference intervals are lacking in China, hindering related clinical applications. Methods: A HPLC-tandem mass spectrometry (HPLC-MS/MS) pipeline was developed, rigorously validated, and applied to simultaneously quantify corticosterone, cortisone, cortisol, 18-OH cortisol, androstenedione (A4), 11β-hydroxyandrostenedione (11-OH A4), dehydroepiandrosterone, and dehydroepiandrosterone sulfate in serum samples from 455 reproductive-aged women (18–45 yrs) in Guangxi, China. Age-dependent concentration trends were analyzed, and reference intervals stratified by age (2.5th to 97.5th percentiles) were established. Correlations with body-composition metrics, ethnicity, and the menstrual cycle were investigated. Results: The HPLC-MS/MS method demonstrated high precision (intra- and inter-assay CVs < 15%), accuracy, and sensitivity. All eight hormones exhibited significant age-related declines (P < 0.001 for seven hormones; P = 0.001 for 11-OH A4). Notably, 11-OH A4 levels were significantly lower in the 35–45-yr (3.05 nmol/L) and 25–34-yr (3.09 nmol/L) age groups than in the 18–24-yr (3.57 nmol/L) age group, whereas no significant difference was observed between the 35–45-yr and 25–34-yr age groups. Weak negative correlations were observed between the body mass index and corticosterone and cortisone levels, whereas ethnicity and the menstrual cycle showed no significant associations with hormone levels. Conclusions We developed an HPLC-MS/MS-based method for simultaneously quantifying eight adrenocortical hormones, including 11-OH A4, and defined age-specific reference intervals for reproductive-aged Chinese women. These findings advance the clinical utility of adrenocortical hormones in diagnosing and managing reproductive disorders.

Background Depressive symptoms have become a significant factor affecting the physical and mental health of the occupational population, and workers in petroleum refining enterprises face multiple stressors in their work environment. Objective To explore the impact of occupational noise exposure on depressive symptoms among workers in a petroleum refining enterprise. Methods This cross-sectional study was conducted in July 2024 using a questionnaire survey among workers of a petroleum refining enterprise in Hainan Province. Basic information of the subjects was collected. The Center for Epidemiologic Studies Depression Scale (CES-D) was used to measure depressive symptoms, the Chinese version of the Pittsburgh Sleep Quality Index (PSQI) scale was used to assess sleep quality, and the Chinese version of the Effort-Reward Imbalance (ERI) scale was used to evaluate occupational stress. Chi-square test was employed to compare the differences in reporting depressive symptoms among populations with different characteristics. Binary logistic regression models were used to analyze the impact of occupational noise exposure and other factors on depressive symptoms. Results The overall positive rate of depressive symptoms in the study population was 42.7%. The results of the multifactor analysis indicated that compared with the control group, employees in both the low-exposure and high-exposure groups had elevated odds of depressive symptoms, with OR (95%CI) of 2.244 (1.131, 4.454) and 1.970 (1.009, 3.850), respectively. This association remained robust after adjusting for potential confounders, including gender, age, work tenure, and other occupational exposures. Additionally, female [OR (95%CI)=1.483 (1.039, 2.118)], exposure to benzene, toluene, or xylene [OR (95%CI)=1.621 (1.208, 2.174)], sleep disturbance [OR (95%CI)=3.772 (2.942, 4.838)], and occupational stress [OR (95%CI)=2.018 (1.575, 2.585)] were also significantly associated with higher odds of depressive symptoms. Conclusion The positive rate of depressive symptoms is relatively high among employees in this petrochemical enterprise, and occupational noise exposure may be a risk factor for depressive symptoms.

To explore the advantages of traditional Chinese medicine （TCM） and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications （DMC）， refine key pathophysiological insights and treatment principles， and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine， hosted by the China Association of Chinese Medicine， was held in Beijing， 2024. Experts in TCM， Western medicine， and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis， diagnostic and treatment challenges， and mechanism research related to DMC， ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development， research prioritization， and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM， strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.

ObjectiveTo investigate the role of DEAD-box helicase 3 X-linked （DDX3X） in immune-mediated liver injury （ILI）， and to clarify its mechanism by regulating endoplasmic reticulum stress （ERS）-dependent apoptotic pathway and its association with the clinical progression of hepatitis B. MethodsMice were given injection of concanavalin A （ConA） via the caudal vein to establish a model of ILI， PBS （control group） and different concentrations of ConA were injected into the tail vein of hepatocyte-specific DDX3X-knockout mice （DDX3X^ΔHep and DDX3X-flox mice （DDX3X^fl/fl）， respectively.. The log-rank survival analysis， measurement of the serum levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）， and HE staining of liver tissue were performed to assess liver injury， and qRT-PCR and Western Blot were used to measure the mRNA and protein expression levels of glucose-regulated protein 78 （GRP78）， CCAAT/enhancer-binding protein homologous protein （CHOP）， and DDX3X in liver tissue. Intraperitoneal injection of 4-phenylbutyric acid （4-PBA， 100 mg/kg） was performed to inhibit ERS. Serum samples （n=30） and liver tissue samples （n=6） were collected from healthy controls， chronic hepatitis B （CHB） patients， and hepatitis B virus-associated liver failure （HBV-LF） patients； ELISA was used to measure the serum level of DDX3X， and qRT-PCR/Western Blot was used to analyze the expression of targets in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group of mice， the expression of DDX3X in the liver of mice induced by ConA was significantly increased after liver injury （P<0.05）， and hepatocyte-specific DDX3X knockout increased the 72-hour survival rate of mice by 55% （compared with 20% in the DDX3X^fl/fl group）， with significant reductions in the serum levels of ALT and AST （P<0.000 1） and the expression levels of the ERS markers GRP78 and CHOP （P<0.05）. After ERS was inhibited by 4-PBA， there was alleviation of liver injury （with reductions in ALT and AST， P <0.001） and a reduction in DDX3X expression （P<0.01）. The analysis of clinical samples showed that the mRNA and protein expression levels of liver DDX3X in CHB patients and HBV-LF patients were significantly higher than those in healthy controls （all P<0.01）， and there was a significant increase in the serum level of DDX3X in HBV-LF patients （P<0.000 1）. ConclusionDDX3X exacerbates ILI by regulating the ERS-dependent apoptotic pathway （GRP78/CHOP）， and its expression is associated with the progression of hepatitis B. Therefore， it can be used as a potential therapeutic target.

ObjectiveTo reveal the compatibility rules of the core compound formula and traditional Chinese medicine （TCM） components for prevention and treatment of chronic atrophic gastritis （CAG） by Huazhuo Jiedu （turbidity-resolving and detoxifying） therapy through uniform design in view of the potential regulatory pathway of endogenous turbid-toxin retention in CAG， elucidate the potential pathway by which the compound formulas regulate the Wnt/β-catenin-connexin 43 （CX43） signaling axis， demonstrated the efficacy enhancement and toxicity attenuation among the components， thus providing experimental evidence for the scientific application of multi-component， multi-target compatibility in TCM compound formulas. MethodsA mouse model of CAG was established with N-methyl-N-nitrosourea （MNU）. Sixty mice were randomly allocated into 5 groups （n=12 per group）： blank control， model， low-dose Huazhuo Jiedu core compound formula （HXFF-L， 4.004 g·kg^-1）， high-dose Huazhuo Jiedu core compound formula （HXFF-H， 8.008 g·kg^-1）， and positive control （folic acid at 11 mg·kg^-1 + vitamin B₁₂ at 14 mg·kg^-1）. The therapeutic efficacy and molecular mechanism of the core compound formula were evaluated by hematoxylin-eosin （HE） staining， Alcian blue-periodic acid-Schiff （AB-PAS） staining， enzyme-linked immunosorbent assay （ELISA）， transmission electron microscopy （TEM）， immunofluorescence assay， and Western blot. The chemical components of the core compound formula were detected and effective components were screened by ultra-high performance liquid chromatography coupled with quadrupole-orbitrap high-resolution mass spectrometry （UHPLC-Q-Orbitrap HRMS）. Another 120 mice were randomized into blank control， model， and 8 HXFF component-combination groups with different dosages （uniform design groups 1-8， n=12 per group）. A uniform design was adopted， with pathological scores of mouse gastric tissue as well as serum levels of pepsinogen Ⅰ （PGⅠ）， pepsinogen Ⅱ （PGⅡ）， and gastrin 17 （GAS17） set as dependent variables. Multivariate linear regression analysis was performed to predict the optimal proportion of core components in the compound formula and explore its compatibility rules. ResultsPharmacodynamic validation showed that compared with the blank control group， the model group had decreased gastric weight index （P<0.01） and increased pathological damage score and inflammatory factor levels （P<0.01）. Compared with the model group， these indicators were reversed in the HXFF-L， HXFF-H， and positive control groups （P<0.05， P<0.01）. Immunofluorescence results showed that CX43 expression was downregulated in the model group compared with the blank control group （P<0.01） and upregulated in the HXFF-H and positive control groups compared with the model group （P<0.05， P<0.01）. Western blot results showed that compared with the blank control group， the model group demonstrated upregulated protein levels of Wnt and β-catenin in the gastric tissue （P<0.01）. Compared with those in the model group， the protein level of Wnt was downregulated in the HXFF-L， HXFF-H， and positive control groups （P<0.05， P<0.01）， while that of β-catenin was downregulated only in the HXFF-H group （P<0.05）. A total of 25 compounds were identified in the core compound formula， including 9 from Coptidis Rhizoma， 10 from Pogostemonis Herba， 2 from Scorpio， and 4 from Lilii Bulbus. According to the category and origin of compounds， Coptidis Rhizoma alkaloids， Pogostemonis Herba volatile oils， Scorpio peptides， and Lilii Bulbus alkaloids were finally selected for component research. The predicted optimal combination of TCM components was as follows： Coptidis Rhizoma alkaloids at 75 mg·kg^-1， Pogostemonis Herba volatile oils at 2.25 mg·kg^-1， Scorpio protein extract at 90 mg·kg^-1， and Lilii Bulbus alkaloids at 9 mg·kg^-1. Coptidis Rhizoma alkaloids， Scorpio protein extract， Pogostemonis Herba volatile oils， and Lilii Bulbus alkaloids can ameliorate pathological lesions of the gastric tissue. Scorpio protein extract exerted positive regulatory effects on the gastric function， while Pogostemonis Herba volatile oils and Lilii Bulbus alkaloids showed negative correlations with partial gastric function indicators. Further investigations are required to clarify their dose-effect relationships and safety profiles. ConclusionThis study， for the first time， predicts the optimal proportion and synergistic regulatory mechanism of the core compound formula of Huazhuo Jiedu therapy， revealing the synergistic compatibility logic of its components. It provides a novel strategy for the precise TCM treatment of CAG and the modern development of compound formulas.

ObjectiveTo reveal the compatibility rules of the core compound formula and traditional Chinese medicine （TCM） components for prevention and treatment of chronic atrophic gastritis （CAG） by Huazhuo Jiedu （turbidity-resolving and detoxifying） therapy through uniform design in view of the potential regulatory pathway of endogenous turbid-toxin retention in CAG， elucidate the potential pathway by which the compound formulas regulate the Wnt/β-catenin-connexin 43 （CX43） signaling axis， demonstrated the efficacy enhancement and toxicity attenuation among the components， thus providing experimental evidence for the scientific application of multi-component， multi-target compatibility in TCM compound formulas. MethodsA mouse model of CAG was established with N-methyl-N-nitrosourea （MNU）. Sixty mice were randomly allocated into 5 groups （n=12 per group）： blank control， model， low-dose Huazhuo Jiedu core compound formula （HXFF-L， 4.004 g·kg^-1）， high-dose Huazhuo Jiedu core compound formula （HXFF-H， 8.008 g·kg^-1）， and positive control （folic acid at 11 mg·kg^-1 + vitamin B₁₂ at 14 mg·kg^-1）. The therapeutic efficacy and molecular mechanism of the core compound formula were evaluated by hematoxylin-eosin （HE） staining， Alcian blue-periodic acid-Schiff （AB-PAS） staining， enzyme-linked immunosorbent assay （ELISA）， transmission electron microscopy （TEM）， immunofluorescence assay， and Western blot. The chemical components of the core compound formula were detected and effective components were screened by ultra-high performance liquid chromatography coupled with quadrupole-orbitrap high-resolution mass spectrometry （UHPLC-Q-Orbitrap HRMS）. Another 120 mice were randomized into blank control， model， and 8 HXFF component-combination groups with different dosages （uniform design groups 1-8， n=12 per group）. A uniform design was adopted， with pathological scores of mouse gastric tissue as well as serum levels of pepsinogen Ⅰ （PGⅠ）， pepsinogen Ⅱ （PGⅡ）， and gastrin 17 （GAS17） set as dependent variables. Multivariate linear regression analysis was performed to predict the optimal proportion of core components in the compound formula and explore its compatibility rules. ResultsPharmacodynamic validation showed that compared with the blank control group， the model group had decreased gastric weight index （P<0.01） and increased pathological damage score and inflammatory factor levels （P<0.01）. Compared with the model group， these indicators were reversed in the HXFF-L， HXFF-H， and positive control groups （P<0.05， P<0.01）. Immunofluorescence results showed that CX43 expression was downregulated in the model group compared with the blank control group （P<0.01） and upregulated in the HXFF-H and positive control groups compared with the model group （P<0.05， P<0.01）. Western blot results showed that compared with the blank control group， the model group demonstrated upregulated protein levels of Wnt and β-catenin in the gastric tissue （P<0.01）. Compared with those in the model group， the protein level of Wnt was downregulated in the HXFF-L， HXFF-H， and positive control groups （P<0.05， P<0.01）， while that of β-catenin was downregulated only in the HXFF-H group （P<0.05）. A total of 25 compounds were identified in the core compound formula， including 9 from Coptidis Rhizoma， 10 from Pogostemonis Herba， 2 from Scorpio， and 4 from Lilii Bulbus. According to the category and origin of compounds， Coptidis Rhizoma alkaloids， Pogostemonis Herba volatile oils， Scorpio peptides， and Lilii Bulbus alkaloids were finally selected for component research. The predicted optimal combination of TCM components was as follows： Coptidis Rhizoma alkaloids at 75 mg·kg^-1， Pogostemonis Herba volatile oils at 2.25 mg·kg^-1， Scorpio protein extract at 90 mg·kg^-1， and Lilii Bulbus alkaloids at 9 mg·kg^-1. Coptidis Rhizoma alkaloids， Scorpio protein extract， Pogostemonis Herba volatile oils， and Lilii Bulbus alkaloids can ameliorate pathological lesions of the gastric tissue. Scorpio protein extract exerted positive regulatory effects on the gastric function， while Pogostemonis Herba volatile oils and Lilii Bulbus alkaloids showed negative correlations with partial gastric function indicators. Further investigations are required to clarify their dose-effect relationships and safety profiles. ConclusionThis study， for the first time， predicts the optimal proportion and synergistic regulatory mechanism of the core compound formula of Huazhuo Jiedu therapy， revealing the synergistic compatibility logic of its components. It provides a novel strategy for the precise TCM treatment of CAG and the modern development of compound formulas.

Objective:To construct a method for hepatitis B virus (HBV) DNA detection based on recombinase-mediated isothermal amplification (RAA)-clustered regularly interspaced short palindromic repeats and their associated protein 13a (CRISPR-Cas13a).Methods:Through the alignment and screening of HBV DNA sequences, a positive plasmid was constructed, and recombinase-aided amplification (RAA) primers and CRISPR RNA (crRNA) were designed. A method for detecting HBV DNA based on the RAA-CRISPR-Cas13a system was developed, and the specificity and sensitivity were evaluated. Utilizing the CRISPR-Cas13a system, 70 clinical samples from HBV DNA-positive patients with various viral loads collected at Beijing You′an Hospital from 2019 to 2021 were analyzed. The detection results were further compared with those results using real-time quantitative polymerase chain reaction (qPCR).Results:The optimal RAA amplification primers and crRNA were first screened using the RAA-CRISPR-Cas13a method, with the sensitivities for detecting HBV DNA standards and for clinical samples at 1 IU/ml and<10 IU/ml, respectively, demonstrating specificity for HBV DNA detection. Compared with qPCR (the gold standard), the detection consistency between the two methods was 100% (70/70).Conclusion:This study established a method for detecting HBV DNA by integrating recombinase-aided amplification (RAA) technology with CRISPR/Cas13a technology.

Objective To explore the clinical and imaging manifestations of posterior tibial tendon dysfunction(PTTD).Methods A retrospective selection was conducted on 61 patients with clinically diagnosed PTTD.According to the classification criteria of Johnson and Strom,8 patients were classified as stage 1,17 as stage 2,20 as stage 3,and 16 as stage 4.The clinical manifestations and imaging features of patients were summarized at each stage.Results Among the 8 patients with stage 1 PTTD,no abnormalities were found on X-ray examination of the weight-bearing foot,while MRI revealed posterior tibial tendon(PTT)tendinitis in 4 cases(50.0％).For stage 2 patients(n=17),X-ray examination of the weight-bearing foot showed a decrease in the longitudinal arch of the foot in 15 patients(88.2％).MRI mainly revealed PTT degeneration in 12 cases(70.6％),PTT tears in 2 cases(11.8％),and other structural injuries in 5 cases(29.4％).For stages 3-4 patients(n=36),X-ray examination of the weight-bearing foot showed flatfoot in all cases.MRI mainly revealed PTT degeneration in 12 cases(33.3％),PTT tears in 24 cases(66.7％),spring ligament injuries in 33 cases(91.7％),and deltoid ligament injuries in 23 cases(63.9％).Conclusion Patients with stage 1 PTTD often have a normal foot longitudinal arch and may present with PTT tendinitis.Middle-aged females are more commonly affected by advanced PTTD,which is characterized by gradual decrease and collapse of the foot arch,accompanied by varying degrees of PTT degeneration and tears.The spring ligament injuries and deltoid ligament are frequently involved in structural injuries associated with PTTD.

Objective:To select low-dose radiation-activated genes with intrinsic radiation protection by developing a model for adaptive responses to low-dose ionizing radiation, in order to explore the mechanisms behind the radiation resistance of the candidate genes.Methods:The cells were divided into adaptive response induction group and whole transcriptome sequencing group. The level of DNA damage was assessed using the γ-H2AX immunofluorescence assay. The low-dose radiation-activated candidate genes with radiation protection were selected through whole transcriptome sequencing and quantitative reverse transcription PCR (RT-qPCR)-based validation. The anti-radiation effect of candidate gene CCND1 was assessed based on CCK-8 cell proliferation and γ-H2AX immunofluorescence assay. After up- and down-regulation of CCND1 expression, the anti-radiation mechanism of CCND1 was preliminarily explored through transcriptome sequencing analysis.Results:A model for low-dose ionizing radiation-induced adaptive responses of lymphocytes was constructed. Using this model, six candidate genes with radiation protection, including CCND1, ZMAT3, MGAT3, DFFB, CYP4F2, ITGA6, were selected. Compared to the control group, overexpressed CCND1 led to significantly enhanced proliferation ability of AHH-1 cells ( t = 7.92-14.76, P < 0.05) and distinctly lowered level of DNA damage ( t = 2.79-9.68, P < 0.05) after 2 Gy of X-ray irradiation. Furthermore, compared to the control group, the CCND1 knockdown caused significantly decreased cell proliferation ability ( t = 13.58-26.25, P < 0.05) and notably elevated level of DNA damage of cells ( t = 2.87-7.61, P < 0.05). Transcriptome sequencing revealed that up- and down-regulation of CCND1 expression resulted in the activation of pathways related to cell growth, death, and damage repair. Conclusions:By selecting six low-dose-activated candidate genes with radiation protection and revealing the function of CCND1 in radiation protection, this study provides a new perspective for the development of radiation protection agents from the perspective of adaptive responses to low-dose radiation.

Objective:To determine the prevalence, risk factors, and longitudinal associations of loneliness during mid- to late pregnancy with anxiety and depressive symptoms in late pregnancy.Methods:In this prospective cohort study, 1 107 pregnant women at 24-28 weeks' gestation were enrolled between June 2021 and December 2022. Psychological status was assessed during mid-pregnancy (24-28 weeks) and late pregnancy (≥32 weeks) using standardized electronic questionnaires, including the Revised University of California Los Angeles Loneliness Scale (UCLA) Loneliness Scale-Short Form (Cronbach's α=0.82), Patient Health Questionnaire-9 ( α=0.86), and Generalized Anxiety Disorder-7 ( α=0.88). Multivariate logistic regression identified independent risk factors for loneliness. Cross-lagged path models analyzed the longitudinal predictions between loneliness and anxiety/depressive symptoms. Results:The prevalence of loneliness decreased significantly from 10.8% (120/1 107) in mid-pregnancy to 4.8% (37/777) in late pregnancy ( χ2=21.81, P<0.001). Multivariate analysis identified independent risk factors for loneliness: age <30 years ( OR=1.70, 95% CI: 1.15-2.50), annual household income <50 000 CNY ( OR=2.53, 95% CI: 1.28-5.02), unemployment during pregnancy ( OR=1.57, 95% CI: 1.03-2.39), history of alcohol consumption ( OR=1.63, 95% CI: 1.03-2.56), and the presence of mid-pregnancy depressive ( OR=2.76, 95% CI: 1.51-5.04) and anxiety symptoms ( OR=1.65, 95% CI: 1.01-2.71) (all P<0.05). Cross-lagged path models indicated bidirectional associations between loneliness and both anxiety ( β=0.32, P<0.01) and depressive symptoms ( β=0.28, P<0.01). However, the predictive effect of loneliness on subsequent depressive and anxiety symptoms ( β=0.28-0.32) was substantially stronger than the reverse prediction (mid-pregnancy anxiety on late-pregnancy loneliness: β=0.12; mid-pregnancy depression on late-pregnancy loneliness: β=0.11). Loneliness demonstrated high temporal stability (autoregressive effects β=0.29-0.32). Conclusion:Loneliness in mid-pregnancy exhibits a symmetric bidirectional association with anxiety and depressive symptoms in late pregnancy, suggesting it may be a core driver in the development of these emotional symptoms. Younger maternal age (<30 years), low household income (<50 000 CNY/year), unemployment during pregnancy, and a history of alcohol consumption were associated with a higher risk of loneliness and should be prioritized for psychological screening and intervention.