ObjectiveProtein-protein interactions (PPIs) are fundamental to the execution of biological functions within living cells. However, traditional biochemical methods, such as co-immunoprecipitation (Co-IP), often fail to capture transient, weak, or membrane-associated interactions due to the stringent detergent requirements for cell lysis. Proximity labeling (PL) has emerged in recent years as a transformative technology for mapping the proteomes of specific subcellular compartments and identifying dynamic interactomes in situ. Golgi protein 73 (GP73, also known as GOLPH2), a resident type II Golgi transmembrane protein, is a well-recognized clinical biomarker for liver diseases, including hepatocellular carcinoma (HCC). Despite its clinical significance, the comprehensive physiological and pathological functions of GP73 remain partially understood. This study aims to establish an APEX2-mediated proximity labeling system specifically targeting GP73 to map its interactome in a living cellular environment, thereby providing new insights into its molecular roles and regulatory mechanisms. MethodsTo achieve spatial specificity, we first constructed a stable cell line expressing a fusion protein consisting of GP73 and the engineered soybean peroxidase APEX2. The localization of the GP73-APEX2 fusion protein was validated to ensure it correctly targeted the Golgi apparatus. The proximity labeling reaction was initiated by incubating the cells with biotin-phenol (BP) for 30 min, followed by a brief (1 min) treatment with1 mmol/L hydrogen peroxide (H₂O₂). This catalytic reaction converts BP into highly reactive, short-lived biotin-phenoxyl radicals that covalently attach to endogenous proteins within a small labeling radius of the GP73-APEX2 enzyme. Subsequently, the cells were quenched, and biotinylated proteins were enriched using high-affinity streptavidin-coated magnetic beads. The captured “neighbor” proteins were subjected to on-bead digestion and analyzed via liquid chromatography-tandem mass spectrometry (LC-MS/MS) for high-throughput identification. Rigorous bioinformatics analysis, including Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and protein-protein interaction network mapping, was performed to interpret the biological significance of the identified candidates. ResultsOur results demonstrate the successful establishment of a robust and sensitive APEX2-based proximity labeling system for GP73. We identified a total of 95 high-confidence interacting proteins that were significantly enriched in the GP73 proximity proteome compared to control groups. Bioinformatics analysis revealed that these interactors were predominantly associated with biological processes such as vesicular transport, protein localization, and, most notably, molecular functions related to “ribosome binding” and “translation regulation”. This suggested an unexpected role for the Golgi-resident GP73 in the cellular translation machinery. To validate these findings, we performed targeted biochemical assays which confirmed a direct interaction between GP73 and the subunits of the eukaryotic translation initiation factor 3 (eIF3) complex, specifically EIF3G and EIF3I. Furthermore, functional validation using the surface sensing of translation (SUnSET) assay—a non-radioactive method to monitor protein synthesis—revealed that the overexpression of GP73 significantly promoted global protein translation levels in the cell, whereas its depletion or inhibition resulted in reduced translation efficiency. ConclusionThis study successfully utilized APEX2-mediated proximity labeling to provide the first systematic map of GP73 interactome in living cells. Our findings uncover a novel, unconventional function of GP73 as a regulator of cellular protein translation, likely mediated through its interaction with the eIF3 complex. This discovery significantly broadens our understanding of the biological roles of GP73 beyond its traditional function in the Golgi apparatus and suggests that it may act as a bridge between Golgi-related trafficking and the protein synthesis machinery. Furthermore, the technical framework established in this study provides a valuable template for investigating other complex organelle-associated protein networks and resolving transient macromolecular interactions in various physiological and pathological contexts.

Objective To explore the prevalence of depressive symptoms in postoperative patients with ovarian cancer and to analyze its influencing factors from multiple dimensions, including clinical characteristics, psychological factors, and laboratory indicators. Methods A cross-sectional study was conducted, which enrolled 235 postoperative patients with ovarian cancer. Depressive status was assessed using the patient health questionnaire, and the demographic, pathological, and medical record data of the patients were collected using the generalized anxiety disorder scale, Pittsburgh sleep quality index, European organization for research and treatment of cancer quality of life questionnaire core 30, and ECOG performance status score. Peripheral blood tumor marker (CA125), routine blood test, lymphocyte subsets, and serum cytokine levels were measured. Univariate and multivariate binary logistic regression analysis were used for statistical analysis. Results The prevalence of depression in postoperative patients with ovarian cancer was 39.15% (92/235). Univariate analysis showed that ECOG score ≥ 2 points, pain, anxiety, poor sleep quality, low quality of life, low life satisfaction, tumor recurrence, six or more cycles of chemotherapy, as well as higher levels of CA125, NLR, and NAR, and lower hemoglobin levels were significantly associated with depression (all P<0.05). Multivariate binary Logistic regression analysis showed that anxiety (OR=1.975, 95%CI: 1.231-3.170), sleep efficiency (OR=4.181, 95%CI: 1.211-14.43), sleep latency (OR=34.806, 95%CI: 4.258-284.542), ECOG performance status score, cognitive function (OR=0.918, 95%CI: 0.868-0.97), and life satisfaction were independent risk factors for depression (all P<0.05). Laboratory indicators were not independent influencing factors in the multivariate Logistic regression model. Conclusion Depression in postoperative patients with ovarian cancer is influenced by physiological, psychological, and social factors. Clinical management should focus on patients with anxiety, sleep disorders, poor physical condition, and low life satisfaction, and a comprehensive prevention and treatment strategy centered on psychological intervention and taking into account symptom management and social support should be implemented.

BACKGROUND: Biomarkers-based prediction of long-term risk of acute coronary syndrome (ACS) is scarce. We aim to develop a risk score integrating clinical routine information (C) and plasma biomarkers (B) for predicting long-term risk of ACS patients. METHODS: We included 2729 ACS patients from the OCEA (Observation of cardiovascular events in ACS patients). The earlier admitted 1910 patients were enrolled as development cohort; and the subsequently admitted 819 subjects were treated as validation cohort. We investigated 10-year risk of cardiovascular (CV) death, myocardial infarction (MI) and all cause death in these patients. Potential variables contributing to risk of clinical events were assessed using Cox regression models and a score was derived using main part of these variables. RESULTS: During 16,110 person-years of follow-up, there were 238 CV death/MI in the development cohort. The 7 most important predictors including in the final model were NT-proBNP, D-dimer, GDF-15, peripheral artery disease (PAD), Fibrinogen, ST-segment elevated MI (STEMI), left ventricular ejection fraction (LVEF), termed as CB-ACS score. C-index of the score for predication of cardiovascular events was 0.79 (95% CI: 0.76-0.82) in development cohort and 0.77 (95% CI: 0.76-0.78) in the validation cohort (5832 person-years of follow-up), which outperformed GRACE 2.0 and ABC-ACS risk score. The CB-ACS score was also well calibrated in development and validation cohort (Greenwood-Nam-D'Agostino: P = 0.70 and P = 0.07, respectively). CONCLUSIONS CB-ACS risk score provides a useful tool for long-term prediction of CV events in patients with ACS. This model outperforms GRACE 2.0 and ABC-ACS ischemic risk score.

Objective:To examine frailty in residents aged over 50 in the Chengdu community, investigate the risk factors linked to frailty, and analyze the impact of exercise habits on frailty.Methods:This retrospective study utilized a stratified whole-sample method to select Chengdu residents aged over 50 from 6 communities.Relevant information was collected through face-to-face interactions using a self-developed frailty questionnaire between May 2022 and May 2023.Exercise habits were defined as engaging in physical activity more than three times a week for at least 30 minutes per session, consistently maintained for a minimum of five years prior to the survey.The frailty index(FI)was established based on the multiple-deficit cumulative model.Respondents were classified into categories of no frailty(FI<0.1), mild frailty(0.1 ≤ FI<0.2), and moderate-severe frailty(FI≥0.2).Univariate and multivariate logistic regression analyses were conducted to investigate the factors influencing frailty and to compare the prevalence and progression of frailty among individuals with and without exercise habits across different age groups.Results:A total of 999 participants, comprising 483(48.3%)males and 516(51.7%)females, were enrolled in the study.The average age of the participants was 65.70±9.61 years.Among them, 346(34.6%)were classified as having mild frailty, while 107(10.7%)had moderate-severe frailty.Univariate analysis revealed that age, gender, marital status, education, smoking, alcohol consumption, and exercise habits were potential factors influencing frailty( P<0.05).Multifactorial logistic regression models indicated that individuals aged 70-79 and 80-85 were predictors of mild and moderate-severe frailty, respectively.Being female was identified as an independent risk factor for both mild frailty( OR: 1.196; 95% CI: 1.262-3.073)and moderate-severe frailty( OR: 2.721; 95% CI: 1.282-5.774).Additionally, not engaging in regular exercise was associated with an increased risk of mild frailty( OR: 1.588; 95% CI: 1.149-2.193)and moderate-severe frailty( OR: 3.186; 95% CI: 1.819-5.583).Statistical analysis revealed significant differences in frailty distribution and frailty index changes between individuals with and without exercise habits over the age of 60( P<0.05). Conclusions:Age and gender were found to be closely associated with frailty, with the impact of exercise habits on frailty becoming more pronounced after the age of 60.Encouraging middle-aged and older individuals to adopt regular exercise routines can play a significant role in promoting healthy aging in China.

Biomolecular condensates are formed through phase separation of biomacromolecules such as proteins and RNAs. These condensates exhibit liquid-like properties that can futher transition into more stable material states. They form complex internal structures via multivalent weak interactions, enabling precise spatiotemporal regulations. However, the use of inconsistent and non-standardized terminology has become increasingly problematic, hindering academic exchange and the dissemination of scientific knowledge. Therefore, it is necessary to discuss the terminology related to biomolecular condensates in order to clarify concepts, promote interdisciplinary cooperation, enhance research efficiency, and support the healthy development of this field.

Objective To explore the effect of a shared care applied to pregnancy care among pregnant women with hypertensive disorder of pregnancy(HDP).Methods A total of 88 pregnant women with HDP who received ob-stetric check-ups in a tertiary A-level matemity hospital in Nanjing City from January to October 2023 were conve-niently selected as the research subjects.According to the order of obstetric examinations,44 pregnant women with HDP from June to October 2023 were selected as an experimental group,and 44 pregnant women with HDP from January to May 2023 were selected as a control group.The experimental group received the shared care clinic management model,and the control group received the routine management.The scores of the rate of blood pres-sure attainment,the incidence of disease progression,the questionnaire on knowledge of HDP,the Hypertension Self-Management Scale,the Pregnancy-Related Anxiety Scale,and the Patient Satisfaction Questionnaire were compared between the 2 groups.Results 41 patients in the experimental group and 39 patients in the control group com-pleted the study.After the intervention,the blood pressure compliance rate of pregnant women with HDP in the experimental group was 95.12％,higher than 79.49％in the control group.The incidence of disease progression was 7.32％,lower than 25.64％in the control group.HDP Knowledge Questionnaire score(25.24±4.07),Hypertension Self-Management Scale score(114.39±14.89),and Patient Satisfaction Questionnaire score(19.56±1.14)were higher than that of the control group.The score of Pregnancy-Related Anxiety Scale was(18.02±2.15),lower than(23.18±9.58)in the control group.The score of Patient Satisfaction Questionnaire was(19.56±1.14),higher than that(17.26±2.25)in the control group.The differences were statistically significant(P＜0.001).Conclusion The shared care clinic model can improve HDP pregnant women's blood pressure control compliance,HDP knowledge,and self-management,reduce the incidence of disease progression,pregnancy-related anxiety,and improve satisfaction,which provides a reference for optimising the HDP management model.

Objective To investigate the correlation between dual-energy computed tomography angiography(CTA)parameters of carotid plaques and acute stroke events.Methods A retrospective analysis was conducted on the clinical and imaging data of patients who underwent dual-energy head and neck CTA and brain MRI scans.Utilizing the Siemens workstation(Syngo.Via VB40B),region of interest(ROI)were placed on the thickest slice of the carotid plaque in the axial plane to obtain parameters such as fat fraction(FF),virtual non-contrast(VNC)value,iodine concentration(IC),electron density(Rho),effective atomic number(Zeff),dual energy index(DEI),spectral curve,and corresponding CT values at 40 keV(40 keVHU)and 90 keV(90 keVHU).The slope of the energy spectrum curve(λ)was calculated within the 40 keV-90 keV range.Patients with acute cerebral infarction(ACI)in the ipsilateral anterior circulation territory were classified into the ACI group,while those without were classified into the non-acute cerebral infarction(NACI)(NACI group).Qualitative data were analyzed using the x2 test,and quantitative data were analyzed using the t-test.The predictive performance was assessed using the area under the curve(AUC)of the receiver operating characteristic(ROC)curve,and the differences between different ROC curves were compared using the DeLong test.Results A total of 72 patients were included,with 21 in the ACI group and 51 in the NACI group.The mean values of FF,Zeff,and 40 keVHU in the ACI group were greater than those in the NACI group.Statistically significant differences were observed between the groups for Zeff,DEI,40 keVHU,and λ(P＜0.05).40 keVHU demonstrated the highest predictive performance,and the AUC,sensitivity,and specificity was 0.789,81.0％,and 74.5％,respectively.A combined variable constructed through logistic regression analysis yielded an AUC,sensitivity,and specificity of 0.796,85.7％,and 70.6％,respectively,with no significant statistical differences compared to single factor variables.Conclusion Dual-energy CTA parameters of carotid plaques may aid in predicting intraplaque hemorrhage(IPH)and the occurrence of acute stroke events.

The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose-catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16 ℃ in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni-NTA affinity column.Size-exclusion chromatography and SDS-PAGE analysis demonstrated that the puri-fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD+as a coenzyme to NADP+.The optimal temperature and pH of the TeXDH were 40 ℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26％of the initial enzyme activity,while the divalent metal ions Mg2+or Ca2+could recover the enzyme activity of TeXDH,reaching 103.32％and 110.69％of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn2+,Ni2+,Cu2+,Zn2+,Co2+,and Cd2+significantly inhibited the activity of TeXDH.ICP-MS and molecular doc-king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn2+ions and 1 mol/L Mg2+ion.Further-more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.

Aim To investigate the protective effect of liraglutide(LIRA)on acetaminophen(APAP)-in-duced hepatotoxicity at the in vivo level and to reveal the underlying mechanism.Methods Forty SPF grade male C57BL/6J mice were randomly divided into the Control,LIRA(200 μg·kg-1),APAP(500 mg·kg-1),LIRA+APAP,LIRA+APAP+3-methylade-nine(3-MA,30 mg·kg-1)groups,with eight mice in each group.The mice were administered for three con-secutive days,and the materials were taken after 24 h.The general condition and body weight of mice in each group were recorded,and liver morphology was ob-served.Serum ALT and AST levels,as well as SOD ac-tivity,MDA,and GSH content in liver homogenates,were measured using biochemical assay kits.The levels of inflammatory cytokines IL-6,TNF-α,and IL-1β in serum were detected by ELISA.Liver pathological changes were assessed by HE staining,while mitochon-drial and autophagosome structures in liver tissues were observed using transmission electron microscopy.The number of PCNA-positive cells in liver tissues was e-valuated using immunohistochemical staining.The pro-tein expression levels of LC3Ⅱ,p62,Bax,Bcl-2,PC-NA,and CyclinD1 in liver tissues were determined by Western blot.Results LIRA pretreatment can im-prove the general condition of mice with acetamino-phen-induced liver injury(AILI),reduce serum ALT and AST levels,and effectively ameliorate the appear-ance and morphology of the liver as well as the patho-logical damage to liver tissue.Simultaneously,the lev-els of inflammatory cytokines IL-6,TNF-α,and IL-1βare significantly decreased;SOD activity and GSH con-tent are significantly increased,while MDA content is significantly reduced.Transmission electron microsco-py observations reveal the presence of numerous auto-phagosomes in the cytoplasm of liver tissue.Immuno-histochemical staining results indicate a significant in-crease in the number of PCNA-positive cells.Further-more,the expression of LC3Ⅱ,Bcl-2,PCNA,and Cy-clinD1 proteins in liver tissue is significantly upregulat-ed,while the expression of p62 and Bax proteins is significantly downregulated.However,after interven-tion with the autophagy inhibitor 3-MA,the aforemen-tioned protective effects of LIRA are significantly.Conclusions LIRA pretreatment can significantly im-prove liver injury in AILI mice.Its protective mecha-nism may be related to enhancing autophagy in hepato-cytes,thereby reducing oxidative stress,inflammatory response and apoptosis in liver of AILI mice.

Aim To investigate the effect of p-AMPK activity on autophagy in different subtypes of MDA-MB-231(triple-negative breast cancer cells)and MCF-7(estrogen receptor-positive cells)and its regulatory mechanism.Methods MDA-MB-231 cells were trea-ted with EBSS,Baf-A1,and EBSS+Baf-A1 for four hours,and MCF-7 cells for eight hours.The effects of autophagy on cell proliferation and apoptosis were ob-served,mitochondrial morphology was examined,and the expression of autophagy markers LC3B,P62,LAMP1,TOM20,AMPK,p-AMPK,ULK1,and Bec-lin1/VPS34 proteins was detected.The autophagy pathway was validated by inhibiting AMPK activity.Results Breast cancer cells underwent autophagy af-ter starvation induction(EBSS),with inconsistent au-tophagy processes observed in different subtypes of breast cancer cells.Autophagy inhibited cell prolifera-tion.In MDA-MB-231 cells,autophagy led to an in-crease in p-AMPK levels and a decrease in ULK1 lev-els,initiating autophagy through p-AMPK activation of ULK1.In MCF-7 cells,both p-AMPK and ULK1 levels decreased after autophagy,suggesting that autophagy might not be mediated by p-AMPK activation.Conclu-sions MDA-MB-231 cells primarily initiate autophagy by directly activating ULK1 by p-AMPK,independent of the MTOR pathway.In MCF-7 cells autophagy might be triggered by inhibiting MTOR through AMPK activity or directly activating MTOR through other up-stream factors.Regulating p-AMPK activity based on the autophagy pathways in different cell subtypes could enable more precise targeting and treatment of different types of breast cancer.