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 analyze the incidence of surgical site infection (SSI) in patients undergoing colorectal cancer (CRC) surgery and to identify risk factors associated with SSI in an attempt to provide a reference for clinical prevention strategies.Methods:A retrospective cohort study was conducted. Clinical data were retrospectively collected from a total of 2,248 patients who underwent surgery for pathologically confirmed CRC between 2017 and 2022 at two centers: Huangshan Shoukang Hospital ( n=649) and Northern Jiangsu People's Hospital ( n=1 599). Inclusion criteria consisted of the following: (1) age >18 years; (2) pathologically confirmed CRC treated with curative resection, including extended resections (e.g. pelvic exenteration); (3) no surgical incisions other than abdominal or perineal; and (4) no use of prosthetic implants. The incidence of SSI was analyzed, and multivariate logistic regression was used to identify independent its risk factors. Results:A total of 121 patients (5.4%) developed SSI. Among them, 68 cases (56.2%) were organ/space infections, 35 cases (28.9%) were deep incisional infections, and 18 cases (14.9%) were superficial incisional infections. The median postoperative hospital stay was significantly longer in patients with SSI compared to those without (21.0 days vs. 13.0 days, U=65,754, P<0.001). The median hospitalization cost was also significantly higher in the SSI group (56,550 yuan vs. 43,645 yuan, U=72,008, P<0.001). Multivariate logistic regression analysis identified body mass index (BMI) ≤ 20 kg/m 2 (OR=4.25, 95%CI: 3.38-5.34, P<0.001), diabetes mellitus (OR=3.44, 95%CI: 1.89-6.24, P<0.001), open surgery (OR=4.23, 95%CI: 2.37-7.56, P<0.001), and colostomy or ileostomy (OR=1.67, 95% CI: 1.04-2.69, P=0.034) as independent risk factors for SSI. Conclusion:To prevent SSI following CRC surgery, attention should be given to optimizing body weight and glycemic control, promoting minimally invasive surgical approaches when feasible, and cautiously considering the necessity of colostomy or ileostomy.

Protein-protein interactions play an extremely important role in the biochemical functions of cells,and in-depth analysis of protein interactions is the key to understanding cellular life activities.In this study,we systematically mined the interacting proteins of Golgi protein 73(GP73)using classical immunoprecipitation combined with mass spectrometry,and sought to further analyze the molecular func-tion of GP73.Hepatocellular carcinoma cell line HepG2 was selected,and a stable cell line overexpress-ing GP73-3Flag was constructed using lentiviral infection technology.A total of 78 high-confidence GP73 interacting proteins were identified by immunoprecipitation coupled with mass spectrometry.Bioinformat-ics analyses suggested that GP73 interacted with nearly 40 cytosolic proteins and participated in the bio-logical processes of RNA transport,splicing,and translation.Further immunofluorescence and cytosolic protein isolation experiments confirmed the cytosolic localization of GP73 in a variety of tumor cells.Based on the 78 interacting proteins,we further screened protein interaction networks related to mRNA splicing and verified the existence of interactions between GP73 and seven proteins,including HNRN-PH3,SMN1,RBM14,andNCBP1,by co-immunoprecipitation experiments.In addition,minigene spli-cing assay results indicated that GP73 inhibited the splicing efficiency of pre-mRNA by cells.This study contributes to the expansion of knowledge regarding the function of GP73 and aids in elucidating its criti-cal role in cell biology and its potential association with diseases.

Objective:To analyze the incidence of surgical site infection (SSI) in patients undergoing colorectal cancer (CRC) surgery and to identify risk factors associated with SSI in an attempt to provide a reference for clinical prevention strategies.Methods:A retrospective cohort study was conducted. Clinical data were retrospectively collected from a total of 2,248 patients who underwent surgery for pathologically confirmed CRC between 2017 and 2022 at two centers: Huangshan Shoukang Hospital ( n=649) and Northern Jiangsu People's Hospital ( n=1 599). Inclusion criteria consisted of the following: (1) age >18 years; (2) pathologically confirmed CRC treated with curative resection, including extended resections (e.g. pelvic exenteration); (3) no surgical incisions other than abdominal or perineal; and (4) no use of prosthetic implants. The incidence of SSI was analyzed, and multivariate logistic regression was used to identify independent its risk factors. Results:A total of 121 patients (5.4%) developed SSI. Among them, 68 cases (56.2%) were organ/space infections, 35 cases (28.9%) were deep incisional infections, and 18 cases (14.9%) were superficial incisional infections. The median postoperative hospital stay was significantly longer in patients with SSI compared to those without (21.0 days vs. 13.0 days, U=65,754, P<0.001). The median hospitalization cost was also significantly higher in the SSI group (56,550 yuan vs. 43,645 yuan, U=72,008, P<0.001). Multivariate logistic regression analysis identified body mass index (BMI) ≤ 20 kg/m 2 (OR=4.25, 95%CI: 3.38-5.34, P<0.001), diabetes mellitus (OR=3.44, 95%CI: 1.89-6.24, P<0.001), open surgery (OR=4.23, 95%CI: 2.37-7.56, P<0.001), and colostomy or ileostomy (OR=1.67, 95% CI: 1.04-2.69, P=0.034) as independent risk factors for SSI. Conclusion:To prevent SSI following CRC surgery, attention should be given to optimizing body weight and glycemic control, promoting minimally invasive surgical approaches when feasible, and cautiously considering the necessity of colostomy or ileostomy.

Objective To investigate the functional brain characteristics of the frontal and bilateral tempo-ral regions during a verbal fluency task(VFT)in patients with stable schizophrenia using functional near-infrared spectroscopy(fNIRS),and to evaluate its potential clinical application in assisting the diagnosis of schizophrenia.Methods A total of 45 stable schizophrenia patients and 30 healthy controls were enrolled in the study.fNIRS was employed to assess brain activation patterns in the frontal and bilateral temporal lobes during the VFT.The integral values,centroid values,and slopes of the frontal and bilateral temporal lobes were measured.Differences in demographic characteristics,word count,and fNIRS-derived parameters between the two groups were analyzed.A logistic regression model and receiver operating characteristic(ROC)curve analysis were conducted to evaluate the diagnostic performance of the fNIRS measures in distinguishing schizophrenia patients from healthy controls.Results During VFT,the patient group demonstrated reduced word count and decreased integral values and slopes in the frontal lobe,as well as lower integral values in the bilateral temporal lobes,compared to the healthy control group(all P＜0.05).However,no statistically significant differences were observed between the two groups in terms of frontal lobe centroid value,temporal lobe centroid value,or bilateral temporal lobe slope(all P＞0.05).The logistic regression model indicated that the temporal lobe integral value was significantly associated with the diagnosis of schizophrenia(OR=0.979,P=0.005).ROC curve analysis revealed that the frontal lobe integral value,frontal lobe slope,and temporal lobe integral value exhibited acceptable discriminatory power for disease classification,with area under the curve(AUC)values of 0.681,0.644,and 0.779,respectively(all P＜0.05).Conclusion In stable schizophrenia,the integral values of the frontal and temporal lobes,as well as the slope of the frontal lobe,decrease during the VFT,suggesting potential clinical utility in differentiating individuals with schizophrenia from healthy controls.

Protein-protein interactions play an extremely important role in the biochemical functions of cells,and in-depth analysis of protein interactions is the key to understanding cellular life activities.In this study,we systematically mined the interacting proteins of Golgi protein 73(GP73)using classical immunoprecipitation combined with mass spectrometry,and sought to further analyze the molecular func-tion of GP73.Hepatocellular carcinoma cell line HepG2 was selected,and a stable cell line overexpress-ing GP73-3Flag was constructed using lentiviral infection technology.A total of 78 high-confidence GP73 interacting proteins were identified by immunoprecipitation coupled with mass spectrometry.Bioinformat-ics analyses suggested that GP73 interacted with nearly 40 cytosolic proteins and participated in the bio-logical processes of RNA transport,splicing,and translation.Further immunofluorescence and cytosolic protein isolation experiments confirmed the cytosolic localization of GP73 in a variety of tumor cells.Based on the 78 interacting proteins,we further screened protein interaction networks related to mRNA splicing and verified the existence of interactions between GP73 and seven proteins,including HNRN-PH3,SMN1,RBM14,andNCBP1,by co-immunoprecipitation experiments.In addition,minigene spli-cing assay results indicated that GP73 inhibited the splicing efficiency of pre-mRNA by cells.This study contributes to the expansion of knowledge regarding the function of GP73 and aids in elucidating its criti-cal role in cell biology and its potential association with diseases.

Objective:To compare the application of Micronutritional Risk Assessment(MNA),Universal Screening Tool for Malnutrition(MUST)and Nutritional Risk Screening 2002(NRS2002)in nutritional risk assessment among patients with digestive tract tumors during perichemotherapy,based on the Global Leadership Initiative on Malnutrition(GLIM)standard.Methods:A prospective cross-section study was conducted,including 114 patients with digestive tract tumors hospitalized by Department of General Surgery,Huangshan Shoukang Hospital from January 2020 to December 2021.All patients were evaluated by GLIM assessment,the correlation between GLIM and MNA,MUST and NRS 2002 screening results was compared,and the consistency among different methods was compared.Patients were divided into malnourished group(nutritional risk group)or normal nourished group according to the results of the three tools.The differences in single anthropometric or test indicators between the groups were compared.Results:According to GLIM,the proportion of malnutrition was 36.8%.The proportion of malnutrition evaluated by MNA,MUST,and NRS2002 were 63.2%,47.4%,and 32.5%,respectively.The sensitivity and negative predictive value of MNA in assessing nutrition-related risks were the highest,while the specificity,Jorden index,Kappa value and positive predictive value of NRS2002 were the highest.There were statistical differences in levels of body mass index,hemoglobin(Hb),triglyceride,total cholesterol,albumin,prealbumin(P-ALB),blood creatinine,lymphocyte counts,and hospitalization costs between two groups assessed by three different tools(P<0.05).Levels of Hb and P-ALB were statistically different between the two groups of the three screening tools.Conclusion:Based on GLIM evaluation results,MNA and other nutritional screening tools are applicable to the assessment of nutritional risks of patients with gastrointestinal cancer during perichemotherapy due to the joint evaluation of measurement indicators.The MNA is more recommended with the highest detection rate and sensitivity for nutritional risks assessment.

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 investigate the effects of evolocumab combined with atorvastatin on post-percutaneous coronary intervention （PCI） efficacy and coronary microcirculation in patients with acute ST-segment elevation myocardial infarction （STEMI）. METHODS This retrospective cohort study included 194 hospitalized patients with acute STEMI who underwent PCI in the First Affiliated Hospital of Nanyang Medical College from Jan. 2022 to Dec. 2024. The patients were divided into control group （100 cases） and combination therapy group （94 cases） according to the different therapy plans. The control group was given Atorvastatin calcium tablets orally， at a dose of 20 mg， once a day. On the basis of the control group， the combination therapy group received an initial injection of Evolocumab injection 140 mg within 24 hours after PCI， followed by subsequent injections every 2 weeks. Both groups received continuous treatment for at least 30 days. Blood lipid indicators [total cholesterol （TC）， triglycerides （TG）， low-density lipoprotein cholesterol （LDL-C） and high-density lipoprotein cholesterol]， inflammation indicators [C-reactive protein （CRP） and interleukin-6 （IL-6）]， vascular Δ 基金项目河南省医学科技攻关计划项目（No.LHGJ20230971） endothelial function and microcirculation indicators [fibrinogen， ankle-brachial index （ABI） and nitric oxide （NO）]， cardiac function indicators [left ventricular ejection （LVEF）， left ventricular end-diastolic diameter and left ventricular end-systolic diameter] before treatment and after 30 days of treatment， as well as the occurrence of adverse reactions during the treatment， were compared between the two groups. RESULTS Before treatment， there were no statistically significant differences in the levels of the indicators of blood lipid， inflammation， vascular endothelial function and microcirculation， and cardiac function between the two groups （P＞0.05）. After 30 days of treatment， the levels of TC， TG， LDL-C， CRP， IL-6 and fibrinogen in both groups were significantly reduced compared to those before treatment within the same group. The levels of NO and LVEF in both groups， as well as ABI in the combination therapy group， were significantly elevated compared to those before treatment within the same group. Moreover， the improvements in the levels of TC， LDL-C， CRP， IL-6， fibrinogen and NO in the combination therapy group were more pronounced than those in the control group during the same period （P＜0.05）. There were no statistically significant differences in the incidence of liver function impairment， gastrointestinal adverse reactions， or the overall incidence of adverse reactions between the two groups （P＞0.05）. CONCLUSIONS Compared with atorvastatin alone， evolocumab combined with atorvastatin more effectively reduces the blood lipid levels， improves the inflammatory status， vascular endothelial function and coronary microcirculation in patients with acute STEMI after PCI， while the safety is comparable to that of atorvastatin alone.

Objective To investigate the functional brain characteristics of the frontal and bilateral tempo-ral regions during a verbal fluency task(VFT)in patients with stable schizophrenia using functional near-infrared spectroscopy(fNIRS),and to evaluate its potential clinical application in assisting the diagnosis of schizophrenia.Methods A total of 45 stable schizophrenia patients and 30 healthy controls were enrolled in the study.fNIRS was employed to assess brain activation patterns in the frontal and bilateral temporal lobes during the VFT.The integral values,centroid values,and slopes of the frontal and bilateral temporal lobes were measured.Differences in demographic characteristics,word count,and fNIRS-derived parameters between the two groups were analyzed.A logistic regression model and receiver operating characteristic(ROC)curve analysis were conducted to evaluate the diagnostic performance of the fNIRS measures in distinguishing schizophrenia patients from healthy controls.Results During VFT,the patient group demonstrated reduced word count and decreased integral values and slopes in the frontal lobe,as well as lower integral values in the bilateral temporal lobes,compared to the healthy control group(all P＜0.05).However,no statistically significant differences were observed between the two groups in terms of frontal lobe centroid value,temporal lobe centroid value,or bilateral temporal lobe slope(all P＞0.05).The logistic regression model indicated that the temporal lobe integral value was significantly associated with the diagnosis of schizophrenia(OR=0.979,P=0.005).ROC curve analysis revealed that the frontal lobe integral value,frontal lobe slope,and temporal lobe integral value exhibited acceptable discriminatory power for disease classification,with area under the curve(AUC)values of 0.681,0.644,and 0.779,respectively(all P＜0.05).Conclusion In stable schizophrenia,the integral values of the frontal and temporal lobes,as well as the slope of the frontal lobe,decrease during the VFT,suggesting potential clinical utility in differentiating individuals with schizophrenia from healthy controls.