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 effects and mechanism of Jianpi Shuyi Decoction in improving pancreatic fibrosis in chronic pancreatitis(CP)based on network pharmacology and animal experiments.Methods TCMSP was used to screen the active components and targets of Jianpi Shuyi Decoction.GeneCards was used to obtain the disease targets of pancreatic fibrosis,and the intersection of drug and disease targets was used to construct the protein interaction network and the drug-component-target network,and the core target genes were screened out.GO and KEGG pathway enrichment analysis was performed on the intersecting targets.Caerulein was used to induce CP mouse model,and Jianpi Shuyi Decoction was given for gavage.HE and Sirius red staining were used to observe pancreatic tissue inflammation and collagen deposition,respectively.RT-qPCR was used to observe the mRNA expression levels of Acta2,COL1A1,PI3K and Akt1 in pancreatic tissue.Immunohistochemistry staining was used to observe the protein expression levels of α-SMA,COL-1,p-PI3K and p-Akt in pancreatic tissues.Results A total of 181 active components were screened from Jianpi Shuyi Decoction,corresponding to 284 targets,with 240 targets overlapping between drugs and disease and the core targets were PTGS2,HSP90AA1,etc.193 signaling pathways were obtained from KEGG pathway enrichment analysis,primarily involving lipids and atherosclerosis,chemical carcinogenic-receptor activation,PI3K-Akt signaling pathway and others.The results of animal experiments showed that,compared with the normal group,the model group showed a large number of inflammatory cell infiltration and collagen deposition in pancreatic tissue,the mRNA expression of Acta2,COL1A1,PI3K and Akt1 in pancreatic tissue significantly increased(P<0.01),and the protein expression of α-SMA,COL-1,p-PI3K,p-Akt significantly increased(P<0.01);Jianpi Shuyi Decoction significantly reduced the inflammation and collagen deposition in pancreas of mice,reduced the mRNA expression of Acta2,COL1A1,PI3K and Akt1(P<0.05),and attenuated the protein expression of α-SMA,COL-1,p-PI3K and p-Akt in pancreatic tissue(P<0.05).Conclusion Jianpi Shuyi Decoction may exert a therapeutic effect on CP pancreatic fibrosis by regulating the PI3K/Akt signaling pathway,attenuating inflammation and collagen deposition in the pancreas,and reducing the levels of α-SMA and COL-1.

Objective To explore the effects and mechanism of Jianpi Shuyi Decoction in improving pancreatic fibrosis in chronic pancreatitis(CP)based on network pharmacology and animal experiments.Methods TCMSP was used to screen the active components and targets of Jianpi Shuyi Decoction.GeneCards was used to obtain the disease targets of pancreatic fibrosis,and the intersection of drug and disease targets was used to construct the protein interaction network and the drug-component-target network,and the core target genes were screened out.GO and KEGG pathway enrichment analysis was performed on the intersecting targets.Caerulein was used to induce CP mouse model,and Jianpi Shuyi Decoction was given for gavage.HE and Sirius red staining were used to observe pancreatic tissue inflammation and collagen deposition,respectively.RT-qPCR was used to observe the mRNA expression levels of Acta2,COL1A1,PI3K and Akt1 in pancreatic tissue.Immunohistochemistry staining was used to observe the protein expression levels of α-SMA,COL-1,p-PI3K and p-Akt in pancreatic tissues.Results A total of 181 active components were screened from Jianpi Shuyi Decoction,corresponding to 284 targets,with 240 targets overlapping between drugs and disease and the core targets were PTGS2,HSP90AA1,etc.193 signaling pathways were obtained from KEGG pathway enrichment analysis,primarily involving lipids and atherosclerosis,chemical carcinogenic-receptor activation,PI3K-Akt signaling pathway and others.The results of animal experiments showed that,compared with the normal group,the model group showed a large number of inflammatory cell infiltration and collagen deposition in pancreatic tissue,the mRNA expression of Acta2,COL1A1,PI3K and Akt1 in pancreatic tissue significantly increased(P<0.01),and the protein expression of α-SMA,COL-1,p-PI3K,p-Akt significantly increased(P<0.01);Jianpi Shuyi Decoction significantly reduced the inflammation and collagen deposition in pancreas of mice,reduced the mRNA expression of Acta2,COL1A1,PI3K and Akt1(P<0.05),and attenuated the protein expression of α-SMA,COL-1,p-PI3K and p-Akt in pancreatic tissue(P<0.05).Conclusion Jianpi Shuyi Decoction may exert a therapeutic effect on CP pancreatic fibrosis by regulating the PI3K/Akt signaling pathway,attenuating inflammation and collagen deposition in the pancreas,and reducing the levels of α-SMA and COL-1.

OBJECTIVE To establish the HPLC fingerprint of Xintong granules and the quantitative analysis of multi-components by single-marker method(QAMS)to determine the contents of 7 components,so as to provide a scientific basis for their quality control.METHODS HPLC method was used to establish the fingerprints for 10 batches of Xintong granules(No.S1-S10),and similarity evaluation,cluster analysis(CA)and partial least squares-discriminant analysis(PLS-DA)were performed.At the same time,the contents of seven components,including puerarin,daidzin,calycosin-7-O-β-D-glucoside,stilbene glycoside,naringin,icariin and tanshinone ⅡA,were determined by QAMS method,and were compared with the results of external standard method.RESULTS A total of 18 common peaks were marked and 7 peaks were identified in the HPLC fingerprints for 10 batches of Xintong granules,namely puerarin(peak 4),daidzin(peak 7),calycosin-7-O-β-D-glucoside(peak 9),stilbene glycoside(peak 10),naringin(peak 12),icariin(peak 17),and tanshinone ⅡA(peak 18);the similarities among them were more than 0.990,and CA and PLS-DA results showed that S4-S5,S8-S10,S1-S3 and S6-S7 were clustered into three categories,respectively.Using naringin as the internal standard,the contents of puerarin,daidzin,calycosin-7-O-β-D-glucoside,stilbene glycoside,icariin and tanshinone ⅡA were determined to be 7.868 1-10.181 2,1.709 2-2.374 1,0.285 2-0.326 3,1.024 1-1.523 9,0.140 2-0.290 4,and 0.077 1-0.219 4 mg/g,respectively,by the QAMS.These results showed no significant differences compared to those obtained by the external standard method.CONCLUSIONS Established HPLC fingerprint and QAMS method are convenient,stable and accurate,which can provide a basis for the quality evaluation of Xintong granules.

Objective To discuss the application of partial antiplatelet drug genotype detection and thromboelastography-platelet mapping(TEG-PM)in guiding the selection of antiplatelet regimens in patients with intracranial aneurysms(IAs)after receiving stenting.Methods A total of 106 patients with IAs in the First Affiliated Hospital of Kunming Medical University,who underwent implantation of stent and received the testings of platelet-endothelial aggregation receptor 1(PEAR 1)and clopidogrel-related gene-cytochrome P450 enzyme 2C19(CYP2C19),and some of whom received TEG-PM testing from January 2019 to August 2022,were collected for this study.The patients were divided into group A(gene detection group,according to the drug-related gene detection results to adjust the medication)and group B(combination group,according to the two testing results to guide the medication).The patient's gender,age,testing data were collected,and the occlusion of IAs,stent intimal hyperplasia,drug-related hemorrhagic and ischemic complications during follow-up period were recorded.Results A total of 123 IAs lesions in 106 patients were treated.The patient's mean age was(53.67±6.66)years,67 patients were female.Group A had 41 patients and group B had 65 patients.No statistically significant differences in the baseline data,IAs features,stent types used,and medication regimen existed between the two groups(all P＞0.05).In Group A,the ischemic complications and hemorrhagic complications occurred in two patients each.In Group B,no ischemic complications occurred and 4 patients developed hemorrhagic complications.The difference in the incidence of related complications between the two groups was not statistically significant(P=0.287 and P=0.782 respectively).There were no statistically significant differences in the postoperative one-month and 3-month intimal hyperplasia grade and the aneurysm occlusion rate between the two groups(all P＞0.05).The postoperative 6-month overall intimal hyperplasia grade in Group A was slightly higher than that in Group B,and the difference was statistically significant(P=0.034).Conclusion In order to improve the precision and individualized treatment of antiplatelet therapy,it is suggested that clinicians should adopt TEG-PM-guided conventional double-antibody therapy first when making selection of testing items.For patients with insufficient inhibition rate indicated by TEG-PM,testing of the genes associated with antiplatelet drugs should be used.Based on the genetic test results it is necessary to determine the reasons for the insufficient inhibition rate as well as to adjust the medication promptly according to the specific situation of the patient,so as to ensure the effectiveness of antiplatelet therapy and achieve the purpose of individualized precision therapy.

Objective To explore the the detection of MMP-2,-7,-9,and-12 enzymatic activity using the CEACAM1-derived fluorescent peptide substrate Site 84,investigating the application of substrate Site 84 to distinguishing between MMP-2 and MMP-9 in the gelatinase spectrum of MMPs.Methods The fluorescent enzymatic method was employed to observe the detection of MMP-2,-7,-9,and-12 enzymatic activity using substrate Site 84;further observations were made on the sensitivity and specificity of substrate Site 84 to enzymatic activity of MMP-2 and MMP-9 within the gelatinase spectrum;the kinetic parameters(Km and Kcat)of the enzymatic reaction between substrate Site 84 and MMP-2 were obtained.Results Using Site 84 as a substrate,enzymic kinetics curves for MMP-12,-7,-2 were obtained,but no enzymatic activity curve for MMP-9 was observed.Furthermore,Site 84 specifically detected the enzymatic activity of MMP-2 within the gelatinase spectrum,capable of detecting low concentration(0.6 μM)of MMP-2 enzymatic activity,but no obvious enzymatic reaction was observed for high concentration(6 μM)of MMP-9;the kinetics parameters for the enzymatic reaction between Site 84 and MMP-2 were Km=315 μM,Kcat/Km=2 565/MS.Conclusion The CEACAM1-derived substrate Site 84 serves as a novel fluorescent peptide substrate,enabling the acquisition of enzymatic activity curves for MMP-12,-7 and-2,and specifically detecting the enzymatic activity of MMP-2 within the MMP gelatinase spectrum.

Objective To explore the effect of project-based learning on the training of sharp injury prevention skills in the pre-job training for nursing interns and its impact on the incidence of sharp injuries during internship.Methods Two classes(class A and B),majoring in nursing at Medical College of Nanchang Institute of Technology in March 2023 were selected as the research subjects by convenience sampling method.Coin flipping method was used to designate class A as the routine group and class B as the intervention group.The routine group received rou-tine pre-job training which mainly focused on retrospective intensive training on nursing operation skills.On this ba-sis,the intervention group integrated project-based learning as compensation education on sharp injury prevention skills.Kirkpatrick's four level training evaluation model was adopted to comprehensively evaluate the educational effectiveness at the four progressive levels of"reaction,learning,behavior,and outcome"at corresponding stages.Results 56 nursing interns were included in the class A routine group and class B intervention group,respectively.The course evaluation score(128.67±4.39 vs 117.28±6.55),needlestick protection knowledge cognition score(109.11±4.38 vs 96.44±6.72),safe injection behavior score(38.45±4.91 vs 32.30±5.62),occupational iden-tity score(58.02±8.55 vs 51.77±15.86),and job competency score(82.59±13.35 vs 75.61±15.09)of nursing interns in the intervention group were all higher than those in the routine group,differences were all statistically sig-nificant(all P＜0.05).The incidence of sharp injuries(19.64％ vs 57.14％)and the average frequency of occu-rrence(1.45 vs 2.13)in nursing interns in the intervention group were both lower than those in the control group.The case intervention rate(87.50％ vs 45.59％)and case reporting rate(93.75％vs 32.35％)after sharp injury were both higher than those in the routine group,and the differences were both statistically significant(both P＜0.05).Conclusion Introducing project-based learning in pre-job training for nursing interns can effectively improve their mastery of protection skills,reduce the incidence of sharp injuries during internships,and have important prac-tical value in cultivating their occupational protection abilities.

Objective:To explore the risk fall death associated with compound hot extremes.Methods:This study collected data on fall deaths in Guangdong, Hunan, and Zhejiang Provinces from 2013 to 2018 and matched their exposure to meteorological data. Based on a time-stratified case-crossover design, a conditional logistic regression model embedded with a cross-basis function of the distributed lag nonlinear model was applied to estimate the risk of fall to death due to compound hot extremes.Results:Compared with regular days, compound hot extremes significantly increased the risk of death from falls ( OR=1.19, 95% CI: 1.09-1.30), and women ( OR=1.27, 95% CI: 1.11-1.45) and the elderly age 65 and above ( OR=1.24, 95% CI: 1.12-1.39) were more sensitive to compound hot extremes. The maximum duration of compound hot extremes was 7 days, and the maximum intensity was 6.2 ℃, and the duration and intensity were proportional to the risk of death from falls. The risk of death from falls increased by 12% ( OR=1.12, 95% CI: 1.06-1.18) each day, increasing in duration after linearization. The risk of death from falls increased by 16% ( OR=1.16, 95% CI: 1.10-1.22) for each 1 ℃ increase in linearized intensity. Conclusion:Compound hot extremes increase the risk of death cases from falls.

Objective:To investigate the predictive value of color Doppler ultrasound combined with electrocardio-gram for right heart dys function in patients with pulmonary heart disease(PHD).Methods:A total of 100 PHD patients admitted in Dongcheng Branch of First Affiliated Hospital of Anhui Medical University between January 2020 and December 2023 were retrospectively analyzed.According to results of 6min walking test(6MWT),pa-tients were divided into good right heart function group(n=64,≥350m)and right heart dysfunction group(n=36,＜350m).The indexes of cardiac color ultrasound[isovolumic relaxation time(IVRT),isovolumetric contraction time(IVCT)and right ventricular Tei index],ECG[24h mean R-R interval standard deviation(SDNN),normal R-R interval standard deviation per 5min(SDANN)and the ratio of low frequency components to high frequency components(LF/HF)]were compared between two groups.Receiver operating characteristic(ROC)curve was drawn to analyze the diagnostic value of color Doppler ultrasound,ECG and their combination for right heart dys-function in PHD patients.Spearman correlation coefficient was used to analyze the association of color Doppler ul-trasound,ECG and their combination with right heart dysfunction in PHD patients.Results:Compared with those in good right heart function group,patients in right heart dysfunction group had significant higher IVRT[(120.64±14.08)ms vs.(97.87±10.93)ms],IVCT[(84.28±12.33)ms vs.(71.92±10.61)ms]and Tei index[(0.85±0.11)vs.(0.63±0.07)](P＜0.001 all),and significant lower SDNN[(75.52±12.58)ms vs.(85.58±11.75)ms],SDANN[(63.86±10.92)ms vs.(76.75±11.71)ms]and LF/HF[(1.33±0.19)vs.(1.84±0.27)](P＜0.001 all).ROC curve indicated that the AUC of color Doppler ultrasound combined ECG in diagnosing right heart dysfunction in PHD patients was 0.911(95％CI 0.838～0.959),which was significantly higher than those of color Doppler ultrasound[0.775(95％CI 0.681～0.853),Z=2.404,P=0.016]and ECG[0.688(95％CI 0.588～0.777),Z=3.968,P=0.001]alone.Spearman correlation analysis indicated that there was a significant positive correlation of color Doppler ultrasound(r=0.547),ECG(r=0.375)and their combination(r=0.810)with right heart dysfunction in PHD patients(P＜0.001 all),and the correlation between combined detection and right heart dysfunction in PHD patients was significantly higher.Conclusion:Color Doppler ultrasound combined with ECG possesses high diagnostic performance for right heart dysfunction in PHD patients.

Objective:To investigate the effect of oleanolic acid(OA)on inflammatory injury of pancreatic acinar cells in-duced by lipopolysaccharide(LPS)by regulating stromal cell-derived factor-1(SDF-1)/CXC chemokine receptor 4(CXCR4)signal axis.Methods:Rat pancreatic acinar cells AR42J were treated with 5,10,20 and 40 μmol/L OA,and the cell viability was detected using CCK-8 method to determine the optimal dosage for subsequent experimental OA treatment;AR42J cells were randomly divided into control group(NC),LPS group(10 mg/L LPS),OA low-dose group(OA-L group,10 mg/L LPS+5 μmol/L OA),OA medium dose group(OA-M group,10 mg/L LPS+10 μmol/L OA),OA high-dose group(OA-H group,10 mg/L LPS+20 μmol/L OA),Rr-SDF-1 activator(SDF-1)group(10 mg/L LPS+20 ng/ml Rr-SDF-1)and OA-H+Rr-SDF-1 group(10 mg/L LPS+20 μmol/L OA+20 ng/ml Rr-SDF-1).CCK-8 method and EdU staining were applied to detect proliferation of AR42J cells;flow cytometry was applied to detect apoptosis of AR42J cells;activity of superoxide dismutase(SOD)and content of malondialdehyde(MDA)in cell supernatant were detected by spectrophotometry;ELISA was applied to detect levels of IL-6 and TNF-α in cell supernatant;Western blot was applied to detect expressions of CyclinD1,Bcl-2 associated X protein(Bax),SDF-1 and CXCR4 proteins in cells.Results:5,10 and 20 μmol/L OA were selected as the low,medium and high doses for subsequent treatment of AR42J cells;compared with NC group,A450 value,EdU positive cell rate,SOD activity,and CyclinD1 protein expression in LPS group decreased,while apoptosis rate,MDA content,IL-6,TNF-α levels,Bax,SDF-1 and CXCR4 protein expressions increased(P<0.05);compared with LPS group,A450 value,EdU positive cell rate,SOD activity,and CyclinD1 protein expression in OA-L group,OA-M group and OA-H group increased,while apoptosis rate,MDA content,IL-6,TNF-α levels,Bax,SDF-1 and CXCR4 protein expressions reduced,the change trends of corre-sponding indicators in Rr-SDF-1 group was opposite to the above(P<0.05);compared with OA-H group,A450 value,EdU positive cell rate,SOD activity,and CyclinD1 protein expression in OA-H+Rr-SDF-1 group decreased,while apoptosis rate,MDA content,IL-6,TNF-α levels,Bax,SDF-1 and CXCR4 protein expressions increased(P<0.05).Conclusion:OA may alleviate LPS induced inflam-matory damage to pancreatic acinar cells in rats by inhibiting the SDF-1/CXCR4 pathway.