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.

Aortic regurgitation and mitral regurgitation are more common in elderly heart valve disease,and both may be present in some patients.Severe aortic regurgitation complicated with severe mitral regurgitation often requires surgical valve replacement,but in patients at high risk of surgery,the risk of perioperative mortality is significantly increased.Therefore,for such patients,minimally invasive interventions can significantly improve long-term patient outcomes while reducing surgical risk.This article report a case of transcatheter aortic valve replacement combined with transcatheter edge-to-edge repair in the treatment of severe aortic regurgitation combined with mitral valve prolapse,in order to explore new treatment ideas for similar cases.

Objective To design a performance testing platform to evaluate the working status and performance characteristics of the ventilator proportional solenoid valve.Methods The performance testing platform had its hardware including a high-pressure gas source,a pressure regulating valve,sensors and etc,and its software designed based on PyQt5 and composed of several modules for data acquisition,parameter setting,image display,indicator computation,result output and etc.Two kinds of proportional solenoid valves(Valve 1、Valve2)were selected for static and dynamic tests to verify the performance of the platform.Results The platform developed facilitated the proportional solenoid valve to carry out accurate computation of static and dynamic indicators at real time and time domain and waveform feature extraction of sensor data by precision control and data acquisition for the proportional solenoid valve.Static tests showed that Valve 1 gained advantages over Valve 2 in static flow characteristics involving in lowered repeatability,return error and offset while enhanced stability;dynamic tests indicated Valve 2 had rapid flow variations and significant flow fluctuation impacts,Valve 1 showed smooth dynamic response changes,and Valve 2 behaved better than Valve 1 in dynamic performance.Conclusion The testing platform developed comprehensively demonstrates the performance characteristics and working performance of the ventilator proportional solenoid valve,which is of great significance to enhance the reliability and safety of the ventilator.[Chinese Medical Equipment Journal,2025,46(1):13-19]

Objective:To explore the influence of initial high-risk cytogenetic abnormalities on the outcomes of children with acute myeloid leukemia (AML) after post-transplant Cyclophosphamide (PTCy)-based allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:A retrospective cohort study.AML children who underwent PTCy-based allo-HSCT after the first complete remission at Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science and Technology between April 2017 and April 2024 were enrolled.Patients were divided into intermediate-risk and high-risk groups based on their initial cytogenetic features.These patients were further divided into complex karyotype, 11q23 rearrangement, and other karyotype groups.Clinical characteristics and survival outcomes were compared among these groups.Measurement and count data were analyzed using Wilcoxon rank-sum/Kruskal-Wallis and χ2 tests, respectively.Survival and risk factor analyses were performed using Kaplan-Meier and Cox proportional hazards methods, respectively. Results:A total of 51 AML children who underwent allo-HSCT were included in this study.The median age at transplantation was 3.2 years and the median follow-up time was 4.6 years.There were 26 cases in the intermediate-risk group and 25 cases in the high-risk group; 8 cases in the complex karyotype group, 14 cases in the 11q23 rearrangement group, and 29 cases in the other karyotype groups.By the end of the follow-up on November 30, 2024, 11 patients relapsed, 8 patients died, and 13 patients developed grades Ⅱ-Ⅳ acute graft-versus-host disease (GVHD).The 3-year overall survival (OS), relapse-free survival (RFS), and grades Ⅱ-Ⅳ acute GVHD-free and relapse-free survival (GRFS) were 84.0% (95% CI: 74.4%-94.8%), 74.5% (95% CI: 63.4%-87.5%), and 58.8% (95% CI: 46.7%-74.0%), respectively.The 3-year OS of the high-risk group was significantly lower than that of the intermediate-risk group (71.8% vs.96.2%, P=0.022), while differences in 3-year RFS and GRFS between the 2 groups were not statistically significant (68.0% vs.80.8%, P=0.400; 52.0% vs.65.4%, P=0.420).The 3-year OS, RFS and GRFS of the complex karyotype group were significantly lower than those of 11q23 rearrangement and other karyotype groups (50.0% vs.85.7%, 93.1%, P=0.009; 37.5% vs.85.7%, 79.3%, P=0.022; 25.0% vs.64.3%, 65.5%, P=0.049).Multivariate analysis showed that a complex karyotype was an independent prognostic factor affecting 3-year OS and GRFS [OS: HR=6.79 (95% CI: 1.13-43.80), P=0.044; GRFS: HR=3.72(95% CI: 1.13-12.20), P=0.030]. Conclusions:High-risk cytogenetic features are significant predictors of survival outcomes in pediatric AML patients undergoing PTCy-based allo-HSCT.

Objective To construct a glioma grading evaluation model based on magnetic resonance diffusion tensor imaging(DTI)parameters,systemic immune-inflammation index(SII),and albumin to globulin ratio(AGR),and analyze its evaluation effect.Methods A total of 120 patients with brain glioma treated in our hospital from January 2022 to January 2024 were selected and divided into a high-grade glioma(HGG)group of 53 cases and a low-grade glioma(LGG)group of 67 cases according to the WHO tumor classification.The magnetic resonance DTI parameters,SII,and AGR of patients between the two groups were compared.A binary Logistic regression analysis model was constructed to analyze the risk factors for the occurrence of HGG;the receiver operating characteristic(ROC)curve was drawn,and the area under the curve(AUC)was calculated to analyze the evaluation value of the glioma grading model.Results Compared with the LGG group,the HGG group had lower fractional anisotropy(FA)value,apparent diffusion coefficient(ADC),and AGR of the tumor body and the peritumoral edema area(P<0.05),and higher SII(P<0.05).Binary Logistic regression analysis showed that decresed ADC value of the tumor body and the peritumoral edema area,decreased AGR,and increased SII were all the risk factors for the occurrence of HGG(P<0.05).The ROC curve analysis showed that the AUC of the combined assessment of FA value and ADC of the tumor body,FA value and ADC of the peritumoral edema area,SII and AGR for glioma grading was higher than that of their individual assessment(P<0.05).Conclusion The occurrence of HGG is related to the ADC of the tumor body and peritumoral edema area,SII,and AGR.The model constructed based on magnetic resonance DTI parameters,SII and AGR has high evaluation value for the grading of glioma.

Aimed to find a safe and effective drug to replace nitroimidazole drugs in aquaculture pro-duction for the prevention and treatment of Trichomoniasis in pigeons,which can improve the eco-nomic benefits of meat pigeon breeding and ensure food safety.Firstly,Trichomonas was isolated and cultured from the crop of diseased pigeons and identified.After stable passage,a quantitative method for in vitro detection of Trichomonas was established by combining an automated cell counter and quantitative real-time PCR technology.To prepared the drug,powders of Sophora fla-vescent and Philodendron were made into herbal water extracts(SFPA)and mixed with copper sulfate(CS)solution.Then added the drug to the culture medium of Trichomonas to determine the effective concentration of it.A total of 135 pairs each of Silver King and Mimas breeding pigeons in the same laying period were selected and randomly divided into three groups,with 6 replicates in each group and 15 pairs of breeding pigeons in each replicate.Four days before brooding,the three groups were fed with 200 mL of pure water,0.5 g/L metronidazole(MDZ)solution,and a mixed solution of 30 g/L SFPA and 0.5 g/L CS,respectively.The feeding experiment lasted for 26 d.Results showed that the mixed solution of SFPA and CS had a significant killing effect on Trichomonas in vitro(P＜0.05).Feeding the drug to breeding pigeons significantly reduced the in-fection rate of breeding pigeons by Trichomonas(P＜0.05).The drug had no significant effect on the serum biochemical indexes,antioxidant properties,immunoglobulin levels of breeding pigeons,the average cage weight,immune organ indexes,meat quality and slaughter performance of squabs(P＞0.05).The results suggested that adding SFPA and CS to pigeons can effectively prevent and treat Trichomoniasis and improve production performance.It can replace nitroimidazole drugs without affecting the immune level of breeding pigeons and the weight,immune level,slaughter performance and meat quality of squabs,thereby reduce drug residues in poultry products and en-hance the food safety.

Objective:To study the effects of astragalus polysaccharide on the proliferation of osteoblasts and the effects of astraga-lus polysaccharide on bone repair in rabbits with apical periodontitis.Methods:CCK8 assay was used to detect of astragalus poly-saccharide on the proliferation of MC3T3-E1 cells.The expression of osteogenic marker factor BMP-2 and Runx-2 mRNA and pro-tein of MC3T3-E1 cells were detected by RT-PCR and Western blot respectively.The animal models of mandibular incisors with apical periodontitis in rabbits was established and verified by roentgenogram and HE staining.The repair ability of bone tissue was detected by roentgenogram.The expression of BMP-2 in the rabbit apical tissue was detected by immunohistochemistry.The results were compared between the treated and control rabbits(n=3).Results:Astragalus polysaccharide promoted cell proliferation after 24 h,48 h and 72 h exposure(P＜0.05).Roentgenogram and HE staining confirmed that the rabbit apical periodontitis was successfully constructed.The bone defect volume in the treated rabbits apical tissue was less and the expression of BMP-2 was high-er than that in the control rabbits(P＜0.05).Conclusion:Astragalus polysaccharide promotes the proliferation ability of osteoblasts and promotes the bone repair of rabbits with apical periodontitis.