1.Establishment and Preliminary Analysis of GP73 Interactome Using Proximity-dependent Labeling Technology
Mu-Yi LIU ; Chang ZHANG ; Meng-Xin YANG ; Xin-Long YAN ; Lu-Ming WAN ; Cong-Wen WEI
Progress in Biochemistry and Biophysics 2026;53(3):711-723
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.
2.Establishment of HPLC characteristic chromatograms and content determination of nine constituents for Yixin Fumai Granules
Xin-ru CHI ; Zheng-wei CHEN ; Jie LI ; Ai-ying WU ; Li-hua YIN ; Hong-bing LIU ; Jing-guang LU
Chinese Traditional Patent Medicine 2025;47(1):1-6
AIM To establish the HPLC characteristic chromatograms for Yixin Fumai Granules,and to determine the contents of sodium danshensu,protocatechualdehyde,chlorogenic acid,calycosin-7-O-β-D-glucoside,ferulic acid,rosalinic acid,salvianolic acid A,salvianolic acid B,schisandrol A.METHODS The analysis was performed on a 35 ℃ thermostatic Acutfex PA-C18 column(4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 210,250,280,320 nm.Subsequently,cluster analysis and principal component analysis were performed.RESULTS There were 11 characteristic peaks in the characteristic chromatograms for 15 batches of samples with the similarities of more than 0.980.Nine constituents showed good linear relationships within their own ranges(r≥0.999 6),whose average recoveries were 97.60%-107.02%with the RSDs of 0.78%-1.87%.Various batches of samples were clustered into 4 categories,2 principal components demonstrated the accumulative variance contribution rate of 89.454%.CONCLUSION This sensitive and reproducible method can provide a reference for the quality evaluation and control of Yixin Fumai Granules.
3.Glycosylated ceramide-related metabolic enzymes and atherosclerosis
Xin TAN ; Qiang WEI ; Yuan-hong LIAO ; Jing-kun LU
Chinese Pharmacological Bulletin 2025;41(6):1005-1010
Atherosclerosis is an arterial lesion involving abnor-mal lipid metabolism and an inflammatory response,and is the initiating factor of many cardiovascular and cerebrovascular dis-eases.Glycosylated ceramides are derivatives of ceramide mole-cules with a glycosylated group attached,which not only affect the structural integrity of cell membranes,but also regulate apop-tosis,inflammation and lipid metabolism,and disorders of glyco-sylated ceramide metabolism are closely related to the occurrence and progression of atherosclerosis.This review focuses on the specific functions of the glycosylated ceramide-related metabolic enzymes glucose ceramide synthetase,glucosylceramide syn-thetase,lactose ceramide synthetase,and galactosidase in athero-sclerosis,as well as their important effects on the development of atherosclerosis.Targeted therapeutic strategies for these meta-bolic enzymes,related drug development and significant potential in atherosclerosis prevention and treatment are also reviewed.
4.Effects of shaving or segmental bowel resection on intestinal function in patients with bowel endometriosis:a 10-year follow-up study
Qi TIAN ; Yun CHEN ; Xin-xiang LI ; Wei-qi LU ; Jiang-feng YE ; Ke-qin HUA ; Xiao-fang YI
Fudan University Journal of Medical Sciences 2025;52(3):349-357
Objective To investigate the clinical characteristics of preoperative intestinal symptoms in patients with bowel endometriosis and to compare the effects of shaving versus segmental bowel resection on postoperative intestinal function.Methods A total of 105 patients diagnosed with bowel endometriosis and treated by the same surgical team at the Obstetrics and Gynecology Hospital,Fudan University between Aug 1,2013 and Dec 30,2017 were prospectively enrolled in this study.Clinical data were collected via outpatient visits and telephone follow-ups at four time points:preoperative(T0)and postoperative(T1:Nov 2018;T2:Nov 2020;T3:Apr 2024).The primary outcome was bowel symptoms and gastrointestinal function scores;secondary outcome was pain scores.A generalized estimating equation(GEE)model was used to analyze the interaction effect of surgical approach and follow-up time on outcomes.Results Ultimately,a total of 89 patients were included(15.24%loss to follow-up),among whom 79 patients(88.76%)underwent shaving excision.Preoperatively,46 patients(51.68%)presented with bowel symptoms,primarily anus bulge(21 cases,46.65%)and diarrhea(15 cases,32.61%)during menstruation.Postoperatively,there was a significant increase in constipation rates(T1:71.43%;T2:50.00%;T3:72.00%).Both surgical groups exhibited significant improvements in dysmenorrhea,gastrointestinal discomfort scores as well as gastrointestinal quality of life index(P<0.000 5).However,the segmental resection group had significantly higher scores for low anterior resection syndrome,constipation compared with the shaving excision group(P=0.02 and P=0.05).Conclusion Approximately half of the patients with bowel endometriosis exhibit typical bowel symptoms preoperatively,such as anus bulge and diarrhea.Both shaving excision and segmental resection effectively alleviate pain;however,shaving excision demonstrates an advantage regarding preservation of bowel function,whereas segmental resection may elevate risks associated with postoperative constipation or altered defecation patterns due to structural changes.The selection of surgical approach should carefully balance lesion removal and functional preservation,moreover,be sure that potential risks are thoroughly informed to patients prior to surgery.
5.The effect of adenoidectomy on the prognosis of children with refractory chronic sinusitis and adenoid hypertrophy
Lang XU ; Lu LIU ; Wei ZHANG ; Pengpeng WANG ; Xin NI
Chinese Archives of Otolaryngology-Head and Neck Surgery 2025;32(8):529-533,537
OBJECTIVE To analyze the distribution of the nasopharyngeal microbiota in children with refractory chronic sinusitis and adenoid hypertrophy using 16S rRNA next-generation high-throughput sequencing technology,and to compare the effects of adenoidectomy on the nasopharyngeal microbiota in such patients,with the aim of providing microbiological evidence for the clinical treatment of refractory chronic sinusitis with adenoid hypertrophy.METHODS Children diagnosed with chronic sinusitis with adenoid hypertrophy who visited Chengdu Women and Children's Central Hospital from January 2024 to March 2025 and whose symptoms were not controlled after drug treatment were selected.They were divided into a non-surgical group and a surgical group based on whether they underwent adenoidectomy.The VAS scores,Lund-Kennedy scores,and microbial community detection rates between the two groups were compared.RESULTS The final VAS scores and Lund-Kennedy scores for surgical group were both lower than those for non-surgical group,with statistically significant differences(tVAS score=3.491,P<0.001;tLund-Kennedy score=4.166,P<0.001).Comparisons of microbial α diversity analysis and β diversity at the phylum and genus levels between the two groups showed no statistically significant differences(tα diversity analysis=0.524,P=0.604;Rβ diversity at the phylum level=0.075,P=0.454;Rthe genus level=0.117,P=0.538).The dominant bacterial groups at the phylum level were the same in both groups,including Proteobacteria,Bacillales,Actinobacteria,Bacteroidetes,and Firmicutes,but the average relative abundance differed between the two groups.Species comparison analysis showed that at the phylum level,the abundance of Proteobacteria and Thermodesulfovibrio in surgical group was higher than that in non-surgical group;At the genus level,the abundance of the genus Moraxella was higher in non-surgical group than in surgical group,while the abundance of the genera Acidovorax,Stenotrophomonas,Delftia,Burkholderia,Caballeronia,Paraburkholderia,Lactobacillus,Devosia,norank_f_Muribaculaceae,Paracoccus,and Kocuria was lower in non-surgical group than in surgical group.CONCLUSION Children with refractory sinusitis and adenoid hypertrophy all have rich microbial communities in the nasopharynx.Adenoid surgery has no significant effect on the diversity of the nasopharyngeal microbial community.However,the composition of the microbial community has changed,and there are significant species differences,which may be related to the resolution of local obstruction after adenoidectomy and changes in the survival environment of the nasopharyngeal microbial community.
6.Genomic characterization of group A Streptococcus of different emm-type in Tianjin City from 2011 to 2024
Xiaohui LU ; Wei ZHANG ; Wen LI ; Aiping YU ; Guangwen LIU ; Baolu ZHENG ; Xuan CHEN ; Xin GAO ; Xiaoyan LI
Chinese Journal of Preventive Medicine 2025;59(5):702-709
To characterize the genomes of different emm-type group A Streptococcus (GAS), their virulence genes and drug resistance profiles in Tianjin City from 2011 to 2024. After PCR, a total of 42 strains with different years and emm types were selected for whole genome sequencing and multi-locus sequence typing (MLST), and the core genomes were used to generate a phylogenetic tree, after which the virulence genes and resistance genes were identified and analyzed, followed by the drug susceptibility test. In this study, the GAS strains were dominated by emm1 (50.0%) and emm12 (40.4%), and the MLST phenotypes were categorized into six types: ST36 (40.4%), ST1274 (26.1%), ST28 (23.8%), ST921 (4.7%), ST46 (2.3%), and ST403 (2.3%). There was a high consistency between their emm-types and ST types. A total of 68 virulence genes were detected in the genomes of 42 GAS strains, involving functional genes encoding exotoxin, bacterial adhesion, extracellular enzymes, etc. The virulence genes they carried were significantly different between emm1-type and emm12-type strains, such as speA. At the same time, the carrying rates of some virulence genes in the same emm-type strains changed with time, such as hyl. The resistance genes were basically the same among different emm-type strains except for the vanSE gene detected in all emm12 strains. The results of drug sensitivity showed that the GAS strains isolated in Tianjin City from 2011 to 2024 were sensitive to penicillin, cefazolin, chloramphenicol, vancomycin, and levofloxacin, while the resistance rates to erythromycin, azithromycin, clarithromycin, and clindamycin ranged from 88.5% to 100.0%, and there was a certain degree of consistency between the resistance phenotypes and the detected resistance genes. Overall, the main emm types and evolutionary features of GAS in Tianjin City from 2011 to 2024 were consistent with the dominant types in China, and the carrying rate of virulence genes and drug resistance genes differed significantly among different emm-type strains, and there were continuous evolution and variation in the prevalence of virulence genes in GAS.
7.Arthroscopic LUtarjet procedure to treat recurrent anterior shoulder dislocation combined with a glenoid bone defect
Qi CHEN ; Jicheng GONG ; Xin XU ; Wei LU
Chinese Journal of Orthopaedic Trauma 2025;27(8):681-688
Objective:To observe the efficacy of arthroscopic LUtarjet procedure in the treatment of recurrent anterior shoulder dislocation combined with a glenoid bone defect.Methods:A retrospective study was conducted to analyze the data of 15 patients with recurrent anterior shoulder dislocation combined with a glenoid bone defect who had been treated by the arthroscopic LUtarjet procedure at Department of Orthopedics, The 928th Hospital of PLA Joint Logistics Support Force from September 2022 to November 2023. All patients were male, aged (25.0±1.9) years, involving 10 left and 5 right sides. The anterior fear test for the affected shoulder was positive in all cases before surgery, and the dislocations averaged (13.8±5.4) times. By comparing the three-dimensional CT scans of both shoulders before surgery, the severity of the glenoid bone defect was calculated as over 20% wider than the width of the healthy glenoid. The shoulder function was evaluated at 6 and 18 months after operation using the American Shoulder and Elbow Surgeons score (ASES), Rowe score, Walch-Duplay score and shoulder active range of motion. Comparisons were made between preoperation and postoperation. CT plain scan and three-dimensional reconstruction were used to evaluate the location and resorption of the transferred bone block of the coracoid process.Results:All patients were followed up for (19.8±1.1) months. The operation time for the 15 patients was (117.6±13.8) min. Re-dislocation occurred in none of the patients during postoperative follow-up. There was no significant difference in the flexion elevation, lateral external rotation, internal rotation or abduction 90° external rotation between preoperation, 6 months or 18 months after operation ( P>0.05). Respectively, the ASES, Rowe and Walch-Duplay scores were (93.3±2.3) points, (90.8±2.6) points, and (95.9±1.0) points at 18 months after operation, and (89.6±3.8) points, (84.5±2.7) points, and (87.2±3.9) points at 6 months after operation, showing significant improvements compared with the preoperative values [(70.9±6.1) points, (42.9±6.0) points, and (66.2±11.0) points] ( P<0.05). In the 3 patients whose immediately postoperative CT scan showed that their bone blocks were slightly lower in position, the bone blocks grew and shaped well at the last follow-up. The last follow-up observed partial bone resorption in one patient. Conclusion:In the treatment of recurrent anterior shoulder dislocation combined with a glenoid bone defect, the arthroscopic LUtarjet procedure can restore satisfactory stability of the shoulder joint, leading to good short- and mid-term efficacy.
8.Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury (version 2025)
Aijun XU ; Shuixia LI ; Bo CHEN ; Mengyuan YE ; Lejiao LANG ; Ning NING ; Lin ZHANG ; Changqing LIU ; Zhonglan CHEN ; Weihu MA ; Weishi LI ; Xiaoning WANG ; Dongmei BIAN ; Jiancheng ZENG ; Xin WANG ; Yuan GAO ; Yaping CHEN ; Jiali CHEN ; Yun HAN ; Xiuting LI ; Yang ZHOU ; Xiaojing SU ; Qiong ZHANG ; Tianwen HUANG ; Ping ZHANG ; Hua LIN ; Xingling XIAO ; Ruifeng XU ; Fanghui DONG ; Bing HAN ; Luo FAN ; Yanling PEI ; Suyun LI ; Xiaoju TAN ; Rongchen GUO ; Yefang ZOU ; Xiaoyun HAN ; Junqin DING ; Yi WANG ; Shuhua DENG ; Jinli GUO ; Yinhua LIANG ; Yuan CEN ; Xiaoqin LIU ; Junru CHEN ; Haiyang YU ; Lunlan LI ; Ying REN ; Yunxia LI ; Jianli LU ; Ying YING ; Lan WEI ; Yin WANG ; Qinhong XU ; Yanqin ZHANG ; Yang LYU ; Shijun ZHANG ; Sui WENJIE ; Sanlian HU ; Shuhong YANG ; Guoqing LI ; Jingjing AN ; Baorong HE ; Leling FENG
Chinese Journal of Trauma 2025;41(6):530-541
Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.
9.Relationship between Helicobacter pylori infection and disease severity and pathological type of inpatients with intestinal polyps
Wei YOU ; Dalin LU ; Yan CHEN ; Xin WANG ; Yizheng FANG ; Lunshan WANG
Journal of Public Health and Preventive Medicine 2025;36(5):85-88
Objective To investigate the relationship between Helicobacter pylori (Hp) infection and disease severity and pathological type of intestinal polyps in inpatients. Methods The data of 303 inpatients with intestinal polyps in the hospital from August 2022 to February 2025 were collected and analyzed. The clinical characteristics of patients were analyzed, and the influencing factors of pathological types of polyps were explored. Results Among the 303 inpatients with intestinal polyps, there were 135 Hp positive cases and 168 Hp negative cases. The number of polyps, maximum polyp diameter, number of ileocecus/ascending colon/transverse colon polyps, number of descending colon/sigmoid colon/rectal polyps and adenomatous polyps in the Hp-positive group were higher than those in the Hp-negative group (P<0.05). Multivariate logistic regression analysis indicated that age [OR (95%CI)=1.03 (1.01-1.05)] and positive Hp[OR (95%CI)=2.61 (1.62-4.20)] were independent risk factors of occurrence of adenomatous polyps (P<0.05). ROC curve results revealed that the AUCs of age, positive HP and combination in the diagnosis of adenomatous polyps were 0.574, 0.608 and 0.646, and the 95%CI values were 0.509-0.638, 0.545-0.672 and 0.584-0.708 respectively. The efficiency of combination of the two indexes was higher than that of single diagnosis. Conclusion Hp infection is associated with disease severity in inpatients with intestinal polyps, and it may be involved in the occurrence and development of adenomatous polyps.
10.Formulation and Analysis of Clinical Pharmacist Teacher Training Standard
Ping LIN ; Jiancun ZHEN ; Wei ZHANG ; Zhuo WANG ; Yangui XU ; Pinfang HUANG ; Xin HUANG ; Qingchun ZHAO ; Ying ZHOU ; Jin LU ; Jing LIU ; Li YOU
Herald of Medicine 2025;44(3):404-407
Clinical pharmacist teacher training is an important mean to improve the quality of clinical pharmacy talent cultivation and ensure the service ability and level of the clinical pharmacist team.The Pharmacy Administration and Pharmacy Practice in Healthcare Institutions-Part 4-8-2:Pharmacy Administration-Pharmacy Training Management-Clinical Pharmacist Teacher Training was based on the newly revised management document for clinical pharmacist teacher training of the Chinese Hospital Association.After sorting out relevant materials,such as standards,policies and regulations,technical specifications,liter-ature,documents of the Chinese Hospital Association,expert opinions,and the current situation of clinical pharmacist teacher training in China,the standard was formulated.In the standard,12 key elements,which can be divided into 3 parts of base manage-ment,training process and assessment,quality management and evaluation improvement,were standardized.This article aimed to introduce the construction method and content of the standard,to facilitate the understanding of the standard content for medical institutions which joined or willing to join the clinical pharmacist teacher training base,and to provide a reference for other medi-cal institutions to carry out related work.


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