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 introduce the causes of accidents and the diagnosis and treatment of two patients with radiation-induced skin injury admitted to our hospital in 2023, and to provide a reference for the clinical treatment of subsequent radiation-induced skin injury. Methods The clinical treatment process of two patients with acute skin injury caused by external radiation exposure were summarized and analyzed. Results The exposure history of the two patients was reconstructed, the flaw detection scenario was simulated, the biological dose and hand skin exposure dose were estimated, and the infrared thermal imaging device was used for dynamic monitoring. A comprehensive analysis was conducted based on clinical manifestations and other data. The diagnosis of “Xie” was excessive exposure combined with acute radiation-induced skin injury on both hands (Grade IV for the right hand palm, index finger, and middle finger and Grade II for the left hand little finger). The diagnosis of “Hao” was acute radiation-induced skin injury on both hands (Grade I). The two patients received different clinical treatment measures: “Xie” was treated with both local and systemic therapies, while “Hao” was mainly treated with systemic therapy. Conclusion After systematic and effective treatment, the radiation-induced skin injuries healed in both patients.

ObjectiveCerebral ischemic injury triggers a complex pathological cascade characterized by excessive reactive oxygen species (ROS) accumulation, persistent oxidative stress, and sustained neuroinflammation in the injured brain microenvironment. These events collectively drive mitochondrial dysfunction, microglial overactivation, pro-inflammatory cytokine release, and progressive neuronal apoptosis, ultimately leading to severe and irreversible neurological deficits. However, conventional therapeutic strategies face critical limitations, including poor blood-brain barrier penetration, insufficient local drug concentration, uncontrolled drug release, and off-target systemic side effects. To address this pathological process, we rationally designed and fabricated an injectable ROS-responsive hydrogel loaded with polydopamine nanoparticles (PDA NPs) for spatiotemporally controlled antioxidation, anti-inflammation, and neuroprotection in the ischemic injury microenvironment. The present study aimed to systematically characterize the physicochemical properties, ROS-responsive drug release behavior, biocompatibility, and neuroprotective efficacy of this composite hydrogel system in vitro. MethodsPDA NPs were fabricated via oxidative self-polymerization. The ROS-responsive hydrogel was cross-linked using N1-(4-boronobenzyl)-N3-(4-boronophenyl)-N1,N1,N3,N3-tetramethylpropane-1, 3-diaminium (TSPBA) and polyvinyl alcohol (PVA). Morphology, particle size, Zeta potential, and structure of PDA NPs were characterized by dynamic light scattering (DLS), Zeta potential analysis, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Microstructure, rheological properties, shear-thinning behavior, and ROS-triggered release profiles of the hydrogel were examined by SEM and rheometry. Biocompatibility was evaluated using HT22 mouse hippocampal neurons with CCK-8 and live/dead staining. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was established to simulate ischemic injury in vitro. ROS levels and neuronal apoptosis were detected by DHE staining and TUNEL assay. Microglial polarization and pro-inflammatory cytokine expression were analyzed using immunofluorescence and RT-qPCR in BV-2 microglia. Transwell co-culture was used to verify the indirect neuroprotection mediated by modulated microglia. ResultsCharacterization results confirmed that the as-prepared PDA NPs were monodispersed spherical nanoparticles with uniform diameter and negative surface potential, demonstrating favorable dispersibility and robust ROS-scavenging activity. The TSPBA-PVA hydrogel exhibited a highly porous interconnected network, suitable mechanical strength, and obvious shear-thinning behavior, supporting its application as an injectable implant. More importantly, the hydrogel displayed typical ROS-responsive degradation and on-demand PDA NP release in a ROS-concentration-dependent manner. In vitro cellular experiments demonstrated that the PDA NP-loaded hydrogel possessed excellent biocompatibility with HT22 cells. In the OGD/R model, the hydrogel significantly reduced intracellular ROS accumulation and markedly suppressed neuronal apoptosis. Furthermore, the composite hydrogel effectively redirected BV-2 microglia from the pro-inflammatory M1 toward the anti-inflammatory M2 phenotypes, downregulated the expression of pro-inflammatory cytokines including TNF-α, IL-1β, and IL-6, and reduced inflammatory damage. Transwell co-culture assays further validated that M2-polarized microglia mediated by the hydrogel significantly enhanced the survival of OGD/R-injured HT22 neurons and attenuated apoptosis. ConclusionIn this study, we successfully developed a novel injectable ROS-responsive hydrogel loaded with PDA NPs for synergistic antioxidative and anti-inflammatory neuroprotection. This intelligent hydrogel system enables ROS-triggered on-demand release of PDA NPs, efficiently scavenges excessive ROS, inhibits oxidative stress injury, modulates microglial polarization, and suppresses neuroinflammation, thereby exerting robust neuroprotective effects in vitro. This biomaterial platform provides a promising strategy for the targeted and controlled delivery of bioactive nanomaterials in the central nervous system diseases and establishes a solid experimental foundation for the development of in situ injectable therapies for ischemic brain injury.

In the context of"Artificial Intelligence(AI)+Education"era,this study introduces the teaching reform and devel-opment path of the course"Fundamentals of Immunology and Pathogenic Biology"by Professor Lu Fangguo's team at Hunan University of Traditional Chinese Medicine.After twenty years of exploration,the course has successfully transitioned from traditional teaching to intelligent teaching,achieving a comprehensive upgrade in educational concepts,teaching methods,and resources.The reform process is divided into three stages:Early exploration,comprehensive reform,and deepening development.It encompasses the construction of a smart teaching platform,the development and promotion of new forms of digital teaching resources,and the deep integration of ideological and political education.This reform has significantly enhanced teaching quality and students'overall competencies,showcasing the innovative spirit of educators.It has gained nationwide recognition and promotion,providing valuable references for the innovation of medical education in the new era.

OBJECTIVE: Hepatocellular carcinoma (HCC) is sensitive to ferroptosis, a new form of programmed cell death that occurs in most tumor types. However, the mechanism through which ferroptosis modulates HCC remains unclear. This study aimed to investigate the oncogenic role and prognostic value of FANCD2 and provide novel insights into the prognostic assessment and prediction of immunotherapy. METHODS: Using clinicopathological parameters and bioinformatic techniques, we comprehensively examined the expression of FANCD2 macroscopically and microcosmically. We conducted univariate and multivariate Cox regression analyses to identify the prognostic value of FANCD2 in HCC and elucidated the detailed molecular mechanisms underlying the involvement of FANCD2 in oncogenesis by promoting iron-related death. RESULTS: FANCD2 was significantly upregulated in digestive system cancers with abundant immune infiltration. As an independent risk factor for HCC, a high FANCD2 expression level was associated with poor clinical outcomes and response to immune checkpoint blockade. Gene set enrichment analysis revealed that FANCD2 was mainly involved in the cell cycle and CYP450 metabolism. CONCLUSION To the best of our knowledge, this is the first study to comprehensively elucidate the oncogenic role of FANCD2. FANCD2 has a tumor-promoting aspect in the digestive system and acts as an independent risk factor in HCC; hence, it has recognized value for predicting tumor aggressiveness and prognosis and may be a potential biomarker for poor responsiveness to immunotherapy.
Humans ; Carcinoma, Hepatocellular/diagnosis* ; Liver Neoplasms/diagnosis* ; Immunotherapy ; Fanconi Anemia Complementation Group D2 Protein/metabolism* ; Prognosis ; Male ; Female ; Middle Aged ; Biomarkers, Tumor/metabolism*

Objective:To investigate the characteristic distribution of vaginal microbiota in infertile women.Methods:We collected the results of vaginal microbiological examinations from 19322 initial visit infertile women who visited the Reproductive Medicine Center of the Department of Obstetrics and Gynecology,the Second Affili-ated Hospital of Air Force Medical University from March 1,2023 to July 31,2024.The vaginal microbiota infection status of patients was compared in different age groups(＜25 years old,25-＜30 years old,30-＜35 years old,35-＜40 years old,≥40 years old)and different seasons(spring,summer,autumn,winter).Results:①Among 19322 women,6027 cases(31.19％)showed abnormal vaginal microecology.Pathogenic microorganisms were detected in 3093 cases,including 2882 cases of single vaginitis,211 cases of mixed vaginitis,3764 cases with vagi-nal cleanliness grade Ⅲ-Ⅳ,and 3965 cases with abnormal lactobacilli.Among patients with single vaginitis,1349 cases(46.81％)were diagnosed with vulvovaginal candidiasis(VVC),which was the highest proportion.Aerobic vaginitis(AV)followed with 752 cases(26.09％),and bacterial vaginosis(BV)had 671 cases(23.28％),trichomonal vaginitis(TV)with 110 cases(3.82％).Among patients with mixed vaginitis,AV+BV was the most common with 96 cases(45.49％).②The detection rates of cleanliness grade Ⅲ-Ⅳ,abnormal lactobacilli,abnor-mal microorganisms(unclear pathogen),single vaginitis(BV,VVC and TV),and mixed vaginitis showed statisti-cally significant differences across different seasons(P＜0.05).Specifically,the detection rates of cleanliness grade Ⅲ-Ⅳ and abnormal microorganisms(unclear pathogens)were significantly higher in autumn than in other seasons(P＜0.05),while the detection rate of abnormal lactobacilli was higher in spring than in other seasons(P＜0.05).③The detection rates of abnormal lactobacilli,abnormal microorganisms(unclear pathogen),single vaginitis(BV,VVC and AV),and mixed vaginitis showed significant differences among different age groups(P＜0.05).Specifically,the detection rate of abnormal microorganisms(unclear pathogen)was higher in the age group＜25 years than in other age groups(P＜0.05),while the detection rate of BV among single vaginitis cases was higher in the age group≥40 years than in other age groups(P＜0.05).Conclusions:The vaginal microecol-ogy of infertile women varies in terms of infection rates across different age groups and seasons.Patients with simple vaginitis have the highest rate of VVC,while those with mixed vaginitis have the highest proportion of AV+BV infection.

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.

AIM To establish a rapid quantitative analysis model for saponins in Paris polyphylla var.yunnanensis(PPY)by near infrared spectroscopy.METHODS The contents of polyphyllins Ⅰ,Ⅱ,Ⅶ and there total content in PPY were determined by HPLC,while spectral data within the range of 10 000 to 4 000 cm-1 were collected.A quantitative analysis model was established by combining these data with partial least squares regression(PLSR).Multivariate scatter correction(MSC)and vector normalization(SNV)were applied prior to further preprocessing the spectra with original,first-order derivative(1stD),or second-order derivative(2ndD)treatments.Lastly,the model was optimized through non-smoothing(NS),Norris Derivative filtering(Nd),and Savitzky-Golay filtering(S-G)method.Model stability was evaluated based on correlation coefficients and variance.The predicted contents of each saponin component in the validation set samples were calculated.RESULTS The contents of polyphyllins Ⅰ,Ⅱ,Ⅶ were 0.42-17.98,0.46-10.44,0.23-3.86 mg/g,respectively.The total content ranged from 2.91 to 22.1 mg/g.The optimal parameters of three saponins were achieved when selecting the MSC+2ndD+S-G pretreatment method.The corresponding ratio of line segment length to segment gap was 13∶5,15∶5,11∶5,with correlation coefficients of 0.982,0.930,0.958,respectively.The root mean square errors of calibration(RMSEC)were 0.702,0.797,0.238,and the root mean square errors of prediction(RMSEP)were 1.120,0.835,0.304,respectively.The optimal parameters for the total content were obtained when selecting the MSC+2ndD+NS pretreatment method,with a correlation coefficient of 0.970,a RMSEC of 1.090,and a RMSEP of 1.740.CONCLUSION This accurate and rapid method can be used for detection of saponin contents in P.Polyphylla.

Objective:To explore the relationship between sperm DNA integrity, semen parameters, seminal plasma neutral α-glucosidase activity and in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) outcomes. Methods:The clinical data of 300 male infertile patients who underwent routine semen analysis in the Second Affiliated Hospital of Air Force Medical University from October 2023 to April 2024 and whose spouses received IVF/ICSI treatment were retrospectively analyzed. The results of sperm DNA integrity [sperm DNA fragmentation index (DFI)], semen parameters (sperm percentage, sperm density, sperm motility, normal morphology sperm rate) and seminal plasma neutral α-glucosidase activity at admission were recorded. According to their spouses′ IVF/ICSI outcomes, the 300 patients were divided into the successful pregnancy group (those with successful intrauterine pregnancy detected by ultrasound 30 days after transplantation) and the unsuccessful pregnancy group (those without successful intrauterine pregnancy detected by ultrasound 30 days after transplantation). The differences in sperm DNA integrity, semen parameters and seminal plasma neutral α-glucosidase activity at admission between the two groups were compared. The receiver operating characteristic (ROC) curve was used to analyze the predictive efficacy of sperm DNA integrity, semen parameters and seminal plasma neutral α-glucosidase activity in male infertile patients at admission for their spouses′ IVF/ICSI outcomes.Results:According to the IVF/ICSI outcomes of the spouses of male infertile patients, 169 cases (56.3%) had successful intrauterine pregnancy detected by ultrasound 30 days after transplantation; 131 cases (43.7%) had unsuccessful intrauterine pregnancy. At admission, there was no statistically significant difference in clinical data between the two groups (all P>0.05). The DFI and sperm density in the successful pregnancy group were lower than those in the unsuccessful pregnancy group, while the sperm percentage, sperm motility, normal morphology sperm rate and seminal plasma neutral α-glucosidase activity were higher than those in the unsuccessful pregnancy group (all P<0.05). The area under the ROC curve (AUC) values of single indicators including DFI, sperm percentage, sperm density, sperm motility, normal morphology sperm rate, and seminal plasma neutral α-glucosidase level in predicting the IVF/ICSI outcomes of spouses of male infertile patients were 0.719, 0.718, 0.812, 0.779, 0.769, and 0.736, respectively; the sensitivities were 70.42%, 77.46%, 69.01%, 70.42%, 69.01%, and 77.46%, respectively; the specificities were 66.38%, 55.02%, 81.22%, 73.80%, 77.29%, and 62.88%, respectively; the Youden indexes were 0.368, 0.325, 0.502, 0.442, 0.463, and 0.404, respectively; all differences were statistically significant (all P<0.05). Conclusions:Sperm DNA integrity, semen parameters and seminal plasma neutral α-glucosidase activity can assist in predicting IVF/ICSI outcomes, providing an important reference for the treatment outcomes of male infertile patients.

Objective To investigate the effect and mechanism of M2 macrophage polarization induced by HCT116 with highly metastatic colorectal cancer cells on immune escape in colorectal cancer.Methods After the co-cul-ture of colorectal cancer cells conditioned medium(CM)and PMA-induced M0 macrophages,the polarization of M2 macrophages was observed by flow cytometry,real-time polymerase chain reaction(qPCR)and enzyme-linked immunosorbent assay(ELISA)experiments.The CMM0,CMSW480-Mφand CMHCT116-Mφ were co-cultured with HCT116 cells,and the expression of programmed death-ligand 1(PD-L1)was detected by Western blot and qPCR.At the same time,the stimulated HCT116 cells were co-cultured with human T lymphocytes to detect the survival of HCT116 cells and the levels of tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ).The difference of kruüppel-like factor 4(KLF4)expression between SW480 and HCT116 cells was detected by Western blots,qPCR and ELISA.After pretreatment of HCT116 cells with KFL4 inhibitor Kenpaullone(Ken),the CMHCT116 and CMHCT116+Ken were co-cultured with M0 macrophages and the polarization of M2 macrophages was observed by flow cytometry,qPCR and ELISA.The CMHCT116-Mφ and CM(HCT116+Ken)-Mφ was co-cultured with HCT116 cells,and the PD-L1 expression of HCT116 cells was detected by Western blot and qPCR.Results After the stimulation of M0 macrophages with CM,the proportion of CD11b+CD206+cells in HCT116-Mφ cells was higher,and the expres-sion of M2 macrophage markers interleukin(IL-10)and transforming growth factor-β(TGF-β)were higher.Com-pared with the CMSW480-Mφ group,the PD-L1 protein expression level was higher in the CMHCT116-Mφ group.After co-culture with T lymphocytes,the cell survival rate are the most in CMHCT116-Mφ group,while the levels of TNF-α and IFN-γ were the lowest.After the addition of Ken,the polarization ratio and markers of M2 macrophages decreased.Compared with CMHCT116-Mφ group,the expression of PD-L1 in HCT116 cells of the CM(HCT116+Ken)-Mφ group de-creased.Conclusion Highly metastatic colorectal cancer cells induce polarization of M2 macrophages by secreting KLF4,promote PD-L1 expression in colorectal cancer cells,facilitate tumor immune escape,and provide potential targets for clinical immunotherapy.