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.

Abstract To estavlish a more comprehensive theoretical framework for the weight management of children and adolescents, the study systematically expounds on the two core strategies for dietary and exercise intervention. It explores the mechanism of diet, physical activity, breating exercise, functional movement correction and multi dimensional integrated exercise modalities in preventing weight rebound after weight loss in overweight and obesity children and adolescents. Future advancements in research methodology are expected to improve the evidence system of collaborative interventions, so as to provide precise strategies for obesity management in children and adolescents.

Objective: To explore the association between daytime outdoor physical activity (OPA) and oral health indicators among primary school students, aiming to provide evidence for developing oral health intervention strategies based on natural exposure. Methods: In October 2023, based on the "Tianchang Children s Light Exposure and Growth Development Cohort", 799 second and third grades children were recruited from two primary schools in Tianchang, Chuzhou City, Anhui Province. Physical activity intensity and light exposure were objectively monitored for 24 hours over 5 consecutive days using triaxial accelerometers synchronized with portable illuminance meters. Standardized oral examinations were performed to record dental caries, gingivitis, and malocclusion. Demographics, lifestyle variables, and household socioeconomic data were collected via questionnaires. Multiple linear regression (for the number of carious teeth) and Logistic regression (for gingivitis risk) were used to analyze the relationship between daytime outdoor moderate to vigorous physical activity (MVPA) and oral health outcomes. Results: The average daytime outdoor MVPA was (0.76±0.35)h, with (0.95±0.40)h on weekdays and (0.49±0.47)h on weekends. The detection rates for dental caries, gingivitis, and malocclusion were 31.0%, 4.6%, and 59.7%, respectively. Compared with children with good oral health, the duration of outdoor MVPA on school days was reduced in children with caries or gingivitis ( Z =-11.4, -5.01, both P <0.01). Multiple regression analysis showed that after adjusting for factors such as gender, age, body mass index, oral hygiene behaviors, an increase in daytime outdoor MVPA duration was associated with a decrease in the number of dental caries ( β=-0.64, 95%CI =-0.93 to -0.35) and a reduced risk of gingivitis ( OR= 0.58 , 95%CI =0.34-0.98) in primary school students (both P <0.05). The association was primarily observed on school days ( β=-0.72, 95%CI = -1.07 to -0.37; OR=0.42, 95%CI =0.21-0.85) (both P < 0.05). Conclusions Daytime outdoor MVPA on weekdays is significantly associated with a lower number of carious teeth and a reduced risk of gingivitis in primary school students. Increasing daytime outdoor activities on weekdays may serve as a promising and potential strategy for promoting children s oral health.

Objective: To investigate the intervention effects and mechanisms of function-rehabilitation-oriented physical training on the physical health of higher vocational college students, so as to provide replicable strategies for integrating physical education with health education in such institutions. Methods: In March 2024, a cluster random sampling method was employed to select 312 first and second year non sports major students from Chongqing Water Resources and Electric Engineering College as research subjects, and the selected classes were randomly divided into an intervention group ( n =151) and a control group ( n =161) by using a random number table. Both groups received a 16 week intervention program consisting of 2 academic hours per week from Macrh to July 2024. The control group followed the standard teaching curriculum for public physical education courses in higher vocational education, while the intervention group incorporated a function-rehabilitation-oriented physical training module into the regular physical education curriculum. For the module, stratified functional correction was implemented based on pre class functional movement screening results, with dedicated 10-15 minute intervention sessions per lesson accompanied by personalized exercise prescriptions delivered three times weekly. Before and after the intervention, Functional Movement Screen, the Exercise Attitude Scale, and the National Student Physical Health Standards (2014 Revision) were used for pre and post intervention assessments. Main effect of the intervention was examined via repeated measures analysis of variance, while chain mediating effect was analyzed by using the Hayes Process macro program. Results: After the intervention, the scores for functional movement quality (15.82±2.35), physical exercise attitude (237.64±17.22), and physical health level (76.45±7.82) in the intervention group were significantly higher than those before intervention (13.10±2.41, 199.51±15.78, 70.30±8.34) and those in the control group during the same period (13.08±2.81, 199.02±18.67, 71.42±7.56)( t =10.07, 20.34, 6.58; 9.21, 19.17, 5.57, all P < 0.01 ). Function-rehabilitation-oriented physical training directly and positively predicted physical health status ( β =0.18). Functional movement quality (effect size=0.21) and physical exercise attitude (effect size= 0.15 ) each served as independent mediators, their chain mediating effect was statistically significant (effect size=0.10)(all P < 0.01 ), with the total indirect effect accounting for 71.88% of the overall effect. Conclusion Function-rehabilitation-oriented physical training significantly improves the physical health of vocational college students through both direct effects and chain pathways, making it suitable for widespread implementation in vocational education institutions.

This article explores the pathogenesis ， evolution and the prevention-treatment strategies of hypertension-related coronary microvascular dysfunction （H-CMD） from the collateral disease theory. Long-term hypertension may lead to coronary microvascular vasomotor imbalance， endothelial injury， vascular remodeling， microthrombosis， reduced perfusion reserve， and microvascular rarefaction， suggesting a progressive pathological process from functional disturbance to structural injury， collateral obstruction， and insufficient repair. Collateral disease theory emphasizes the network-like distribution， gradual branching， and permeating and nourishing functions of collaterals， as well as the concept that prolonged disease enters the collaterals， providing a reference for understanding coronary microcirculatory dysfunction and malnourishment of heart collaterals. Accordingly， the traditional Chinese medicine （TCM） pathogenesis of H-CMD can be summarized into four interconnected stages： Collateral Qi stagnation， scorching injury of collateral vessels， phlegm-stasis obstruction of collaterals， and collateral deficiency with malnourishment. Collateral Qi stagnation is the initiating stage， mainly corresponding to abnormal coronary microvascular vasomotor regulation and endothelial dysfunction. Scorching injury of collateral vessels represents the developmental stage and is closely associated with inflammation， oxidative stress， and endothelial barrier injury after Qi stagnation transforms into fire. Phlegm-stasis obstruction of collaterals represents the prolonged stage and may aggravate vascular wall thickening， luminal stenosis， microthrombosis， and insufficient microcirculatory perfusion. Collateral deficiency with malnourishment is the late-stage outcome， reflecting deficiency of healthy Qi， collateral damage， Qi-blood malnourishment， and weakened repair capacity， mainly manifested as microvascular rarefaction， occlusion， and microcirculatory homeostatic imbalance. Therapeutically， stage-based syndrome differentiation and the combined strategy of dredging and nourishing collaterals should be emphasized. Specifically， promoting Qi movement， resolving stagnation， soothing the liver， and harmonizing collaterals should be adopted in the early stage. Clearing heat， pacifying the liver， calming and protecting collaterals should be adopted in the developmental stage. Resolving phlegm， activating blood， and dredging collaterals should be implemented in the prolonged stage. Supplementing Qi， nourishing blood， tonifying the kidney， regulating balance， strengthening healthy Qi， and nourishing collaterals should be carried out in the late stage. Related formulas and medicinals may act by regulating the nitric oxide/endothelin-1（NO/ET-1） balance， inhibiting inflammation and oxidative stress， improving microcirculatory perfusion， and promoting angiogenesis and endothelial repair. Collateral disease theory may provide a theoretical basis for understanding the dynamic pathogenesis of H-CMD and for stage-based TCM intervention.

Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by progressive cognitive decline, functional impairment, and neuropsychiatric symptoms. It represents the most prevalent form of dementia among the elderly population. Accumulating evidence indicates that oxidative stress plays a pivotal role in the pathogenesis of AD. Notably, elevated levels of oxidative stress have been observed in the brains of AD patients, where excessive reactive oxygen species (ROS) can cause extensive damage to lipids, proteins, and DNA, ultimately compromising neuronal structure and function. Amyloid β‑protein (Aβ) has been shown to induce mitochondrial dysfunction and calcium overload, thereby promoting the generation of ROS. This, in turn, exacerbates Aβ aggregation and enhances tau phosphorylation, leading to the formation of two pathological features of AD: extracellular Aβ plaque deposition and intracellular neurofibrillary tangles (NFTs). These events ultimately culminate in neuronal death, forming a vicious cycle. The interplay between oxidative stress and these pathological processes constitutes a core link in the pathogenesis of AD. The signaling pathways mediating oxidative stress in AD include Nrf2, RCAN1, PP2A, CREB, Notch1, NF‑κB, ApoE, and ferroptosis. Nrf2 signaling pathway serves as a key regulator of cellular redox homeostasis, exerts important antioxidant capacity and protective effects in AD. RCAN1 signaling pathway, as a calcineurin inhibitor, and modulates AD progression through multiple mechanisms. PP2A signaling pathway is involved in regulating tau phosphorylation and neuroinflammation processes. CREB signaling pathway contributes to neuroplasticity and memory formation; activation of CREB improves cognitive function and reduce oxidative stress. Notch1 signaling pathway regulates neuronal development and memory, participates in modulation of Aβ production, and interacts with Nrf2 toco-regulate antioxidant activity. NF‑κB signaling pathway governs immune and inflammatory responses; sustained activation of this pathway forms “inflammatory memory”, thereby exacerbating AD pathology. ApoE signaling pathway is associated with lipid metabolism; among its isoforms, ApoE-ε4 significantly increases the risk of AD, leading to elevated oxidative stress, abnormal lipid metabolism, and neuroinflammation. The ferroptosis signaling pathway is driven by iron-dependent lipid peroxidation, and the subsequent release of lipid peroxidation products and ROS exacerbate oxidative stress and neuronal damage. These interconnected pathways form a complex regulatory network that regulates the progression of AD through oxidative stress and related pathological cascades. In terms of therapeutic strategies targeting oxidative stress, among the drugs currently used in clinical practice for AD treatment, memantine and donepezil demonstrate significant therapeutic efficacy and can improve the level of oxidative stress in AD patients. Some compounds with antioxidant effects (such asα-lipoic acid and melatonin) have shown certain potential in AD treatment research and can be used as dietary supplements to ameliorate AD symptoms. In addition, non-drug interventions such as calorie restriction and exercise have been proven to exerted neuroprotective effects and have a positive effect on the treatment of AD. By comprehensively utilizing the therapeutic characteristics of different signaling pathways, it is expected that more comprehensive multi-target combination therapy regimens and combined nanomolecular delivery systems will be developed in the future to bypass the blood-brain barrier, providing more effective therapeutic strategies for AD.

Objective: To investigate the weight loss effect of a comprehensive intervention model combining caloric restriction (CR), physical activity (PA), behavioral therapy (BT), breathing exercise (BE), and functional movement corrective training (FMCT)-referred to as the "CPBBF" model in overweight and obese female college students, so as to provide a reference for scientific weight loss interventions for college students. Methods: From March to May 2022, 46 overweight and obese female college students from Chongqing Water Resources and Electric Engineering College were recruited and randomly divided into an experimental group (24 participants) and a control group (22 participants). The control group received CR (prohibiting ad libitum snacking), PA in the first week, high intensity interval training (HIIT) for 30 s, and moderate intensity continuous training (MICT) for 1-5 min alternate 4 sets, duration 15-20 min. From the second week, adjust to HIIT and MICT alternating 3 min each for 5 sets, totaling 30 min, 4 times/week, 70 min/time and BT (60-90 min/session, 3 times/week). The experimental group incorporated FMCT (10-15 min of focused training per session, integrated with PA and daily life) and BE (advocating a gradual transition to proper breathing methods in daily life and low intensity training, 5 sessions/day, 10 min each). Body oxygen level test (BOLT), Functional Movement Screen (FMS), sports exercise attitude, and body composition indicators were measured at baseline (T0), after 12 weeks of intervention (T1), and after one year of follow up (T2). The differences were analyzed between groups through generalized estimation equations, and mixed effect model analysis was employed to explore predictive relationships among variables. Results: The results of the generalized estimation equation showed that time main effects of BOLT values, FMS scores, and exercise attitude among female college students were statistically significant ( Wald χ 2=18.75, 14.89, 12.45, all P <0.01); further intragroup comparisons revealed that BOLT， functional motor screening (FMS) scores, and physical exercise attitudeof female college students in the experimental group increased compared to T0, while the control group only showed an increase at T1 (all P <0.05). The group main effects for the aforementioned three indicators were statistically significant ( Wald χ 2=6.33, 5.21, 4.88), and the time by group interactions of BOLT values and FMS scores were also statistically significant ( Wald χ 2=4.56, 3.97) (all P <0.05). The time main effects of body weight, body mass index (BMI), and body fat ratio(BFR) in female college students were statistically significant ( Wald χ 2=44.27, 13.90, 82.33); further intragroup comparisons revealed that the experimental group of female college students showed a decrease in body weight, BMI and BFR at T1 and T2 compared to T0, while the control group only showed a decrease in these indicators at T1 (all P <0.05). The group main effects of weight and BFR were statistically significant ( Wald χ 2= 4.11 , 6.46), and the time by group interaction of BFR was statistically significant ( Wald χ 2=8.73) (all P <0.05).The results of mixed effect model analysis showed that BOLT ( β =1.52) and FMS ( β =1.81) could both positively predict physical exercise attitude, and physical exercise attitude had statistically significant negative predictive effects on weight, BMI, and BFR ( β =-0.08, -0.03 , -0.03) (all P <0.01). Conclusion The "CPBBF" comprehensive intervention effectively maintains weight loss effects by modulating the energy compensation mechanism with strong robustness.