Objective To construct glucose oxidase（GOx）–loaded nanogels （GONGs）, optimize their formulation, and evaluate their capacity to inhibit the Warburg effect in glioma cells. Methods A responsive polymer （HAM） was synthesized and used to self-assemble GONGs, which were then characterized. Encapsulation efficiency and drug loading were determined using fluorescence spectrophotometry. Biocompatibility was tested by measuring cytotoxicity and hemolytic activity. Western blotting was used to evaluate the effects of GONGs on the expression of proteins associated with the Warburg phenotype and oxidative damage in glioma cells. Results GONGs prepared at a drug–to–polymer ratio of 1∶10 exhibited a particle size of 140.3 nm and a zeta potential of −27.2 mV. Compared with free GOx, GONGs markedly reduced cytotoxicity, increased the IC₅₀ in hUVEC cells from 2.150 nmol/L to 74.86 nmol/L, and significantly decreased hemolysis. At a GOx concentration of 2 nmol/L, GONGs effectively downregulated glycolysis-related proteins, such as HK2 and LDHA, and inhibited glutamine metabolism in glioma cells. Conclusion GONGs exhibited high GOx loading capacity, significantly reduced GOx-induced cytotoxicity, inhibited the Warburg effect in glioma cells and induced oxidative damage.

Hepatic fibrosis is a reversible pathological process in various chronic liver diseases and is closely associated with the development and progression of severe liver diseases such as liver cirrhosis and hepatocellular carcinoma， and it has emerged as a significant global health challenge. In recent years， studies have shown that histone lactylation， a newly discovered epigenetic modification， actively participates in regulating the progression of hepatic fibrosis. This article systematically reviews the core regulatory effect of histone lactylation modification in the interaction between inflammatory microenvironment and hepatic fibrosis， in order to clarify the cascade regulatory mechanism of “inflammation-hepatic fibrosis” and provide new insights for early diagnosis， targeted intervention， and prevention of malignant transformation in hepatic fibrosis.

Liver fibrosis is a critical pathological process in the progression of chronic liver diseases and is mainly driven by the activation of hepatic stellate cells， and it is characterized by excessive deposition of extracellular matrix. Programmed cell death is widely observed in liver fibrosis and plays diverse roles in different types of cells. It is not only a mechanism for maintaining cellular homeostasis through metabolism， but it can also regulate the development and progression of diseases. Ferroptosis has recently emerged as a research focus due to its close association with lipid metabolism. This article systematically reviews the related concepts of lipid metabolism， ferroptosis， and liver fibrosis， analyzes the bridging role and dual regulatory function of ferroptosis in lipid metabolism and liver fibrosis， and further highlights the clinical application value of targeting ferroptosis in hepatic stellate cells for the treatment of liver fibrosis， in order to provide new perspectives for the mechanistic studies on liver fibrosis and the optimization of related clinical intervention strategies.

Objective To explore the molecular mechanism of autophagy mediated by the protein kinase B(AKT)/mammalian target protein of rapamycin(mTOR)pathway in the rehabilitation of muscle atrophy associated with rotator cuff tears(RCTs).Methods Forty male C57BL/6J mice were randomly assigned to the following four groups:sham group,RCTs group,RCTs+exercise group,and RCTs+exercise+rapamycin group,with 10 mice in each group.On the eighth week after grouping,healing of the bone-tendon interface and muscle cell atrophy were analyzed by histology.The mRNA expression levels of muscle-atrophy-related genes(Atrogin-1,Bnip 3,MuRF-1)in supraspinatus muscle tissue were measured by real-time quantitative reverse transcription polymerase chain reaction.The expression of LC3 and AKT/mTOR signal pathway proteins in the supraspinatus muscle tissue of the groups was detected by Western blot,and the degree of autophagy in each group was analyzed by transmission electron microscope.Results Compared with the sham operation group,in RCTs group's maturity score for the bone-tendon interface at the supraspinatus tendon anchorage and the cross-sectional area of the supraspinatus muscle fibers decreased significantly(P＜0.001),while muscle loss and the expression of Atrogin-1,Bnip 3,and MuRF-1 increased significantly(P＜0.001).Compared with the RCTs group,the RCTs+exercise group showed a significant increase in bone-tendon interface maturity score and cross-sectional area of the supraspinatus muscle fibers(P＜0.01)and a decrease in muscle loss and the expression of Atrogin-1,Bnip 3,and MuRF-1(P＜0.01).Compared with the sham group,the RCTs group's LC3Ⅰ/LC3Ⅱ and degree of autophagy in the supraspinatus muscle increased significantly(P＜0.001),while p-AKT/AKT and p-mTOR/mTOR expression decreased significantly(P＜0.01).Compared with RCTs group,the RCTs+exercise group's LC3Ⅰ/LC3Ⅱ and degree of autophagy decreased significantly(P＜0.01)and p-AKT/AKT and p-mTOR/mTOR expression increased significantly(P＜0.001).The addition of rapamycin significantly reversed the rehabilitation effect of exercise in the RCTs group.Conclusions This study confirmed the anti-atrophy effect of exercise rehabilitation in RCT diseases and showed that its mechanism is related to AKT/mTOR signal activation,which inhibits autophagy.

OBJECTIVES: The mechanism of the odontogenic differentiation of apical papillary cells (APCs) stimulated by bioactive glass 45S5 is still unclear. This study aims to investigate the effect of autophagy on the odontogenic differentiation of APCs stimulated by bioactive glass 45S5. METHODS: APCs were isolated and cultured in vitro, and the cell origin was identified by flow cytometry. The culture medium was prepared with 1 mg/mL 45S5, and its pH and ion concentration were determined. The experiments were divided into control, 45S5, and 3-methyladenine (3-MA) 45S5 groups. In the 45S5 group, APCs were induced to culture with 1 mg/mL 45S5. In the 3-MA 45S5 group, the autophagy inhibitor 3-MA was added to 1 mg/mL 45S5. Protein immunoblotting assay (Western blot) was used to detect the expression of autophagy-associated proteins of microtubule-associated protein 1 light-chain 3β (LC3B) and P62 after 24 h of induction culture in each group. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of bone sialoprotein (BSP), Runt-related transcription factor 2 (Runx2), dentin sialophosphoprotein (DSPP), and dentin matrix protein-1 (DMP-1) after 7 d of induction culture. Cellular alkaline phosphatase (ALP) staining analyzed cellular ALP activity at 7 d of induction, and alizarin red staining evaluated the formation of mineralized nodules at 21 d of induction. RESULTS: The pH of the 45S5 extract culture medium was 8.65±0.01, which was not significantly different from that of the control group (P>0.05). The silicon ion concentration of the 45S5 induction culture medium was (1.56±0.07) mmol/L, which was higher than that of the control group (0.08±0.01) mmol/L (P<0.05). The calcium ion concentration of the 45S5 induction culture was (1.57±0.15) mmol/L, which was not significantly different from that of the control group (P>0.05). Western blot results showed that LC3B-Ⅱ/Ⅰ ratio increased and P62 expression decreased in the 45S5 group compared with those in the control group (P<0.05). By contrast, the ratio decreased and the expression increased in the 3-MA 45S5 group compared with those in the 45S5 group (P<0.05). RT-qPCR results showed that the expression of BSP, Runx2, DMP-1, and DSPP enhanced in the 45S5 group compared with that in the control group (P<0.05), but the expression decreased in the 3-MA 45S5 group compared with that in the 45S5 group (P<0.05). Semi-quantitative analysis of ALP staining and alizarin red staining showed that the ALP activity was enhanced, and the formation mineralized nodule increased in the 45S5 group compared with those in the control group. The ALP activity weakened, and the formation mineralized nodules were reduced in the 3-MA 45S5 group compared with that those in the 45S5 group. CONCLUSIONS Cell autophagy participates in the odontogenic differentiation of APCs induced by 1 mg/mL 45S5 in vitro.
Autophagy/drug effects* ; Cell Differentiation/drug effects* ; Odontogenesis/drug effects* ; Dental Papilla/cytology* ; Humans ; Microtubule-Associated Proteins/metabolism* ; Glass/chemistry* ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Extracellular Matrix Proteins/metabolism* ; Ceramics/pharmacology* ; Adenine/pharmacology* ; Sialoglycoproteins/metabolism* ; Phosphoproteins/metabolism* ; Integrin-Binding Sialoprotein/metabolism* ; Alkaline Phosphatase/metabolism* ; RNA-Binding Proteins

Objective To explore the molecular mechanism of autophagy mediated by the protein kinase B(AKT)/mammalian target protein of rapamycin(mTOR)pathway in the rehabilitation of muscle atrophy associated with rotator cuff tears(RCTs).Methods Forty male C57BL/6J mice were randomly assigned to the following four groups:sham group,RCTs group,RCTs+exercise group,and RCTs+exercise+rapamycin group,with 10 mice in each group.On the eighth week after grouping,healing of the bone-tendon interface and muscle cell atrophy were analyzed by histology.The mRNA expression levels of muscle-atrophy-related genes(Atrogin-1,Bnip 3,MuRF-1)in supraspinatus muscle tissue were measured by real-time quantitative reverse transcription polymerase chain reaction.The expression of LC3 and AKT/mTOR signal pathway proteins in the supraspinatus muscle tissue of the groups was detected by Western blot,and the degree of autophagy in each group was analyzed by transmission electron microscope.Results Compared with the sham operation group,in RCTs group's maturity score for the bone-tendon interface at the supraspinatus tendon anchorage and the cross-sectional area of the supraspinatus muscle fibers decreased significantly(P＜0.001),while muscle loss and the expression of Atrogin-1,Bnip 3,and MuRF-1 increased significantly(P＜0.001).Compared with the RCTs group,the RCTs+exercise group showed a significant increase in bone-tendon interface maturity score and cross-sectional area of the supraspinatus muscle fibers(P＜0.01)and a decrease in muscle loss and the expression of Atrogin-1,Bnip 3,and MuRF-1(P＜0.01).Compared with the sham group,the RCTs group's LC3Ⅰ/LC3Ⅱ and degree of autophagy in the supraspinatus muscle increased significantly(P＜0.001),while p-AKT/AKT and p-mTOR/mTOR expression decreased significantly(P＜0.01).Compared with RCTs group,the RCTs+exercise group's LC3Ⅰ/LC3Ⅱ and degree of autophagy decreased significantly(P＜0.01)and p-AKT/AKT and p-mTOR/mTOR expression increased significantly(P＜0.001).The addition of rapamycin significantly reversed the rehabilitation effect of exercise in the RCTs group.Conclusions This study confirmed the anti-atrophy effect of exercise rehabilitation in RCT diseases and showed that its mechanism is related to AKT/mTOR signal activation,which inhibits autophagy.

Objective:To construct the rpoN gene knockout strain (Δ rpoN) and the complemented strain (CΔ rpoN) of Vibrio vulnificus ( V. vulnificus), and investigate the role of the rpoN gene in regulating bacterial motility and biofilm formation. Methods:The Δ rpoN strain of V. vulnificus was constructed using homologous recombination. Bacterial motility was assessed via swimming assays, and flagellar morphology was observed by transmission electron microscopy. Biofilm formation capacity was evaluated using crystal violet and Congo red staining assays, as well as colony morphology analysis. Real-time quantitative RT-PCR (qRT-PCR) was used to detect mRNA levels of target genes associated with flagellar synthesis and biofilm formation in Δ rpoN and the wild-type strains. Results:The V. vulnificus genome harbored a single rpoN gene, encoding a protein with high amino acid sequence similarity to RpoN homologs in other bacterial species. The Δ rpoN strain was successfully constructed. Compared with the wild-type strain, the Δ rpoN strain exhibited complete loss of motility on soft agar plates, absence of flagellar, and downregulated mRNA levels of flagellar synthesis-related genes. Conclusions:In V. vulnificus, RpoN regulates flagellar assembly by modulating the expression of flagellar synthesis genes, thereby controlling bacterial motility and biofilm formation.

Objective:To evaluate the effectiveness of a rotation training program for primary care general practitioners (GPs) in Taizhou, China, designed around three core competencies.Methods:A longitudinal study was conducted. A total of 111 primary care GPs participating in the 1st to 5th sessions of the Taizhou GP rotation training program at Taizhou Hospital (the designated training center) in Zhejiang Province from July 2022 to December 2024 were included. Based on the World Organization of Family Doctors (WONCA) competency model, combined with literature review and expert consultation, 15 potential training components were identified. A needs assessment survey among primary care GPs was then conducted, leading to the finalization of three core competencies for the training: ①Evidence-based management of common diseases, frequently-occurring diseases, and common symptoms; ②Early identification, management, and referral of critically ill patients; ③Chronic disease management and health promotion capabilities. A 4-month full-time immersion training program was structured around these competencies, comprising three phases: theoretical training (1 week), clinical comprehensive skills training (14 weeks), and primary care practice training (1 week). Assessments included a pre-and post-training theoretical knowledge test and a final clinical assessment. The final assessment utilized a three-station Objective Structured Clinical Examination (OSCE) evaluating patient consultation, clinical reasoning & decision-making, and clinical procedural skills. A self-administered satisfaction survey was also distributed post-training.Results:The 111 participants were predominantly aged 30-49 years (72.1% (80/111)), held bachelor′s degrees (87.4% (97/111)), were licensed physicians (78.4% (87/111)), and had junior or intermediate professional titles (87.4% (97/111)). Post-training theoretical scores were significantly higher than pre-training scores (76.26±7.00 vs. 69.94±6.40, t=-10.45, P<0.001). All participants 100%(111/111) passed the final OSCE assessment. The mean scores for the OSCE stations were: patient consultation 85.99±7.30, clinical reasoning & decision-making 82.72±7.61, and clinical procedural skills 89.60±5.65. Satisfaction rates were 100.0% (111/111) for the overall program, the three-phase design, the core competency training content, theoretical training, clinical skills training, and clinical rotation departments. Satisfaction was 98.2% (109/111) for the 4-month full-time duration, 99.1% (110/111) for the "2+X" clinical rotation model (2 weeks each in General Practice and Emergency Medicine+elective rotations in 4 other departments), and 97.3% (108/111) for the primary care practice base. Conclusions:The competency-based rotation training program for primary care GPs in Taizhou effectively enhanced participants′ theoretical knowledge and clinical practical skills, and achieved high levels of participant satisfaction. This model offers valuable insights for optimizing primary care GP training in similar settings.

With the acceleration of societal aging, heart failure in the elderly—characterized by a complex metabolic imbalance—has emerged as a pivotal challenge in public health.This condition severely compromises the physical well-being and quality of life among senior populations, primarily due to a pronounced imbalance between cardiac energy supply and demand.Among the various interventions studied, photosynthesis in plants, the principal energy source for aerobic organisms, presents a novel avenue for exploration.By effectively converting solar energy into chemical energy, photosynthesis sustains plant life.Introducing its principles into heart failure treatment could potentially optimize metabolism for elderly patients while significantly reducing oxidative stress and inflammation.Further investigations suggest that the antioxidants and bioactive byproducts generated during photosynthesis may play critical roles in heart failure treatments, particularly in modulating inflammatory pathways.In summary, emulating the mechanisms of photosynthesis could represent a promising strategy for treating heart failure in the elderly.This review aims to delve deeply into the application of photosynthesis in heart failure treatments for older adults and its underlying mechanisms, with the hope of providing insights and guidance for future endeavors in geriatric medicine.

Objective:To construct the rpoN gene knockout strain (Δ rpoN) and the complemented strain (CΔ rpoN) of Vibrio vulnificus ( V. vulnificus), and investigate the role of the rpoN gene in regulating bacterial motility and biofilm formation. Methods:The Δ rpoN strain of V. vulnificus was constructed using homologous recombination. Bacterial motility was assessed via swimming assays, and flagellar morphology was observed by transmission electron microscopy. Biofilm formation capacity was evaluated using crystal violet and Congo red staining assays, as well as colony morphology analysis. Real-time quantitative RT-PCR (qRT-PCR) was used to detect mRNA levels of target genes associated with flagellar synthesis and biofilm formation in Δ rpoN and the wild-type strains. Results:The V. vulnificus genome harbored a single rpoN gene, encoding a protein with high amino acid sequence similarity to RpoN homologs in other bacterial species. The Δ rpoN strain was successfully constructed. Compared with the wild-type strain, the Δ rpoN strain exhibited complete loss of motility on soft agar plates, absence of flagellar, and downregulated mRNA levels of flagellar synthesis-related genes. Conclusions:In V. vulnificus, RpoN regulates flagellar assembly by modulating the expression of flagellar synthesis genes, thereby controlling bacterial motility and biofilm formation.