Abstract: Objective　To investigate the effect of ring finger protein 130 (RNF130) on myocardial ischemia-reperfusion injury (MI/RI) and its potential mechanism. Methods　Male C57BL/6J mice were divided into four groups (n=6): Sham, MI/RI, MI/RI+Vector, and MI/RI+RNF130 overexpression (MI/RI+RNF130OE). Cardiac function was evaluated by echocardiography 24 hours after ischemia-reperfusion. Pathological changes, oxidative damage, and apoptosis in myocardial tissues were observed via IHC, DHE, and TUNEL staining. Protein expression was detected using Western blot, immunofluorescence, and immunohistochemistry. Proteomic analysis was performed to identify downstream proteins regulated by RNF130, and protein-protein interactions were validated by immunoprecipitation (IP) assay. Results　Compared with the MI/RI+Vector group, RNF130 overexpression significantly improved cardiac function, as indicated by increased left ventricular ejection fraction (EF) and fractional shortening (FS), reduced myocardial infarction area, and decreased expression of NOX-2 and BAX proteins(P<0.05). DHE and TUNEL staining showed that RNF130 overexpression alleviated myocardial oxidative damage and apoptosis(P<0.05). Proteomic analysis and IP assays revealed a significant interaction between RNF130 and PARP1, with PARP1 expression inversely correlated with RNF130. Conclusions　RNF130 may mitigate MI/RI injury by regulating the PARP1 ubiquitination pathway, providing a new target for therapeutic intervention.

Abstract: Objective To observe the effects of four different modeling method on the hypothalamuspituitary-adrenal (HPA) axis, blood rheology, platelet aggregation rate, and myocardial ischemia in rats, and to provide new ideas for the establishment of a rat model of “double heart” disease in line with clinical diagnosis and treatment characteristics. Methods Sixty-nine male Sprague-Dawley rats were divided randomly into a Control group (unstimulated), chronic unpredictable mild stimulation (CUMS) group, isoproterenol (ISO) group (intraperitoneal injection of ISO), high-fat diet (HFD) group (fed high-fat chow), and composite model (CUMS+ISO+HFD) group ( n= 12 rats in the Control and HFD groups; n = 15 rats in the other three groups, respectively). Modeling procedures were carried out for a total of 8 weeks, with ISO injection started from week 6 of the experiment for a total of 3 weeks. At the end of modeling, rats in each group were subjected to absent-field and sugar-water preference behavioral tests. Electrocardiography ( ECG) was performed to observe changes in ECG lead II in each group. Serum levels of adrenocorticotropic hormone (ACTH), cortisol (Cor), corticosterone (CORT), endothelin-1 (ET-1), and soluble intercellular adhesion molecule-1 ( sICAM-1) were detected by enzyme-linked immunosorbent assay. Myocardial histopathological changes were detected by hematoxylin / eosin ( HE ) staining. Serum total cholesterol ( TC ), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were measured using an enzyme labeling instrument. Whole-blood high-cut viscosity (200 V/ S), whole-blood low-cut viscosity (10 I/ S), plasma viscosity, and fibrinogen were assessed using an automatic blood rheology analyzer. The maximum platelet aggregation rate (MAR) and average platelet aggregation rate (AAR) induced by arachidonic acid and adenosine diphosphate were detected using a whole-blood platelet aggregometer. Results Compared with the Control group, all four model groups had significantly lower absenteeism distance and number of entries into the central region in the absent-field test, and a lower sugar-water preference ratio (P<0. 01). ECG revealed ST-segment elevation in the ISO and CUMS+ISO+HFD groups, tachycardia in the CUMS group, and mild ST-segment elevation in the HFD. Serum ACTH, Cor, CORT, ET-1, and sICAM-1 were all significantly elevated in the four model groups (P<0. 01). HE staining showed that myocardial tissue was severely damaged in rats in the ISO and CUMS+ISO+HFD groups, with pathological changes such as localized fibrosis and inflammatory infiltration of the myocardium, while mild cardiomyocyte disarrangement and fracture was seen in the CUMS and HFD groups. Rats in the HFD group had increased serum TC and LDL (P<0. 01) and decreased HDL contents (P<0. 01). Compared with the Control group, vwhole-blood high-cut viscosity (200 V/ S), whole-blood low-cut viscosity (10 I/ S), plasma viscosity, and fibrinogen were all increased in the CUMS, HFD, and CUMS+ISO+HFD groups (P<0. 01, P<0. 05), while whole blood highcut viscosity (200 V/ S), whole blood low-cut viscosity ( 10 I/ S), plasma viscosity, and fibrinogen levels were decreased in rats in the ISO group (P< 0. 01, P<0. 05). MAR and AAR were significantly higher in rats in the CUMS, HFD, and CUMS+ISO+HFD groups (P<0. 01), while the platelet aggregation rate was decreased in the ISO group compared with the Control group (P<0. 01, P<0. 05). Conclusions These result showed that the rat CUMS+ISO+HFD model better reflected the complexity of clinical double heart disease than the other three models.

Abstract: Objective　To investigate the regulatory effects of astragaloside IV on the hypoxia-inducible factor 1-α (HIF-1α) signaling pathway in diabetic nephropathy and to explore its potential therapeutic mechanisms. Methods　We established a diabetic nephropathy mouse model by injecting streptozotocin, and assessed the effects of astragaloside IV on the expression of Collagen I and α-smooth muscle actin by quantitative reverse transcription polymerase chain reaction and Western blot. Datasets of patients with diabetic kidney disease downloaded from the Gene Expression Omnibus (GEO) database were subjected to differential gene enrichment analysis, and activated pathways were screened out. The effects of astragaloside IV in reducing renal fibrosis and the inflammatory response and regulating macrophage polarization were evaluated by immunohistochemistry and flow cytometry. The key role of the HIF-1α pathway in diabetic nephropathy was further validated using the HIF-1α inhibitor LW6. Results Analysis of the GEO database showed that the HIF-1α/nuclear factor-κB signaling pathway was activated in patients with diabetic nephropathy. Astragaloside IV treatment significantly inhibited the expression of HIF-1α and its downstream fibrosis-related molecules in the diabetic nephropathy mouse model, reducing renal fibrosis and inflammatory responses. Astragaloside IV also promoted M2 macrophage polarization while suppressing M1 macrophage activation. The critical role of the HIF-1α pathway in the pathology of diabetic nephropathy was confirmed by experiments using the HIF-1α inhibitor LW6. Conclusions　This study demonstrated that astragaloside IV can significantly mitigate fibrosis and inflammation in diabetic nephropathy by regulating the HIF-1α signaling pathway and promoting favorable macrophage polarization.

Abstract: Objective　To examine the anti-central-fatigue function of maca and its underlying mechanism. Methods　Forty Sprague-Dawley rats were divided randomly into a negative control group, model control group, and low-, medium-, and high-dose maca groups (0.6, 1.2, and 2.4 g/kg·body weight). Rats in all groups except the negative control group were subjected to multi-factor stimulation, including cold-water swimming, sleep deprivation, restraining, and tail-clamping, to establish central fatigue rat models. Rats in the low-, medium-, and high-dose maca groups received 0.6, 1.2, or 2.4 g/kg maca, respectively, by gavage for 35 days. Behavioral testing was carried out using the Morris water-maze, sucrose-preference, and tail-suspension tests. Markers of oxidative stress in the hippocampus, including superoxide dismutase (SOD), malondialdehyde (MDA), and catalase (CAT), were detected using test kits. Proteins connected with the AMP-activated protein kinase (AMPK)/sirtuin 1 (SIRT1)/ peroxisome proliferator-activated receptor γ coactivator 1-α (PGC-1α) signaling pathway in the hippocampus were detected by Western blot. Results　Rats in the low-, medium-, and high-dose maca groups spent significantly more time in the target quadrant compared with the model control group (P<0.05 or P<0.01), but there was no significant dose-effect relationship. Rats in the medium- and high-dose maca groups showed decreased escape latency (P<0.05), increased time crossing the platform location (P<0.05), increased sucrose preference (P<0.05), decreased tail suspension time (P<0.05), increased the activities of CAT (P<0.01) and SOD (P<0.05), and decreased MDA content (P<0.01). Rats in the low-, medium-, and high-dose maca groups also showed significantly increased protein expression levels of AMPK and nuclear respiratory factor 1 (P<0.01 or P<0.05), but no significant dose effect relationship was observed. Rats in the medium- and high-dose maca groups showed increased protein expression of PGC-1α (P<0.05 or P<0.01), and rats in the high-dose maca group showed increased protein expression of SIRT1 and mitochondrial transcription factor A (P<0.05 or P<0.01). Conclusions　Maca can improve the indicators of central fatigue in rats, determined by behavioral testing and oxidative stress-related factors. The underlying mechanism may be related to its regulatory effects on the AMPK/SIRT1/PGC-1α signaling pathway.

Abstract: Objective　To elucidate the effects and mechanisms of Biejiajian pill (BJJP)-containing serum on the malignant biological behaviors of hepatocellular carcinoma Huh7 cells. Methods　This research knocked down CKLF-like MARVEL transmembrane domain containing 6 (CMTM6) expression using a CMTM6-specific small interfering RNA (siRNA). Healthy Sprague-Dawley rats were used to prepare normal rat serum and low- (0.55 g/ kg), medium- (1.1 g/kg), and high- (2.2 g/kg) BJJP-containing. Huh7 cells were cultured with normal fetal bovine serum (BC), normal rat serum (NC), and low-, medium-, and high-dose BJJP serum (LBJJP, MBJJP, and HBJJP, respectively). BJJP-containing serum and si-CMTM6 were applied to Huh7 cancer cells, and the proliferation, migration, and invasion abilities were evaluated by CCK-8 and Transwell assays, respectively. Protein expression levels of proliferating cell nuclear antigen (PCNA), epithelial-mesenchymal transition (EMT) markers, and CMTM6 were detected by Western blot. Results　CMTM6 knockdown significantly reduced the mRNA and protein expression level of CMTM6 in Huh7 cells (P<0.05). There were no significant differences between the BC and NC groups in terms of cell proliferation, migration, invasion, expression levels of PCNA, EMT markers, and CMTM6 (all P>0.05). BJJP-containing serum markedly inhibited Huh7 cell proliferation, migration, and invasion (P<0.05), downregulated PCNA, CMTM6, N-cadherin, and Vimentin expression, and upregulated E-cadherin compared with the NC group (all P<0.05). CMTM6 knockdown suppressed malignant behaviors, with reduced PCNA, Vimentin, and N-cadherin and elevated E-cadherin expression (all P<0.05). Conclusions　BJJP containing serum can significantly inhibit Huh7 cell growth, invasion, migration, and EMT progression, potentially mediated via CMTM6 suppression.

Abstract: Objective　To investigate the molecular mechanism of hepatic fibrosis (HF) regulation by liver sinusoidal endothelial cells (LSECs) via endothelial mesenchymal transition (EnMT), and to predict the natural active components using bioinformatics, machine learning, and cellular experiments. Methods　HF and EnMT gene matrices were obtained and the intersecting genes were extracted and enriched using Limma difference analysis and weighted gene co-expression network analysis (WGCNA). The diagnostic genes were screened using a combination of random forest method, support vector machine-recursive feature elimination and network topology analysis, and immune infiltration analysis and prediction of natural active ingredients were performed. The expression of diagnostic genes and the pharmacological effects of the predicted ingredients were finally verified by cellular experiments. Results　Differentialanalysis yielded 3034 EnMT-associated and 4133 HF-associated differential genes. WGCNA analysis yielded 4589 EnMT-associated Hub genes and 763 HF-associated Hub genes. Thirty-eight intersecting genes were extracted, which were mainly enriched in the pathways of basement membrane and extracellular matrix receptor interaction. Four diagnostic genes, CFP, COL4A2, ITGA1, and GRPEL1, were screened by multidimensional analysis. Immune infiltration analysis showed that the diagnostic genes were closely associated with mast cell resting state, memory B cells, and memory CD4⁺ T cells. Reverse transcription-polymerase chain reaction analysis showed significantly increased mRNA expression levels of the four diagnostic genes in the Jagged1-induced model group (P<0.05). The predicted components, sterol, kaempferol, and quercetin, all had good binding activities with the diagnostic genes. Enzyme-linked immunosorbent assay result confirmed that all three active components significantly reduced the expression of collagen type IV α2 chain protein in Jagged1-induced LSECs, with quercetin having the most significant effect (P<0.01). Conclusions　This study elucidated the molecular mechanism of hepatic sinusoidal endothelial cells involved in the pathological process of HF through mesenchymal transition. We also propose a diagnostic marker system including CFP, COL4A2, ITGA1, and GRPEL1 as core genes. The result also suggest that natural active ingredients, such as quercetin, may exert anti-HF pharmacological effects by targeting these diagnostic genes.

Abstract: Objective　To examine the effects of tryptophan hydroxylase 2 (Tph2) gene knockout on social play behavior during adolescence and on aggressive behavior in adult male rats, in order to elucidate the role of central 5-hydroxytrypatamine (5-HT) synthesis deficits in social behavior during postnatal development. Methods　Male Tph2-knockout (Tph2^KO) and wild-type (WT) rats were used in this study. Locomotor activity and anxiety levels during adolescence were evaluated after 4 weeks of age using an open-field test and adolescent social play behavior was assessed at 5 and 6 weeks of age using the adolescent social-interaction test. Locomotor activity and anxiety levels during adulthood were evaluated at 8 weeks of age by open-field test, and adult aggressive behavior was assessed at 10 and 11 weeks of age using the resident-intruder test. Results　Adolescent Tph2^KO male rats exhibited a significantly shorter total travel distance in the open-field test compared with WT male rats (P<0.05), but there was no significant difference in central zone exploration time or central zone entries between the two groups (P>0.05). Adolescent Tph2^KO rats also displayed significantly increased social play behavior (P<0.05) and reduced neutral social behaviors (P<0.01) in the social-interaction test compared with WT controls, but there was no significant difference in overall social behaviors (P>0.05). Adult Tph2^KO male rats exhibited a significantly reduced total travel distance (P<0.05), a trend towards decreased central zone exploration time (p=0.100), and significantly fewer central zone entries (P<0.05) in the open-field test compared with WT rats. There were no significant differences in overall social behavior, aggressive behavior, or neutral social behaviors in the resident-intruder test between the two genotypes (P>0.05). Notably, Tph2^KO rats exhibited increased same-sex mounting behavior during both adolescence and adulthood (P<0.05, p=0.076, respectively). Conclusions　Tph2 gene knockout significantly increased social play behavior during adolescence and elevated same-sex mounting behavior during adolescence and adulthood in male rats, but had no significant effect on adult aggressive behavior. These findings suggest that central 5-HT synthesis deficiency selectively modulates adolescent social behavior and impacts same-sex mounting behavior across developmental stages.

Abstract: Objective　To examine the role and mechanism of clopidogrel in the development of salt-sensitive hypertension. Methods　8-week-old Dahl salt-sensitive (Dahl SS) rats and control salt-resistant (SS13^BN) rats were divided randomly into six groups and fed for 8 weeks with normal salt (0.4% NaCl, NS), high salt (8% NaCl, HS), or high salt combined with clopidogrel gavage (8% NaCl+10 mg/(kg·d)) clopidogrel, HS+CLO). Arterial systolic blood pressure was measured continuously over 8 weeks by the tail-cuff method, and systolic blood pressure was measured by carotid cannulation after 8 weeks (56 days). Renal histopathology was observed by hematoxylin and eosin staining, and renal inflammatory cell infiltration was detected by immunohistochemistry. Peripheral blood platelet activation and platelet-leukocyte aggregation were analyzed by flow cytometry, and the renal inflammationrelated proteins tumor necrosis factor-α, interleukin (IL)-1β, IL-6, and key proteins in the p38MAPK/nuclear factor (NF)-κB signaling pathway were detected by Western blot. Results　Compared with the NS group, Dahl SS HS rats had significantly increased blood pressure(P<0.05), aggravated renal tissue damage, increased inflammatory cell infiltration, increased expression of inflammatory cytokines(P<0.05), elevated peripheral blood platelet activation(P<0.05)and platelet-leukocyte aggregation(P<0.05), and increased expression of p38MAPK/NF-κB signaling pathway proteins(P<0.05). Clopidogrel effectively alleviated these phenotypes induced by high salt in Dahl SS rats. Conclusions　Clopidogrel alleviated high-salt-induced salt-sensitive hypertension and decreased renal inflammatory responses and dysfunction in Dahl SS rats by inhibiting platelet activation and the p38MAPK/NF-κB signaling pathway.

Abstract:Animal surgery courses are a critical component of medical training; however, teaching practices have demonstrated problems, such as the difficulty of interdisciplinary teaching, students’ weak concepts of sterility, poor clinical thinking, insufficient teamwork, shallow emotional investment, and ineffective value guidance. By integrating the characteristics of student learning and utilizing virtual simulation experiments alongside peer role models, an emotional-guidance teaching model has been established to enhance the effectiveness of “life education”, through strengthened emotional identification and improved value guidance. The result indicate that this teaching model forms a closed-loop teaching process, aligns with students’ cognitive patterns and cultivates their comprehensive abilities, and enhances teaching effectiveness.

Abstract:Myocardial hypertrophy is characterized by the deleterious response of the heart to various stressors, leading to an increase in cardiomyocyte size and subsequent cardiac dysfunction. Autophagy, as a catabolic degradation process necessary for maintaining cellular homeostasis, has recently become an important mechanism in relation to the development of myocardial hypertrophy. Drawing on insights from recent molecular, cellular, and pharmacological studies, this review comprehensively explores the dual roles of autophagy compounds in myocardial hypertrophy. We investigated the contribution of autophagy flux in cardiomyocytes under physiological and pathological conditions, highlighting the complex interactions among autophagy and cardiomyocyte growth, survival, and function. In addition, we discuss the potential of modulating autophagic activity using drugs for the treatment of myocardial hypertrophy and heart failure. A critical examination of established models and the exploration of new autophagy regulators will further our understanding of the complex mechanisms that control myocardial hypertrophy and facilitate the development of targeted therapies. Ongoing research is expected to elucidate the definitive role of autophagy regulators, providing insights into their therapeutic potential and implications for clinical interventions in patients with myocardial hypertrophy and related pathologies.

Abstract:The glymphatic system is a recently discovered essential waste-clearance pathway in the central nervous system that plays a pivotal role in maintaining brain homeostasis. Growing evidence suggests that dysfunction of this system is closely associated with the pathogenesis of various neurodegenerative disorders; however, its precise mechanistic involvement in Parkinson’s disease is poorly understood. This review systematically summarizes the structural and functional characteristics of the glymphatic system, provides an in-depth exploration of its potential role in Parkinson’s disease pathophysiology, and synthesizes current research evidence on glymphatic system-targeted therapeutic strategies, with the aim of establishing a theoretical foundation for advancing the understanding of PD pathogenesis and developing novel diagnostic and therapeutic approaches.

Abstract:Migraine is a common neurological disorder with a complex pathogenesis that is currently not fully understood; however, the role of mitochondrial function in migraine pathogenesis has recently attracted widespread attention. This review considers the latest research progress on the relationship between mitochondrial dysfunction and migraine, including mitochondrial energy metabolism, oxidative stress, calcium homeostasis, and neuroinflammation. We introduce the epidemiological and clinical characteristics of migraine, and provide a detailed exploration of the key role of mitochondria in these processes. Mitochondrial dysfunction may lead to increased neuronal excitability, abnormal vasoconstriction, and inflammatory responses, thereby inducing migraine. Based on the evidence of mitochondrial involvement in the pathogenesis of migraine, we propose future research directions and potential treatment strategies, with the aim of providing new ideas for the prevention and treatment of migraine.

Abstract:Renal cancer is a common and increasingly prevalent malignancy with a complex pathogenesis influenced by genetics, smoking, and obesity. Current treatment mainly involves surgery with adjunctive chemotherapy, radiation, and immunotherapy, but high rates of recurrence and metastasis indicate its limited effectiveness, emphasizing the need for better therapeutic targets. Growing evidence indicates that epigenetic modifications, particularly RNA modifications, play a critical role in renal cancer development and progression. This review highlights recent advances in renal cancer epigenetics, focusing on RNA modifications such as N6methyladenosine (m6A), N7-methylguanosine (m7G), 5-methylcytosine (m5C), N1-methyladenosine (m1A), adenosine-to-inosine (A-to-I), N6,2’-O-dimethyladenosine (m6Am), and N4-acetylcytidine (ac4C), along with their regulatory factors. It also explores the diagnostic and therapeutic potential of targeting RNA modifications and associated proteins.

Abstract:Colorectal cancer (CRC) is the third most common malignant tumor worldwide after lung cancer and breast cancer, and is the second leading cause of cancer-related death. The causes of CRC are complex and treatment efficacy varies according to disease stage. Animal models thus play a critical role in research into CRC prevention and treatment. This review critically assesses the pathogenesis and characteristics of CRC from the perspectives of traditional Chinese medicine and Western medicine. We also discuss recent method and mechanisms of modeling in existing animal models of CRC. Current CRC animal models are largely based on modern medical modeling techniques and the evaluation criteria are also mainly based on Western medicine diagnostic indicators. These models are thus limited by the lack of combined traditional Chinese and Western medicine approaches in terms of disease diagnosis and treatment, making it difficult to elucidate the mechanisms responsible for the development of CRC. In addition, reliable large-animal models are rarely reported. A better understanding of the mechanisms of CRC integrating traditional Chinese medicine and Western medicine may provide useful insights to improve the predictability of animal models.

Abstract:Gastric cancer is one of the most common malignant tumors worldwide and presents many treatment challenges, especially in relation to late-stage treatment and its high recurrence rate. Angiogenesis is a key process in the growth and metastasis of gastric cancer, and studies aimed at researching and developing antiangiogenesis therapies are therefore important for the treatment of gastric cancer. Exosomes are nanoscale vesicles released by cells, which serve as important messengers for intercellular communication. They regulate local and distant cell communication by transporting specific exosomes components, and it can also promote or inhibit the development and progression of gastric cancer by regulating the growth, proliferation, and angiogenesis of tumor cells. Exosomes transport bioactive molecules from donor cells to recipient cells, leading to reprogramming of target cells and cascade molecular reactions. This review summarizes the relationship between exosomes and tumor angiogenesis in gastric cancer, and the current research status of exosomes in anti-angiogenic therapy. The importance of the physiological structure and function of exosomes in gastric cancer angiogenesis indicates the potential importance of combining exosomes with anti-angiogenic therapy for the treatment of gastric cancer.

Abstract:Rapid developments in biotechnology have led researchers to seek new method to improve the efficiency and accuracy of biomedical research and drug development, promoting interdisciplinary integration. Recent advancements in artificial intelligence (AI) technologies have brought unprecedented opportunities to this field. The integration of various AI models allows researchers to better utilize multi-omics data, identify disease phenotypes, interpret animal behavior, assess treatment effects, improve experimental designs, reduce the use of experimental animals, enhance animal facility management, and improve animal welfare. This article reviews the advancements in AI biomedical research over the past decade and discusses its contributions to disease phenotype identification, the selection and design of experimental animal models, animal behavior analysis, and animal facility management. It also points out the challenges related to data standardization, AI model selection and interpretability, the extrapolation process from AI models to animal experiments and clinical practice, as well as ethical considerations in using AI in sensitive areas involving human genetics and personalized medicine. This review aims to help researchers and practitioners in relevant fields understand the current state and opportunities of AI development, thus providing support for its broader application.