Abstract: Objective To explore the therapeutic mechanism of ursolic acid against silicosis fibrosis based on network pharmacology, molecular docking and in vitro experiments. Methods Ursolic acid targets were obtained from databases, including GeneCards and PubChem. Disease-related databases (GeneCards, OMIM) were searched for targets related to silicosis fibrosis and epithelial-mesenchymal transition ( EMT). A micro-biotech platform was used to screen for intersecting targets, and a protein-protein interaction network was constructed using the STRING database and Cytoscape to screen the core targets. The David database was used for GO and KEGG enrichment analysis. AutoDock was used for molecular docking validation. Key targets were validated using beas-2B cells. ResultsWe obtained 179 ursolic acid targets, 8023 silicosis fibrosis targets, 6809 EMT-related targets, and 133 intersecting targets. Nine core targets, including AKT, STAT3, and MMP9, were identified, among which MMP9 and AKT had the highest connectivity in the protein-protein interaction network. Molecular docking showed that ursolic acid had strong binding activity with MMP9 ( binding energy - 8. 4 kJ/ mol) and AKT ( binding energy - 7. 9 kJ/mol). KEGG analysis indicated the PI3K-AKT signaling pathway to be a key regulatory pathway. In vitro experiments showed that ursolic acid significantly inhibited the decrease in cell viability induced by SiO2 (CCK8). Ursolic acid also reduced p-AKT expression (Western blot,P<0. 05); downregulated expression of the fibrosis marker α-SMA and the mesenchymal marker Vimentin, while upregulated expression of the epithelial marker E-cadherin (immunofluorescence / Western blot,P<0. 05). Conclusions Ursolic acid may play an anti-silicosis fibrosis role by inhibiting AKT phosphorylation, reducing MMP9 levels and regulating EMT.

Abstract: Objective To investigate the core targets and mechanisms by which isorhamnetin prevents and ameliorates liver fibrosis. Methods Bioinformatic data were integrated to identify liver fibrosis-related targets via differential gene analysis and weighted gene co-expression network analysis (WGCNA). These targets were compared with those that mediate isorhamnetin’s action to identify common targets. Machine learning optimized core targets that were validated for causal association using Mendelian randomization. Molecular docking and dynamics simulations assessed target function. ResultsWe identified 113 interactive targets of liver fibrosis and isorhamnetin, which were primarily enriched in phosphatidylinositol 3 kinase-protein kinase B(PI3K-AKT), tumor necrosis factor(TNF), and other signaling pathways. Machine learning combined with Mendelian randomization pinpointed aryl hydrocarbon receptor(AHR), caspase3(CASP3), and mitogen-activated protein kinase 14(MAPK14) as core targets. Multidataset validation confirmed their consistent expression and significant diagnostic efficacy ( area under the curve >0. 7). Molecular simulations demonstrated stable binding of isorhamnetin to these targets (binding energy<-7. 0 kcal /mol). Conclusions Isorhamnetin inhibits liver fibrosis by targeting AHR, CASP3, and MAPK14 to regulate inflammation, apoptosis, and metabolic pathways. This study provides novel insights into the anti-fibrotic mechanisms of traditional Chinese medicine components.

Abstract: Objective To investigate the protective effect of Yulangsan Chalcone ( YLSC) on H9c2 cardiomyocyte injury induced by angiotensin II ( Ang II ) and its possible mechanism. Methods H9c2 cardiomyocytes were divided into blank control, model, and low-, medium-, and high-dose YLSC groups. After intervention with YLSC, the cells were induced with Ang II (1 μmol / L) to form an injury model. Cell viability was detected by cell counting kit-8 assay, autophagosomes were detected by monodansylcadaverine staining, reactive oxygen species were detected using the dichlorodihydrofluorescein diacetate fluorescence probe method, apoptosis was detected by Hoechst 33342 staining, and the ATP content was detected by enzyme-linked immunosorbent assay. Bcl-2 interacting protein 3 ( Bnip-3) and autophagy-related protein 5 ( Atg5) gene levels were detected by reverse transcription-polymerase chain reaction, and expression levels of the autophagy marker proteins p62, Beclin1, and microtubule-associated protein light chain 3 (LC3) II/ I were detected by Western blot. ResultsYLSC alleviated Ang II-induced H9c2 cardiomyocyte injury by enhancing cardiomyocyte viability(P<0. 05 or P<0. 01), promoting the formation of autophagosomes, and reducing the generation of reactive oxygen species(P<0. 01). YLSC also reduced cell apoptosis(P<0. 01), increased ATP levels in cells(P<0. 05 or P<0. 01), promoted the gene expression of Bnip-3 and Atg5(P<0. 05 or P<0. 01), and reduced the expression of p62 and increased the expression of Beclin1 and LC3II/ LC3I(P<0. 05 or P<0. 01). Conclusions YLSC can effectively inhibit Ang II-induced H9c2 cardiomyocyte injury and apoptosis, and its mechanism may be related to the initiation of myocardial autophagy, activation of autophagy-related factors, and clearance of apoptotic cells.

Abstract: Objective To investigate the effect of osteopontin (OPN) interference on the Warburg effect in hepatoma cells and to explore the related molecular mechanism. Methods Small interfering ( si) RNA targeting OPN (si-OPN) and negative Control siRNA (si-NC) were transfected into HepG2 and SK-HEP-1 cells. The glucoseuptake capacity of hepatoma cells was detected using a fluorescent probe ( 2-NBDG), and lactate production in HepG2 and SK-HEP-1 cells was evaluated using a lactate-detection kit. Expression levels of the glycolysis-related genes and proteins glucose transporter type 1 ( GLUT1), hexokinase 2 ( HK2), and lactate dehydrogenase A (LDHA) were determined by quantitative reverse transcription-polymerase chain reaction and Western blot. We then silenced the expression of galectin-3-binding protein (LGALS3BP) in HepG2 and SK-HEP-1 cells, and overexpressed LGALS3BP in OPN-silenced HepG2 and SK-HEP-1 cells, and evaluated the expression levels of LGALS3BP,GLUT1, HK2, and LDHA, and the glucose-uptake capacity and lactate production in the above cells. ResultssiOPN significantly reduced glucose uptake and lactate production in HepG2 and SK-HEP-1 cells compared with the Control and si-NC groups, and significantly reduced GLUT1, HK2, and LDHA gene and protein expression levels (P<0. 05). si-OPN reduced LGALS3BP protein expression in HepG2 and SK-HEP-1 cells, and silencing LGALS3BP reduced glucose uptake, lactate generation, and the expression levels of GLUT1, HK2, and LDHA ( P<0. 05).Overexpression of LGALS3BP rescued the suppression of the above glycolytic metabolism-related indicators ( P<0. 05). These result suggest that OPN stimulated glycolytic metabolism in hepatocellular carcinoma cells via LGALS3BP. Conclusions OPN can affect the expression of glycolysis-related genes via LGALS3BP, thereby regulating the Warburg effect in hepatoma cells.

Abstract: Objective To examine the regulatory relationship between the tensin 1 (TNS1) gene and miR-574-5p in cardiac arrest, assess its clinical significance, and verify the therapeutic potential of targeted inhibition of miR-574-5p. Methods Oxygen-glucose deprivation /reoxygenation (OGD/ R) cardiomyocyte and mouse asphyxia cardiac arrest / cardiopulmonary resuscitation (ACA/ CPR) models were established. Expression levels of miR-574-5p and TNS1 were detected by RT-qPCR. Protein expression levels of TNS1 in cardiomyocytes were detected by Western blot. The targeting relationship between miR-574-5p and TNS1 was verified by dual-luciferase reporter gene assay. Cell viability was detected by Cell Counting Kit-8 assay, and apoptosis was measured by flow cytometry. Serum levels of the cardiac injury marker cardiac troponin I ( cTnI ) and the oxidative stress markers malondialdehyde and 4-hydroxynonenal were detected by enzyme-linked immunosorbent assay. The cardiac function indices dp / dtmin and dp /dtmax were evaluated using a hemodynamic monitoring system, and cardiac function parameters, including left ventricular ejection fraction and left ventricular fractional shortening, were determined by echocardiography. ResultsOGD/ R treatment significantly upregulated the expression of miR-574-5p and inhibited the mRNA and protein expression of TNS1 in cardiomyocytes, while inhibition of miR-574-5p improved cardiomyocyte survival and alleviated oxidative stress injury (P<0. 05). In the ACA/ CPR model, cardiac function indices were significantly improved (P<0. 05), cardiac injury and oxidative stress markers were reduced (P<0. 05), and the upregulation of miR-574-5p and downregulation of TNS1 expression patterns in myocardial tissues were reversed in the miR-574-5p antagonist group (P<0. 05). Conclusions This study confirmed that targeted inhibition of miR-574-5p can improve cardiac function by upregulating the expression of TNS1, providing a new therapeutic target for myocardial protection after cardiac arrest.

Abstract: Objective To investigate the effect of pinocembrin ( Pin) on the apoptosis and migration of mouse melanoma B16 cells, and its relationship with the disheveled-associated activator of morphogenesis 2 (DAAM2) / Wnt / β-catenin signaling axis. Methods CCK8 method was used to screen the Chinese medicine monomers that could inhibit the viability of B16 cells in vitro. Cell apoptosis and migration of melanoma B16 cells were detected by flow cytometry and wound healing assay, respectively. Possible core genes of Pin were analyzed by RNA-seq sequencing combined with GEPIA database analysis. Expression levels of DAAM2 and the Wnt / β-catenin signaling pathway-related genes and proteins Axin2, phospho ( p)-glycogen synthase kinase (GSK)-3β, and p-β-catenin were detected by real-time fluorescence quantitative polymerase chain reaction ( qPCR) and Western blot.Reversion experiments were performed by transfection with a DAAM2-overexpression plasmid. A mouse melanoma model was established in vivo, and the mice were then treated with low and high doses (10 mg / kg, 30 mg / kg) of Pin by gavage and melanoma growth was observed. The effects of Pin on DAAM2, Axin2, β-catenin, and p-GSK3β expression in melanoma tissues were detected by Western blot, to clarify its effect on melanoma progression. ResultsPin significantly promoted apoptosis (P<0. 001) and inhibited the migration of B16 cells ( P<0. 01, P<0. 001).RNA-seq and GEPIA analysis suggested that DAAM2 was a key target. Pin significantly reduced DAAM2 mRNA and protein expression, as shown by qPCR and Western blot ( P<0. 05, P<0. 01, P<0. 001), and down-regulated expression levels of the Wnt / β-catenin signaling pathway-related proteins p-GSK3β, p-β-catenin, and Axin2 (P<0. 001). High-dose Pin significantly inhibited melanoma growth and weight in mice in vivo, and reduced DAAM2,axin2, β-catenin, and p-GSK3β expression in tumor tissues (P<0. 05, P<0. 01), while DAAM2 overexpression reversed the effects of Pin on the DAAM2 / Wnt / β-catenin signaling axis. Conclusions Pin can promote the apoptosis of melanoma cells, inhibit their migration, and inhibit melanoma growth in mice by down-regulating the DAAM2 / Wnt / β-catenin signaling axis.

Abstract: Objective To establish a method for detecting mRNA expression levels of the fatty acid-binding protein 4 gene (FABP4) in crab-eating macaques, Macaca fascicularis. Methods Six pairs of primers for real-tim quantitative polymerase chain reaction (RT-qPCR) were designed according to the mRNA sequence of the FABP4 gene. Adipose tissue was collected from Macaca fascicularis, and total RNA was extracted and reverse transcribed into cDNA. RT-qPCR was carried out using the designed primers, and the primers with the most sensitive detection and without non-specific amplification were selected according to the amplification curve and melting peaks. A standard curve for gene expression was constructed using cDNA diluted at 10P>0, 10^-1, 10^-2, 10^-3, and 10^-4. Macacafascicularis were divided into young and old groups to validate the fluorescence RT-qPCR detection method for FABP4expression in Macaca fascicularis adipose tissue. ResultsA pair of primers with the highest detection sensitivity was screened out, and a fluorescence RT-qPCR method for detecting FABP4 expression in Macaca fascicularis adipose tissue was established. Relative expression levels of the FABP4 gene in adipose tissue were higher in elderly compared with young macaques. Conclusions We successfully established a RT-qPCR method for detecting transcription levels of the FABP4 gene in Macaca fascicularis.

Abstract: Objective To develop a mouse cerebral edema / pulmonary edema model under different conditions and compare method of calculating water content. Methods Fifty male BALB/ c mice were divided randomly into a Control group and four experimental groups: 6000 m/ 48 hours, 6000 m/ 72 hours, 8000 m/ 48 hours,and 8000 m/ 72 hours (n= 10 mice per group). The baseline mass was recorded in each group. Mice in all groups except the Control group were placed in a simulated plateau environment at an altitude of 6000 or 8000 m for 48 or 72 hours. After exposure, the body mass was measured, and the lung wet weight, brain wet weight, lung dry weight, and brain stem weight were determined. Three different method were used to calculate the model water content, and pathological changes in the brain and lung were observed by hematoxylin / eosin staining. ResultsThe body mass decreased significantly in all groups except the control group (P<0. 01), with the greatest decrease after 72 hours at the same altitude. In the pulmonary edema model, pathological analysis showed slight pulmonary edema in the 6000m/ 48 hour group, but there was no significant difference from the control group (P>0. 05). Using the calculation result from method 2 or 3, the water contents were significantly higher in the 6000 m/ 72 hour, 8000 m/ 48 hour, and 8000 m/ 72 hour groups, compared with the Control group (P<0. 01), and pathological analysis indicated significant edema in the lungs in these three groups; however, method 1 found no significant difference in lung water contents in these three groups compared with the control group (P>0. 05). In terms of cerebral edema, method 1 showed that the brain water content in each experimental group was significantly lower than that of the Control group (P<0. 01), while method 2 and 3 showed consistent results, with significantly higher brain water contents in all the experimental groups compared with the Control group ( P<0. 01). Pathological analysis showed significant cerebral edema in all four experimental groups. Conclusions Method 1 is less reliable for calculating water contents, while method 2 and 3 showed similar results, including the ability to correct body mass changes. These method are thus recommended as the standard calculation method for the plateau cerebral edema / pulmonary edema model.

Abstract:This article examines the current status of Zhejiang Province’ s laboratory animal industry following the implementation of license separation reform. We find that while the reform has lowered market entry barriers and increased the number of market entities, it has also introduced new challenges to the regulatory system.Both the production and use of laboratory animals have expanded, with quality improvements observed, yet issues of insufficient standardization persist. Although scientific research and innovation capabilities have improved, regional development disparities remain evident. To address these challenges, we propose countermeasures, such as refining the regulatory framework, strengthening industry self-regulation, and enhancing scientific research and innovation capabilities, to foster the healthy development of Zhejiang Province’s laboratory animal industry.

Abstract:Artificial intelligence (AI) has also been widely applied in laboratory animal science teaching.This paper analyzes the application status of AI-based virtual simulation technology in the teaching of laboratory animal science at home and abroad, the application status of AI in the interaction of laboratory animal science teaching, the application status of AI in the teaching of laboratory animal models, the advantages and disadvantages of AI in the teaching of laboratory animal science, and lists the commonly used AI tools. It can be expected that AI technology will play an important role in the future teaching of laboratory animal science.

Abstract:Laboratory animal facilities play an important role in scientific research in hospitals. Their construction standards and specifications, design principles, functional layout, equipment configuration, and environmental control, as well as their operational management, quality control, safety management, personnel training, and other aspects should all follow unified standards and requirements. The complex and systematic nature of the construction of laboratory animal facilities, however, mean that the design concept, purpose, geographical location, construction requirements, personnel composition, and other aspects of the facilities differ among research institutions and hospitals. Different problems are also encountered in the specific construction and operation practice of laboratory animal facilities. Here, we discuss the construction and operation of laboratory animal facilities at the Central China Subcenter of the National for Cardiovascular Disease. We summarize the experience of our unit in the construction and operation of laboratory animal facilities, to provide theoretical guidance and a practical reference for the construction and operation of laboratory animal facilities in the hospital system.

Abstract:The primary isolation and culture of penile corpus cavernosum cells are key techniques for studying the pathological mechanisms of erectile dysfunction (ED). This review summarizes the isolation and culture method of the major cell types in the penile corpus cavernosum, including smooth muscle cells, endothelial cells, pericytes, and fibroblasts. We discuss commonly used techniques, including enzymatic digestion, tissue explantation, immunomagnetic bead separation, and Matrigel induction, and compare their advantages and disadvantages. We also summarize the method for identifying different cell types. This systematic review of cell culture and identification techniques thus provides technical support and references for further research into the roles of penile corpus cavernosum cells in the onset, progression, and treatment of ED.

Abstract:Acute kidney injury (AKI) is a prevalent clinical syndrome characterized by a rapid decline in renal function over a short time and accompanied by symptoms of abnormal urine output and azotemia. The pathological mechanisms underlying AKI are highly complex, primarily involving inflammatory responses, oxidative stress, and cell death. In recent years, the global incidence of AKI has risen annually, making it an urgent public health challenge. Emerging research indicates that macrophages play a dual role in the progression and recovery of AKI by dynamically modulating iron metabolism. This review systematically discusses the mechanisms by which dysregulated macrophage iron metabolism influences AKI progression: iron overload exacerbates inflammation by promoting M1 polarization, lipid peroxidation, and ferroptosis, whereas iron restriction may enhance antioxidant capacity via activation of pathways, such as Nrf2, that facilitate M2 polarization and renal tissue repair. Based on

Abstract:The abuse of addictive substances can lead to legal and public health problems, causing great harm to individuals, families, and society. Addictive substances cause behavioral changes and cognitive dysfunction by affecting the release of neurotransmitters and causing neuroplastic changes in the brain. Repetitive transcranial magnetic stimulation(rTMS)is a non-invasive neuromodulation method for treating substance use disorders, which can improve craving behavior and cognitive impairment of patients. This article reviews recent studies of this treatment and discusses new avenues for the treatment of substance use disorders.

Abstract:Ischemic stroke is one of the most prevalent cerebrovascular diseases. The HIF-1α/ VEGF signaling pathway is as a crucial mediator of angiogenesis and has demonstrated bidirectional effects in ischemic stroke treatment through multifaceted mechanisms involving angiogenesis, neurogenesis, oxidative stress, the inflammatory response, and autophagy. Emerging evidence indicates that traditional Chinese medicine can exert therapeutic effects on ischemic stroke by targeting the HIF-1α/ VEGF pathway, effectively ameliorating oxidative stress, inflammatory reactions, cellular autophagy, and apoptosis, while promoting vascular regeneration in ischemic regions. Here, we review the dual-phase regulatory mechanisms and functional characteristics of the HIF-1α/ VEGF signaling pathway in ischemic stroke pathogenesis and progression. Furthermore, we summarize recent advances in traditional Chinese medicine-based interventions that target this pathway to provide evidence-based theoretical support and clinical references that will help to