Abstract: Objective　To explore the efficacy and underlying mechanism of Astragalus in the treatment of viral pancreatitis using network pharmacology, with confirmation of its efficacy and mechanism in cell experiments. Methods　Astragalus and viral pancreatitis targets obtained from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) and GeneCards databases were combined to obtain intersection targets. GO functional enrichment and KEGG signaling pathway enrichment analyses of therapeutic targets were conducted using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) database. The interactions between therapeutic targets were analyzed using the STRING database and Cytoscape 3.10.2, and the core therapeutic targets were screened. Molecular docking between the most effective therapeutic components and the core targets was performed using PyMOL 3.0 and AutoDock Tools 1.5.7. CVB3 was used to construct a viral pancreatitis cell model for verification of the core targets. Results　Seventy-eight therapeutic targets were identified. Enrichment analyses revealed the possible involvement of pathways related to cancer, lipids and atherosclerosis, and PI3K-AKT signaling. AKT1, TP53, HIF1A, CASP3, IL-6, and MMP9 were identified as possible core targets. The result of cell experiments showed that the expression level of AMY was significantly increased in the model group(P<0.05). The Astragalus injection group exhibited significantly decreased expression levels of AMY, AKT1, TP53, HIF1A, CASP3, IL-6, and MMP9 compared with the model group(P<0.05). Conclusions　Astragalus injection effectively treated viral pancreatitis, and its therapeutic mechanism may involve reduced expression levels of AKT1, TP53, HIF1A, CASP3, IL-6, and MMP9.

Abstract: Objective　To investigate the impact of enterovirus 71 (EV71) on skeletal muscle injury and explore its mechanism in relation to the caspase-1/interleukin (IL)-1β signaling pathway in EV71-induced skeletal muscle damage. Methods　One-day-old BALB/c suckling mice were divided randomly into three groups: normal control (NC) ( n=60), EV71 infection model ( n=60), and caspase-1 inhibitor (EV71+VX765) ( n=15) groups. The NC and EV71 model groups were further subdivided into four subgroups (5, 7, 10, and 14 days) ( n= 5 mice per group). An EV71-infected model was established by intraperitoneal injection of 25×10³ μL/kg EV71 viral solution for 3 consecutive days. Mice in the caspase-1 inhibitor group received VX765 (20 mg/kg) intraperitoneally 6 hours post-viral inoculation, continued daily for 10 days until sample collection. Mice in the NC group received an equivalent volume of saline containing 5% dimethylsulfoxide and 10% PEG300, followed by 2% cell maintenance solution after 6 hours. Post-modeling body weight and clinical disease scores were recorded. Pathological skeletal muscle damage was observed by hematoxylin-eosin (HE) staining, and expression levels of EV71 VP-1 (viral capsid protein), pro-caspase-1, cleaved-caspase-1, IL-1β, α-smooth muscle actin (SMA), and Collagen I were detected by Western blot and immunofluorescence. Results　Compared with the NC group at the same time points, mice in the EV71 model group exhibited reduced body weight, elevated disease scores, and skeletal muscle pathology characterized by inflammatory cell infiltration, myofiber dissolution, and decreased cross-sectional area (HE staining). Western blot showed significantly increased levels of EV71 VP-1, IL-1β, α-SMA, and Collagen I in skeletal muscle homogenate from EV71 mice at 5, 7, and 10 days post-infection (P<0.001). In contrast, mice in the VX765 group showed improved body weight, reduced clinical scores (P<0.01), and significant downregulation of EV71 VP-1(P<0.01), pro-caspase-1, cleaved-caspase-1, IL-1β, and Collagen I compared with the EV71 model group (P<0.01). These findings were confirmed by immunofluorescence, indicating that inhibition of caspase-1 alleviated EV71-induced skeletal muscle injury. Conclusions　EV71 may induce skeletal muscle injury by activating the caspase-1/IL-1β signaling pathway.

Abstract: Objective　To systematically investigate the effects of Dangmu extract syrup on the growth and development of 4-day-old (postnatal day 4,PND4 ) Sprague-Dawley (SD) rats and its toxic reactions. Methods According to the whole litter design method, 128 young mice (PND2 ) were randomly divided into negative control group and low, medium and high dose groups. From PND4 , the animals were orally given pure water, 31 g/kg, 93 g/kg and 280 g/kg (calculated as raw herb material) of Dangmu extract syrup, respectively, once daily for 18 consecutive days, with a 15 d of recovery phase. During the study period, the general state, growth and development, nerve reflex function, spontaneous behavior, hematology, coagulation, blood biochemistry, immune function, growth hormone and histopathology of the animals in each group were observed or examined. Results　After 18 d of continuous administration, compared with the negative control group, GLU (male and female) in the medium and high dose groups increased (P<0.05 or P<0.01), LDH (male and female) and AST (male) in the medium and high dose groups, ALT, AST (female) in the high dose group decreased (P<0.05 or P<0.01), RET and percentage of RET (male and female) in the low and high dose groups, RET (male) in the medium dose group increased (P<0.05 or P<0.01), spleen mass and the organ-to-body mass ratio (male and female) in the low and high dose groups and female in the medium dose group increased (P<0.05 or P<0.01). Splenic nodule structures were formed in all dose groups with large size and number, and there was a dose relationship in the degree of changes. After 15 d recovery period, compared with the negative control group, GLU (female) in the low dose group increased (P<0.05), ALT, AST, ALP, TG (female) in the medium and high dose groups, GGT, TG, TCHO (male) in the medium dose groups, AST, ALP, TG, LDH (male) in the high dose group decreased (P<0.05 or P<0.01), RET (female) and percentage of RET (male) in the high dose group increased (P<0.05). compared with the 18 d of continuous administration, the spleen structures of the animals in each group were more completely developed and the splenic nodule structures were obvious, but no significant difference was noted in the comparison between groups. No significant drug-related changes were observed in other test result. Conclusions　Dangmu extract syrup advanced the development of complete spleen structure in 4-day-old SD rats, accompanied by the enhancement of its hematopoietic function, and at the same time, it caused the animals, blood glucose to rise, the enhancement of glucose metabolism function led to the increase of related enzyme consumption, and led to the decrease of some liver function parameters, and showed a dose correlation. There was no gender difference in the changes, which were reversible after stop administration, and the mechanism of the changes needs to be further explored and confirmed. In the clinical trials, attention should be paid to the control of the dose of the test article, and regular monitoring of the spleen and related blood and clinical chemistry parameters.

Abstract: Objective　To systematically investigate the effects of Dangmu extract syrup on the growth and development of 4-day-old (postnatal day 4,PND4 ) Sprague-Dawley (SD) rats and its toxic reactions. Methods According to the whole litter design method, 128 young mice (PND2 ) were randomly divided into negative control group and low, medium and high dose groups. From PND4 , the animals were orally given pure water, 31 g/kg, 93 g/kg and 280 g/kg (calculated as raw herb material) of Dangmu extract syrup, respectively, once daily for 18 consecutive days, with a 15 d of recovery phase. During the study period, the general state, growth and development, nerve reflex function, spontaneous behavior, hematology, coagulation, blood biochemistry, immune function, growth hormone and histopathology of the animals in each group were observed or examined. Results　After 18 d of continuous administration, compared with the negative control group, GLU (male and female) in the medium and high dose groups increased (P<0.05 or P<0.01), LDH (male and female) and AST (male) in the medium and high dose groups, ALT, AST (female) in the high dose group decreased (P<0.05 or P<0.01), RET and percentage of RET (male and female) in the low and high dose groups, RET (male) in the medium dose group increased (P<0.05 or P<0.01), spleen mass and the organ-to-body mass ratio (male and female) in the low and high dose groups and female in the medium dose group increased (P<0.05 or P<0.01). Splenic nodule structures were formed in all dose groups with large size and number, and there was a dose relationship in the degree of changes. After 15 d recovery period, compared with the negative control group, GLU (female) in the low dose group increased (P<0.05), ALT, AST, ALP, TG (female) in the medium and high dose groups, GGT, TG, TCHO (male) in the medium dose groups, AST, ALP, TG, LDH (male) in the high dose group decreased (P<0.05 or P<0.01), RET (female) and percentage of RET (male) in the high dose group increased (P<0.05). compared with the 18 d of continuous administration, the spleen structures of the animals in each group were more completely developed and the splenic nodule structures were obvious, but no significant difference was noted in the comparison between groups. No significant drug-related changes were observed in other test result. Conclusions　Dangmu extract syrup advanced the development of complete spleen structure in 4-day-old SD rats, accompanied by the enhancement of its hematopoietic function, and at the same time, it caused the animals, blood glucose to rise, the enhancement of glucose metabolism function led to the increase of related enzyme consumption, and led to the decrease of some liver function parameters, and showed a dose correlation. There was no gender difference in the changes, which were reversible after stop administration, and the mechanism of the changes needs to be further explored and confirmed. In the clinical trials, attention should be paid to the control of the dose of the test article, and regular monitoring of the spleen and related blood and clinical chemistry parameters.

Abstract: Objective　To compare the effects of Tripterygium wilfordii polyglycosides, cyclophosphamide, and cisplatin on the establishment of a mouse model of diminished ovarian reserve (DOR). Methods　Mice were randomly divided into the following treatment groups: control (Ctrl), Tripterygium wilfordii polyglycosides (TWP), cyclophosphamide (CTX), and cisplatin (DDP). Mice in the TWP group received a 50 mg/kg suspension of Tripterygium wilfordii polyglycosides by gavage for 14 days, mice in the CTX group received a 20 mg/kg cyclophosphamide suspension by intraperitoneal injection for 14 days, and mice in the DDP group received a 1.5 mg/ kg cisplatin solution by intraperitoneal injection for 14 days. The body weight, uterine index, and ovarian index were recorded, the estrous cycle was monitored using the vaginal smear method , and the levels of anti-Mullerian hormone (AMH), estradiol (E2 ), follicle stimulating hormone (FSH), and luteinizing hormone (LH) were detected using ELISA. Hematoxylin and eosin staining was used to detect ovarian follicle development. The rates of oocyte maturation and fertility were analyzed. Results　The three treatment groups of mice all showed the following: significantly decreased body weight and ovarian index (P<0.05); apparent disorder of the estrous cycle; significantly decreased levels of AMH and E2 (P<0.05); decreased and increased rates of developing follicles and atretic follicles, respectively (P<0.05); and significantly decreased rates of oocyte maturation, pregnancy, and live birth (P<0.05). Conclusions　DOR mouse models were successfully constructed using Tripterygium wilfordii polyglycosides, cyclophosphamide, or cisplatin, as evidenced by decreased body weight and ovarian index, disordered estrous cycle and hormones, and DOR function, resulting in reduced rates of oocyte maturation, pregnancy, and total number of live births. These DOR effects were most appropriate in the cyclophosphamide group.

Abstract: Objective　To observe the effects of growth differentiation factor-15 (GDF-15) on collateral circulation and cardiac function in rats with acute myocardial infarction (AMI) in relation to the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP)/protein kinase G (PKG) signaling pathway. Methods　An AMI rat model was constructed by ligating the left anterior descending coronary artery. After modeling, the rats were divided randomly into Sham, Model, and GDF-15 groups ( n=12 rats per group). Rats in the GDF-15 group were injected intraperitoneally with recombinant GDF-15 protein, and the other two groups were injected with the same amount of normal saline twice a week for 8 consecutive weeks. Cardiac function was detected by echocardiography. Pathological damage to rat myocardial tissue was detected by hematoxylin and eosin staining and the collateral circulation was observed by CD31 immunohistochemical staining. Vascular endothelial growth factor (VEGF) mRNA expression was detected by quantitative polymerase chain reaction. Transcriptomic sequencing of heart tissues in the model and GDF15 groups was performed and differentially expressed genes (DEGs) were screened. Pathway enrichment analysis of the DEGS was carried out according to the Kyoto Encyclopedia of Genes and Genomes (KEGG). Nitric oxide (NO), reactive oxygen species (ROS), and cGMP were detected using kits, and VEGF, endothelial nitric oxide synthase (eNOS) monomer, p-eNOS^ser1177 monomer, eNOS dimer, and PKG protein were detected by Western blot. Results Left ventricular end-systolic diameter (LVEDs) and left ventricular end-diastolic diameter (LVEDd) were increased (P<0.001), and left ventricular ejection fraction (LVEF) and the short-axis shortening rate (FS) were decreased in the Model group compared with the Sham group(P<0.001). Myocardial cell necrosis was more severe, vascular density in the infarcted area was decreased(P<0.05), but VEGF mRNA and protein levels were no change(P>0.05), and levels of NO, eNOS dimer, cGMP, and PKG protein were decreased(P<0.05), and expression levels of ROS, eNOS monomer, and p-eNOS^ser1177 monomer were increased (P<0.05). LVEDs and LVEDd decreased (P<0.05), LVEF and FS increased(P<0.01), myocardial cell necrosis was relieved, vascular density in the infarcted area increased significantly(P<0.0001), and VEGF mRNA levels increased(P<0.0001), compared with the Model group. Transcriptomic sequencing identified 324 DEGs, including 230 up-regulated and 94 down-regulated genes. According to KEGG enrichment analysis, the cGMP-PKG signaling pathway showed the most significant difference in the T20 pathway. VEGF, NO, eNOS dimer, cGMP, and PKG protein levels were all increased(P<0.05), while ROS, eNOS monomer, and p-eNOS^ser1177 monomer were decreased in the GDF-15 group (P<0.05). Conclusions GDF-15 can promote collateral circulation in ischemic myocardium and improve cardiac function by inhibiting eNOS decoupling and activating the NO/cGMP/PKG pathway.

Abstract: Objective　To investigate the characteristics and differences between two type 2 diabetic model mice, db/db and KK-Ay, as models of diabetic nephropathy (DN), and to verify them with irbesartan treatment. Methods　A total of 102 db/db and 102 KK-Ay mice (12 weeks old) were screened based on the albumin-tocreatinine ratio (ACR) and 24-hour urinary total protein (UCFP), with 78 mice of each strain selected and divided into 6 groups. The db/db + irbesartan group and KK-Ay + irbesartan group were administered irbesartan (40 mg/kg), the db/db group and KK-Ay group received an equal volume of drinking water for experimental animals, and the db/db + test article A groups and KK-Ay + test article A groups were given different doses of test article A (only the result of mortality data in the test articles groups were presented in the study, while other result were not shown). Wild-type C57BL/6J (WTa /WTb group) served as the normal control and were given experimental animal drinking water. Daily clinical observations were made, body mass was measured weekly, and ACR and UCFP were monitored periodically. At the end of administration, the mice were dissected, and the brain, liver, lungs and kidneys were collected, and the organ coefficients (organ mass×1000/body mass) were calculated. Blood was collected to measure serum total protein (TP), albumin (Alb),blood urea nitrogen (BUN), glucose (GLU), triglycerides (TG), and cholesterol (CHO). Kidney tissues were stained with HE, MASSON, and PAS to observe histopathological changes. Results　Among the 78 KK-Ay mice, 5 died over 8 weeks, while among the 78 db/db mice, 21 died over 6 weeks. ACR increased with age in both the db/db and KK-Ay groups but decreased in the db/db+irbesartan and KK-Ay+irbesartan groups, though without statistical significance (P> 0.05). UCFP increased with age in the KK-Ay group but significantly decreased after irbesartan treatment (P<0.05). Compared with the WT groups, the kidney coefficients in the db/db and KK-Ay groups showed no significant changes (P>0.05), while the brain liver coefficients decreased significantly (P<0.01) and the liver coefficients increased significantly (P<0.01), but they did not change after irbesartan treatment (P>0.05). Compared with the WT groups, GLU and TG levels were significantly increased in the db/db and KK-Ay groups (P<0.01). Compared with the KK-Ay group, GLU and TG levels were significantly reduced in the KK-Ay+irbesartan group (P<0.01). In the db/db and KK-Ay groups, kidney tissues showed thickening of the glomerular mesangial matrix and mesangial cell proliferation, which were alleviated after irbesartan treatment. Conclusions　Both 12-week-old db/db and KK-Ay mice can serve as models for DN research. Compared with db/db mice, KK-Ay mice exhibited smaller individual differences in ACR, fewer animal deaths after urine collection, and significant improvements in GLU, TG and UCFP after irbesartan treatment.

Abstract:Traditional laboratory animal breeding is costly and time-consuming, thus limiting experimental zoology teaching practices. Zebrafish have unique biological characteristics and have become widely used as an experimental animal model; however, teaching examples related to zebrafish in experimental zoology courses are still scarce. This review explores the application of zebrafish by incorporating them into the “Tissue Sectioning Techniques” and “Immunological Techniques” modules in the “Medical Laboratory Animal Science and Technology” course. The aim is to utilize the advantages of zebrafish in experimental zoology teaching and to explore their potential applications. This study provides a detailed introduction to the experimental procedures, experimental design, and teaching content involving zebrafish, with the aim of offering undergraduate students a more efficient, cost-effective, and practically meaningful experimental teaching platform.

Abstract:The incidence of leukemia, a malignant cancer originating from the hematopoietic system, is increasing annually. Although traditional treatment method such as chemotherapy and hematopoietic stem cell transplantation have improved patient survival rates to some extent, serious side effects, drug tolerance, and high recurrence rates remain. In recent years, studies have shown that the gut microbiota and its metabolites play an important role in the occurrence, development, and complications of leukemia. Imbalance of the gut microbiota can lead to decreased immune function and an intensified inflammatory response, which is a key factor driving disease progression. Some metabolites, such as short-chain fatty acids, enhance immune function and improve patient prognosis through intestinal barrier repair, while others, such as hydrogen sulfide and bile acids, show potential antitumor effects exerted through regulation of tumor cell apoptosis and immune balance. Traditional Chinese medicine aimed at regulating the structure of the gut microbiota and its metabolites has shown great potential in alleviating the side effects of chemotherapy for leukemia. This review covers the role of the gut microbiota and its metabolites in the occurrence, development, and complications of leukemia, and explores treatment strategies for regulating the microbiota, including fecal microbiota transplantation, probiotics, and traditional Chinese medicine intervention. We anticipate that this review will serve as a reference for improving the treatment and prognosis of leukemia.

Abstract:Chronic kidney disease (CKD) comprises a group of clinical syndromes affecting kidney structure and function, with various causes, high treatment costs, and a poor prognosis. Epigenetic modification of gene expression and cell function has been shown to play a key regulatory role in the occurrence and development of CKD. Homocysteine (Hcy) is a common amino acid-containing thiol group in the body. Hyperhomocysteinemia (HHcy) is a damaging condition involving many organs, and is an independent predictor of end-stage renal disease morbidity and mortality. This review considers the relationship between HHcy and chronic renal injury, and examines research progress in the role of epigenetic modification in the mechanism of Hcy-mediated chronic renal injury, with the aim of furthering our understanding of the occurrence and development of CKD. This process and its mechanism provide new ideas and a theoretical basis for further research into CKD.

Abstract:Cinnamomum cassia Presl belongs to the camphor family of evergreen trees, and its bark is used as a medicine. It contains volatile oils, polyphenols, polysaccharides, flavonoids, flavanols, and other chemical constituents that can inhibit the neuroinflammatory response, oxidative stress, and neuronal apoptosis, and provide neuroprotection, as well asimproving cerebral ischemia-reperfusion. Progress in modern medicine has demonstrated that cinnamon has unique advantages in terms of the prevention and treatment of central nervous system diseases (CNSD), and cinnamon and its preparations have been widely used to treat neurodegenerative diseases such as Alzheimer’s disease and Parkinson’s disease. The role of cinnamon for the treatment of CNSD has thus become a hot research topic; however, its mechanism of action in the nervous system has not been comprehensively outlined and summarized. This review therefore summarizes the progress in experimental research worldwide, regarding the use of cinnamon to treat CNSD, to provide an evidence base for clinical research and to develop the use of cinnamon.

Abstract:Alzheimer’s disease (AD) is an age-related neurodegenerative disease that mainly manifests clinically as progressive functional impairments in cognition, memory, and language. With the accelerated transition toward an older population in China, the number of people suffering from AD in China is increasing. The exact pathogenesis of AD remains unclear, with current therapeutic strategies mainly limited to symptomatic treatments. Animal models are important tools for preclinical research, enabling explorations of molecular mechanisms, behavioral functions, and treatment strategies of diseases. Future mechanistic research and drug development of AD should involve the establishment of animal models that are consistent with clinical pathological characteristics. This review summarizes the AD animal models commonly used in research, providing details on the strains, age, modeling method and doses. It also discusses research on traditional Chinese medicine (TCM) components and their pharmacodynamic mechanisms in related AD animal models, aiming to provide references for the development of new animal models and in-depth exploration of the specific pharmacological activities, targets, metabolic pathways, and clinical applications of each TCM component.

Abstract:Copper is an indispensable trace element that participates in numerous metabolic and signaling processes in both its oxidized and reduced states. It is intimately associated with several facets of tumor development, and alterations in copper homeostasis can substantially influence processes such as tumor cell growth, metastasis, modulation of the tumor microenvironment, oxidative stress, cell signaling, and the evasion of tumor cells from immune surveillance. Copper metabolism in tumor cells predominantly promotes immune escape by regulating the expression of programmed death ligand 1. In view of its crucial role in tumor immunity, modulating copper metabolism has emerged as a prospective therapeutic approach. This paper reviews the regulatory mechanisms of copper within the human body and investigates how disruptions to copper metabolism impact tumorigenesis and progression, along with the immune and tumor microenvironments. We also discuss the research value of copper as a target for tumor immunotherapy, thus providing a theoretical basis for future research and clinical applications.

Abstract:Osteosarcoma (OS) is a common primary malignant bone tumor with high mortality, disability, metastasis, and recurrence rates and a complex pathogenesis, Resulting in serious effects on patient quality of life and huge economic burdens on families and society. Traditional Chinese medicine (TCM) has “multi-target, multi component and multi-pathway”characteristics. Recent studies using animal and cell models demonstrated that the mechanism of OS progression was related to Notch, mitogen-activated protein kinase, Wnt/β-catenin, phosphatidylinositol 3-kinase/AKT, Hedgehog and nuclear factor-κB, transforming growth factor-β/Smad and signal transducer and activator of transcription pathways. TCM can exert anti-tumor effects by influencing biological processes such as cell proliferation, migration, invasion, apoptosis, and autophagy via interfering with the above signaling pathways. This review considers the roles of these signaling pathways in OS and summarizes the current research status of TCM interventions in the prevention and treatment of OS, with the aim of providing a reference for future studies of TCM treatments of OS and to provide new ideas for its clinical treatment.

Abstract:Laryngopharyngeal reflux disease (LPRD) occurs in the upper respiratory tract and the upper digestive tract. Its pathogenesis is complex and there a lack of effective treatments. Suitable animal models and standard evaluation method are needed to explore the pathogenesis of this disease and to develop new therapeutic drugs. New Zealand rabbits are commonly used to model LPRD, via balloon dilatation. The standard evaluation method for LPRD include symptom observation and evaluation, imaging evaluation, and pH monitoring. This review considers the animal models and evaluation indexes currently used for pharyngeal reflux with reference to the global literature, to provide a reference for future research.

Abstract:Tumor-associated macrophages (TAM) are a current focus in the study of the tumor microenvironment. To achieve their functionally distinct roles, macrophages undergo phenotypic polarization resulting in two major subgroups: M1 macrophages with pro-inflammatory and anti-tumor effects, and M2 macrophages with anti-inflammatory and pro-tumor effects. Of these, M2 polarization, as the main manifestation of TAMs, has been associated with a poor prognosis in various cancers and has been shown to support malignancies. Gastric carcinoma has a low early-diagnosis rate, late disease stage, and poor prognosis, with biological behavioral characteristics of easy recurrence and metastasis. Drug resistance and toxic side effects currently limit the application and effectiveness of treatments, and there is thus an urgent need to explore new therapeutic drugs and targets. This review summarizes recent progress in studies of TAM in relation to the occurrence, development, and drug resistance of gastric carcinoma, providing new ideas for clinical treatment and prognosis prediction in patients with gastric carcinoma.