Abstract: Objective　A rat model of cerebral small vessel disease (CSVD) was established by unilateral injection of a single dose of sodium laurate into the internal carotid artery. The effectiveness of the model was assessed by behavior scoring and analysis of serum-related indicators, cerebral infarction volume, cerebral microvascular density, hemodynamics, brain histopathology and the expression of blood-brain barrier (BBB)-related proteins. Methods　SPF-grade male SD rats were divided randomly into a control group and a model group ( n= 6 per group). The model group received a single injection of 100 μL of sodium laurate (2 g/L) via the internal carotid artery, while the control group underwent the same surgical procedure but received an equal volume of saline. Neurobehavioral assessments were conducted using the Longa score and postural reflex test. Serum homocysteine (HCY) levels were measured by enzyme-linked immunosorbent assay. Cerebral infarction volume was detected by magnetic resonance imaging and changes in cerebral vascular density were observed by cerebrovascular imaging. The resistance index (RI) and perfusion index (PI) were measured by ultrasonography. Histopathological changes in brain tissue were evaluated by hematoxylin and eosin (HE) staining. Expression of the cerebral microvascular marker CD31 and tight junction proteins ZO-1 and Occludin in brain cortex tissue were detected by immunohistochemical staining. Results　The Longa score, postural reflex score (P<0.05), and cerebral infarction volume were significantly increased (P<0.05) while the cerebral vascular density was decreased in the model group compared with the control group. Serum HCY levels, carotid RI, and PI values were all significantly increased in the model group (P<0.05). HE staining revealed solidified neuronal nuclei and enlarged perivascular spaces in the brain cortex in the model group. Immunohistochemical staining revealed that CD31, ZO-1, and Occludin expression were significantly reduced in the brain cortex in the model group compared with the control group (P<0.05). Conclusions　A rat model of CSVD can be established rapidly and effectively by a single unilateral injection of high concentration sodium laurate via the internal carotid artery. This model is characterized by neurobehavioral abnormalities, cerebral infarction, insufficient cerebral blood supply, reduced vascular density, and disruption of the BBB, suggesting that it may serve as an effective rat model for the study of CSVD.

Abstract: Objective　To investigate the effect and mechanism of Taxilli Herba decoction in preventing miscarriage in rats with threatened abortion. Methods　Seventy-two SPF-grade SD rats were co-housed at a male-tofemale ratio of 2∶ 1. Forty-eight pregnant rats were subsequently divided into a normal group, a threatened abortion model group, a positive control group, as well as Taxilli Herba decoction low/medium/high dose groups (n= 8 rats per group). Rats in the positive control group received 3.02 mg/kg dydrogesterone intragastrically, and rats in the low-, medium-, and high-dose Taxilli Herba decoction groups received 2.5, 5, and 10 g/kg Taxilli Herba decoction intragastrically, respectively. The normal and model groups received intragastric distilled water once a day for 10 consecutive days. On day 10 after administration of the corresponding treatment, rats in all groups except the normal group received intragastric administration of 3.75 mg/kg mifepristone suspension. The effects of Taxilli Herba on the vaginal bleeding rate and abortion rate were observed. Serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estrogen (E), progesterone (P), vascular endothelial growth factor (VEGF), testosterone (T), and aromatase and E and P levels in the ovary in pregnant rats were detected by enzyme-linked immunosorbent assay. Expression levels of E receptor (ER), P receptor (PR), VEGF receptor 2 (VEGFR2), and platelet endothelial cell adhesion molecule-1 (CD31) in the uterine decidua in pregnant rats were detected by immunofluorescence staining, and semi-quantitative analysis was performed. Results　Mifepristone significantly increased the vaginal bleeding rate and abortion rate in pregnant rats, but these were effectively reduced by treatment with Taxilli Herba water decoction. Taxilli Herba water decoction also increased serum E, P, and VEGF, and ovary levels of E and P (P<0.05). The treatment also decreased serum LH, maintained the secretion of FSH, and enhanced ER, PR, VEGFR2, and CD31 expression in decidual tissue. Conclusions　Taxilli Herba demonstrated effective miscarriage-prevention effects in a rat model of threatened abortion. Its mechanism of action involves regulating the normal physiological function of the hypothalamic-pituitary-gonadal axis upstream of the uterus, and enhancing the expression of the downstream P/PR/ VEGF/VEGFR2 and E/ER/VEGF/VEGFR2 pathways.

Abstract: Objective　To investigate the dynamic characteristics of intestinal pathological development at different time points in a mouse model of colitis-associated colon cancer. Methods　A colitis-cancer model was established in C57BL/6 mice using azoxymethane (AOM) combined with dextran sulfate sodium (DSS). Samples were collected at 7, 10, and 14 weeks post-modeling and the spleen index, colon length, mass, and colon mass per unit length were measured. Histopathological changes in the colon were observed by hematoxylin and eosin and Masson staining. Expression levels of the cancer stem cell marker CD44 and Wnt signaling pathway genes Wnt2b, Lrp5, Axin2, and Znrf3 at different pathological stages were detected by reverse transcription quantitative real time PCR. Cancer-associated fibroblasts (FAP), CD44, the proliferation marker Ki67, and goblet cell MUC2 protein were detected by multiple immunofluorescence histochemistry (mIHC) and immunofluorescence. In addition, colon organoids were isolated from model mice at ten and fourteen weeks and cultured in vitro to observe changes in organoid morphology and marker expression. Results　AOM/DSS-induced mice showed reduced, distorted, and branched colon crypt structures with a few collagen fibers at 7 weeks, and varying degrees of colon intraepithelial neoplasia, with an increased proportion of high-grade intraepithelial neoplasia over time and increased collagen fiber staining at ten and fourteen weeks. mRNA levels of CD44 and Wnt2b in the colon were significantly increased (P<0.05) and Axin2 was decreased (P<0.01) in model mice compared with control mice at fourteen week, and levels of Wnt2b, Lrp5, and Znrf3 were increased compared with seven-week mice (P<0.01,P<0.05,P<0.01), and Axin2 was decreased (P<0.01). mIHC staining showed increased expression of FAP and CD44 in the colon in model mice at ten and fourteen weeks, with decreased MUC2 expression. Colon organoids showed cystic dilation, especially at fourteen weeks, with more prominent expression of Ki67 and CD44. Conclusions　The AOM/DSS-induced mouse model exhibited chronic colonic inflammation, low-grade intraepithelial neoplasia, and high-grade intraepithelial neoplasia at seven, ten, and fourteen weeks, respectively. The pathological microenvironment was characterized by fibroblast activation and abnormal proliferation of epithelial cells.

Abstract: Objective　To prepare a stable rat model of chronic liver failure to provide a tool for basic research. Methods　Sixty-six SPF SD rats were divided into a normal group (n= 18) and a modeling group (n=48). Rats in the modeling group received an intraperitoneal injection of 50% CCl₄ olive oil solution (1.5 mL/kg, twice a week). Multidimensional assessment was performed at 8, 16, and 24 weeks, respectively, including ultrasonic examination of liver morphology, hardness, portal vein diameter, and ascites, and collection of serum, plasma, and liver tissue to detect liver function, coagulation function, and blood ammonia levels. Liver tissue injury and fibrosis were observed by hematoxylin-eosin(HE) and Masson staining. Cognitive function was assessed using the water maze test. Survival were recorded simultaneously. Results　Rats in the model group showed decreased activity and appetite, yellow urine, and increased abdominal circumference compared with the normal group. Ultrasound showed enhanced liver parenchyma echo in the model group that thickened with time, secondary ascites formation, portal vein dilation, and portal hypertension. Water maze and blood ammonia tests confirmed cognitive decline (memory and orientation loss) and hepatic encephalopathy in the model group. Gross observation showed that the liver in the model group was atrophied and appeared rough and uneven. HE staining showed hepatocyte swelling, steatosis, and necrosis, and Masson staining confirmed fibrosis progression with pseudolobule formation. The liver function indexes AST, ALT, TBIL and blood ammonia continued to increase, and coagulation dysfunction (prolonged PT and increased INR) gradually increased with the modeling process. Conclusions　Intraperitoneal injection of 50% CCl₄ olive oil solution (1.5 mL/kg,every week) for 24 weeks can stably simulate persistent chronic liver injury in rats and lead to the typical pathological changes and complications of chronic liver failure, based on the decompensation stage of cirrhosis. This model replicates the pathological evolution of human hepatitis from liver fibrosis → liver cirrhosis compensation → decompensation → chronic liver failure, providing a reliable modeling reference for the study of the mechanism of chronic liver failure.

Abstract: Objective　To investigate changes in the pro-inflammatory mediator S100A8/9 and NLRP3/ Caspase-1/IL-1β pathway in a rat kidney-aging model induced by D-galactose. Methods　Twelve SD rats were divided into control and D-galactose groups, and injected subcutaneously in the back of the neck with D-galactose (150 mg/kg) to establish a rat model of kidney aging. Kidney samples were collected under anesthesia after 8 weeks. Kidneys were stained for senescence-associated beta-galactosidase (SA-β-Gal), mRNA expression levels of the aging related genes p21, p16, and p53 were detected by quantitative reverse transcription-polymerase chain reaction(qRT PCR), and histopathological changes were observed by hematoxylin-eosin(HE) and Masson staining. Serum urea nitrogen and creatinine, and catalase (CAT), glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), and malondialdehyde (MDA) levels in the kidney tissues were detected. Reactive oxygen species (ROS) were detected by dihydroethdium staining and protein expression levels of collagen Ⅲ, α-smooth muscle actin (α-SMA), Protein expression of S100A8/9 was detected by immunofluorescence, and transforming growth factor (TGF)-β1 levels in kidney tissues and key factors in the NLRP3/Caspase-1/IL-1β inflammatory pathway were detected by Western Blot. A renal senescence model using HK-2 cells was constructed using H₂ O₂ in vitro, and expression levels of the senescence proteins p21 and p16 and mRNA expression levels of the inflammatory factors IL-18 and tumor necrosis factor-α(TNF-α) were detected. Cell senescence was observed by SA-β-Gal staining. The effects of the S100A8/9 inhibitor paquinimod on expression levels of S100A8/9 and NLRP3/Caspase-1/IL-1β pathway-related proteins in the aging model were also detected. Results　mRNA levels of the aging genes p21, p16, and p53 in kidney tissues were significantly increased in rats in the D-galactose group compared with the control group (P<0.01), and SA-β-Gal staining showed a significant increase in senescent cells (P<0.01). Serum blood urea nitrogen and creatinine levels increased (P<0.05), CAT, GSH-PX, and SOD activities decreased (P<0.01), while MDA activity increased in the D-galactose group (P<0.01). Collagen Ⅲ, α-SMA, and TGFβ1 expression and the ROS content in tissues increased (P<0.05). Glomeruli were atrophied or absent in the D-galactose group, the lumens of the renal sacs and renal tubules were enlarged, the nuclei were deeply stained and constricted, and numerous collagen fibers were deposited. Levels of S100A8 and S100A9 protein (P<0.01), as well as NLRP3, Caspase-1, and IL-1β increased (P < 0.05). Paquinimod alleviated HK-2 cell senescence and decreased expression levels of the senescence proteins p21 and p16, and mRNA levels of the inflammatory factors IL-18 and TNF-α (P<0.05, P<0.01). The number of senile cells was also decreased, shown by SA-β-Gal staining (P<0.01). Paquinimod also inhibited the protein expression of S100A8 and S100A9 (P<0.01) and NLRP3, Caspase-1, and IL-1β (P<0.05 or P<0.01). Conclusions　S100A8/9 participates in the chronic inflammatory response by activating the NLRP3/Caspase-1/IL1β pathway, thereby promoting D-galactose-induced renal aging.

Abstract: Objective　We explored the anti-depressant activity and mechanism of arecoline in vivo in a mouse model of depression induced by chronic unpredictable mild stress. The aim was to explore the possible mechanisms of action, providing experimental evidence for further research into the health benefits of arecoline and theoretical support for the scientific development and utilization of this resource. Methods　Sixty quarantine-qualified SPF C57BL/6J mice were divided randomly into a control group, model group, fluoxetine group (20 mg/kg), and arecoline low-, medium-, and high-dose groups (10, 20, and 40 mg/kg, respectively) according to body mass (n=10 mice per group). The effects of arecoline on the behavior of the mice were evaluated by open-field, tail suspension, and forced-swimming tests. Serum corticosterone and serum and brain levels of superoxide dismutase (SOD) were detected by enzyme-linked immunoassay. Malondialdehyde (MDA), catalase (CAT), 5hydroxytryptamine (5-HT), and norepinephrine (NE) levels in brain tissue, and dopamine (DA), gamma aminobutyric acid (GABA), tumor necrosis factor (TNF-α), interleukin (IL)-10, IL-1β, brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), and cAMP-response element binding protein (CREB) were detected by Western Blot. Results　Arecoline significantly reduced the total distance and average speed of the model mice in open field tests and increased activities, and significantly reduced the immobility time in the tail suspension and forced swimming tests. Arecoline also significantly decreased serum corticosterone levels, increased SOD and CAT, and decreased MDA levels. 5-HT, DA, NE, and GABA levels were significantly increased, and the cytokines TNF-α, IL-6, and IL-1β were significantly decreased. Expression levels of BDNF, TrkB, and CREB in the brain tissue were significantly increased. Conclusions　Research has found that arecoline has a significant antidepressant ability, and its mechanism may be achieved by reducing oxidative stress damage, inhibiting neuroinflammation, regulating neurotransmitter balance, and regulating the BDNF/TrkB/CREB signaling pathway. . This study explored the antidepressant efficacy of arecoline and preliminarily revealed its possible regulatory mechanism, which can provide data support for the neuroactivity of arecoline and lay a theoretical foundation for the development of arecoline as medicine.

Abstract: Objective　The progression of pulmonary fibrosis is a common clinical issue after acute lung injury (ALI). We aimed to construct a model simulating clinical ALI-induced pulmonary fibrosis by repeated challenges with lipopolysaccharide (LPS). We then observed the development from ALI to pulmonary fibrosis, to explore the possible mechanisms mediating the transition from inflammatory injury to fibrosis. Methods　Mice were treated with LPS (1, 2, 4, 8 mg/kg) intranasally to induce ALI. At 7 d, 14 d, 21 d, 28 d, 35 d, and 42 d after modeling respectively, α-smooth muscle actin (SMA), collagen 1 (Col-1), and hydroxyproline levels in lung tissue, and collagen fiber deposition were observed by Masson staining and compared to determine the process and degree of fibrosis formation in different modeling method. Expression changes in interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and transforming growth factor (TGF)-β1 in lung tissue at each time point were detected to explore the mechanisms of fibrosis formation. Results　Treatment of mice with 1, 4, and 4 mg/kg LPS for 3 consecutive days (M-1 group) result ed in a stable ALI-induced pulmonary fibrosis model. Masson staining showed that α-SMA, Col1, hydroxyproline, and collagen fiber deposition in the lung tissue began to increase in M-1 group mice in a time dependent manner after 7 d. Collagen deposition in the lung tissue interstitium was significantly increased at 21 d post modeling, and fibrosis indicators were significantly increased at 28 d, compared with control mice. Collagen deposition continued to increase until 42 d. Hydroxyproline and collagen fibers in the lung tissue in the other model groups with different doses and hit times did not increase significantly compared with the control group. TGF-β1 expression detected by Western blot began to increase gradually 14 d after modelling in the M-1 group, and was significantly higher than in the control group at 28 d. The pro-inflammatory cytokines TNF-α and IL-1β increased significantly on day 7 (acute phase), and TNF-α expression continued to increase until 28 d, while IL-1β gradually decreased after day 7. TNF-α and IL-1β in the lung tissue both continued to decrease after the acute phase in the other model groups without fibrosis. Conclusions　LPS 1, 4, and 4 mg/kg for 3 consecutive days can be used to construct an ALI/acute respiratory distress syndrome-induced pulmonary fibrosis model, via a mechanism that may be related to the sustained high expression of TNF-α regulating TGF-β1 to induce fibroblast activation and proliferation.

Abstract: Objective　To compare the hematological parameters, biochemical profiles, histopathological characteristics, and cytokines associated with lipid metabolism in 8-weeks-old specific pathogen-free (SPF) grade and germ-free(GF) golden hamsters. Methods　Twenty 8-weeks-old SPF grade and GF golden hamsters, with equal numbers of males and females, were utilized in this study. Serum cytokines associated with routine blood parameters, blood biochemistry, and lipid metabolism were quantified using automated hematology and biochemical analyzers, and enzyme-linked immunosorbent assay kits. The cecum, small intestine, lung, spleen, forestomach, and glandular stomach were examined by histopathology. Results　We compared the result between SPF grade and GF golden hamsters at 8 weeks of age. Regarding hematological parameters, females showed significant differences (P<0.05) in white blood cell count, hemoglobin concentration, mean corpuscular hemoglobin (MCH), and mean platelet volume between the two groups, and highly significant differences (P<0.001) in MCH concentration (MCHC), platelet distribution width (PDW), and lymphocyte count (LYM). Males showed significant differences (P<0.05) in mean corpuscular volume, MCH, LYM, neutrophil count, basophil count, and basophil percentage, and highly significant differences (P<0.001) in MCHC, PDW, lymphocyte percentage, and neutrophil percentage. In terms of biochemical parameters, females showed significant differences (P<0.05) in low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol, fasting insulin (FINS), and glycosylated serum protein (GSP), and highly significant differences (P<0.001) in triglycerides (TG). Males showed significant differences (P<0.05) in GSP levels and highly significant differences (P<0.001) in total cholesterol, TG, LDL-C, FINS, and C-peptide levels. For serum cytokines related to lipid metabolism, females showed significant differences (P<0.05) in interleukin (IL)-10, adiponectin (ADP), and IL-6 levels, and highly significant differences (P<0.001) in high sensitivity C-reactive protein (hs-CRP). Males showed significant differences (P<0.05) in hs-CRP levels and highly significant differences (P<0.001) in ADP and tumor necrosis factor-alpha levels. Hematoxylin/eosin staining showed that the cecal muscle layer was thinner and the number of crypts attached to the mucous membrane was reduced in GF compared with SPF grade golden hamsters. In addition, the ileocele was enlarged and the number of goblet cells was increased, the alveolar septum was widened, immune cells in the white pulp of the spleen were increased, and the blood content in the blood sinuses was increased. There was also thinning of the anterior gastric mucosa and the basophilic strength of the glandular gastric tube gland was weakened. Conclusions　This study established the differences in routine blood parameters, blood biochemistry indicators, histopathological features, and cytokines associated with lipid metabolism between 8-weeks-old SPF grade and GF golden hamsters, to establish preliminary reference ranges.

Abstract:Animal disease models are important biological tools for basic medical research. Establishing an ideal animal model is a critical prerequisite for acquiring reliable experimental data. By enabling molecular-level characterization, omics technologies can enhance the precision of animal model assessments, thereby improving the evaluation criteria. This review summarizes the current applications of omics in evaluating animal disease models, discusses their potential for quality control implementation, and proposes novel frameworks for standardized model validation.

Abstract:Chronic ulcers on the body surface are non-healing wounds. Establishing a suitable animal model of chronic wounds will provide an important tool for research aimed at preventing and understanding the complexity of chronic wound formation and related pathological mechanisms in the human body. Animal wound models are usually constructed by inducing molecular abnormalities via external injury interventions to induce wound formation. Common modeling method include surgical resection, pressure ischemia, drugs, and radiation treatment. The success of model construction can then be evaluated by various monitoring method, such as natural recovery of the wound without intervention, measurement of wound size observation of physical signs, measurement of body mass, organ index, and infrared imaging. Despite the existence of numerous modeling and evaluating method, however, there is currently a lack of unified standards for animal chronic wound models. Researchers should thus choose appropriate modeling and model-evaluation method based on their actual needs, to obtain the best experimental result.

Abstract:The development of bio-psycho-social medical models has focused attention on the impact of chronic psychological stress on diseases. Chronic psychological stress falls within the category of emotional etiology in traditional Chinese medicine. This paper systematically reviews research progress into the chronic psychological stress related combination of disease and syndrome, and analyzes the advantages and limitations of different models. We also provide insights into the direction and improvements of future research into chronic psychological stress-related disease pattern models.

Abstract:Diamond-Blackfan anemia (DBA), also known as congenital pure red cell aplasia, is a rare genetic disorder characterized by bone marrow failure, congenital anomalies, and severe red blood cell abnormalities. The rarity of the condition, and consequently limited patient pool and scarcity of research models, means that the pathogenic mechanisms associated with genetic mutations in DBA remain uncertain, and the clinical treatment options are limited. This review synthesizes the findings from zebrafish, mouse, and human cellular models of DBA mutations. We clarify the pathogenic mechanisms and monitor the progression of drugs into clinical trials, thereby aiding further in-depth explorations into the etiology and therapeutic advancements for DBA.

Abstract:Atherosclerosis (AS) is a chronic inflammatory disease involving disorders of lipid and iron metabolism. The establishment of suitable animal models is required to further the study of the etiology, pathogenesis,prevention, and therapeutic measures of AS. The main animal models of AS related to iron and lipid metabolism are mice and miniature piglets, especially male ApoE^-/-mice. Single-factor high-fat diet-induced iron and lipid metabolism disorders are a common type of AS model, manifesting as elevated blood lipid levels, large plaques and iron deposition in the aorta, and significant increases in serum and liver iron levels. This review compares the effects of different intervention factors on iron and lipid metabolism in AS animal models, and summarizes the method of establishing AS animal models using dietary induction, chemical intervention, and gene modification, to provide references and inspiration for future research into AS and metabolic diseases and the development of new drugs.

Abstract:Cancer-induced bone pain (CIBP) causes substantial suffering for cancer patients and also diminishes their quality of life and self-esteem. The mechanisms underlying CIBP are complex and evolve progressively with cancer advancement. Current treatment options show limited efficacy and are often accompanied by adverse effects. Traditional Chinese medicine demonstrates potential advantages in managing CIBP; however, the mechanisms of action remain poorly understood and require further investigation. The development of a standardized, stable, and reproducible animal model is crucial to advancing research on disease pathogenesis and verifying the effectiveness of therapeutic interventions. This review considers recent method for modeling CIBP in animals and summarizes the application of these models in studies of traditional Chinese medicine mechanisms, with the aim of guiding future research directions in CIBP.