Abstract: Objective　To explore the establishment of a subcutaneously transplanted tumor model of hepatocellular carcinoma in mice with Qi stagnation and blood stasis syndrome. Methods　Forty male C57BL/6 mice were randomly divided into 4 groups: NC group, QZXY group, Tumor group, and QZXY + Tumor group. They were categorized based on the modeling of Qi stagnation and blood stasis syndrome (7 days) combined with the modeling of subcutaneous transplantation of hepatocellular carcinoma tumor (20 days). Observations were conducted of the syndrome manifestations as well as the tumor size and weight of the mice after modeling. Results　(1) Body weight: on the 7th day of modeling, the weights of the QZXY group and QZXY + Tumor group were significantly lower than that of the NC group (P<0.05). (2) Body temperature: on the 7th day of modeling, body temperature significantly decreased in the QZXY group (P<0.05), while it increased in the Tumor group (P<0.05) compared with the NC group. On the 27th day of modeling, the temperature of the QZXY + Tumor group was significantly lower than that of the NC group (P<0.05). (3) Syndrome manifestations: according to the syndrome scoring table, mice in both the QZXY group and QZXY + Tumor group exhibited Qi stagnation and blood stasis syndrome on the 7th day of modeling (P<0.05). As modeling time extended, the score of mice in the Tumor group increased with the formation of the tumor, and the score of mice in the QZXY + Tumor group was significantly higher than that of the other three groups (P<0.05). (4) Claw petechiae: the number of claw petechiae significantly increased in all three groups of modeled mice compared with the NC group (P<0.05), with the QZXY + Tumor group showing the highest number. (5) Claw r value: the r value of the claw was significantly lower in all three groups of modeled mice than that in the NC group (P<0.05). Additionally, the r value of the claw in the QZXY + Tumor group was consistently lower than that of the other three groups. (6) Open field activity: the vertical and horizontal activity of mice in the QZXY + Tumor group decreased significantly compared with that of the NC group (P<0.05). (7) Coagulation indexes: APTT, TT, and FIB were significantly increased in the QZXY + Tumor group (P<0.05 or P<0.01) compared with those in the NC group. (8) Tumor size and weight: compared with the Tumor group, the QZXY + Tumor group showed significantly increased tumor size and weight (P<0.05). Conclusions　This study successfully established a subcutaneous transplanted tumor model of hepatocellular carcinoma in mice with Qi stagnation and blood stasis syndrome. The findings indicated that Qi stagnation and blood statsis syndrome may occur during the course of live cancer. Besides, the causes inducing the Qi stagnation and blood stasis syndrome will further accelerate the progression of liver cancer.

Abstract: Objective　To compare the effects of intratracheal instillation by lumbar spinal needle and intratracheal atomization on bleomycin-induced pulmonary fibrosis modeling in mice, to determine the optimal modeling method. Methods　Seventy-two C57BL/6J mice were divided randomly into control, lumbar spinal needle and aerosolization groups, according to body weight (n= 24 mice per group). Mice in the control and lumbar spinal needle groups received intratracheal instillation of saline or bleomycin, respectively, and mice in the aerosolization group received aerosolized bleomycin intracheally by microsprayer aerosolizer. Micro-computed tomography (CT), histopathological changes, hydroxyproline(HYP) levels, Collagen Ⅰ(COLⅠ) and α-smooth muscle actin (α-SMA) protein expression were examined on days 14 and 21 to evaluate the degree of pulmonary fibrosis in each group. Results　Mice in the two model groups showed listlessness, slow responses, and decreased body weights on days 14 and 21, compared with the control group (P<0.001). Micro-CT showed white shadows surrounding the trachea in the lumbar spinal needle group, while the shadows were more diffuse in the aerosol group. The degrees of alveolitis and pulmonary fibrosis were highest in the aerosolization group, with a time-dependent trend. The hydroxyproline contents were significantly increased in the two model groups on days 14 and 21 after modeling (P<0.05), with the increase on day 21 being more significant and stable (P<0.001). COLⅠ expression was significantly increased in both the lumbar spinal needle group and aerosolization group on days 21 after modeling, especially in the aerosolization group(P<0.001). Expression levels of α-SMA were significantly higher in the lumbar spinal needle group and aerosolization group compared with the control group on days 21 (P<0.001); however, there was no significant difference between the two model groups. Conclusions　intratracheal atomization of bleomycin is the optimal method for establishing a mouse model of pulmonary fibrosis.

Abstract: Objective　To observe the expression characteristics of green fluorescent protein (GFP)-positive myeloid cells (mainly monocytes, macrophages, and neutrophils) and their pattern of response to diphtheria toxin (DT) in Lyz2 internal ribosome entry site (IRES) DT receptor/enhanced GFP (DTREGFP) mice. Methods　Transgenic Lyz2 IRES DTREGFP mice (6 ~ 8 weeks old) and C57BL/6J mice with the same genetic background were selected randomly and injected intraperitoneally with DT (25 ng/g) for 3 consecutive days. Bone marrow, peripheral blood, and spleen cells were separated and made into single-cell suspensions with GFP and CD11b as markers. Changes in the proportion of GFP⁺ cells to total myeloid cells (CD11b⁺) was detected in these tissues at different time points before and after drug administration, using flow cytometry. Results　Peripheral blood CD11b⁺GFP⁺ cells accounted for (24.62 ± 5.655) % of CD11b⁺ cells before DT injection, and this proportion gradually decreased after injection and reached a minimum of (7.982 ± 2.729) % on the third day. The proportions in spleen and bone marrow were (13.73 ± 2.994) % and (65.23 ± 4.261) % before DT injection, respectively, decreased most significantly on the first day after injection to (3.468 ± 0.5862) % and (50.98 ± 7.957) %, respectively, and then gradually recovered. Conclusions　DT effectively reduced the proportions of monocytes, macrophages, and neutrophils in Lyz2 IRES DTREGFP mice, but the pattern of change varied among tissues.

Abstract: Objective　To evaluate the effects of fecal status and transplantation method on the intestinal flora structure after fecal microbiota transplantation in germ-free mice. Methods　Thirty-six C57BL/6 mice were divided randomly into three groups: fresh fecal gavage transplantation (group A), frozen fecal gavage transplantation (group B), and frozen fecal rectal transplantation groups (group C). Feces were collected at 2, 4 and 6 weeks after transplantation. All mice were sacrificed at 6 weeks to obtain the contents of the small and large intestines. The structure and function of the gut microbiota and dynamic trends in microbial changes were analyzed by 16S rRNA sequencing. Results　The diversity of the small intestine microbes in both the group A and group C were similar to those in the group B, according to α-diversity analysis (P>0.05), but the diversity of large intestine microbes was significantly increased (P<0.001). According to β-diversity analysis, small intestine samples from the group A and group B clustered in the same area, indicating that the microbial community compositions were similar (P>0.05), but samples from the large intestine were distributed in different areas, showing significant differences (P<0.001). Small and large intestine samples from the group B and group C were distributed in different areas, with significant differences (P<0.001). Linear discriminant analysis effect size showed that Bacteroidota were relatively dominant in the group A, while Verrucomicrobiota and Proteobacteria were relatively dominant in the group B and Firmicutes were relatively dominant in the group C. Functional prediction using PICRUSt2 software showed that neither fecal status nor the method of transplantation affected the functions of the microbial community. Conclusions　Both fresh fecal gavage and frozen fecal rectal transplantation can enhance the microbial diversity of the large intestine, compared with frozen fecal gavage transplantation. Fecal status does not affect the gut function and colonization trends of the microbiota, whereas the method of transplantation affects the colonization trends but not the functions of the microbiota.

Abstract: Objective　Transcriptome sequencing technology (RNA-seq) was used to analyze the mechanism of compound Dancao granules as an intervention for high-fat feed combined with carbon tetrachloride (CCl₄ )-induced nonalcoholic steatohepatitis. Methods　45 male C57BL/6J mice were split into two groups at random: normal control group, model control group, obeccholic acid group 10 mg/(kg·d), and compound Dancao granules low- and high-dose groups 3.74g/(kg·d) and 7.48g/(kg·d), with 9 mice in each group. Normal diet was made available to the control group,and the mice in the model group were given a high-fat diet combined with the subcutaneous injection of CC1₄ , with 100% CC1₄ solution (4 mL/kg) in the first application, and 40% CC1₄-olive oil solution (2 mL/kg) in the second application, twice a week for a total of 6 weeks. Each drug group was administered the respective drug from week 3 for a total of 4 weeks. 12 h after the last administration, the serum and liver tissues of mice in each group were collected, and a biochemical kit was used to detect serum liver function. Hematoxylin-eosin (HE), sirius scarlet, and oil red O staining were used to examine histopathological changes to the liver. The levels of IL-6, IL-10, TNF-α and TGF-β in mice liver were detected via ELISA, and the expression of α-SMA was observed by immunohistochemistry. Differential gene expression was analyzed by RNA-seq and functional enrichment analysis. To verify the differential expression of mRNA, quantitative reverse transcription PCR(qRT-PCR) was used. TDT-mediated dUTP nick-end labeling (TUNEL) staining was employed to identify apoptosis. Results　The model control groups had significantly higher levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), and triglycerides (TG) than normal control group (P<0.01). Additionally, there was obvious inflammatory cell infiltration in the liver tissue, collagen deposition in the sink and interlobule areas, and a significant increase in lipid droplet area (P<0.01). The levels of IL-6 and TNF-α in liver tissue were significantly increased (P<0.01), the levels of IL-10 and TGF-β were decreased (P<0.01), and the expression of α-SMA was significantly increased (P<0.01). The levels of TC, TG, ALT, and AST were significantly lower in groups that received compound Dancao granules and obeccholic acid than the model control group (P<0.01), and inflammatory cell infiltration, collagen deposition, and fat accumulation in the sink and interlobule areas were improved (P<0.01). The levels of IL-6 and TNF-α in liver tissue were significantly decreased (P<0.01), the levels of IL-10 and TGF-β were increased (P<0.05, P<0.01), and the expression of α-SMA was significantly decreased (P<0.01). RNA-seq sequencing result showed that 2819 genes in the normal control group were differentially expressed compared with the model control group, with 543 up-regulated and 2276 down-regulated genes. In a comparison of the model control group and compound Dancao granules group, 240 genes were differentially expressed, including 206 upregulated genes and 34 down-regulated genes. There were 221 genes with overlapping expression in the 2 groups and functional enrichment highlighted cell cycle(Cdt1, Plk1, Bub1b, Ttk, Knl1, Esco2, Cdc6, Ndc80, Cdc25b, Sgo1, Ccnb2, Espl1, Ccne1, Mcm4, Mcm5, Fbxo5, Bub1, Mcm2), apoptosis(Caspase3, Bax, P53, Apaf1, Bak, Caspase8), the P53 signaling pathway (P53, Ccnb2, Apaf1, Bak, Bax, Gtse1, Caspase3, Ccne1), arachidonic acid metabolism (Hpgds, Cyp2c54, Cyp2b10, Tbxas1, Cyp2c50), galactose metabolism (Hk3, Gla, Hk2, Akr1b7) and other signaling pathway genes. RNA-seq sequencing analysis showed that compound Danicao granules mainly regulated the apoptosis signaling pathway, and qRT-PCR confirmed that the mRNA expression of Caspase3, Bax, P53, Apaf1, Bak and Caspase8 in the liver tissue of the model control group was increased compared with that of the normal control group (P<0.01). Compared with the model control group, the compound Dancao granules group showed decreased mRNA expression of Caspase3, Bax, P53, Apaf1, Bak and Caspase8 in liver tissue (P<0.01). TUNEL staining showed that the number of cells showing nuclear shrinkage and apoptotic bodies decreased in the compound Dancao granule administration group. Conclusions　Compound Dancao granules had a significant protective effect against non-alcoholic steatohepatitis induced by high-fat feed combined with CCl₄ , and its mechanism might be connected to the control of genes linked to apoptosis.

Abstract: Objective　To construct an accurate clinical model of lower limb varicose veins in rats through surgery that provides theoretical support for evaluating drug therapy. Methods　30 SD rats, 15 males and 15 females, were randomly divided into a control group and surgical group. In the surgical group, the rats lower limb veins (including the small saphenous vein and femoral vein) were ligated via improved lower limb vein ligation, i.e., the small saphenous vein was completely ligated with the femoral vein, and the thrombosis result ed in a lasting increase in the internal pressure of the deep veins of the lower limb, causing varicose symptoms. On the 6th week after surgery, the varicose veins of the rats in the surgical group were scored to select those thatwere successfully modeled. Then, the successfully modeled rats were randomly divided into a model group and Maizhiling group. Maizhiling 62.5 mg/kg was orally administered to the treatment group once a day, while the control and model group received an equal volume of physiological saline orally every day for 20 consecutive days. On the day before administration and 7 d, 14 d and 20 d after administration, macro photography and scoring were performed on the lower limbs of the rats. After completion, an approximately 1 cm long saphenous vein above the ankle joint of the lower limb on the surgical side was removed from the model group and Maizhiling group rats, while from the control group, the corresponding saphenous vein of the lower limb on the same side was removed. Pathological tissue observation was performed using HE staining, Masson staining, and immunohistochemical examination for interleukin-2(IL-2)and tissue inhibitor of metalloproteinases(TIMP-1). Results　Of the 22 rats in the surgical group, 20 were successfully modeled, with a success rate of 91%. According to the manifestations of venous dilation, varicose veins, and redness in the lower limbs of rats, the varicose vein score of the model group increased significantly compared with that of the control group (P<0.01). After the therapeutic dose of Maizhiling was administered, the varicose vein score in the Maizhiling group decreased significantly compared with that of the model group (P<0.01). Pathological examination showed significant varicose-vein-like changes and mild inflammation in the model group. The Maizhiling group showed reduced varicose veins and inflammation. Conclusions　A rat model of lower limb varicose veins was successfully established, providing a new research method for the study of drugs and treatment method related to lower limb varicose diseases.

Abstract: Objective　To investigate the dose-dependence of inflammation, oxidative stress and epithelial barrier disruption in C57BL/6 mice using different concentrations of PM_2.5 . Methods　A total of 36 male C57BL/6 mice were randomly divided into control group, PM_2.5 5.0 group, PM_2.5 7.5 group and PM_2.5 10.0 group. The trachea of the control group was instilled with normal saline, and the animals were sacrificed after the last poisoning. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung tissue. The ELISA kit was used to detect the levels of IL-4, IL-1β and TNF-α inflammatory cytokines in the serum of four groups of mice. Biochemical kits and ELISA kits were used to detect the levels of oxidative stress nitrogen monoxide (NO), malondialdehyde (MDA) and superoxide dismutase (SOD) in four groups of mice. Deoxyribonucleotide terminal transferase-mediated nick end labeling was used by TUNEL staining to observe the level of apoptosis of epithelial cells. Immunohistochemistry was used to detect the expression of epithelial barrier tight junction proteins. Results　Acute exposure to PM_2.5 led to widening of alveolar septum and exudation of inflammatory cells, increased serum levels of inflammatory cytokines IL-4, IL-1β and TNF-α and NO and MDA in lung tissue, and decreased SOD (P<0.01 or P<0.05). increased apoptosis of epithelial cells; The expression of epithelial barrier tight junction protein was dose-dependent (P<0.01 or P<0.05). Conclusions　Acute exposure to PM_2.5 particulate matter can lead to acute lung injury in mice by promoting the release of inflammation in lung tissue and inducing oxidative and antioxidant imbalance. In addition, PM_2.5 exposure led to epithelial cell apoptosis, especially at a modeled dose of 10 mg/kg, which disrupted the tight junctions of the epithelial barrier and further exacerbated lung injury in mice.

Abstract: Objective　To establish and optimize an animal model of atopic march (AM), skin and lung tissue sensitization under single or combined modeling of ovalbumins (OVA)and calcipotriol (MC903)was compared. Methods 40 SPF BALB/c mice aged 6 ~ 8 weeks were randomly divided into a control group, model group A, model group B and model group C. The 3 of AM models were established with MC903, MC903 + OVA and OVA, respectively, through skin sensitization ( twice) and respiratory sensitization (once). Skin sensitization severity scoring system, immunohistochemistry, and enzyme-linked adsorption assay were used to compare skin lesion morphology, skin or lung histopathology, and immunophenotype. Results　Compared to OVA or MC903 modeling alone, MC903 + OVA modeled mice showed more significant changes in skin morphology, a higher score for skin sensitization severity, more severe skin and airway inflammatory cell infiltration, and more significant changes in the expression of the related inflammatory factors thymic stromal lymphopoietin (TSLP), interleukin 4 (IL-4), IL-13 and IL-10 (P<0.05). Conclusions　An AM animal model optimized by MC903 combined with OVA was successfully constructed that provides a good method ological basis for AM mechanism research.

Abstract:Peptidylarginine deiminases 2 (PADI2) is an enzyme that catalyzes the conversion of arginine residues to citrulline on protein peptides. Aberrant activation of PADI2 can induce excessive tissue inflammation and immune responses, thereby exacerbating the progression of autoimmune diseases (ADs). Through citrullination, PADI2 modifies protein structure and immunogenicity, influencing the production of autoantibodies and regulating the activity of immune cells such as neutrophils and macrophages. This review provides an overview of PADI2’s functions and its pivotal role in ADs, with the Objective of elucidating the mechanisms underlying ADs pathogenesis and identifying novel therapeutic targets and strategies for related diseases.

Abstract:Laboratory animals provide an important experimental resource for life science research, and their uniformity plays a key role in the accuracy and reliability of the experimental result. Genetic quality testing of laboratory animals is thus essential to evaluate the genetic quality of laboratory animals. Progress in modern biotechnology has led to improvements in the method of genetic testing of experimental animals. Single nucleotide polymorphisms, as molecular markers, have been widely used in genetic testing of laboratory animals, and method for their detection in laboratory animals have been updated, in line with biotechnology advances. In this paper, we review current research progress and the application of SNP detection method for genetic quality testing of laboratory animals, and discuss the advantages and disadvantages of various detection method, with a view to providing a reference basis for genetic quality testing of laboratory animals.

Abstract:Non-specific lower back pain is a common clinical disease whose pathogenesis and causes are still unclear, and the advantages and disadvantages of various therapeutic programs are controversial. Current research on this disease is mostly limited to clinical studies, and there is an urgent need to invest in a large number of animal experiments to analyze its underlying mechanisms. The construction of animal models is an important means to study the pathogenesis of non-specific lower back pain and to explore therapeutic method, but there are certain limitations and delays in the establishment of models for this disease. Therefore, this paper reviews the selection of animals, construction method, and evaluation method of relevant indexes of animal models of non-specific lower back pain, and the advantages and disadvantages of various models, to clarify existing problems in current basic research of this disease. We provide new research ideas and aim to lay a theoretical foundation for studies into the mechanisms of non-specific lower back pain and improved therapeutic strategies.

Abstract:Age-related ovarian aging is significantly influenced by oxidative stress, which has intricate underlying mechanisms. Studies conducted recently have demonstrated that oxidative stress is a mediator of several pathological processes that lead to ovarian dysfunction in aging. These processes include telomere shortening, chronic inflammation, apoptosis, and mitochondrial dysfunction. Under oxidative stress, antioxidant treatment can assist in enhancing ovarian function.

Abstract:Non-alcoholic fatty liver disease (NAFLD) is a metabolic syndrome caused by a variety of pathogenic factors and characterized by the excessive deposition of fat in liver cells. At present, the pathogenesis of NAFLD is not fully understood, and there is a lack of specific drugs for NAFLD in clinical practice. Therefore, the establishment of an ideal animal model is extremely important for clarifying the pathogenesis of NAFLD and developing specific drugs and treatments. In addition to NAFLD animal models, an increasing number of disease-syndrome combined models related to NAFLD has emerged with the continuous renewal and development of traditional Chinese medicine over the past five years. This article summarizes the NAFLD animal models commonly used within the past five years, as well as their preparation and evaluation method, to provide a reference for future studies in NAFLD animal model preparation and modeling a combination of disease and syndrome.