Abstract: Objective　The aim of this study was to establish a rat model of inflammation-cancer transformation of inflammatory bowel disease (IBD) and to explore the associated characteristics of the intestinal flora. Methods　Adult male Wistar rats were divided randomly into control and model groups (M1, M2, M3) with different dextran sulfate sodium (DSS) intervention cycles. Colitis-cancer transformation was induced in all rats in the model group by a single intraperitoneal injection of azoxymethane (AOM) combined with free drinking of DSS in different cycles, and disease activity index (DAI) scores were recorded. Rats in the M1, M2 and M3 groups were killed at the end of the first, second, and third cycles of DSS, respectively, and spleen and colon tissues and colon contents were collected. Histological damage and colon carcinogenesis were evaluated in each group using hematoxylin and eosin staining and transmission electron microscopy. Characteristic changes in the intestinal flora were analyzed by 16S rRNA sequencing. Results AOM/DSS administration significantly increased the DAI score, shortened the colon, and increased the spleen index. The intestinal mucosal barrier was progressively destroyed from groups M1 to M3, and the pathological score was gradually increased. Abnormal crypt foci, polyps, low- and high-grade intraepithelial neoplasia, and mucosal carcinoma appeared in turn, while the pathological process showed similar characteristics to carcinogenesis in human inflammatory bowel disease. Screening using 16S rRNA sequencing with two differential abundance testing tools, Wilcoxon and ALDEx2, indicated that changes in flora abundance represented by Bacteroidetes and Monoglobus may be involved in the progression of colitis cancer transformation. The functions of the differential flora were mainly enriched in metabolic pathways, such as lipid and carbohydrate metabolism. Conclusions　The current rat model induced by AOM/DSS can dynamically simulate the pathological characteristics of colitis-cancer transformation, accompanied by changes in the abundance of specific intestinal flora, which may be closely related to the metabolic pathways mediated by the flora.

Abstract: Objective　To investigate changes in the intestinal flora and function in rats with antibiotic-associated diarrhea(AAD) treated with GeGen QinLian Decoction (GQD). Methods　Sixty male or female SPF-grade SD rats were fed for 7 days and then divided randomly into Control(Con) and modeling groups (1∶ 5 ratio). Rats in the modeling group received clindamycin 250 mg/kg by gavage once a day for 7 consecutive days. After successful modeling, the rats were divided randomly into model(Mod), high-dose GQD (GQD-H, 10.08 g/kg), medium-dose GQD (GQD-M, 5.04 g/kg), low-dose GQD (GQD-L, 2.52 g/kg), and live Bifidobacterium power (LBP, 0.15 g/kg) groups (n=10 rats per group). GQD and LBP were administered by gavage, and the Con and Mod groups were given an equal volume of saline by gavage once a day. Feces were collected after 7 consecutive days of administration for macrogenomics sequencing analysis. Results　α diversity and β diversity suggested that intestinal microbial diversity differed between the Mod and GQD-treated groups. GQD increased the abundance of thick-walled bacteria and decreased the abundance of Aspergillus at the phylum level, and increased the relative abundances of the intestinal mucus bacteria Blautia, Bacteroides, Thomasclavelia, and Mediterraneibacter, and decreased the relative abundances of Adlercreutzia, Muribaculum, and Escherichia at the genus level. GQD also up-regulated the amino acid, carbohydrate, and immune disease pathways. Conclusions　GQD improves the abundance ratio of beneficial and pathogenic intestinal bacteria in rats with antibiotic-associated diarrhea, which in turn reduces the intestinal inflammatory response and repairs the intestinal immune system.

Abstract: Objective　The CRISPR/Cas9 system was utilized to generate an Apoe knockout mice model to support further investigations of the role of Apoe in lipid metabolism and atherosclerosis. Methods　Two single guide RNAs designed for Apoe in C57BL/6J mice were co-injected with Cas9 mRNA into fertilized eggs, followed by transplantation into ICR recipient mice to obtain F₀ generation mice. KO mice were identified by polymerase chain reaction (PCR) screening of tail DNA. Apoe mRNA expression in various tissues was assessed by quantitative real-time PCR and lipid indexes were measured in serum samples. Lipid accumulation in the inner lining of aortic vessels was detected by oil red O staining. Results　PCR and sequencing confirmed the successful construction of Apoe KO mice (C57BL/6-Apoe^em1 /Nifdc). Apoe mRNA levels were significantly reduced in the liver, brain, spleen, kidney, and lung tissues of Apoe KO homozygous mic (Apoe^-/-), as shown by reverse transcription quantitative real-time PCR. Serum total cholesterol and low-density lipoprotein cholesterol levels were increased in Apoe^-/- mice, and high-density lipoprotein cholesterol levels were decreased in male Apoe^-/- mice. Extensive lipid plaques were observed in the inner lining of the arteries in Apoe^-/- mice compared with WT mice, under normal chow consumption conditions. Conclusions　This study successfully established an Apoe KO mice model exhibiting a typical abnormal lipid metabolism phenotype with arterial lipid accumulation, even without a highfat diet intervention. This work provides background data for the Apoe KO mice resource and a new model for the study of abnormal lipid metabolism.

Abstract: Objective　To explore the construction method, evaluation system, and related indicators of the Qi Yin deficiency syndrome model of type 2 diabetes (T2DM) in Guizhou miniature pigs. Methods　Twenty-six 6-month-old Guizhou miniature pigs were divided into 2 groups: pigs fed a basic diet for 10 months (normal group, n= 6), and pigs fed a high-sugar, high-fat diet for 10 months followed by Qingpi Fuzi decoction for 3 months (Qi Yin deficiency in T2DM model group, n= 20). Qi Yin deficiency in the T2DM model was created in Guizhou miniature pigs. Traditional Chinese medicine syndrome types were evaluated using five scales, including urination, defecation, movement and mental state, response to the surrounding environment, and color of the skin, hair, and nasal disc. Serum CD4 ⁺ and CD8 ⁺ and their ratios, cAMP and cGMP and their ratios, as well as tongue RGB color values were used to evaluate Qi Yin deficiency syndrome. Fasting blood glucose (FBG), glycosylated hemoglobin (HbA1c), glucose tolerance test (OGTT), homeostasis model assessment of insulin resistance(HOMA-IR), triglycerides (TG), total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), prothrombin time (PT), activated partial thromboplastin time (APTT), and other indicators were used to evaluate T2DM. Results　Physically, the model group pigs were overweight and showed fatigue, with a tendency to lie down, lethargy, overeating and drinking, and dull fur. The defecation, movement and mental state, response to the surrounding environment, and color of the skin, fur, and nasal disc scores were relatively low (P<0.05). The tongue appeared red with no or little coating. Pigs in the model group showed significantly lower of FBG, HbA1c, HOMA-IR, TC, LDL-C, cAMP, and cGMP, compared with the normal group. The RGB values of the tongue images were significantly reduced. The PT, APTT, CD4 ⁺, and CD4 ⁺ /CD8 ⁺ values were also significantly reduced, and TG, HDL-C, and cGMP were all elevated (all P>0.05). Pathological staining showed destruction of pancreatic tissue. Conclusions　The method of using a high-sugar and high-fat diet combined with traditional Chinese medicine for breaking Qi and damaging Yin can be used to establish a small pig model of T2DM with Qi Yin deficiency. The established model shows the main symptoms of fatigue, thirst, shortness of breath and lethargy, redness and dryness of the tongue, lack of coating on the tongue, and a weak pulse in traditional Chinese medicine syndromes, as well as concurrent symptoms including yellow and transparent urine, dry stools, slow response, curling up, dull fur, and a gray and dull nasal disc, as well as clinical manifestations of abnormal glucose and lipid metabolism and insulin resistance.

Abstract: Objective　This study aimed to evaluate the bone integration performance of antibacterial carbon dot (CD)-modified polyether ether ketone (PEEK) in infectious bone defect environments. Methods　Guanidine-based CDs (G-CDs) prepared by the melting method combined with dialysis purification were used to modify PEEK implants using polyvinyl butyraldehyde (PVB) by the soaking-drying method (PEEK/PVB-G-CDs). SD rats were divided into the following groups: (1) PEEK-implanted uninfected (PEEK(-)), (2) PEEK/PVB-G-CDs-implanted uninfected (PEEK/ PVB-G-CDs(-)), (3) PEEK-implanted infected (PEEK(+)), and (4) PEEK/PVB-G-CDs-implanted infected (PEEK/PVB-G-CDs(+)). A hole (diameter 2 mm, depth 5 mm) was drilled at the lateral condyle of the vertical femur in all rats to simulate a bone defect. Rats in the PEEK(-) and PEEK/PVB-G-CDs(-) groups without infection were injected with 30 μL physiological saline into the bone marrow cavity, and rats in the PEEK(+) and PEEK/PVB-G-CDs (+) groups with infection were injected with 30 μL MRSA bacterial suspension (1.5 × 10⁴ colony-forming units/mL) into the bone marrow cavity. The implantation site was observed using animal-specific X-ray examination at 0, 2, and 4 weeks after implantation, and the bone tissue characteristics of the implantation site were evaluated by micro computed tomography (CT) at 6 weeks after surgery. The bone implantation sites in each group of rats were examined by bacterial culture of bone marrow and hematoxylin and eosin staining, Toluidine blue, Goldner trichrome, and immunohistochemical staining. Results　X-ray, Micro-CT, bacterial culture of bone marrow, and histopathological analysis confirmed no signs of infection in the PEEK(-) and PEEK/PVB-G-CDs(-) groups and the implants were integrated with the bone defects. Rats in the PEEK/PVB-G-CDs(+) group showed signs of antibacterial activity that effectively controlled the osteomyelitis caused by MRSA and achieved bone integration, while rats in the PEEK(+) group failed to achieve bone integration because of persistent infection. Immunohistochemical staining confirmed lower levels of anti-inflammatory factors such as IL-4 and IL10 in the PEEK(+) group, and stronger expression of pro-inflammatory factors such as IL-6 and TNF-α compared with the other three groups, indicating that G-CD-modified PEEK inhibited MRSA infection, regulated inflammation levels in the local microenvironment, and promoted bone integration at the site of bone defects. Conclusions　Antibacterial G-CDs modified PEEK exhibits excellent bone integration performance, providing a candidate strategy for future clinical treatment of infectious bone defects.

Abstract: Objective　Use of silencing information regulatory factor 1 (SIRT1)/high mobility group protein B1 (HMGB1)/nuclear transcription factor-κB (NF-κB) to investigate the inhibitory effect and mechanism of Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea on hepatocyte inflammatory apoptosis in mice with carbon tetrachloride (CCl₄ )-induced acute liver injury. Methods　C57BL/6 mice were randomly divided into a control group; model group; resveratrol group; Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea low-, medium-, and high-dose groups; and a positive drug group. The mice were given continuous intragastric administration of the treatments for 14 days. An acute liver injury model was established by intraperitoneal injection of 0.5% CCl₄ olive oil solution (5 mL/kg). The levels of alanine transferase (ALT), aspartate transferase (AST), lactate dehydrogenase (LDH) in serum and hydroxyproline (Hyp), malondialdehyde (MDA), and superoxide dismutase (SOD) in the liver were determined by biochemical method. Serum levels of inflammatory tumor necrosis factor-α(TNF-α), interleukin (IL)-6, and IL-1β were determined by enzyme-linked immunosorbent assay. Eosin-hematoxylin and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) staining were used to examine the pathological morphology and apoptosis in liver tissues. The protein expression of SIRT1, HMGB1, and NF-κB were detected by Western Blot. Results　Compared with those of the control group, the model group’s serum ALT, AST, and LDH levels and liver tissue Hyp activity were significantly increased; MDA and SOD activities in liver tissue were significantly decreased; the levels of inflammatory factors TNF-α, IL-6, and IL-1β in liver tissue were significantly increased; and there were obvious signs of pathological injury and hepatocyte apoptosis in the liver tissue. The expression of SIRT1 protein decreased significantly, while the expression of HMGB1 and NF-κB proteins increased, in liver tissue. Compared with those of the model group, the Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea high group and resveratrol group serum levels of ALT, AST, and LDH and liver tissue Hyp activity were significantly decreased; MDA and SOD activities in liver tissue were significantly increased; the levels of serum inflammatory factors TNF-α, IL-6, and IL-1β were decreased; and pathological injury to liver tissue and the apoptosis of liver cells were significantly improved. The expression of SIRT1 protein in liver tissue was increased, while the expression of HMGB1 and NF-κB protein were decreased in the Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea high-dose group and resveratrol group(P<0.05 or P<0.01). Conclusions　Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea effectively protects against acute liver injury, and its mechanisms may be related to the regulation of the SIRT1/HMGB1/NF-κB signaling pathway and the alleviation hepatocyte inflammatory apoptosis.

Abstract: Objective　To investigate the effect of bear bile powder (BBP) and ursodeoxycholic acid (UDCA) on preneoplastic lesions of hepatocarcinoma, using short-term carcinogenesis bioassay models. Methods　Forty 6-week-old male SD rats were divided randomly into control, diethylnitrosamine (DEN), DEN + BBP (200 mg/kg), and DEN + UDCA (30 mg/kg) groups. All rats, except for the control group, were injected intraperitoneally with 100 mg/kg DEN once a week for 3 weeks. Rats in the DEN + BBP and DEN + UDCA groups also received oral BBP 200 mg/kg or UDCA 30 mg/kg suspended solution, respectively, daily from the beginning to the end of the experiment. Results　There were no 　　　　　　significant differences in body or liver weights between the DEN, DEN + BBP, and DEN + UDCA groups. DEN treatment increased the accumulation of malondialdehyde (MDA), decreased superoxide dismutase (SOD), and reduced glutathione (GSH) activities in liver tissue, while UDCA enhanced SOD and GSH activities and decreased MDA accumulation in liver tissue. In contrast, BBP exerted these antioxidant effects in serum. The number and area of glutathione S-transferase placental(GST-P) type-positive lesions and the Ki-67-positive cell ratio were significantly lower in the DEN + BBP and DEN + UDCA groups than in the DEN group, especially in the DEN + UDCA group. UDCA significantly increased Caspase-9 mRNA expression compared with the model group. Conclusions　BBP and UDCA have significant inhibitory effects on preneoplastic lesions of hepatocarcinoma induced by DEN, and both have antioxidant effects on DEN-induced oxidative stress. The antioxidant mechanisms of BBP and UDCA differ, however, and further research is needed to determine the roles of the antioxidant effects in their anticancer mechanisms.

Abstract:The incidence of chronic fatigue syndrome (CFS) has gradually increased in recent years. Although CFS can severely affect patient quality of life, its clinical diagnosis is easily overlooked. Elucidating the pathogenesis of CFS and treating it from both etiological and symptomatological perspectives are crucial for patient recovery; however, its unclear pathogenesis means that etiological treatment options are limited, and current treatments mainly focus on improving clinical symptoms. In this context, there is a need to establish and evaluate a suitable animal model. This article comprehensively integrates the diagnostic criteria of CFS and progress in related basic research. We also summarize the behavioral experimental method used to evaluate CFS models in basic research in the past 5 years and discuss these evaluations from the perspectives of general condition, fatigue state, cognitive function, emotional state, and pain degree. This review aims to present the current situation, expose problems, trigger reflection, and promote improvements in the behavioral evaluation of animal models of CFS.

Abstract:Viral myocarditis (VMC), as the most common type of myocarditis, can progress to chronic myocarditis and even heart failure, ultimately leading to death. Nuclear factor κB (NF-κB) is a multifunctional transcription factor involved in regulating a wide range of biological processes. Existing evidence suggests that the balance between proinflammatory and anti-inflammatory factors plays a decisive role in the prognosis of VMC. The NF-κB pathway mediates inflammatory responses and regulates pathways such as cell apoptosis, energy metabolism, oxidative stress, and insulin resistance to participate in the pathological progression of VMC. This article analyzes and summarizes the molecular mechanism of NF-κB signaling pathway regulation in VMC from the above five aspects, to provide a reference for future basic research and for the clinical diagnosis and treatment of VMC.

Abstract:Mink are small, fur-bearing mammals with significant economic value. They have also recently demonstrated immense potential as novel experimental animal models owing to their unique biological characteristics and similarities with humans in terms of their respiratory systems, immune responses, and other characteristics. This article comprehensively reviews applied research on the use of mink as an experimental animal model, encompassing their use as animal models for influenza virus infection, COVID-19, animal behavior, canine distemper, vomiting, enzyme digestion, testicular degeneration, and self-injurious behavior. The importance of animal welfare is also emphasized, and the broad prospects for the use of mink as an experimental animal model in scientific research are proposed, offering valuable insights and a reference for the extensive application of mink as a novel experimental animal model in the future.

Abstract:Hearing loss is a disease with complex pathogenic factors and unclear pathogenesis, which can seriously affect patient health and quality of life. There is thus a need for an animal model that can simulate human hearing loss, to allow research into the pathogenesis of hearing loss and provide a basis for its treatment. Existing classical modeling method involve drug injection, including intraperitoneal injection of cisplatin, gentamicin, mitochondrial toxin, D-galactose, furosemide and kanamycin, and neomycin, and congenital cytomegalovirus infection. Physical modeling method include noise, cervical spinal injection of sclerosing agent, ischemia reperfusion, and vasopressin injection. Other novel modeling method also exist, such as genetic modification. In this article, we review the advantages and disadvantages of the above modeling method, with the aim of providing a basis for further research on the modeling method of hearing loss.

Abstract:Pancreatic ductal adenocarcinoma (PDAC) is a common type of pancreatic cancer that is insidious, develops rapidly, and is highly malignant. Traditional treatment strategies are ineffective for PDAC because of its rich extracellular matrix (ECM). Cancer-associated fibroblast (CAF) are the most important component of the ECM, and interact with other immune components in the tumor microenvironment (TME) by secreting numerous effector molecules to form an immunosuppressive TME, which may then allow cancer cells to evade immune system surveillance, promote tumor growth, invasion, and metastasis, and induce ECM remodeling and drug resistance. This review summarizes research progress on the application of targeted CAF in PDAC immunotherapy. We focus on exploring research strategies that promote the transition of TME from an immunosuppressive to an immune-activated state through depleting CAF, inhibiting effector molecules secreted by CAF, reprogramming CAF, and limiting CAF-induced ECM remodeling. This review aims to support the production of more effective therapeutic strategies and provide new method for the immunotherapy of PDAC.

Abstract:Dilated cardiomyopathy (DCM) is a common disease leading to heart failure, arrhythmia, and sudden death. The etiology of DCM is complex and diverse, and the mechanisms have not been fully elucidated. Conventional interventions have a limited ability to improve the prognosis of patients, who have a 10-year survival rate of less than 25%. This study aimed to summarize the construction and characteristics of a DCM animal model and evaluate the clinical compatibility of the model with traditional Chinese and Western medicines. Analysis was based on domestic and overseas research into DCM animal models, Western clinical diagnostic criteria, and traditional Chinese medicine syndrome 　　　　　　differentiation. The DCM modeling method mainly involved gene editing, drug induction, immune induction, viral infection, and rapid pacing induction. Experimental animals included muroids, zebrafish, Drosophila, and pigs, of which mice and rats were most commonly used. Gene editing was the most commonly used method for modelling DCM, followed by doxorubicin-induction. In the literature, the experimental animals, drugs, single or cumulative doses, administration method, and modeling period used varied among studies involving DCM animal models. The level of clinical anastomosis according to traditional Chinese and Western medicines varied considerably, being generally lower in traditional Chinese medicine than Western medicine in the same model. In addition, the modeling standards for DCM animal models were mostly based on Western medicine theories. The differentiation of syndrome models and information collection for the four diagnoses have not been standardized and unified. In the future, stable and homogeneous animal models of high clinical consistency combining both disease and syndrome need to be established to provide a basis for DCM mechanism research and drug development.

Abstract:Systemic lupus erythematosus (SLE) models are divided into four major categories: spontaneous, induced, humanized and gene knockout models. Lupus animal models are of great significance for studying the etiology, pathogenesis, and treatment of SLE. By reviewing and comparing various lupus animal models, we aim to elucidate the main features, advantages, and disadvantages of different types of SLE models. This review provides a reference for researchers aiming to select appropriate models for the exploring genetic and environmental factors, pathogenic mechanisms, and therapeutic modalities of SLE.