Abstract: Objective　This study preliminarily investigated the potential mechanisms of the Huoxue Tongluo prescription (HXTLP) in treating spinal cord injury (SCI) through a combination of network pharmacology, molecular docking technology, and in vivo experimental verification. Methods　The traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) were utilized to select the active ingredients, targets of action were obtained from Swiss target prediction database, and an“active ingredients-targets”network was constructed. SCI-related targets were obtained by accessing online mendelian inheritance in man (OMIM) and human gene database (GeneCards), and a protein interaction network of the common targets of HXTLP and SCI was established based on the search tool for the retrieval of interacting genes/protein (STRING) database. The Metascape database was used in KEGG pathway enrichment and GO analyses of the common targets. Molecular docking of active ingredients and key targets was performed through Autodock 1.5.7 software, and the Results were visualized with Pymol 2.4.0 software. Finally, the effect of HXTLP on SCI was verified by animal experiments. Results　A total of 184 intersection targets were obtained, and the key targets were serine/threonine kinase (AKT1), signal transducer and activator of transcription 3(STAT3), heat shock protein 90 kDa alpha,class A member 1 (HSP90AA1), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), harvey ras (HRAS), estrogen receptor 1 (ESR1), mitogen-activated protein kinase 1 (MAPK1), and epidermal growth factor receptor (EGFR). Molecular docking result showed strong binding abilities between the core active components and key targets. In the animal experiments, the behavioral scores of mice in the HXTLP group increased (P<0.05), the motor function of hind limbs was improved, and the histological morphology of the injured area was more complete compared with those of the model group. Western Blot result revealed that HXTLP effectively inhibited the key target protein (HSP90AA1) and the expression of phospho-STAT3 (P-STAT3) and promoted the expression of phospho-phosphatidylinositol-3-kinase (P-PI3K) and phospho-AKT1 (PAKT1). Conclusions　This study verified that HXTLP has multi-component, multi-target, and multi-pathway synergistic effects in the treatment of SCI and has provided experimental and theoretical bases for further clinical medication research for SCI.

Abstract: Objective　The purpose of this study was to provide a more effective method for researching the prevention and treatment of Graves’ disease by comparing the effects of two plasmid vectors expressing the human thyrotropin receptor (TSHR) A subunit gene in inducing an animal model of Graves’ disease via electroporation. Methods Plasmids pcDNA3.1-THSR A, and pTriEx1.1-THSR A expressing the TSHR A subunit were constructed and used to induce Graves’ disease by intramuscular injection with immediate electroporation once every 3 weeks for a total of 4 times. Mice in the control group were injected with PBS. One week after the second electroporation, blood was collected to measure serum thyrotropin receptor antibody (TRAb). Three weeks following the last electroporation, echocardiography was performed on the mice. Mice were sacrificed 4 weeks after the last electroporation; blood, thyroid, and orbital tissues were collected; serum total thyroxine (TT4) was measured; and histological examination was performed. Results　The average concentrations of serum TRAb in the pcDNA3.1-TSHR A group (n= 15) and the pTriEx1.1-TSHR A group (n= 13) were (6.9 ± 2.0) U/L and (7.5 ± 2.2) U/L, respectively. The latter was significantly higher than that in the control group (4.9 ± 0.5) U/L (p= 0.033). The average concentrations of serum TT4 in the pcDNA3.1-TSHR A group and pTriEx1.1-TSHR A group were (41.4 ± 23.8) ng/mL and (63.2 ± 53.7) ng/mL, respectively, both higher than that in the control group: (20.2 ± 4.0) ng/mL (P<0.01). Thyroid pathology showed thyroid follicular epithelial hyperplasia with T-cell infiltration in the model group. Echocardiography showed that the left ventricle mass in the pTriEx1.1-TSHR A group was higher than those in the control group (p= 0.007) and pcDNA3.1-TSHR A group (p= 0.012). Orbital pathology showed fibrotic changes in the extraocular muscles of mice in the model groups. Conclusions　Both pcDNA3.1 and pTriEx1.1 expressing the TSHR A subunit were able to induce Graves’ disease in mice by electroporation, and the efficiency of the two plasmids in inducing hyperthyroidism and Graves’ ophthalmopathy was similar. The efficiency of pTriEx1.1-TSHR A in inducing thyrotoxic heart disease was better than that of pcDNA3.1-TSHR A.

Abstract: Objective　Based on apolipoprotein a-I (Apoa-I) gene knockout mice, the role and mechanism of phosphotidylcholine (PC) in improving cholesterol reverse transport were explored. Methods　Thirty Apoa-I^-/- mice were randomly divided into an Apoa-I^-/- group, Apoa-I^-/- + HFD group, and Apoa-I^-/- + HFD + PC group using the random number table method ; 30 C57BL/6J mice were randomly divided into a WT group, WT + HFD group, and WT + HFD + PC control groups, with 10 mice in each group. The Apoa-I^-/- group and WT groups were fed basic feed, while the other groups were fed high-fat feed for 8 weeks to establish a hyperlipidemia model. From the 9th week, the WT + HFD + PC group and Apoa-I^-/- + HFD + PC group were given PC 2.5 g/(kg·d), while the remaining mice were given physiological saline by gavage for a total of 4 weeks of intervention. The serum lipid levels of the mice were detected using a fully automated analyzer. Hematoxylin and eosin and Oil red O staining were used to observe pathological and morphological changes, and the COD-PAP method was used to detect cholesterol levels in mouse liver tissue. The ELISA method was used to detect LCTA levels in mouse serum, and RT-qPCR and Western Blot method were used to detect the mRNA and protein expression of cholesterol ATP binding cassette transporter A1 (ABCA1), ATP binding cassette transporter G1 (ABCA1), lecithin cholesterol acyltransferase (LCAT), hepatic lipase (HL), scavenger receptor class B type I (SR-B1), and low-density lipoprotein receptor (LDL-R) in liver tissue. Results　Compared with the WT group, the serum lipid levelsof WT + HFD group mice were significantly increased (P<0.01), LCAT levels were significantly reduced (P<0.05), hepatic fat vacuoles were obvious, hepatic lipid deposition was significant, and liver tissue TC levels were significantly increased (P<0.01). The mRNA and protein expression of ABCA1, ABCG1, LCAT, SR-B1, HL, and LDL-R were significantly reduced (P<0.05, P<0.01). Compared with the WT + HFD group, serum lipid levels in the WT + HFD + PC group were significantly reduced (P<0.05, P<0.01), LCAT levels were significantly increased (P<0.05), hepatic fat vacuoles were significantly reduced, hepatic lipid deposition was alleviated, and liver tissue TC levels were significantly reduced (P<0.05); mRNA and protein expression of ABCA1, LCAT, SR-B1, HL and LDL-R were significantly increased (P<0.05, P<0.01). The serum levels of TC, TG, and LDL-C were significantly increased, while the levels of LCAT、HDL-C were significantly reduced (P<0.05, P<0.01) in the Apoa-I^-/- + HFD group mice. Hepatocytes underwent balloon-like transformation, liver lipid deposition was significantly aggravated, and liver tissue TC levels were significantly increased (P<0.05). The mRNA and protein expression of ABCA1, LCAT and HL were significantly reduced (P<0.05, P<0.01). Compared with the WT + HFD + PC group mice, the Apoa-I^-/- + HFD + PC group mice showed a significant increase in serum lipid levels (P<0.05, P<0.01), LCAT levels were significantly reduced (P<0.05), significant hepatic lipid vacuoles, significant hepatic lipid deposition, and a significant increase in TC levels in liver tissue (P<0.05). Their mRNA and protein expression of ABCA1, ABCG1, LCAT, SR-B1, and HL were also significantly reduced (P<0.05, P<0.01). Conclusions　Phosphatidylcholine can improve dyslipidemia by interfering with Apoa-I and thus regulating cholesterol reverse transport.

Abstract: Objective　To compare the serological and pathological characteristics of 3 nonalcoholic steatohepatitis (NASH) models: high-fat diet (HFD) with carbon tetrachloride (CCl₄ ) injection, methionine and choline deficient diet (MCD), and Aymlin liver NASH (AMLN) diet-induced NASH models. Methods　3 NASH models were established by feeding mice an HFD with CCl₄ injection for 10 weeks, MCD for 8 weeks and NASH for 26 weeks. After feeding, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), glucose (GLU), liver triglyceride (TG), total cholesterol (TC), and malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity were measured. Insulin levels were measured by enzyme-linked immunosorbent assay (ELISA) and the homeostasis model assessment of insulin resistant (HOMA-IR) index was calculated. Hematoxylineosin (HE), Sirius red, and oil red staining were used to indicate pathological changes to the liver. The NAS score was used to grade the pathology. Results　Compared to each normal control (NC) group mice, all mice in the 3 model groups had an obvious increase in serum transaminase and liver TG, TC, MDA levels and SOD activity. The levels of serum FINS, GLU and the HOMA-IR index were significantly increased in the AMLN and CCl₄ + HFD model groups but decreased in the MCD model group. According to the HE, oil red staining and NAS score, mice in all 3 groups had NASH phenotypic changes. Liver collagen deposition was most obvious in mice in theCCl₄ + HFD model group. Liver lipid droplets were most abundant in the AMLN model group. Conclusions　All the above 3 animal models can stably simulate the serological and pathological changes of NASH in human. The AMLN model can simulate the progress and mechanism of the disease, as well as systemic metabolic disorders such as insulin resistance and oxidative stress. However, it is time-consuming and the fibrosis progression rate is slow. The MCD diet can simulate the serological and pathological features of NASH in 8 weeks, but no obesity or insulin resistance occurred. The CCl₄ combined with HFD model can induce NASH model in 10 weeks, which can simulate its serological and pathological changes, and the liver has obvious fibrous deposition and oxidative stress damage.

Abstract: Objective　To investigate the characteristics of Chinese medicine syndromes and the possible metabolic substance basis of spontaneously hypertensive rat (SHR). Methods　10-week-old SPF SHR and WKY of the same strain were divided into SHR group and WKY group with 8 rats in each group. The general state, temperament, peripheral vascular filling, tongue characteristics, diet, water intake, urine and feces volume and characteristics, blood pressure, heart rate, respiratory rate, pain threshold, and open field behavior of SHR rats were observed and tested comprehensively to identify the possible syndrome types of Chinese medicine. At the same time, liquid chromatography tandem mass spectrometry was used to analyze non-targeted serum metabolites to preliminarily reveal the material basis of blood pressure elevation and Chinese medicine syndrome manifestations. Results Compared with WKY group, the scores of dark yellow hair color, irritable degree and peripheral capillary filling were higher in SHR group (P<0.0001). Red tongue color, dry tongue, little body fluid; 24h diet and water intake, urine volume and fecal volume were less (P<0.05), fecal water content was lower (P<0.001); systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), heart rate (HR) and respiratory rate (RR) were significantly higher (P<0.05); Lower pain threshold (P<0.0001); The open field experiment shows that the moving distance and residence time of the edge are longer (P<0.001). Serum non-targeted metabolomics result showed that, compared with WKY group, the SHR group had 114 metabolites with significant differences (P<0.05). These differential metabolites were mainly lipids and lipid-like molecules (40.35%), organic acids and derivatives (22.8%), and organoheterocyclic compounds (15.79%). A total of 25 metabolic pathways were identified by KEGG enrichment analysis. Further differential abundance analysis showed that 16 pathways were activated, only 4 pathways were inhibited, and 5 pathways were not significantly changed. The glutamatergic synapse and GABAergic synapse were activated, while the serotonergic synapse was inhibited. Conclusions　The symptoms of SHR include impatience and irritability, peripheral vascular dilation and collateral circulation formation, bulbar conjunctival congestive swelling, red tongue coloration, a dry tongue, constipation, redyellow urine of low volume, and a rapid heart rate and high respiratory rate. All these suggest that SHR is a syndrome of hypertension with hyperactivity of liver-yang. The material basis of SHR is not only related to lipid, amino acid, and carbohydrate metabolism disorders, but also may be related to metabolic disorders of glutaminergic, GABAergic, and serotonergic neural pathways.

Abstract: Objective　In this study, we aimed to predict the inhibitory mechanism of Shenlingcao oral liquid (SLC) in non-small cell lung cancer (NSCLC) by network pharmacology and verify it by molecular docking and in vivo experiments. Methods　The active ingredients and corresponding targets of SLC and NSCLC were obtained by database and literature search. Targets of SLC common to NSCLC were selected to construct the protein interaction network, and GO and KEGG enrichment analysis and molecular docking were performed. A Lewis lung cancer mouse model was constructed and divided into Model group, SH group, and SL group. The latter two groups were intragastrically administered 8.75 g SLC lyophilized powder/kg and 3.50 g SLC lyophilized powder/kg, respectively. After 14 days of drug intervention, tumor growth, pathological changes in tumor tissue, and apoptosis in tumor tissue were observed in tumor-bearing mice; changes in blood routine indexes and the tumor tissue expression of p-AKT, AKT, p-PI3K, PI3K, and Bcl-2 protein of mice were detected. The result of the KEGG enrichment analysis were verified. Results　Network pharmacological analysis showed that there were 77 active ingredients, 618 potential targets, 1498 potential targets for NSCLC, and 179 drug and disease intersection targets. Target intersection enrichment analysis showed that they were mainly concentrated in the phosphatidylinositol 3 kinase-protein kinase B (PI3K-AKT) signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, and other related pathways. Molecular docking showed that the top 10 core components had good bonding ability with the top 10 core targets. In the animal experiments, compared with the Model group, SH group and SL group had significantly decreased tumor volume and weight (P<0.05, P<0.01), and significantly decreased white blood cell, neutrophil, and monocyte numbers (P<0.01, P<0.001). Red blood cells, platelets, and hemoglobin were significantly increased (P<0.05, P<0.01, P<0.001); apoptotic cells were significantly increased in early tumor tissue (P<0.05, P<0.01), and the protein expression levels of p-PI3K/PI3K, p-AKT/AKT, and Bcl-2/GAPDH were significantly decreased (P<0.05, P<0.01). The expression levels of PI3K, AKT1, and Bcl-2 genes were significantly decreased (P<0.05, P<0.01). Conclusions　The mechanisms of SLC activity against NSCLC may be related to the activation of the PI3K-AKT pathway and the promotion of apoptosis.

Abstract: Objective　To analyze differences in the intestinal microbiota and transcriptomics between Nmethyl-N’-nitro-N-nitrosoguanidine (MNNG) gastric cancer rats and normal rats and to analyze the correlation between the two, so as to provide a reference for related studies using MNNG gastric cancer rats as a model. Methods A total of 12 Wistar rats were randomly divided into normal (NM) and gastric cancer (GC) groups. The GC group was given a concentration of 20 mg/mL of MNNG by gavage with a dose of 100 g/mL once a day, and the NM group was given the same amount of normal saline by gavage. Samples were collected for testing after 16 weeks of continuous intervention. The gastric tissues were collected and stained by HE staining to observe morphological changes in the gastric mucosa of the two groups, and the expression levels of differential genes were detected by transcriptome sequencing. The cecal contents were collected for 16S rRNA sequencing. Results　(1) Visual observation and HE Results showed that the volume of gastric mucosa in the NM group was normal, the surface was glossy, the gastric wall was elastic, the direction of the mucosal folds was regular, there were no hyperplasia or hemorrhagic spots. In the GC group, the volume of gastric mucosa was reduced, the gastric wall was thinned, elasticity was poor, the direction of the folds was disordered and irregular, and there was a bulge accompanied by yellow-black keratotic hyperplasia. In the NM group, the squamous epithelial layer, submucosa, and muscular layer of the gastric mucosa were clear, with no hyperplasia and keratinization. In the GC group, the gastric mucosa had disorganized layers and cell polarity, with different cell morphologies; the squamous epithelial layer was destroyed, and squamous epithelial cells were hyperplasic, keratinized, and had invaded the muscular layer by proliferation. The modeling was considered successful. (2) The Results of intestinal microbiota sequencing showed that the abundance of Akkermansia and Lactobacillus in MNNG gastric cancer rats decreased significantly, and the abundance of the rumen coccaceae Prevonella, and Blauter increased significantly. (3) The three key pathways obtained by transcriptomic sequencing and KEGG pathway enrichment analysis were amebiasis, systemic lupus erythematosus, and the PI3K-Akt signaling pathway, and five genes differentially enriched in these three pathways were those for MCPT8I2, IGH-6, IGHG1, ACTN2, and VEGF-D. (4) Combined analysis of intestinal microbiota and transcriptomics showed that _UCG-005, Prevonella _UCG-003 and Brautella were positively correlated with amebiasis, systemic lupus erythematosus, and the PI3K-Akt signaling pathway. Conclusions　The abundance of intestinal microbiota in gastric cancer rats formed by MNNG gavage is different from that of normal rats. The genes for MCPT8I2, IGH-6, IGHG1, ACTN2 and VEGF-D may be up-regulated in gastric cancer induced by MNNG gavage. Combined analysis of intestinal microbiota and differential genes suggested that the mechanism of MNNG carcinogenesis may be mainly related to the destruction of gastric mucosa and the inflammatory response.

Abstract: Objective　To establish a rat model of type 2 diabetes mellitus with hepatic fibrosis using a highlipid-sugar diet (HLSD) combined with inducement with carbon tetrachloride (CCl₄ ) and streptozotocin. Methods SD rats were randomly divided into a liver fibrosis (LF) group, type 2 diabetes combined with liver fibrosis (DLF) group, and normal group, with 10 rats in each group. All rats in the LF or DLE group were fed an HLSD for 4 weeks; then, all rats were intraperitoneally injected with 20% CCl₄ (once a week) according to body weight (5 mL/kg) to induce hepatic fibrosis for another 4 weeks. During CCl₄ treatment, rats in the DLF

Abstract: Objective　To investigate the changes of inflammatory factors in bronchoalveolar lavage fluid (BALF), lung tissue, and serum of a rat pneumonia model induced by inhalation of lipopolysaccharides (LPS). Methods　Three days after modeling by LPS 4 mg/mL inhalation, 15 min/d, was conducted while monitoring the particle size distribution and aerosol concentration of LPS, the degree of inflammation in lung tissues of rats in each group was observed via HE staining, and neutrophils in BALF were counted by microscope. The contents of interferon gamma (IFN-γ), interleukin-1 beta (IL-1β), IL-4, IL-5, IL-6, IL-10, IL-13, tumor necrosis factor alpha (TNFα), and KC/GRO in lung tissue, serum, and BALF were detected by Meso Scale Discovery. Results　The lung histopathology of model rats displayed focal and diffuse alveolar epithelial necrosis with shedding and the aggregation and infiltration of inflammatory cells. The particle size distribution of atomized LPS was as follows, Dv(10) = 0.6974 μm, Dv(50) = 3.387 μm, Dv(90) = 8.836 μm. The aerosol concentration of LPS was 4.08 g/m³, and the calculated inhalation dose for rats was 47.10 mg/kg. The neutrophil count (P<0.01) and contents of IL-1β, IL-6, and TNF-α (P<0.05, P<0.001, P<0.001) in the BALF, and the contents of IL-1β, IL-6, and KC/GRO in lung tissue (P<0.01, P<0.05, P<0.01), of model rats were significantly increased. No biologically significant changes were observed in inflammatory factor levels in the serum. Conclusions　In the acute pneumonia model induced by inhalation of LPS, significant changes in inflammatory factors such as IL-1β, IL-6, KC/GRO, and TNFα were observed in both lung tissue and bronchoalveolar lavage fluid (BALF), while no notable changes in these inflammatory factors were detected in serum. This indicates that the inflammation responses are primarily localized in the lungs.

Abstract:Precision therapy has become an important approach in modern medicine, with the goal of providing individualized treatment according to the characteristics of individual patients. The successful development of precision medicine depends on the application of preclinical cancer models. Patient-derived organoid (PDO) xenograft models display characteristics of both PDO models and in vivo patient-derived tumor xenograft models. This type of model can not only maintain the heterogeneity of the original tumor, but also has additional advantages, such as large-scale cultivation, high-throughput drug screening in vitro and drug sensitivity testing in vivo. It is an innovative, precise preclinical disease model. In this review, we summarize the basic characteristics of the PDO xenograft model, analyze its construction method and influencing factors, further discuss its application in precision therapy, with the aim of providing a reliable preclinical experimental tool for individualized cancer treatment.

Abstract:Cardiovascular diseases that develop from hypercholesterolemia-induced atherosclerosis have emerged as a significant threat to human health. Recently, probiotics exhibiting cholesterol-lowering properties have emerged as a prominent area of research. Numerous studies have demonstrated that Lactobacillus reuteri can effectively reduce endogenous cholesterol synthesis, regulate cholesterol transport, and promote cholesterol degradation by modulating the expression of key genes, such as sterol-regulatory element binding protein 2, 3-hydroxy-3-methylglutaryl coenzyme A reductase, and cholesterol 7 alpha-hydroxylase, in both the liver and intestinal epithelial cells of the host. This leads to a notable decrease in total cholesterol and low-density lipoprotein cholesterol levels in the host serum. The present paper offers a comprehensive overview of the underlying mechanisms responsible for the cholesterol-lowering effects exerted by L.reuteri, aiming to provide valuable insights into the treatment of hypercholesterolemia and the development of probiotics with cholesterol-lowering properties.

Abstract:Sarcopenia is an age-related skeletal muscle degenerative disease. Physiologically aging mice are the most commonly used animal model for studying sarcopenia. As sarcopenia is characterized by decreased skeletal muscle mass and reduced muscle strength, exercise performance as a reflection of muscle function is widely used to evaluate sarcopenia.Methods of evaluating the motor function of sarcopenia mouse models are generally designed based on muscle endurance, muscle strength, coordination, and balance. The method include tests such as treadmill exhaustion, voluntary wheel use, grip strength, horse grid, bars and balance beam tests. By collating recent publications, we have systematically summarized the method used for evaluating motor function, including the tests’ principles, procedures, evaluation indexes, advantages, and disadvantages. We then propose an operational program for evaluating the sarcopenia phenotype, which will be of help to researchers wishing to choose evaluation method appropriate to their specific research purposes. Further innovative technology for assessing motor function that could be instructive in the evaluation of skeletal muscle function and diagnosis of sarcopenia is summarized.

Abstract:Rheumatoid arthritis (RA) is a chronic autoimmune disease that significantly impacts joints. Experimental models are crucial tools for studying the pathogenesis and pharmaco-toxicological mechanisms of RA and screening RA drugs. Commonly used experimental RA models include animal models and in vitro models. With advancements in biotechnology and biomaterials, experimental RA models have evolved from induced and 2D in vitro models to spontaneous gene modification and 3D in vitro models. Moreover, some progress has been made in the study of traditional Chinese medicine disease and syndrome combination models. This paper summarizes the research progress made in the modeling, monitoring, and evaluation of experimental RA models to provide a reference for related research.

Abstract:As a model organism that bridges the gap between cells and traditional mammals, the zebrafish has broad application prospects in radiation medicine research. Among its unique advantages are its characteristics of a high homology with human genes, high fertility, short embryonic period, and transparent and easy to observe embryos, making it an important tool in radiation medicine research. Recently, remarkable progress has been made in the application of zebrafish to investigate low-dose radiation biological effects, radiation therapy, and radiation damage prevention and treatment, key areas of radiation medicine. In this paper, these applications are reviewed; we explore the value of zebrafish in radiation medicine research and provide a reference for experimental research in related fields.

Abstract:As China has become the second largest market for medical devices in the world, the domestic medical device industry has been growing. As an important part of preclinical evaluation of medical devices, large animal research directly affects the research and application of medical devices. Large animals are widely used in the evaluation of safety and feasibility of medical devices because they are closer to humans in terms of body size, anatomical structure and physiological functions. In large animal experimental research, the selection of suitable experimental animals and the establishment of suitable animal disease models are the basis for ensuring the smooth progress of experiments. In this paper, the selection of experimental animals and the establishment of disease models in medical device large animal experimental research are systematically sorted out, and the existing problems and deficiencies are pointed out.