Abstract: Objective To investigate the protective effects and potential mechanisms of tetrahydrocurcumin (THC) on thoracic aortic aneurysm and dissection (TAAD) in mice. Methods TAAD was induced in 3-week-old C57BL / 6J mice by oral administration of β-aminopropionitrile (BAPN) (diluted in drinking water, 1 g / (kg·d)).Eighty mice were divided randomly into Con, BAPN, BAPN + THC, and BAPN + THC + 3-TYP (SIRT3 inhibitor) groups (n = 20 mice per group). The survival rate of mice in each group was recorded after 4 weeks. The maximum diameter of the aorta was measured and the histomorphology and aortic wall elastin integrity were evaluated by hematoxylin and eosin and elastin van Gieson staining. Macrophage infiltration was detected by immunohistochemical staining and α-smooth muscle actin ( α-SMA ) and osteopontin ( OPN ) expression were detected by immunofluorescence staining. The production of reactive oxygen species (ROS) was measured by dihydroethidium staining and superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels were determined using kits.Protein expression levels of matrix metalloproteinase (MMP) 2, MMP9, interleukin ( IL)-6, tumor necrosis factor (TNF)-α, nuclear factor erythroid 2-related factor 2 (NRF2), NADPH oxidase 2 (NOX2), α-SMA, OPN, sirtuin 3 ( SIRT3), Ac-SOD2, and SOD2 were measured by Western Blot. Results Mice in the BAPN + THC group showed significantly higher survival and a lower incidence of TAAD compared with the BAPN group and the degree of aortic dilatation and morphology and structure were improved (P<0. 05). Infiltration of CD68-positive macrophages and MMP2, MMP9, IL-6, and TNF-α expression levels were lower (P<0. 05), ROS generation, MDA content,and NOX2 expression in aortic tissue were also significantly decreased, while SOD activity and NRF2 expression were increased (P<0. 05). α-SMA expression was also increased, while OPN expression was reduced ( P<0. 05).SIRT3 expression was increased while the Ac-SOD2 / SOD2 ratio was decreased ( P<0. 01). Treatment with the SIRT3-specific inhibitor and silencing of SIRT3 counteracted the ability of THC to resist TAAD via the SIRT3 signaling pathway ( all P<0. 05). Conclusions THC alleviated inflammation and oxidative stress in aortic tissues by activating the SIRT3 signaling pathway, thus inhibiting the phenotypic transformation of vascular smooth muscle cells and resisting the formation of TAAD in mice.

Abstract: Objective To explore the effect of evening primrose oil (EPO) on aortic endothelial damage in rats with polycystic ovary syndrome (PCOS), using network pharmacology and in vivo experiments. Methods The potential targets of EPO for improving aortic endothelial injury in PCOS rats were predicted by network pharmacology,and the selected core targets and renin-angiotensin signaling (RAS) pathway were verified by experiments. Fifty-eight female SD rats were divided randomly into a blank group (n= 10) and a modeling group (n = 48). Rats in the blank group were fed a normal diet and rats in the modeling group received a high-fat diet for 8 weeks. The PCOS model was prepared at week 6 by administration of letrozole (1 mg / (kg·d)) for 21 days. Blood was taken from the tail vein after modeling and serum was collected to detect hormone levels. The model rats were then divided randomly into four groups and treated with the corresponding drugs for 6 weeks. Blood, blood vessels, and ovaries were then collected. Tissue morphology was examined by hematoxylin and eosin staining and serum levels of luteinizing hormone (LH), testosterone (T), follicle-stimulating hormone (FSH), endothelin (ET-1), and tumor necrosis factor (TNF-α) were detected by enzyme-linked immunosorbent assay ( ELISA). Serum levels of nitric oxide ( NO) were determined by spectrophotometry. Protein expression levels of core targets and RAS pathway-related factors were assessed by western blotting and immunohistochemistry. Results Twenty-five intersection targets of EPO and PCOS were identified by network pharmacological analysis. Kyoto encyclopedia of genes and genomes analysis showed that EPO improved vascular injury in PCOS rats via multiple pathways, including RAS. Serum levels of ET-1, FSH, LH,and T measured by ELISA were significantly decreased after EPO treatment, compared with the model group (P<0. 01). EPO significantly decreased the expression levels of AngⅠ, VEGF-B, AT₂R, ET-1, and TNF-α proteins in the aorta (P<0. 01) and significantly increased expression levels of Ang Ⅱ, CD31, and endothelial NO synthase proteins ( P<0. 01). Conclusions EPO may ameliorate vascular endothelial injury in PCOS model rats by inhibiting the RAS signaling pathway and by overactivation of the ACE/ Ang Ⅱ/ AT1 axis.

Abstract: Objective Exploring the behavioral changes induced by quinpirole in obsessive-compulsive disorder (OCD) mouse, investigating the activation of neurons in different brain regions, and identifying differentially expressed genes (DEGs) and enriched biological pathways through transcriptome sequencing technology to elucidate the pathogenesis of OCD. Methods Randomly assign 32 male C57BL / 6J mice, aged two months, to an OCD group and a control group (n= 16). Administering quinpirole (0. 75 mg / kg) via subcutaneous injection to the OCD group mice every other day for a total of 19 injections, while the control group mice received an equivalent volume of saline solution. Following the completion of the model construction, open field testing, elevated plus maze testing, and marble burying tests were conducted. After the completion of behavioral studies, tissue samples were collected. Neuronal damage was assessed using Nissl staining, while the expression of c-Fos and Iba1 proteins was examined through immunofluorescence staining. Transcriptome sequencing technology was utilized to screen for differentially expressed genes and to enrich relevant signaling pathways. The expression of inflammatory cytokines, including TNF-α, NF-κB p65, phosphorylated NF-κB p65 ( p-NF-κB p65), and IL-6, was detected using Western Blot analysis. Results Mouse induced with OCD by quinpirole exhibit anxiety-like behaviors and compulsive-like behaviors. Neurons in the hippocampal and hypothalamic regions exhibit signs of damage. The expression of c-Fos and Iba1 proteins is increased in the cortex, striatum, hypothalamus, and other brain regions. Western Blot result indicate a significant increase in the expression of pro-inflammatory cytokines such as TNF-α, p-NF-κB p65, and IL-6. Conclusions In OCD mouse, neurons in multiple brain regions are abnormally activated, microglia exhibit dysfunction, and neuroinflammation induced by the activation of the NF-κB signaling pathway accompanies the development of OCD.

Abstract: Objective To investigate the effects of aerobic exercise and different exercise habits on skeletal muscle function and the possible molecular mechanisms in colorectal cancer-loaded mice. Methods Thirty-five 5-week-old BABL / c male mice were acclimatized to feeding for 1 week and then divided randomly into the following groups: control (D), tumor (M), exercise preconditioning (QAM), lifetime exercise (AM), and exercise (HAM) groups (n = 7 mice per group). Mice in the QAM and AM groups underwent aerobic exercise regimen 1 from weeks2 ~ 6. At week 7, mice in the experimental groups received 0. 2 mL of CT26 colon cancer cell suspension subcutaneously in the dorsal aspect of the left hind limb, while control mice received 0. 2 mL of saline at the corresponding site. Mice in the AM and HAM groups were subjected to aerobic exercise regimen 2 for weeks 7 ~ 9.The general status and skeletal muscle mass and function were monitored in all mice throughout the experiments. After completion of the experiment, samples were collected and the cross-sectional area of gastrocnemius muscle fibers (CSA) was measured by hematoxylin and eosin staining and expression levels of proteins related to synthesis and catabolism of the gastrocnemius muscle were analyzed by Western Blot. Results ( 1) The weight ratio of the gastrocnemius muscle was significantly lower in mice in the M, QAM, and HAM groups compared with group D, and was significantly higher in AM mice compared with M, QAM, and HAM mice. (2)Grip strength, endurance, skeletal muscle circumference, and CSA were significantly lower in group D mice compared with the other groups, and was most enhanced in group HAM. Endurance and CSA were consistently enhanced in groups QAM, AM, and HAM. (3)Muscle RING-finger protein-1 (MuRF1) expression levels were significantly lower in groups M, QAM, AM, and HAM than in group D, significantly lower in groups AM and HAM than in group M, and significantly lower in group HAM than in group QAM. (4) Fibronectin type Ⅲ domain-containing protein 5 ( FNDC5) expression levels were significantly lower in group M than in groups D and QAM. ( 5) Peroxisome proliferator-activated receptor gamma coactivator 1-alpha expression levels were significantly lower in the QAM and HAM groups compared with group M.(6)Expression levels of phospho (p)-AMP-activated protein kinase (AMPK) / AMPK and p-AMPK were significantly higher in group QAM than in groups D and M, p-AMPK expression significantly lower in groups AM and HAM was than in group QAM, and AMPK expression was significantly lower in groups QAM, AM, and HAM than in group D. Conclusions Exercise preconditioning and continuous aerobic exercise can improve skeletal muscle mass and function in CT26 colon cancer-loaded mice by activating AMPK phosphorylation to stimulate skeletal muscle secretion of FNDC5, thereby regulating the expression of MuRF1 protein.

Abstract: Objective To determine the expression of type Ⅲ interferon ( IFNL) and IFNL receptor 1 (IFNLR1) in the genital tract in rhesus macaques to elucidate the biological characteristics of rhesus macaque IFNL system and promote the use of these monkeys in studies on the pathology, drugs and vaccines for human diseases. Methods IFNL1, IFNL3, and IFNLR1 mRNA levels in normal and simian-human immunodeficiency virus (SHIV) / simian immunodeficiency virus ( SIV)-infected genital tract tissues of rhesus macaques were detected by gene-specific reverse transcription-polymerase chain reaction, and IFNLR1 immunoreactivity was examined by confocal microscopy. Results IFNL1 and IFNL3 mRNA levels in the uterus and vagina in normal rhesus macaques were below the measurement threshold, but mRNA levels of IFNLR1 and its variant lacking exon 6 could be readily detected. IFNLR1 immunoreactivity was primarily detected in the covering or glandular epithelia of the uterus and vagina. IFNL1 and IFNL3 mRNA levels were increased in some SHIV/ SIV-infected macaques, while mRNA levels of IFNLR1 were unchanged and mRNA levels of IFN-stimulated genes ( Mx1, Mx2, and OAS) were significantly increased. Conclusions These data indicate that the type Ⅲ IFN system is expressed in the reproductive tract of rhesus macaques, as in humans, thus providing a scientific basis for the use of rhesus macaques in studies of human infectious diseases, such as AIDS.

Abstract: Objective To investigate the mechanism of action of the Astragalus-Chinese yam combination in treating cancer-related fatigue (CRF) in mice. Methods The active components and related targets of AstragalusChinese yam were obtained from the traditional Chinese medicine systems pharmacology database and analysis platform. CRF-associated targets were identified using the GeneCards database. Intersecting targets were analyzed using the DAVID database for gene ontology and kyoto encyclopedia of genes and genomes enrichment analyses. A network diagram depicting “ drug-active component-intersecting targets-disease” was constructed using Cytoscape software, and a protein-protein interaction network was created to identify the top five core target proteins based on degree values. Molecular docking simulations were performed using Autodock Vina software. Twenty-five mice were divided randomly into a blank group and a modeling group in a 1 ∶ 4 ratio. After successfully establishing the CRF model using Lewis lung cancer cells, mice in the modeling group were further divided into model, Chinese yam (0. 2g / kg), Astragalus (0. 6 g / kg), and Astragalus-Chinese yam combination groups (0. 3 + 0. 1 g / kg) (n= 5 mice per group). The treatments were administered by gavage twice daily for 14 consecutive days. Grip-strength and forcedswimming tests were conducted. The mice were then euthanized and tissues were collected. The gastrocnemius muscles were weighed and stained with hematoxylin and eosin to reveal the muscle fiber morphology. Results A total of 23 effective active components of Astragalus-Chinese yam were identified through network pharmacology analysis,with 199 intersecting drug-disease targets. These targets mainly participated in biological processes such as protein phosphorylation through cellular components ( cytoplasm, membrane, nucleus) and performed molecular functions such as protein binding. A total of 155 signaling pathways, including pathway in cancer and the phosphatidylinositol 3-kinase-protein kinase B signaling pathway, were involved in CRF. The critical targets of Astragalus-Chinese yam for CRF included serine / threonine kinase, tumor necrosis factor, epidermal growth factor receptor, B-cell lymphoma 2,and caspase 3. The active components quercetin and diosgenin interacted with the highest number of targets and demonstrated binding energies < - 5. 0 kJ/ mol with the five core targets, indicating strong ligand-receptor binding affinity. Mice in the Chinese yam and Astragalus groups exhibited increased grip strength and prolonged swimming times compared with the model group. Gastrocnemius muscle volume and mass were increased, with well-organized muscle fibers and clear boundaries, and the effects were even more pronounced in the Astragalus-Chinese yam combination group. Conclusions Astragalus-Chinese yam treats CRF via a multi-target, multi-pathway approach,enhancing muscle strength and endurance in mice, improving gastrocnemius muscle volume and mass, and alleviating muscle atrophy, thereby mitigating the associated symptoms of CRF in mice.

Abstract: Objective To construct an animal model of post-stroke depression (PSD) based on the theory of “depression, stasis, phlegm”, with the aim of developing and validating an Objective assessment system. Methods Rats were divided randomly into five groups: control, depression, stroke, PSD, and Baishile decoction groups. A PSD syndrome-based animal model was established in rats using a combination of middle cerebral artery occlusion (MCAO) and chronic unpredictable mild stress (CUMS). “Depression, stasis, phlegm” were then evaluated in the model rats using the Morris water maze, open field, forced swimming, and sucrose preference tests, and by detection of neurotransmitter levels, brain tissue pathology, tongue and forepaw color RGB values, and blood rheology. Results PSD rats exhibited significantly shorter target quadrant dwelling times, platform crossings, and climbing and rearing frequencies, a significantly lower sucrose preference, and a significantly higher immobility time in the forced swim test compared with control rats. Hematoxylin and eosin and Nissl staining revealed brain tissue damage in PSD rats. Serum and cerebrospinal fluid levels of 5-hydroxytryptamine ( 5-HT) were significantly decreased, glutamate levels were significantly increased, and tongue and forepaw RGB values were all decreased. Blood rheology showed a hypercoagulable state and blood lipid metabolism-related indicators were significantly abnormal. Rats in the Baishile decoction group showed significant improvements compared with the PSD group, including increased target quadrant dwelling times, number of platform crossings, and climbing and rearing frequencies, increased sucrose preference,decreased immobility time in the forced swim test, improved brain tissue pathology, increased serum and cerebrospinal fluid levels of 5-HT, decreased glutamate levels, increased tongue and claw RGB values, and varying degrees of improvement in blood rheology and blood lipid metabolism-related indicators. Conclusions The combination of MCAO and CUMS successfully established a syndrome-based animal model of PSD exhibiting the characteristics of “depression, stasis, phlegm”, with corresponding Objective assessment criteria.

Abstract: Objective To explore the mechanism of NLRP3 gene knockout in relation to the abnormal mucosal barrier and inflammatory factors in ulcerative colitis (UC) mice. Methods Thirty-two NLRP3-knockout (NLRP3^{- / -}) mice and 30 C57BL / 6 wild-type (WT) mice were divided randomly into six groups: NLRP3^{- / -}blank,NLRP3^{- / -} model, NLRP3^{- / -}mesalazine, WT blank, WT model, and WT mesalazine groups. Except for mice in the two blank groups, mice in the other groups were given 3% dextran sodium sulfate to drink freely for 5 days to establish an UC mouse model. After successful establishment of the model, mice in each group underwent intragastric administration of the respective solution for 7 consecutive days. The general condition, body weight, disease activity index (DAI) score, and colon length were observed and evaluated in each group. Histopathological changes in the colon were observed by hematoxylin and eosin staining. ZO-1, claudin-1, occludin, tumor necrosis factor (TNF)-α and interleukin ( IL)-6 expression in colon tissue were detected by immunohistochemistry. Results (1) The DAI score was significantly higher in the NLRP3^{- / -} model group compared with the WT model group on day 12, while colon length was significantly shorter and pathological injury of the intestinal mucosa was more serious. Expression levels of ZO-1, claudin-1, and occludin in colon tissue were lower whereas expression levels of TNF-α and IL-6 were significantly higher in the NLRP3^{- / -} model group compared with the WT model group. ( 2) Regarding the two mesalazine groups, the DAI score was significantly higher and expression levels of ZO-1, claudin-1, and occludin in colon tissue were lower in the NLRP3^{- / -}mesalazine compared with the WT mesalazine group on day 12. Conclusions Specific knockout of the NLRP3 gene makes mice more sensitive to UC. Compared with WT mice, NLRP3^{- / -}UC micehave more severe mucosal barrier injury and release more inflammatory factors. Mesalazine could repair the mucosal barrier and reduce inflammation in NLRP3- / -and WT UC mice. Under the same experimental conditions, mesalazinerepaired the mucosal barrier more effectively in WT compared with NLRP3^{- / -}UC mice.

Abstract: Objective To construct a uricase-deficient mouse model with stable inheritance using the CRISPR/ Cas9 system, and evaluate its ability to simulate the disease characteristics of patients with hyperuricemia. Methods Double single guide RNAs (sgRNAs) were designed on both sides of exon 2 ~ 4 of the Uox gene. SgRNA and Cas9 mRNA for gene knockout were microinjected into the fertilized eggs of mice. After culture for 2 ~ 4 h, the embryos were transferred to surrogate mother mice to produce an F0 generation. Uox-knockout mice were identified by polymerase chain reaction and sequencing analysis. Positive mice were then mated with wild-type ( WT) mice to produce an F1 generation, and heterozygous female and male F1 mice were then selected to obtain homozygous F2 mice. Serum and urine levels of uric acid, creatinine, and urea, and serum alanine aminotransferase (ALT) and aspartate aminotransferase ( AST) levels were detected and compared between homozygous and wild-type mice.Pathological changes in kidney and liver tissues were observed by hematoxylin and eosin and Masson staining. Results Urine levels of serum uric acid ( male: ( 4116. 8 ± 1928. 1) μmol / L, P<0. 001; female: ( 2998. 0 ± 547. 7) μmol / L, P<0. 01) and serum levels of uric acid (male: (478. 4 ± 114. 6) μmol / L, P<0. 001; female: (507. 7± 129. 6) μmol / L, P<0. 001), creatinine ((91. 8 ± 55. 6) μmol / L, P<0. 001), urea ((28. 6 ± 13. 9) mmol / L,P<0. 05), ALT (( 53. 3 ± 23. 3) U/ L, P<0. 01), and AST (( 203. 3 ± 70. 3) U/ L, P<0. 001) were significantly increased in Uox ^{- / -} mice compared with WT mice. Histopathological examination showed moderate hepatocyte degeneration in the liver, moderate-to-severe tubular cystic dilation, degeneration, and fibrosis in the kidney, glomerular hypertrophy and hyperplasia, small-vessel dilation and congestion, and infiltration of stromal monocytes and lymphocytes in Uox ^{- / -} mice. Conclusions We successfully established a homozygous uricase-deficient mouse strain using CRISPR/ Cas9 technology, as a suitable animal model for research in the field of hyperuricemia.

Abstract: Objective To provide a comprehensive review of the modeling method of the empty bottle stimulation ( EBS ) anxiety model, including commonly used experimental animal strains and genders, animal grouping, modeling procedures, modeling duration, primary behavioral evaluation method, and the underlying pathological mechanisms. This aims to offer a reference for the application of the EBS anxiety model in anxiety disorder research. Methods Searches were conducted in databases such as CNKI and PubMed to collect all literature related to the EBS anxiety model, which were then systematically summarized and organized. Results (1) Male adult SD or Wistar rats are predominantly used as experimental animals; (2) The optimal modeling period is 2 weeks;(3) Behavioral evaluations primarily utilize the open field test, elevated plus maze test, and light-dark box test; (4)Pathological mechanisms involve abnormal neurotransmitter metabolism in brain regions such as the hippocampus,prefrontal cortex, and amygdala. Conclusions The EBS anxiety model exhibits an anxiety-like behavioral phenotype and associated neurobiological mechanisms, validating its utility as an animal model for the study of anxiety disorders.However, further exploration and refinement are required for its standardized construction protocol and the understanding of its mechanistic underpinnings.

Abstract: Objective To establish a rapid and accurate droplet digital PCR (ddPCR) detection method for detecting Staphylococcus aureus (SA) in laboratory animals and the environment. Methods Using the heat-stable nuclease gene (nuc) of SA as the target gene, a pair of specific primers and probes are designed within its conserved region. Optimize the reaction conditions, test the dynamic range, and evaluate the specificity and stability of the method. Using the same template, test reactions were performed with both ddPCR and real-time quantitative PCR (qPCR) method to assess the interchangeability between the two approaches. Finally, the method is applied to the detection of various clinical samples. Results The kinetic range of the established SA ddPCR method is 100 ~15 000 copies/ μL, with a detection limit of 2. 5 copies and a quantification limit of 10 copies; The specificity of this method was tested, and only SA showed positive droplets, while no positive droplets were found for other pathogens;After measuring three parallel samples, the standard deviation and relative standard deviation were calculated. It was found that within the dynamic detection interval of ddPCR, as the target copy number gradually decreased, the relative standard deviation showed an upward trend, but remained below 25%. This result indicates that the detection method has good stability. Conclusions The established ddPCR method for detecting SA has the advantages of high sensitivity, strong specificity, good stability, and good reproducibility. This method can be applied for the detection of SA in laboratory animals.

Abstract:Senile osteoporosis is a growing public health challenge with significant impacts on the daily life of the elderly population as a result of its hidden nature, high prevalence, and high risk of disability. Suitable animal models that simulate senile osteoporosis are crucial for understanding its pathological mechanism and to facilitate the development of anti-osteoporosis drugs and identify new therapeutic targets. This review considers the most commonly used method for creating animal models of senile osteoporosis, analyzes their advantages and limitations, and discusses research progress in animal models in terms of evaluation indicators, to provide references for research using animal models of senile osteoporosis.

Abstract:Prostaglandin D2 (PGD2) is a biologically active substance with important roles in a variety of physiological and pathological processes. PGD2 exerts its biological functions mainly through prostaglandin D2 synthase (PGDS), which is closely related to inflammation and immune regulation. Recent studies have found that PGD2 and its synthase, PGDS, are able to directly inhibit tumor cell proliferation, induce apoptosis, suppress migration and invasion, and further regulate the tumor immune microenvironment to affect the immunotherapy of tumors, demonstrating good tumor therapeutic potential. In this paper, we review the biological properties of PGD2 and its synthase, focusing on its role in the immunotherapy of tumor models. We explore the immunotherapeutic efficacy of PGD2 and its synthase, and their roles in promoting immune cell infiltration in the tumor microenvironment, and discuss their potential as new targets for tumor therapy.

Abstract:In recent years, germ-free rats and mice have received extensive attention and application in the field of biomedical research, particularly in gut microbiota studies. The demand for germ-free rodents has been increasing, requiring many research institutions to establish quality germ-free animal populations. The process of breeding first-generation germ-free animals involves highly specialized and technically challenging work, such as using artificial rearing techniques to feed rodent pups. This review article provides an overview of key aspects of artificial rearing techniques, including nursing method, preparation of artificial milk, and sterilization method. It retrospectively summarizes the key technical points and, based on this foundation, offers prospects for further applications of artificial rearing techniques.