Abstract: Objective To explore the effect of the Trim44 gene in the nervous system, Trim44 conditional knockout rats were used to analyze behavioral abnormalities and pathological changes in the brain. Methods TRIM44 expression in brain tissue was detected by Western blot and immunohistochemistry. Neuronal apoptosis was determined by TUNEL staining. Behavioral changes of knockout rats were assessed by Morris Y, Morris water, and food maze tests. Abnormal expression of apoptosis-related proteins was detected by Western blot. Results TRIM44 was highly expressed in brain tissues of adult rats. The knockout efficiency of Trim44 in brain tissues was 67%. Trim44 knockout decreased learning and memory abilities, increased neuronal apoptosis, and expression of active caspase 3 and 9 proteins in aged rats. Conclusions Trim44 knockout promotes neuronal apoptosis and impairs learning and memory abilities in rats. These result suggest that TRIM44 plays an important role in the nervous system, and the Trim44 knockout rat model can be used for further mechanism exploration.

Abstract: Objective To investigate the effect of hydroxysafflor yellow A (HSYA) on oxidative stress and apoptosis of H9C2 cardiomyocytes after oxygen-glucose deprivation/ reperfusion injury and to explore its possible mechanism. Methods After rat H9C2 cardiomyocytes were stably subcultured to the third passage, they were randomly divided into control, hypoxia/ reoxygenation model (OGD/ R), low-dose HSYA (40 μmol/ L; HSYA40), and high-dose HSYA (80 μmol/ L; HSYA80) groups with 10 duplicate wells in each group. After 6 hours of treatment, the cells were analyzed by CCK-8 assays and aspartate transferase (AST), creatine kinase (CK), and lactate dehydrogenase (LDH) activities in the medium were measured by a biochemical analyzer. Corresponding kits were used to measure the contents of superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) in cells. Flow cytometry was used to assess the apoptosis rate. Western blot was used to detect apoptosis-related proteins B-cell lymphoma/ leukemia-2 (Bcl2) and Bcl2-related X Protein (Bax) in cells. Caspase-3 expression was analyzed semiquantitatively. Results Compared with the normal group, cell proliferation in the OGD/ R group was significantly lower, AST, CK, and LDH activities in the medium were significantly increased, SOD and CAT activities were significantly decreased, and the MDA content was increased. The apoptosis rate was significantly increased, Bcl2 and Bax mRNA expression was significantly increased, Bcl2/ Bax protein expression was significantly decreased, and Caspase-3 protein expression was significantly increased. Compared with the OGD/ R group, cell proliferation in the hydroxysafflor A administration group was significantly increased, AST, CK, and LDH activities in the medium were significantly decreased, SOD and CAT activities were significantly increased, and the MDA content was decreased. The apoptosis rate was significantly reduced and expression of anti-apoptotic gene Bcl-2 was upregulated, while the expression of proapoptotic gene Bax was significantly downregulated, Bcl2/ Bax protein expression was significantly increased, and caspase-3 protein expression was significantly decreased. Conclusions HSYA protects H9C2 cardiomyocytes from oxygen-glucose deprivation/ reperfusion injury by inhibiting oxidative stress injury and apoptosis. Its mechanism may be related to improving the activity of antioxidant enzymes, upregulating Bcl-2 gene expression, downregulating Bax gene expression, increasing Bcl-2/ Bax protein expression, and inhibiting the expression of pro-apoptotic protein Caspase-3.

Abstract: Objective To observe the effect of Huangyangning on oxidative stress in dogs with atrial fibrillation. Methods Fifteen beagle dogs were randomly divided into five groups: blank control, model, sham-operated, Huangyangning (0. 24 mg/ kg), and probenecid (7. 58 mg/ kg) groups. A canine atrial fibrillation model was established by placing bipolar electrodes in the right heart ear through femoral vein puncture and connecting a high frequency pacemaker at the caudal end to provide high frequency stimulation. Model establishment was considered successful when atrial fibrillation was confirmed by an electrocardiogram after 1 week of pacing or when an episode of atrial fibrillation was induced during the programmed control. Gavage was started at 1 day after surgery, atrial pacing was started at 1 week after surgery, and samples were collected after 4 weeks. Myocardial fiber changes were observed by Masson staining. Serum levels of angiotensinII (AngII) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) were measured by ELISA. Reactive oxygen species (ROS) and total antioxidant capacity (TAC) were measured by chemiluminescence. Expression of NADPH oxidase (Nox) subunit genes p22 and p47 was measured by RT-PCR and Western blot. Results Compared with the blank group, serum AngII and NADPH were significantly increased (P< 0. 05) and ROS/ TAC levels were significantly increased (P< 0. 05) in the model group, expression of p22 and p47 genes was significantly increased, and expression of p22 and p47 proteins was increased. There was no significant increase in the serum NADPH level after treatment with Huangyangning or probenecid. ROS/ TAC levels were significantly lower than in the model group (P< 0. 05). Rac-1, p22 and p47 gene expression was significantly lower than in the model group (P< 0. 05). p22 and p47 protein expression was also significantly lower than in the model group (P< 0. 05). Conclusions Huangyangning reduces oxidative stress in dogs with atrial fibrillation, and its mechanism of action may be related to the AngII-NOx-ROS signaling pathway.

Abstract: Objective To investigate the virulence, drug resistance genes, and whole genome information of Aeromonas dhakensis from shrimp and preliminarily evaluate the pathogenicity of Aeromonas dhakensis. Methods Molecular identification and MLST typing of shrimp isolates were performed. The pathogenicity of this strain was examinedby β-hemolysis, extracellular protease, liquid toxicity test, cytotoxicity, and mouse tests, and virulence gene detection. The characteristics of drug resistance were analyzed by a drug sensitivity test and drug resistance gene detection. Eggnog and Prophage Hunter tools were used to analyze the whole genome sequence. Results The 18FX22 strain was identified as Aeromonas dhakensis. The pathogenicity of this strain was characterized by β-hemolysis and extracellular protease activity. It was pathogenic to Caenorhabditis elegans, mouse fibroblasts, and mice. The strain was resistant to amoxicillin/ clavulanic acid, cefazolin, and polymyxin, which carried 10 virulence genes and 12 drug resistance genes. Whole genome sequencing showed that the whole genome size of 18FX22 was 4 811 079 bp and GC content was 61. 33%. COG annotation indicated that 18FX22 had 3850 genes with 20 classes of functions. Prophage prediction revealed that the 18FX22 genome contained two prophage sequences. Cluster analysis of Aeromonas dhakensis worldwide showed that this strain was more closely related to GCA016729585. Conclusions Aeromonas dhakensis was isolated from commercial shrimp and found to be highly toxic, indicating that the control of seafood food safety should be strengthened. This study provides a reference to prevent and control Aeromonas dhakensis infection.

Abstract: Objective To investigate the trend in changes of body weight, ingestion, blood lipids, and pathology of liver tissue in two models of hyperlipidemia in rats, and to compare differences of the hyperlipidemia models of different genders. Methods Rats were fed two kinds of high fat diet for 7~8 weeks to establish models of mixed hyperlipidemia and hypercholesterolemia. The body weight, ingestion, and general state of the animals were recorded during the modeling period. Changes in serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) were detected and analyzed at 0, 1~2 and 7~8 weeks. Results The body weight of the mixed hyperlipidemia high-fat diet group was significantly higher than control group, ingestion was decreased significantly, and total food use was increased, while ingestion of hypercholesterolemia high-fat diet group was increased and there was no significant difference in body weight or total food use. At the end of modeling, TC and TG levels in the mixed hyperlipidemia groups was increased, the TC level in the hypercholesterolemia group was increased, TG was unchanged, and the degree of hepatic steatosis in the mixed hyperlipidemia group was stronger. In the same model, the overall change in blood lipids and the degree of liver steatosis in male rats were more severe than those in female rats. Conclusions There were differences in body weight, ingestion, lipid levels and liver pathology in high fat diet groups of the two models with a stronger degree in male than in female rats of the same model.

Abstract: Objective To subject vaginal smears of female SD rats and C57BL/6J mice to rapid Gram staining, observe the cell morphology, and develop a simple staining method to rapidly identify the estrous cycle stage. Methods Vaginal smears from 12 female SD rats and 12 female C57BL/6J mice at 56~62 days of age were obtained by the vaginal swab method every day at 9:00, dried, and then subjected to rapid Gram staining for continuous observation over 12 days. Cell morphology was observed under a light microscope to determine the estrous cycle stage. Results Rapid Gram staining was performed quickly and simply, bright in the observation field with a clean background, and provided a complete overview of the changes in the estrous cycle of rats and mice. Conclusions The rapid Gram staining method can easily and intuitively judge the estrous cycle stage of rats and mice, staining steps can be optimized, abnormal conditions in the staining can be analyzed stepwise, and the staining time of each smear can be shortened to less than 1 min. The method is easy for novice experimenters to master and can be applied widely.

Abstract: Objective To investigate whether perirenal adipose tissues as an endocrine organ regulate arterial blood pressure in rats via exosomes. Methods Spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats were used in this study. Perirenal fat of normotensive WKY rats and SHRs was cultured in serum-freeidentified by nano-size measurement, transmission electron microscopy, and surface markers, which were consistent with the characteristics of exosomes (Exo) and used for subsequent experiments. SHR and WKY rats were randomly divided into two groups and injected with Exo-SHR or Exo-WKY via the tail vein once a week for 6 weeks. Arterial blood pressure was measured while rats were conscious using a non-invasive caudal cuff system. Acute experiments were conducted at the end of week 6 after the injection of exosome-like vesicles. Carotid artery cannulation was used to directly measure arterial blood pressure. HE and Masson staining was used to observe cardiovascular pathological remodeling. Peripheral sympathetic tone (PST) was expressed as the relative ratio of the depression response induced by intravenous injection of hexamethonium hydrochloride to that in WKY rats treated with Exo-WKY. Exosomes labeled with PKH67 green fluorescent dye were injected into the tail vein for in vivo tracing, and then incubated with primary cultured neurons for in vitro tracing. Results Exosomes secreted from perirenal white adipose tissue were typical exosome semi-cup-like vesicles under transmission electron microscopy. The average diameter of nanoparticles was approximately 100 nm, and surface-specific markers TSG101 and CD9 were expressed. The systolic blood pressure (SBP) of SHRs began to decrease in the second week after injection of Exo-WKY and remained low throughout experiment (P< 0. 05). Exo-WKY also decreased the crosssectional area of cardiomyocytes, aortic media thickening, and PST in SHRs (P< 0. 05). However, after administering Exo-SHR to WKY rats, no significant changes were observed in SBP, the cross-sectional area of cardiomyocytes, media thickness of the aorta, or PST. Green fluorescence was observed in the cardiovascular center, including the PVH, nucleus tracts solitary, rostral ventrolateral medulla, and in cultured neurons at 24 h after application of PKH67-labeled exosomelike vesicles. Conclusions Perirenal adipose tissue-derived exosomes act as transport carriers to remotely regulate arterial blood pressure. Exo-SHR lack contents that inhibit blood pressure elevation. Exo-WKY supplementation reduces arterial blood pressure and sympathetic tension, which improves cardiovascular remodeling.

Abstract: Objective To explore the protective effect and mechanism of curcumin (CUR) on acetaminophen (APAP) -induced acute kidney injury in rats. Methods Forty-eight male SD rats were randomly divided into normal, model, positive, CUR low (50 mg/ kg), medium (100 mg/ kg), and high (200 mg/ kg) dose group. After intragastric administration of N-acetylcysteine ( NAC) and CUR for 10 days, acute kidney injury models were established by intragastric administration of 2 g/ kg APAP. After 24 hours, blood was collected and kidneys were collected, the kidney index was calculated, the levels of serum creatinine (Cr) and blood urea nitrogen (BUN) were measured. Pathological injury of the kidney was evaluated by HE staining, activities of glutathione (GSH), total superoxide dismutase (T-SOD), and catalase (CAT), and the malondialdehyde (MDA) level in kidneys were assessed. Expression of Trx-1, TXNIP, and NLRP3 inflammasome in kidneys was detected by Western blot. Results Compared with the normal group, the kidney index and the levels of serum Cr and BUN in the model group were increased significantly (P< 0. 01), pathological sections showed obvious lesions in the kidney, activities of GSH, T-SOD, and CAT in kidneys were significantly decreased (P< 0. 01), the MDA content was significantly increased (P< 0. 01), Trx-1 expression in the kidney was significantly decreased (P< 0. 05), and expression of TXNIP, NLRP3, ASC, Cleaved caspase-1, and mature IL-1β was significantly upregulated (P< 0. 01). Compared with the model group, the kidney index in CUR medium, and high dose groups was significantly decreased (P< 0. 01), pathological damage had improved, levels of serum Cr and BUN were decreased (P< 0. 01), activities of GSH, T-SOD, and CAT in the kidney were increased, MDA content was decreased (P< 0. 05 or P< 0. 01), Trx-1 expression was significantly increased (P< 0. 05 or P< 0. 01), and expression of TXNIP, NLRP3, ASC, Cleaved caspase-1, and mature IL-1β protein was significantly downregulated ( P< 0. 01). Conclusions CUR pretreatment alleviates acetaminophen-induced acute kidney injury by regulating the Trx-1/ TXNIP/ NLRP3 signaling pathway in rats.

Abstract: Objective To examine the morphological characteristics of the eyeballs of tree shrews and to compare the similarities and differences of the anatomical structure of the eye of tree shrews, monkeys, and humans to provide the necessary histological data to establish ocular disease models using tree shrews. Methods Paraffin-embedded sections of tree shrew and rhesus monkey eyeballs were prepared and analyzed by HE staining. Results The tree shrew eyeball mainly consisted of a lens and eyeball wall. The ocular wall was divided into three layers including the fibrous membrane, vascular membrane, and retina from the outside to inside. The anterior 1/6 of the fibrous membrane was the cornea, and the posterior 5/6 was the sclera. The vascular membrane was the middle layer of the ocular wall. It was composed of loose connective tissue, abundant blood vessels, and pigment cells. It was divided into the iris, ciliary body, and choroid from front to back. The retina was the innermost layer of the eyeball wall and had a typical structure of 10 layers. The histological structure of the tree shrew eyeball was consistent with that of rhesus monkey and human eyeballs, and the proportion of each tissue layer was approximately the same. The main differences were that the sclera epithelium of tree shrews contained melanocytes, and the proportion of each retinal layer was different from that of humans. Conclusions The eye tissue structure of tree shrews is similar to that of non-human primates and humans, and the tissue morphology is approximately the same. There is potential to establish an animal model of human eye-related diseases to clarify the pathogenesis of eye diseases.

Abstract: Objective To integrate the gene expression profiles of heart disease animal models and patients, systematically characterize the differences in gene expression, screen differentially expressed genes, analyze the similarities and differences of gene expression, and provide a basis for animal experiments and comparative medicine research. Methods Microarray and transcriptome data of heart disease animal models and patients were downloaded from a public database. Data were sorted by species and disease type and stage, standardization and removal of batch effects were performed, data were converted in the available format for the database, and an online database was built. Results The comparative transcriptomics database of heart disease animal models realized global searching and queries of genes and functional pathways, and analysis of gene expression in humans and mice, diseases, tissues, and stages. It also interactively displayed sample expression profiles, visualized gene expression, and facilitated comparative analysis. The database allowed online differential gene mining, enrichment and identification of functional pathways, exploration of the biological processes of susceptibility genes involved in regulation, explanation of the susceptibility mechanism, and online data analysis. Conclusions A comparative transcriptomic database of heart disease animal models was established to provide data resources and online analysis tools to study heart disease.

Abstract:Euthanasia of laboratory animals is among the fundamentals of animal experimental ethics. Apart from the widely accepted euthanasia method involving anesthesia, non-anesthesia method are also commonly used. On the basis of the “ 3R” principle of animal experimental ethics and the “ five freedoms” of experimental animal welfare, we systematically discuss the ethical legitimacy of two non-anesthetic euthanasia method, decapitation and cervical dislocation. Additionally, potential ethical problems and possible improvements of air embolism, liquid nitrogen freezing, bloodletting, and apnea as euthanasia method are discussed. We also propose feasible approaches of euthanasia involving integrative method. Our analysis may assist researchers in performing ethical non-anesthesia euthanasia.

Abstract:Ferroptosis is novel iron-dependent programmed cell death characterized by excessive accumulation of reactive oxygen species and lipid peroxidation. In recent years, an increasing number of studies have focused on the roles of immunomodulation in ferroptosis. Immune cells are the most important part of the immune system. This review discusses the current research progress on the relationship between ferroptosis and immune cells. The role of glutathione peroxidase 4 (Gpx4) in immune cell proliferation and the immune response is summarized. Ferroptosis releases various molecules including damage-associated molecular patterns, which influences the differentiation and function of immune cells and thereby affects disease progression. Moreover, immune factors or their metabolites secreted by immune cells affect the occurrence of ferroptosis. This review concludes with a brief summary and outlook for future research, which may be helpful to guide the direction of disease treatment.

Abstract:The rabbit is a classical animal model in biomedical research. The development of genetically modified rabbit models has been severely impeded because of the lack of pluripotent stem cells and the extremely low efficiency of somatic cell nuclear transfer. With the rapid development of novel genome-editing techniques, such as ZFNs, TALENs, and CRISPR/ Cas9, genetic engineering of rabbits has developed rapidly. Genetically modified rabbit models are playing increasingly important roles in the field of biomedicine. In this review, we discuss the development of genetically modified rabbit models and their applications in human disease research to provide a theoretical reference for promoting the development of human disease animal models.

Abstract:Hepatic encephalopathy (HE) is a neuropsychiatric syndrome that frequently occurs in the course of acute or chronic liver diseases or an abnormal portal shunt. Astrocytes play a critical role in the pathogenesis of HE. In recent years, HE has been found to be accompanied by morphological and functional changes of astrocytes and HE is effectively alleviated by drugs that act on astrocytes. Therefore, astrocytes may be a potential therapeutic target for HE. This article reviews the role of astrocytes in HE to provide new clinical ideas for treatment.

Abstract:Ischemic cerebral stroke is a clinical syndrome in which various cerebrovascular diseases cause blood supply disorders in the brain, leading to ischemia and hypoxic necrosis of local brain tissue and corresponding neurological deficits rapidly appear. Astrocyte extracellular vesicles contain various growth factors, proteins, and miRNAs. By participating in communication between brain cells by binding to endothelial cell receptors, transferring to neurons, and being taken up by neurons, astrocyte extracellular vesicles promote neurogenesis and angiogenesis after cerebral ischemia. Thus, astrocyte extracellular vesicles have a protective effect on neurons damaged after cerebral ischemia and are a potential target for treatment and prevention of central nervous system diseases. This article reviews the contents of astrocyte extracellular vesicles and their protective effects against ischemic stroke to provide a theoretical basis for clinical research of novel therapeutic strategies.

Abstract:The classical view is that astrocytes are support cells of neurons and have no direct role in brain information processing. Accumulating evidence indicates that neuron-astrocyte signaling dysfunction may be related to the pathology of various neurological and neurodegenerative diseases. This has gradually become a hot topic in the study of depression pathogenesis in recent years. This review summarizes the evidence that astrocyte-neuron communication is involved in brain oxidative defense, synaptic plasticity, brain energy metabolism, neuroinflammation, and glutamate cycling in depression. Such evidence suggests that dysfunction of astrocyte-neuron communication crosstalk is a major mechanism in the pathogenesis of depression. This review also discusses the recent view that astrocyte-neuron interactions are involved in the pathogenesis of depression, as well as the existing problems in the research, to provide ideas and references for subsequent research.

Abstract:Immunotherapies are a novel anti-tumor treatment after surgery, radiotherapy, and chemotherapy. Among them, the therapeutic effect of immune checkpoint inhibitors is the most promising, but the effect of treatment with a single immune checkpoint inhibitor in cancer patients is limited. Traditional Chinese Medicine has unique advantages as cancer treatment. Its adjuvant immunotherapy benefits patients by improving the deficiency of immunotherapy alone. Thus, in this review, we summarize the current strategies of the combination of cancer immunotherapy with Traditional Chinese Medicine, analyze the mechanism of anti-tumor immunity by combined treatment, and propose the prospect of its clinical application to provide effective strategies for combined immunotherapy of cancer by Traditional Chinese Medicine.

Abstract:We discuss studies aimed to promote a healthy aging process, prevent health problems related to aging, correctly understand the aging mechanism, and develop effective anti-aging treatment measures. Systemic injection of D-galactose has been used for artificial induction of aging in vitro and in vivo, and anti-aging research. This review summarizes the characteristic changes of aging induced by D-galactose from four aspects: oxidative stress, mitochondrial dysfunction, immune aging, and the influence of immune aging on various organs. Furthermore, a theoretical basis for antiaging is provided from various viewpoints.