Abstract:Objective Using the advantages of zebrafish embryo transparency for live imaging, the role of imatinib in the development of blood vessels and nerves in zebrafish was studied. Methods First, we investigated the safe concentration range of imatinib for zebrafish in the blood vessel and neuron development stage using wild type zebrafish embryos. Within the safe concentration range, transgenic zebrafish embryos with a labeled vascular system were treated with imatinib. Moreover, endothelial cell and neuron co-labeled zebrafish were treated with imatinib to study the specificity for angiogenesis inhibition. The molecular mechanism of imatinib-inhibited angiogenesis was investigated. Results Imatinib concentration below 100 μg / mL was not lethal to zebrafish embryos. In this concentration, imatinib significantly inhibited the development of zebrafish intersegmental vessels (ISVs), including the process of ISV lumenization and the proliferation of endothelial cells. Additionally, imatinib had no significant effect on motor neurons and neuronal precursor cells. Moreover, gene expression analysis showed that imatinib was able to significantly inhibit vegfr2 and pdgfrα mRNA expression. Conclusions Imatinib significantly inhibited angiogenesis in zebrafish embryos by regulating VEGF and PDGFR signals.

Abstract:Objective To explore the effects of microRNAs ( miRNAs ) on proliferation, migration and angiogenesis in a miniature pig model of myocardial infarction (MI). The miRNAs differently expressed between MI and control pigs were selected, in particular those potentially targeting the ANGPT-2 gene ( an important growth factor during angiogenesis), and their effects on proliferation, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) were investigated. Methods Bioinformatics software was used to predict the differentially expressed miRNAs targeting ANGPT-2 in the MI and control groups. The effects of miRNAs on the proliferation, migration and angiogenesis of HUVECS were detected by EdU staining proliferation, transwell chamber and in vitro experimental method, respectively. Results miR-144, miR-21-3p, miR-142-5p and miR-27b-3p were found to potentially target ANGPT-2 and were significantly differently expressed between the MI and control groups. In HUVECs, miR-144 had no significant effect on proliferation, but significantly inhibited migration and vessel formation; miR-21-3p inhibited proliferation and migration, but promoted vessel formation; miR-142-5p and miR-27b-3p inhibited proliferation, migration and vessel formation. Of these four miRNAs, miR-142-5p and miR-27b-3p significantly inhibited the expression of ANGPT-2 mRNA. Conclusions After MI, miR-142-5p and miR-27b-3p may inhibit angiogenesis by suppressing the proliferation, migration and vessel formation of vascular endothelial cells, by targeting the 3′ UTR of ANGPT-2 and down-regulating ANGPT-2 mRNA expression.

Abstract:Objective To investigate the underlying mechanisms of the analgesic effect of electroacupuncture (EA), we assessed the effect of EA on microglial activation and the expression of brain-derived neurotrophic factor (BDNF) in the L₄-L₆ spinal cord in rats with diabetic neuropathic pain (DNP). Methods SD rats were divided into a normal group (N group), a model group (M group), and an EA group (EA group). The rats in the EA group were treated with EA at “Zusanli” (ST36) and “Kunlun” (BL60) starting on week 6, with 30 min per session, once every other day for 7 sessions. Fasting blood glucose ( FBG), thermal paw withdrawal latency ( PWL), and paw withdrawal thresholds (PWT) were measured at baseline, 4 weeks, 6 weeks, and 8 weeks. Immunofluorescence was used to measure the expression of microglia (CD11b) and BDNF in samples from the L₄-L₆ spinal cord. Results 1) Compared with that in the N group, FBG was significantly increased in the M group and the EA group, and there was no significant difference in FBG between the M group and the EA group at 6 weeks and 8 weeks.(2) Compared with that in the N group, the PWL and PWT were significantly decreased in the M group at 6 weeks and 8 weeks. After EA treatment, compared with that in the M group, the PWL and PWT in the EA group were significantly increased at 8 weeks. there was no significant difference in PWL and PWT between the M group and the EA group at 6 weeks. (3) Compared with that in the N group, the expression of microglia (CD11b) and BDNF in the M group were increased significantly, and compared with that in the M group, the expression of microglia ( CD11b) and BDNF were significantly decreased in the EA group. Conclusions The result suggest that microglia and BDNF in the spinal cord might be involved in the development of DNP. The analgesic effect of EA might be achieved by inhibition of the expression of microglia and BDNF.

Abstract:Objective To establish a non-alcoholic fatty liver disease (NAFLD) rat model in a new type of hyperlipidemia (WSHc) rat. We observed the pathological characteristics of the NAFLD model and explored possible research applications, as well as new options for animal models of NAFLD. Methods 20 WSHc rats aged 7 – 8 weeks were randomly divided into 2 groups according to whether they would receive a common diet or high-fat diet. Another 20 Wistar rats of the same age were then grouped as a strain control, and received the same treatment. After 12 weeks of continuous feeding, blood samples were taken to detect blood biochemical indicators and elastic ultrasound imaging was performed. After execution of the animals, the liver tissue was subjected to HE, oil red “ O ”, Masson, and immunofluorescent staining, and the general cell morphology, lipid performance, inflammatory response, and liver fibrosis stage in each group were observed. The lesion characteristics of the liver of WSHc rats after a high-fat diet were analyzed. Results Compared with Wistar rats on a high-fat diet, WSHc rats showed a significant increase in serum total cholesterol, similar to that seen in patients with clinical hyperlipidemia. We also found that liver lipids were abnormally deposited to form lipid droplets, and that inflammatory cells positioned around the lipid droplets formed an inflammatory lesion. Further, we observed an increase in macrophages and instances of scattered fibrosis. These pathological changes were not found in the liver tissue of high-fat-exposed Wistar rats. Conclusions Compared with conventional rats, WSHc rats on a high-fat diet represent a more clinically similar NAFLD animal model, which is characterized by blood biochemical indicators, a severe inflammatory response, and sporadic liver fibrosis approaching clinical levels.

Abstract:Objective To use liquid chromatography-mass spectrometry based plasma metabonomics to study the characteristics of human flora-associated (HFA) mice with coronary heart disease. Methods Twenty-eight female germ free C57BL/ 6J mice were divided into healthy control and coronary heart disease (CAD) groups. Each mouse was orally inoculated with a stool suspension from healthy people or patients with CAD to establish the HFA mouse models. At 6 and 10 weeks after inoculation, plasma metabolites were studied and PCA and PLS-DA statistical method were used to identify the characteristic metabolites. Results The PCA, PLS-DA and variable importance in projection analysis identified 30 characteristic metabolites present at 6 and 10 weeks after inoculation; structural identification found that L-carnitine, phenylpyruvic acid, and 1-naphthol, 2-naphthol were significantly higher in the CAD group compared with the control group. These potential biomarkers were related to the bile acid pathway and glycine, serine, threonine pathway. Conclusions The metabolism disorder in HFA mice appears similar to that of patients with CAD.

Abstract:Objective To establish a mouse model of allergic conjunctivitis by comparison of different antigens, routes of administration, and ages of mice. We compared their immune response status and screened for optimal sensitization conditions to provide new ideas and method to establish mouse models of allergic conjunctivitis. Methods Sixty-five SPF Balb / c mice were selected to establish allergic conjunctivitis models using different antigens ( ovalbumin, ragweed pollen, house dust mite ), administration routes ( gastric administration, aerosol inhalation, subcutaneous injection) and ages ( 1, 2, 3 and 8 weeks). Results The expression levels of interleukin-17 ( IL-17) and C-C motif chemokine ligand 5 ( CCL5) mRNA were increased in the ovalbumin, house dust mite and ragweed pollen groups, especially the later, compared with the negative control group. Compared with the negative control group, the expression of IL-17 mRNA was increased in the subcutaneous injection, gavage, inhalation and positive control groups. The expression of CCL5 mRNA in the gavage group was close to that of the negative control group, while its expression level in the subcutaneous injection group was closer to that of the positive control group. Expression levels of CCL5 and IL-17 mRNA were similar in 1 - and 2-week-old mice, but significantly lower than those in 3 - and 8-week-old mice. Conclusions Ragweed pollen provided better sensitization in the preparation of an allergic conjunctivitis mouse model. Subcutaneous injection of antigen achieved satisfactory result in the allergic conjunctivitis mouse model. Although there were no obvious eye symptoms, the whole body and local areas were sensitized. The immune response of 3-week-old Balb / c mice was close to that of adult Balb / c mice.

Abstract:Objective To examine the effect of a compound healthy ear agent (CHEA) from traditional Chinese medicine (TCM) on i) age-related apoptosis of cochlear hair cells (OHC) and spiral ganglion neurons ( SGN) using transmission electron microscopy (TEM), and on ii) localized expression of brain-derived neurotrophic factor (BDNF) and neuron-specific nuclear antigen (NeuN) to investigate mechanisms of CHEA actions. Methods Twenty-two C57BL/ 6J mice were randomly divided into two groups at 1 month of age. Eleven animals were provided tap water daily until 7 months of age; the age-related cochlear degeneration control group (7-month-old control group). Another 11 animals drank CHEA 1. 83 g / (kg·d) until 7 months of age; the TCM-treated group (7-month-old TCM group). Cochlea of euthanized animals were removed and processed into paraffin-embedded slices. Using TEM, ultrastructural changes were examined in the remaining and undisintegrated OHC and SGN associated with OHC at the basal region of cochlea from 7-month-control and 7-month-old TCM groups. Localized expression of BDNF and NeuN in SGN was observed by LCM combined with a morphological multi-overlap immunofluorescence technique, and examined using statistical analyses. Results In the 7- month-old control group, remaining and undisintegrated OHC and SGN associated with OHC in cochlea basal regions exhibited atrophied cytostomes, irregular nuclei and chromatin structure, texture structure presented vague, and high electron density; in particular, SGN were different in size and shape, significantly decreased in number, and exhibited nuclei chromatin that had disintegrated or dissolved to form vacuoles, representing the presence of apoptosis. However, the 7-month-old TCM group had no obvious apoptosis in OHC or SGN of basal cochlear regions, which exhibited more complete morphological structures and nuclei with a uniform chromatin distribution, texture structure presented clear, a lower electron density, and only individual nuclei had dissolved to form vacuoles. The localized expression levels of BDNF and NeuN in the basal cochlear region were higher (P < 0. 05) in the 7-month-old TCM versus the 7-month-control group. Conclusions Ultrastructural changes of nuclei and decreased expression of NeuN in cochlear OHC or SGN were important apoptotic features in aging cochlear neurocytes. Apoptosis in SGN was earlier than in OHC. The TCM reduced the ultrastructural changes in OHC or SGN, and increased NeuN expression in SGN, which suggests that TCM had a protective effect upon SGN, consistent with TEM observations. The mechanism of action may involve promotion of BDNF expression, which exerts a series of biological effects.

Abstract:Objective To observe the effects of taurine on blood hormones and hepatic insulin growth factor (IGF) gene expression in intrauterine growth-restricted pregnant rats. Methods Healthy SD female and male rats were housed 1 ∶2 in cages and pregnant rats were randomly selected into the blank control group (C), model group (M), model + taurine group (MT), and taurine group (T). In the M and MT groups, 10 pregnant rats were fed a low-protein diet. Day 12 of pregnancy, C, and M were provided drinking water, and MT and T groups received taurine 300 mg / (kg·d). Weekly monitoring of weight gain in pregnant rats. At day 21 of gestation, baseline characteristics of fetuses were determined. Serum growth hormone (GH), insulin growth factor 1 (IGF-1), insulin (INS), and thyroxine (T3 and T4) levels were detected by ELISA. Fluorescent quantitative PCR was used to detect the hepatic expression levels of growth hormone receptor (GHR), insulin growth factoR-1 ( IGF-1), insulin growth factor binding protein-1 ( IGFBP-1) and insulin growth factor binding protein-3 ( IGFP-3) mRNA in pregnant rats. Results Taurine markedly improved the baseline characteristics, compared with the M group. Pregnant rats in the MT group had significantly increased (P < 0. 05) weights, blood INS and T4 levels, and liver IGF-1 and IGFBP-3 mRNA expression, whereas IGFBP-1 mRNA expression was significantly reduced (P < 0. 01). Conclusions The addition of taurine can improve the thyroid function and GH/ IGF-1 axis disorder in low- protein diet pregnant rats, and may play an important role in alleviating fetal growth restriction.

Abstract:Objective To explore the effects of two types of type I interferon (IFN-I) receptor deletions upon the reproductive performance of mice. Methods In vitro fertilization (IVF) was performed using two types of IFN-I receptor- deficient mice (dificient in IFN-α and IFN-α/ β receptor signaling) and wild-type background ( C57BL/ 6) mice. Each group contained 5 superovulated mice and the assay was repeated in triplicate. The female and male gametes of two the types of IFN receptor-deficient mice and the wild-type mice were examined using IVF. The IVF conditions were optimized and method to increase the fertilization rate were explored. Results The average IVF rates ( number of 2-cell embryos as a percentage of oocytes) for the two types of IFN receptor-deficient mice were significantly lower (P < 0. 05) than that of wild-type mice. The IVF rate of mice with IFN-α receptor deficiency was higher ( P < 0. 05) than that of mice with IFN-α/ β receptor deficiency. The IVF rates using receptor-deficient sperm and C57BL/ 6 oocytes were significantly higher (P < 0. 05) than those using receptor-deficient oocytes and C57BL/ 6 sperm. The IVF rates were significantly increased (P < 0. 05) by extending the sperm capacitation time from 30 min to 1 h or adding glutathione to the capacitation and fertilization solutions form 0 mmol / L to1 mmol / L. Conclusions Deletion of the IFN-I receptor may reduce the fertility of mice, and the effect on oocytes appears more significant than that upon sperm. Extending the sperm capacitation time or changing the capacitation and fertilization fluid composition improved the fertilization rate.

Abstract:Objective To establish a novel long-term depression ( LTD) model in mice, and to compare this model with the common depression mouse model. Methods Using early maternal separation and persistent chronic stress after weaning to prepare LTD mice, which were then compared with chronic unpredictable mild stress and early life stress mice to observe potential differences in their long-term depression-like behaviors. The changes in hippocampal neurons of LTD mice expressing transgenic eGFP were also observed. Results Compared with chronic unpredictable mild stress and early life stress mice, LTD mice showed a stable state of depression in animal behavior experiments. Degeneration of the hippocampal CA1 and CA3 regions of LTD mice was observed in tissue sections. Conclusions The LTD mouse model was successfully established. LTD mice provide a model suitable for the investigation of long-term simulation of depression, which can lead to long-term depression-like behavior and psychological stress.

Abstract:Objective To study a diabetic rat model induced by high-fat diet ( HFD) combined with streptozotocin (STZ), and to provide a stable diabetic animal model for clinical research. Methods Thirty male SD rats were randomly divided into two groups: normal control group (NG, n = 10) fed a normal diet; type 2 diabetes group (DM, n = 20) fed a HFD for 12 weeks + 25 mg STZ/ kg via tail vein injection. The body weight of rats was measured every week during the experiment. The oral glucose tolerance test, insulin tolerance test and the area change under the blood glucose curve were observed one week before STZ injection. After STZ injection, the general condition ( food intake, water consumption, hair luster, moisture and state of padding ), random blood glucose, fructosamine and pancreatic histopathological sections were examined. Results The body weight was significantly increased ( P < 0. 05) in the DM group compared with the NG group. The blood glucose was significantly higher in the oral glucose tolerance test group compared with that of the NG group ( P < 0. 05) at 60, 90 and 120 min, and the area under the curve was also significantly increased (P < 0. 05); blood glucose in the DM group was significantly higher than that of NG group (P < 0. 05), and the area under the curve was also significantly increased ( P < 0. 05). After STZ intervention, there were significant differences in food intake, diet and body weight between the two groups (P < 0. 05). Conclusions After 12 weeks of a HFD, insulin resistance and glucose tolerance were abnormal in rats. The model of type 2 diabetes induced by low-dose STZ provides a valuable animal model for the study of type 2 diabetes.

Abstract:Objective To find a stable method for establishing a loperamide hydrochloride-induced slow transit constipation (STC) model by comparing the differences among STC mouse models established by different loperamide hydrochloride concentrations and administration times. Methods Healthy male C57BL/ 6 mice were given 0. 025, 0. 25 or 2. 5 mg / mL loperamide hydrochloride by intragastric administration twice a day at 9:00 and 15:00. The small intestinal propulsion rate, defecation quantity and other indicators were used to evaluate the effects of different doses and administration times on the STC mouse model. Results ( 1) Comparing the effects of different dosages of loperamide hydrochloride in the range of 0. 5 mg / kg to 50 mg / kg revealed that the small intestinal propulsion rate and defecation quantity significantly decreased in the 50 mg / kg dosage group compared with those in the control group (P < 0. 05). (2) At day 7 after oral administration of loperamide hydrochloride, the small intestinal propulsion rate significantly decreased compared with that in control group (P < 0. 05), whereas this rate did not significantly change at days 3, 10 and 14 (P> 0. 05). Conclusions The small intestinal propulsion rate and defecation quantity significantly decreased after continuous administration of 50 mg / kg loperamide hydrochloride for 7 days. Hence, this is a relatively stable method for establishing an STC mouse model. This study provides valuable data for the exploration of the loperamide hydrochloride-induced STC model.

Abstract:Objective To explore the mechanism by which docosahexaenoic acid ( DHA) inhibits the accumulation of adipose tissue lipid in high-fat diet ( HFD)-induced body weight gain in C57BL/ 6 mice. Methods Animals were fed a control diet (C57BL/ 6 C group), a 45% HFD (C57BL/ 6 H group) or 45% HFD with 0. 2 g DHA (FAD3C) or 0. 4 g DHA (FAD3H) per g of food for 20 weeks. Resting metabolic rate was measured at 19 weeks. Visceral adipose and brown adipose tissue were collected for RNA extraction. Lipogenesis-related and fat browning-related gene expression was detected by quantitative PCR. Results A HFD significantly increased (P < 0. 05) body weight, body fat, and serum leptin and triglyceride ( TG) levels in the C57BL/ 6 H group compared with the C57BL/ 6 C group. DHA significantly decreased (P < 0. 05) body weight, fat mass, and levels of serum leptin and TG in the FAD3C and FAD3H groups compared with the C57BL/ 6 H group. In visceral adipose tissue, DHA significantly downregulated the expression of lipogenesis-related genes, including the CCAAT/ enhancer-binding protein, peroxisome proliferator-activated receptor-γ, and sterol regulatory element-binding protein-1C in FAD3H groups compared with the C57BL/ 6 C group (P < 0. 05). In visceral adipose and brown adipose tissue, DHA supplementation significantly increased (P < 0. 05) the expression of fat browning-related genes, including uncoupling protein 1, and peroxisome proliferator-activated receptor coactivator-1a in FAD3H groups compared with the C57BL/ 6 H group. Conclusions DHA may be used to combat obesity by regulating the resting metabolic rate, levels of leptin, fat and TG, and the expression of browning-related and lipogenesis-related genes.

Abstract:Objective To explore the therapeutic effect and potential mechanism of action of Astragalus propinquus Schischkin and Panax notoginseng ( A&P ) compound in a mouse model of diabetic nephropathy ( DN). Methods Fifty male C57BL/ 6 mice were randomly divided into normal, model, L-A&P, H-A&P and irbesartan groups. Mice were fed a high-fat, high-sugar diet for 2 months, and streptozotocin 50 mg / (kg·d) was intraperitoneally injected for 5 days to induce DN. Different gavage treatments were given to the groups. After 4 weeks of treatment, mice were killed and materials collected. Renal pathology was observed using hematoxylin-eosin and periodic acid – Schiff stained sections. Renal function and the expression of Arid2-IR lncRNA, inflammatory factors [ tumor necrosis factor-α ( TNF-α), interleukin-1β (IL-1β), IL-6], and NF-κB and its downstream signaling molecules were examined. Results The 24 h urine protein, serum creatinine, and urea nitrogen levels were significantly decreased in A&P-treated mice compared with the DN group, which suggests that renal function recovered to a certain extent. Renal pathology was significantly improved, and IL-1β, IL-6 and TNF-α expression decreased in a dose-dependent manner. Expression levels of Arid2-IR and NF-κB and its downstream molecules ( COX2, IL-6) were down-regulated, indicating that activation of the NF-κB signaling pathway was inhibited. Conclusions A&P reduced kidney inflammation in mice with DN, and its mechanism of action may be related to the regulation of Arid2-IR/ NF-κB signaling.

Abstract:Drug toxicity has, in significant measure, affected the discovery and application of new pharmaceutical drugs. Zebrafish have been increasingly used as a biological model in drug toxicology research. Here, we reviewed the application of the zebrafish model in drug toxicity research in a number of fields ( including cardiotoxicity, hepatotoxicity, nephrotoxicity, neurotoxicity, genetic and intergenerational toxicity, and teratogenic toxicity). The relative advantages and disadvantages were summarized, thus providing a basis for further application of the zebrafish model in drug toxicity research.

Abstract:Parkinson’ s disease ( PD) is a serious neurodegenerative disease. Typical pathological changes associated with PD are cellular necrosis and the loss of dopaminergic neurons in many areas of brain, as well as the appearance of multiple dyskinesias. Although the etiology of PD has not been clarified, the use of animal models plays a vital role in investigating the mechanisms of PD. In this article, we review the characteristics of different animal models of PD, discuss the characteristics of different animal species, and compare the clinical differences between animal models and PD patients to provide fundamental support for future PD research.

Abstract:Multiple sclerosis is a chronic inflammatory demyelinating disease of the central nervous system, and a serious disease in the clinical neurological setting. The basic pathological features of MS include inflammatory infiltration, primary demyelination and glial cell activation. Appropriate animal models are necessary for the study of pathogenesis and pathological processes, and for the screening and evaluation of therapeutic drugs. In this review, we will introduce the pathological features and pathogenesis of animal models, as well as the specific MS problems that different models can be applied to.

Abstract:Miniature swine provide valuable research models for cardiovascular disease because their anatomical structure and physiological features are similar to those of human. Several studies have been published in which the miniature swine models of atherosclerosis have been established successfully. These models included general method of a high-fat diet plus balloon injury, as well as new method, such as gene editing-induced familial hypercholesteremia, and local intravascular delivery of lipid and inflammation factors. However, few of these models have been widely used. Long time consumption, high cost and huge heterogeneity among studies are problems that restrict application of these AS models. This article summarizes the breeds of miniature swine, the method used to establish atherosclerosis models, and the advantages and disadvantages of current models.

Abstract:Tic disorder is a common neurodevelopmental disease in childhood, which is mainly manifested as involuntary repetition, sudden movement and / or phonation, and may be accompanied by attention deficit hyperactivity disorder, learning disorder, obsessive-compulsive disorder, conduct disorder, etc., seriously affecting the children’ s learning, social adaptability and psychological healthy development. However, its etiology, pathogenesis and pharmacological mechanism have not been fully clarified, and these basic studies need to be further explored and verified through animal models. At present, there are a variety of construction method of tic disorder models, mainly based on the possible pathogenesis of the disease, such as neurotransmitter abnormalities, immune dysfunction, genetic factors, etc., but there is no animal model that can completely meet the manifestations of tic disorder. This paper summarizes the commonly used animal models of tic disorder, in order to provide a reference for experimental studies.

Abstract:Hemophilic arthropathy is caused by spontaneous intra-articular hemorrhage and repeated intra-articular hematomas. Chronic pain, joint damage and decreased quality of life are the main clinical manifestations of hemophilic arthropathy. The biological mechanisms are not fully understood. Current translational animal models of hemophilia are of great value in determining the pathophysiology of the disease and its complications. Therefore, a hemophilic arthropathy mouse model was established to further investigate the pathophysiological mechanisms of hemophilic arthropathy as well as potential intervention treatments. A murine model of hemophilia provides advantages as an animal model for research because of easy transportation and easy construction,and its constructed hemophilic arthropathy model also provides the basis for future clinical research. This review will describe the construction and investigation of a mouse model of hemophilia arthropathy.