Abstract:Objective To observe the effect of the folk medicine Wubao capsule on the imbalance of Th1 / Th2, Th17 / Treg, and chemokines (Eotaxin, IL-8, MCP-1) in bronchoalveolar lavage fluid (BALF) of asthmatic mice induced by ovalbumin (OVA). Methods BALB/ c mice were randomly divided into five groups (Control group, Model group, Dexamethasone group, Wubao capsule low-dose group, Wubao capsule high-dose group). The asthma model was replicated by OVA sensitization and challenge. The pathological changes of lung tissue were observed by HE and PAS staining. The number of inflammatory cells in BALF was observed by rapid Wright-Giemsa staining. The levels of IFN-γ, IL-4, IL-17A, IL-10, Eotaxin, IL-8, and MCP-1 in BALF were determined by ELISA. Results Wubao capsule can significantly alleviate inflammatory infiltration and mucus secretion in lung tissue of asthmatic mice, reduce the number of various inflammatory cells and the production of IL-4, IL-17A, Eotaxin, IL-8, and MCP-1 in BALF (P<0. 01, P<0. 05), and significantly increase the levels of IFN-γ and IL-10 (P<0. 01, P<0. 05). Conclusions Wubao capsule may play a role in preventing and treating asthma by regulating the balance of Th1 / Th2 and Th17 / Treg-related cytokines and inhibiting the production of the chemokines Eotaxin, IL-8, and MCP-1 in BALF.

Abstract:Objective To observe the effect of fresh yam extract on the ulcer index, mucosal morphological structure, expression of serum superoxide dismutase ( SOD) and glutathione peroxidase ( GSH-Px ), malondialdehyde (MDA), and nitric oxide (NO) in mice with gastric ulcers and explore the mechanism of its prevention and treatment of gastric ulcers. Methods We randomly divided 100 mice into blank control, model, teprenone, and high and low dose fresh yam extract groups. Normal saline was administered to blank control and model groups, and experimental drugs were administered to the other three groups for 45 days. Then, the gastric ulcer model was established by one intragastric administration of anti-inflammatory pain (2. 5 mg / mL) in the groups except the blank control group. All mice were sacrificed after 7 h. The activities of SOD and GSH-Px, and NO and MDA in serum were detected by blood sampling. The ulcer index was measured, and the ulcer inhibition rate was calculated. Pathological sections of gastric tissue were prepared from the whole stomach and stained with HE, and morphological changes of the mucosal tissue were observed. Results Compared with the model group, high dose, low dose, and teprenone groups had reduced gastric ulcers caused by indolence pain, and inflammatory cell infiltration and destruction of the glands in gastric tissue were less. In the high dose group, MDA and NO were decreased in serum, and expression of SOD and GSH-Px was increased (all P<0. 05). Conclusions Fresh yam extract prevents acute gastric ulcers and protects the gastric mucosa, and its mechanism may be related to reducing the serum contents of MDA and NO, and increasing the expression of SOD and GSH-Px for anti-oxidative stress effects.

Abstract:Objective Transverse aortic constriction (TAC) was used to establish a stable and reliable model of heart failure in mice, and the pathological process of heart failure in mice was analyzed and observed. Methods The heart failure model of C57BL/ 6J mice was established by non-thoracotomy TAC. The heart function and degree of heart failure were evaluated by echocardiography, left ventricular mass index, and histopathological examination. Results The left ventricular mass index gradually increased overtime. Compared with that in the sham-operated group, there were significant differences in left ventricular mass index at 4, 8, and 12 weeks after operation (P< 0. 01). Echocardiography showed that the left ventricular anterior wall thickness, left ventricular corrected weight, heart rate, and cardiac output increased significantly 4 weeks after TAC compared with those in the sham operation group ( P < 0. 01). Left ventricular wall thickness, left ventricular corrected weight, and left ventricular end systolic volume increased significantly ( P< 0. 01), while ejection fraction and fractional shortening decreased significantly (P<0. 01). At 12 weeks after TAC, left ventricular enddiastolic diameter, left ventricular endsystolic diameter, left ventricular enddiastolicvolume, and left ventricular endsystolic volumeincreased significantly ( P< 0. 01), ejection fraction and fractional shortening decreased further ( P < 0. 01), and pathological examination showed that myocardial fibers were abnormal tovarying degrees and lesional. Conclusions The experimental result show that the establishment of a mouse heart failure model by non-thoracotomy TAC is stable and feasible, and can simulate the pathophysiological process from left ventricular hypertrophy to heart failure caused by human pressure overload

Abstract:Objective Although the clinical implementation of antiretroviral therapy ( cART) has significantly prolonged the lifetime of AIDS patients, HIV-associated neurocognitive disorders (HANDs) remain a critical concern in a considerable number of HIV-positive individuals. HIV-1Tat protein is a viral protein released from HIV-infected cells, which may be one of the underlying mechanisms behind HANDs. Methamphetamine (METH) can penetrate the blood-brain barrier, damaging nerve cells and increasing the risk of HANDs. The combination of HIV-1Tat and METH can induce autophagy and apoptosis in nerve cells, but the regulatory mechanism behind this has remained unclear. Methods In this study, the primary midbrain neurons of tree shrews were used to determine the expression of ATG5 / 7 proteins and genes induced by HIV-1Tat and METH via Western blot and real-time PCR. Results HIV-1Tat protein and METH induced autophagy in tree shrews’ primary midbrain neurons and had a synergistic effect. ATG5/7genes were involved in this autophagy. Conclusions ATG5/7 genes are involved in HIV-1Tat protein and METH-induced autophagy of nerve cells, which provides a new theoretical basis for studying the relationship between HANDs and autophagy.

Abstract:Objective To evaluate a real-time fluorescence quantitative PCR of dengue virus by detection of key indicators in the dengue mouse model. Methods Appropriate specific primers and probes were selected, the molecular method was used to prepare the plasmid standard, and the PCR system and reaction conditions were optimized. Then, the sensitivity, specificity, and stability of the real-time fluorescence quantitative PCR method were evaluated. Finally, this method were tested by using serum and tissue samples from dengue virus-infected mice. Results One-step real-time fluorescence quantitative PCR was performed successfully. The method was sensitive, and the lowest detection limit was 49. 6 copies/ μL. The method was specific, and non-specific amplification was not observed. The method repeatability was good, the standard deviation of sample Ct values was less than 0. 5, and the CV was <5%. The test result of samples from the dengue virus-infected mice were in line with experimental expectations. Conclusions The qRT-PCR method can be used for quantitative detection and evaluation of dengue virus in a mouse model.

Abstract:Objective To construct a Luc-GFP-labeled human hepatocellular carcinoma ( HCC) cell line with high metastatic potential. In addition, to establish a spontaneous metastasis and conveniently monitored orthotopic model of hepatocellular carcinoma through a cell suspension injection method or a tumor fragment implantation method in nude mice. Methods The recombinant lentiviral expression vector pCDH-CMV-Luc-EF1-GFP-Puro was constructed and transfected into the human hepatoma cell line HCCLM3. The orthotopic xenograft model was established through the cell suspension injection method and the tumor fragment implantation method. The growth and metastasis of the tumors were observed by in vivo imaging and pathological analysis. Results HCCLM3-Luc-GFP, a highly metastatic HCC cell line with GFP expression and Luc activity, was obtained. The tumorigenic rates of the two method were 100%. However, the former can simulate the microenvironment of liver cancer patients, with host factors having less influence. The tumor size and location of the latter are easier to control, with the lung metastatic rate being lower in the former than in the latter. Conclusions The Luc-GFP-labeled highly metastatic human hepatoma cell line HCCLM3-Luc-GFP was successfully constructed. The orthotopic model of highly metastatic and Luc-GFP-labeled HCC in nude mice was successfully established by the above method. This study provides a continuous and effective means to monitor the growth and metastasis of tumors in vivo, for evaluating the efficacy of anti-metastatic drugs against HCC.

Abstract:Objective To investigate the anti-fatigue effect of hairy antler composition and its mechanism. Methods NIH mice and SD rats were randomly divided into a blank control group, Yihua capsule group (600 mg / kg), and three dosage groups of hairy antler composition ( 135, 270, and 810 mg / kg). The Yihua capsule, hairy antler composition, at various concentrations or water were administered orally to the corresponding groups once a day for 30 days. Serum BUN and blood lactic acid after swimming, loaded swimming time, and hepatic glycogen content were measured at 30 min after the last administration. Results The hairy antler composition significantly increased the loaded swimming time and the hepatic glycogen content in mice (P<0. 05) and decreased blood lactic acid (P<0. 05). Conclusions The hairy antler composition has an anti-fatigue effect.

Abstract:Objective Pass-through box, also called pass-through chambers and pass-through windows, allow for the transfer of parts and equipment into and out of a barrier facility. The common use of pass-through box is to sterilize the materials under the exposure of UV radiation. The traditional UV lamps need long time exposure, but the effect is not stable. A xenon (Xe) arc lamp is a specialized type of gas-discharge lamp that produces light by passing electricity through ionized xenon gas at high pressure and emit radiant energy within the UV-C band. In this study, the bactericidal efficacy of ultraviolet pass box and high-throughput xenon ray pass box was compared at indicated time points. Methods After the exposure of the two types of devices at indicated time points respectively, the effects on the staphylococcus aureus, pseudomonas aeruginosa, escherichia coli, candida albicans and bacillus subtilis were tested. Results The efficiency of the xenon arc lamps was 99. 85% with 1-min exposure and 100% with 5-min exposure; The sterilization efficiency of traditional UV lamps below quality in 5 minutes, and the sterilization efficiency of different bacterias varied greatly. However, the sterilization efficiency is about 99. 91% after 30-min irradiation duration. Conclusions The efficiency of the xenon arc lamps can achieve bactericidal effect in 5 minutes, and could replace the UV lamps in the future

Abstract:Objective To analyze the expression of IL-18 and changes of intestinal flora in tree shrews under the action of D-galactose. Methods Ten male tree shrews were randomly divided into normal and D-galactose groups (n = 5). Intraperitoneally injected D-galactose (0. 6 g / kg) for 8 weeks was applied to the D-galactose group. Tree shrews in the normal group were injected with normal saline in the same manner. The ileum of the small intestines and the cecum and colon of the large intestines were separated, and their contents were collected. Expression of IL-18 was detected by immunohistochemistry. The bacterial 16S rDNA sequence in intestinal contents was analyzed by high-throughput sequencing. Results Immunohistochemistry showed that the expression of IL-18 in the ileum, cecum, and colon was significantly increased in the D-galactose group P<0. 05). The abundances of enterobacteriaceae and aeromonas in the cecum and colon were increased significantly in the D-galactose group. The abundance of bifidobacteriaceae was decreased in the colon of the D-galactose group. Conclusions D-Galactose increases the expression of intestinal inflammatory factor IL-18 and causes intestinal flora disorder, suggesting that the imbalance of intestinal flora in tree shrews caused the increased expression of IL-18 and promoted the intestinal inflammatory response

Abstract:Objective To investigate the effects of umbilical cord-derived mesenchymal stem cells on the expression of sex hormones CYR61 and CTGF in rabbits with premature ovarian failure, and to reveal the treatment effects and associated mechanism. Methods Ten rabbits were randomly divided into a model group and a treatment group. The model of premature ovarian failure ( POF) was established by intraperitoneal injection of CTX [ 50 mg / ( kg·d), 2 consecutive days]. The treatment group was treated with UCMSCs (5×106 / ml, 2 ml each time for 3 days), and the model group was injected with the same amount of sterile water. Venous blood was taken at 0, 7, 14, and 28 days after UCMSC treatment, and analyzed for changes in hormone levels by ELISA. Ovarian tissue samples were taken 28 days after UCMSC treatment to detect CYR61 and CTGF proteins and their associated mRNA expression. Results There were no differences in E2 and FSH levels between the two groups before treatment (P>0. 05). After treatment, E2 level in the treatment group increased and the level of FSH decreased. Compared with the model group, the difference was statistically significant (P< 0. 05), and in the treatment group, the level of sexual hormone fluctuated periodically with time. Meanwhile, the model group rabbits always had a low E2 level and a high FSH level, and they did not show periodic fluctuations over time. Also, compared with the model group, the expression of CYR61 and CTGF in the treatment group was significantly increased, and the difference was statistically significant (t_CYR61 = 8. 688, P<0. 05; t_CTGF = 10. 142, P<0. 05). In addition, compared with the model group, the expression levels of CYR61 and CTGF mRNA in the treatment group were significantly increased, and the differences were statistically significant (t_CYR61= 38. 156, P<0. 05; t_CTGF = 14. 646, P<0. 05). CYR61 and CTGF were mainly expressed in the granulosa cells and follicular cell membranes. Compared with the model group, this expression of the treatment group was significantly increased, and the difference was statistically significant ( t_CYR61 = 3. 981, P<0. 05; t_CTGF = 4. 111, P<0. 05). Conclusions Umbilical cord-mesenchymal stem cell therapy can improve sex hormone levels, and upregulate the expression of CYR61 and CTGF in rabbits with POF, thereby activating sleep follicles, restoring ovarian function, and treating POF.

Abstract:Objective To investigate the effects of thrombopoietin on adriamycin-induced cardiomyocyte apoptosis in rats with heart failure based on the TGF-β1 / Smad3 signaling pathway. Methods Rats were randomly divided into control, model, treatment, and inhibitor groups. Rats in the model, treatment, and inhibitor groups were injected with doxorubicin to establish a rat heart failure model. Rats in the treatment group were intraperitoneally injected with 5 μg / kg thyroperoxidase (TPO) daily and rats in the inhibitor group were intraperitoneally injected with 1. 2 mg daily / kg SB431542. TUNEL staining, immunofluorescence, and western blot were used to detect the cell apoptosis of cardiomyocytes, and the expressions of caspase-3, TGF-β1, and p-Smad3. Results Compared with the control group, the cell apoptosis rate of myocardial tissue cells in the model group was significantly increased and the expressions of TGF-β1, p-Smad3 and caspase-3 were significantly increased. Compared with the model group, the apoptosis rate of myocardial tissue in the treatment and inhibitor groups and the expressions of TGF-β1, p-Smad3, and caspase-3, were significantly decreased. The differences were statistically significant compared with the model group ( P < 0. 05). Conclusions TPO reduced the apoptosis rate of cardiomyocytes induced by doxorubicin in rats. The mechanism may be related to the inhibition of the TGF- β1 / Smad3 signaling pathway

Abstract:Objective To study the protective effect of Ginkgo biloba extract 50 (GBE50) on vascular endothelial cells in a high glucose environment. Methods MTT assays were utilized to examine the HUVECs cell viability after the treatment of GBE50, changes in glutathione content were assessed by using the 7-amino-4-chloromethylcoumarin (CMAC) fluorescent stain technique, the effects of reactive oxygen species and GBE50 on autophagy were investigated by flow cytometry. Results MTT assay result showed that GBE50 significantly improved the viability of HUVECs in a high glucose environment. Flow cytometry showed that ROS-positive HUVECs were increased significantly in the high glucose environment, but GBE50 significantly reduced ROS production. GBE50 significantly increased the GSH content in HUVECs in the high glucose environment and decreased the ratio of MDA-positive cells. Immunofluorescence and western blot result showed that GBE50 significantly decreased the level of autophagy in HUVECs. Conclusions GBE50 reduces the effects of a high glucose environment on the activity of vascular endothelial cells by reducing autophagy and ROS production

Abstract:Objective Animal models of low immunity were established in mice using a simple and rapid method. Methods Cyclophosphamide was administered orally at a dose of 100 mg / Kg, hydrocortisone was administered orally at a dose of 80 mg / Kg, a 60% ethanol solution was administered orally at a dose of 20 ml / kg, and skin was irradiated by UVA (long wave) at a dose of 10 J/ cm2 . Each group was treated with medicine for 3 consecutive days. The body weight of mice was measured before and after the test, and the levels of IgG, IgA, and IgM in the serum of mice was determined. In mice, the proportions of CD19+ B cells in peripheral blood lymphocytes and CD3+ T cells in peripheral blood lymphocytes were measured by flow cytometry. The peripheral blood cells of mice were analyzed by an automatic hematology analyzer. Results Immunosuppressive mouse models using cyclophosphamide or hydrocortisone were established. Immunosuppressive models using 60% ethanol or ultraviolet irradiation were not established, and require further study. The body weight of mice, the levels of IgG, IgA, and IgM in the serum, B and T lymphocytes in peripheral blood, and the routine analysis of blood in mice can be used as reference indexes for the establishment of models of low immunity in mice. Conclusions A model of immunosuppression established in mice by the oral administration of cyclophosphamide at 100 mg / kg for 3 days was established. Regarding the various detection indexes, body weight, serum immunoglobulin content, and routine blood analysis are simple to perform, rapid, and of low cost with a high detection rate. These can be used as indexes for the routine detection of established animal models

Abstract:Objective To evaluate matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for detection of pathogenic bacteria in laboratory animals. Methods In total, 40 standard strains and 384 laboratory animal isolates were identified using the Bruker Biotyper system. Results The accuracy rates of Bruker Biotyper to detect standard strains and isolates were 90. 0% and 80. 0%, respectively. There were biases in the identification of Salmonella typhimurium, Yersinia pseudotuberculosis, Shigella spp., Klebsiella pneumoniae, Pasteurella pneumotropica biotype Heyl, and Bordetella bronchiseptica. Conclusions The MALDI-TOF MS method is highly efficient and accurate. It can be used for rapid identification of pathogenic bacteria in laboratory animals

Abstract:Objective To analyze the modeling elements of asthma animal models and provide a method ological reference to improve the success rate of modeling and evaluating test drugs. Methods Using asthma and animal models as key words, we searched for related literature in the China Knowledge Network ( January 2009 to August 2019), recorded experimental animal species, stimulation method , sensitization method , and detection indicators, established a database, and conducted statistical analysis. Results There were 118 articles in the journals, and the most used experimental animals were BALB/ c mice (43 times, 36. 4%) and SD rats (42 times, 35. 5%). The most exciting mode is atomization (102 times, 86. 4%). The most used sensitization method was sensitization by ovalbumin (OVA) (107 times, 90. 6%). The most detected indexes were lung histopathology ( 92 times, 77. 9%), serum biochemistry indicators ( 49 times, 41. 5%), cell counts in alveolar lavage fluid ( 42 times, 35. 6%), immunohistochemistry of lung tissue ( 40 times, 33. 9%), and biochemical indicators in alveolar lavage fluid (39 times, 33. 1%). Conclusions BALB/ c mice or SD rats were used as experimental animals to establish asthma animal models. The sensitization agent was atomized, and OVA was used as the sensitizing antigen to improve the success rate of the model. Lung histopathology, serum biochemical indicators, and cell counts in lavage fluid were used to effectively evaluate test drugs and provide a basis for better experimental animal research in asthma

Abstract:The management of biosafety laboratories is usually based on both officials and veterinarians, but sometimes the management is not clear enough to continue many facilities, and experiments using various animals have increased in recent years. The new situation has caused difficulties with facility management, animal management, risk control, and care of animals infected with pathogens. In this review, comments and suggestions are provided for discussion

Abstract:Graves’ disease ( GD) is a common organ-specific autoimmune disease, and its pathogenesis and etiology are not very clear. Currently, it is generally believed that the pathogenesis of GD is related to genetic, immunological and environmental factors. In recent years, a variety of immune cells have been found to be involved in the pathogenesis of GD, which provides a new idea for the diagnosis and treatment of GD. Various studies have shown that abnormal expression of helper T cells, regulatory T and B cells as well as abnormal secretion of cytokines, are related to the pathogenesis of GD. Therefore, this article reviews the role of helper T cells, regulatory T cells and regulatory B cells in GD

Abstract:Nrf2, Keap1, p62, NLRP3 inflammasome, autophagy

Abstract:Chronic obstructive pulmonary disease(COPD) is a kind of airway inflammatory disease, which has a high morbidity and mortality, and brings heavy burden to patients and society. And its pathogenesis and treatment methods have not been thoroughly studied at present. Although animal models can only simulate part of the characteristics of the disease, they are indispensable for further research on the physiology, pathology and treatment of COPD. This paper summarizes the animal models, modeling methods and measurement indexes of animal models of COPD in combination with the literatures relevant to COPD modeling in recent three years in PubMed

Abstract:Heart failure is the final stage of cardiovascular disease development, and heart failure treatment strategies need to be explored. Cyclic adenosine monophosphate ( cAMP) and cyclic guanosine monophosphate ( cGMP) are second messengers that regulate signal transduction, which are hydrolyzed by a group of distributed and functional specific phosphodiesterases (PDEs) in cardiomyocytes, which play key roles in the pathogenesis of normal cell activity and heart failure. The efficacy of PDE3 and PDE5 inhibitors has been clinically recognized, and clinical and basic researches are constantly updating for their mechanisms of action. Here we summarize these research advances, which in turn helps us to understand the strategies and research directions of heart failure treatment

Abstract:Frozen shoulder is a chronic, self-limited disease of shoulder joints. However, the etiology and pathogenesis are still unclear. Its main pathological changes are shoulder capsule contracture, thickening, fibrosis, and chronic synovitis. Recently, animal models have had an important role in investigating the pathogenesis, treatment, and prevention of frozen shoulder. Animal models using rats, mice, dogs, and rabbits with different modeling method have been widely used for frozen shoulder research by domestic and foreign scholars. However, each animal model of frozen shoulder has advantages and limitations based on specific research interests. In this paper, the modeling method of several typical frozen shoulder animal models are reviewed and discussed in detail. Research progress in the pathogenesis of frozen shoulder disease based on these method is also discussed. This review is expected to provide useful ideas for basic research and clinical medical exploration of frozen shoulder