Laboratory Animal Science

Expression Levels of Various Interferon mRNA and Their Receptors in CAV-1-infected MDCK Cells

2020, 37(03): 1.

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Objective To understand the expression of IFNs and its receptor molecules in Madin-Darby Canine Kidney (MDCK) cell line infected with canine adenovirus type 1 ( CAV-1 ) . Method Several SYBR real-time quantitative PCR( RT-PCR) method were established to detect IFN-β, IFN-βreceptor. ( IFNAR1 ) IFN-λ1, IFN-λ1 receptor( IFN-λ1R) , IFN-λ3 and IFN-λ3 receptor( IFN-λ3R) in canine. The relative transcriptional levels of IFNs and its receptor molecules in Poly I: C treated-MDCK cells and CAV-1 treated-MDCK cells were analyzed. Result The transcriptional level of IFN-λ3R were significantly higher than that of other IFNs and receptor molecules in normal MDCK cells. The transcriptional levels of IFN-λ1R, IFN-λ3 , IFN-λ3R increased significantly in MDCK cells treated with Poly I: C for 24 h, while those of IFN-β, IFN-λ1, and IFNAR1 remained unchanged. However, the transcriptional level of IFN-α was the most highly than others in MDCK cells infected with CAV-1 for 24 h. Conclusion The result showed that there are different levels of various IFN molecules and their receptor transcripts in MDCK cells, and the express pattern varies after induction by. Poly I ：C or adenovirus.

Establishment of Animal Model of Balloon Injury in Rat Carotid Artery

2020, 37(03): 6.

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Objective To optimize the establishment of rat carotid artery balloon-injured model and to improve success rate in it. Method Twenty -five Sprague-Dawley ( SD) rats were randomly divided into three groups. The rats' left carotid artery in group A were deendothelialized by passing an inflatable balloon catheter ( size 2 mm, Cordis) . The rats' left carotid artery in group B were deendothelialized by passing an inflatable balloon catheter (size 1. 5 mm, Cordis) . The rats' left carotid artery in group C were made an arteriotomy without passing balloon catheter. The effect of balloon injury was assessed via Ivans ’ blue staining. Result The success rate in establishment of rat carotid artery balloon-injured model of group A is 45. 5% (5/11) . The success rate of group B is 90. 9% ( 10/11) ( P = 0. 032) . The difference of injured carotid artery intima area between group A and B is not significant ( P = 0. 426 ) . Conclusion The application of 1. 5 mm inflatable balloon catheter could successfully denude the intima of carotid artery in rats and could significantly improve the successful establish rate of rat carotid artery balloon-injured model.

Asiaticoside Modulates MIR-342-5p/AQP3 Axis Protection Against IL-1-Induced Chondrocyte Injury

2020, 37(03): 10.

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Objective To investigate the effect and mechanism of asiaticoside ( Ass ) on chondrocyte injury induced by interleukin 1p( IL-1p ) . Method After different concentrations of Ass were applied to ( IL-1 ) Ass on chondrocyte injury induced by interleukin 1p( IL-1p ) . echaniccellsapoptosis, enzyme-linked immunosorbent assay was used to detect the levels of interleukin-6 ( IL-6) and tumor necrosis factor-a ( TNF-α ) in cell culture supernatants, real-time quantitative PCR was used to detect the expression levels of miR-342-5p and aquaporin 3 (AQP3 ) mRNA, and Western Blot method was used to detect the expression levels of B lymphoma-2 ( Bcl-2) , B lymphoma-2 related protein ( Bax) and AQP3 protein. The double luciferase reporter experiment verified the targeting relationship between miR-342-5p and AQP3. After miR-342-5p inhibitor or AQP3 over expression vector was transfected into chondrocytes, the same method described above were used to detect the effect of inhibiting miR-342-5 porover expressing AQP3 on IL-1p-induced chondrocyte apoptosis and the expression of IL-6 and TNF-α. Result Ass could inhibit the apoptosis rate of chondrocytes induced by IL-1p and the expression of Bax protein, IL-6 and TNF-a and miR-342-5p ( P<0. 05 ) , and promote the expression of Bcl-2 protein, AQP3 mRNA and protein ( P<0. 05 ) . miR-342-5p negatively regulated AQP3 expression in chondrocytes. After inhibiting miR- 342-5p or over expressing AQP3, the apoptosis rate of chondrocytes induced by IL-1p and the expression of Bax protein, IL-6 and TNF-α were decreased ( P < 0. 05 ) , while the expression of Bcl-2 protein was increased ( P < 0. 05). Inhibiting AQP3 reversed the effect of inhibiting miR-342-5p on IL-1p-induced chondrocyte apoptosis and the expression of Bax, Bcl-2, IL-6 and TNF-α. Conclusion Ass can inhibit the apoptosis and inflammatory response of chondrocytes induced by IL-1rocyte apoptosis and the expression of Bax, Bcl-2, IL-6 and TNF-α. drocyte.

Research of the Establishment of Rat Model of Heart Failure by Abdominal Arterial Constriction

2020, 37(03): 19.

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Objective To establish rat model of heart failure by constriction of abdominal aorta, and to compare the effects of abdominal aorta narrowing with different diameters on hemodynamics and pathophysiology in rats. Method Wistar rats were randomly divided into three groups : sham-operated group ( group A ) ; narrowed abdominal aorta diameter up to 0. 45 mm ( group B ) ; narrowed abdominal aorta diameter up to 0. 7 mm ( group C ) ; observed the survival condition and mental state of the rats after the operation, at 4 and 8 weeks detected the interventricular septal depth ( IVSd )、left ventricular posterior walldepth ( LVPWd )、left ventricular end diastolic diameter (LVEDd )、left ventricular end systolic diameter ( LVEDs )、left ventricular ejection fraction ( LVEF ) by echocardiography, and left ventricular mass index( LVMI)、heart/body ratio、lung/body ratio were investigated after 8 weeks. Result The postoperative survival rate in group A is 100% , in group B is 55%, in group C is 70 % ; After 4 weeks, IVSd, LVPWd, LVEDs, LVEDd and EF in group A were significantly different from those in group B and C ( P 0. 05 ) . While after 8 weeks, IVSD、LVPWd、LVEDd、LVEDs、 LVEF in group B and C both had more significant changes compared with those in group A ( P< 0. 05 ) ;at 8 weeks LVMI、heart/body ratio、lung/body ratio were increased markedly in both group B and group C compared with group A ( P<0. 01) , meanwhile compared with group C , LVWI and heart/body ratio were much higher than those in group B ( P <0. 05 ) , and lung/body ratio increased more obviously ( P <0.01). Conclusion Coarctation of abdominal aorta diameter up to 0. 45 mm required shorter time, and has more efficient than narrowed up to 0. 7 mm in heart failure model establishment, it is an ideal method to establish rat model of heart failure.

Optimal Timing of L-Thyroxine Treatment for Subclinical Hypothyroidism in Rats during Pregnancy

2020, 37(03): 25.

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Objective To investigate the optimal timing of L-thyroxine ( L-T4 ) treatment for subclinical hypothyroidism ( SCH ) in rats during pregnancy. Method One hundred and fifty SPF Wistar female rats were randomly divided into 6 groups in accordance with the random number table, each with 25 cases, namely, control group, SCH group, hypothyroidism ( CH) group, group A, group B and group C. Enzyme-linked immunosorbent assay was used to determine the levels of serum thyroid-stimulating hormone ( TSH ) and total thyroxine ( TT4) in pregnant rats. The Morris water maze test was used to detect the learning and memory ability of rats offspring. Then the optimal timing of the treatment was analyzed. Result At P3 , P7 and P40, the weight of offspring of CH group was lower than that of the control group ( P<0.05). At P3 , the weight of offspring was higher in the SCH group than in the CH group ( P<0. 05 ) . At P7, the weight of offspring of CH group was lower than that of the group A and group B ( P<0. 05 ) . And the weight of offspring of CH group was lower than that of the group B and group C at P40 ( P<0. 05 ) . Compared with other groups, the control group had a significant decrease in the escape latency, which is steady and progressive. The distribution of the cortical and hippocampal nerve cells were disordered with unclear boundaries. Conclusion The established model rats reveal that the offspring of the SCH rats have abnormal weight and weakened learning and memory ability, while the conditions can be ameliorated by the L-T4 treatment before E13.

Effect of Shanzha Decoction on Nonalcoholic Fatty Liver in Rats

2020, 37(03): 31.

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Objective To observe the effect of Shanzha decoction on nonalcoholic fatty liver model in rats. Method Sixty Wistar rats were randomly divided into control group, model control group and Shanzha decoction (lower 14. 85 g shengyao/kg, middle 29. 7 g shengyao/kg, high 59. 4 g shengyao/kg dose group) . In the group, except for the blank control group, the rats were fed with high-fat diet for 8 weeks. The blank control group and the model control group were given distilled water by gavage, and the other groups were given the corresponding drugs for 6 weeks. At the end of the experiment, serum malondialdehyde ( MDA ) , superoxide dismutase ( SOD ), alanine aminotransferase ( ALT) , aspartate aminotransferase ( AST) , and triglycerides in liver tissue ( TG ) were measured in each group. The expression of hepatocyte growth factor and connective tissue growth factor in liver tissue was detected by immunohistochemistry. Result Shanzha Decoction significantly reduced liver index, serum MDA, ALT, AST and liver TG content in rats with fatty liver ( P <0. 05 or P <0. 01) , and significantly increased serum SOD activity in rats with fatty liver ( P <0. 05 or P<0. 01) , Shanqi decoction group can significantly reduce the expression of hepatocyte growth factor and connective tissue growth factor in liver tissue of rats with fatty liver ( P<0. 05) . Conclusion Shanzha decoction can be effectively used for the treatment of nonalcoholic fatty liver.

Comparative Study on the Key Points of Quality Control of Wheat Straw Used as Rodents Bedding

2020, 37(03): 37.

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Objective Compare and study the physical characteristics and sanitary indexes of the moisture content, hygroscopicity, heat preservation and hygienic of wheat straw ( after air explosion treatment) , corncob, shaving and paper, so as to provide scientific basis for the development and application of new type of wheat straw in laboratory animal bedding materials. Method According to the local standards of Beijing, the moisture content, water absorption and hygienic indexes of wheat straw bedding materials and other three bedding materials were tested. Result The moisture content and water absorption of wheat straw, the heavy metal residue of wheat straw and the wheat straw pad meet the standard. Conclusion Wheat straw bedding material has good water absorption and heat preservation performance, has the advantages of wide source, low price, easy preservation, and can reduce the consumption of forest resources, is conducive to environmental protection, and can be used as industrial promotion and development.

Animal Studies of Drug-induced Rhabdomyolysis

2020, 37(03): 43.

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Objective To explore the drug-induced rhabdomyolysis by coadministration of statin and fibrate in mice and rats to provide a reference for rhabdomyolysis caused by other reasons such as Haff disease. Method We administered simvastatin ( SMV) and gemfibrozil ( GF) to Wistar rats and ICR mice once daily for 14 days. The serum levels of Creatine Kinase ( CK ) , Myoglobin ( MYO ) and Alanine aminotransferase ( ALT ) , Aspartate aminotransferase ( AST) were tested before and after administration. Tissue samples from skeletal muscle were stained with hematoxylin and eosin ( HE ) and were observed under a microscope for routine pathological examination. Result The serum levels of CK, ALT and AST after SMV and GF administration to rats 10 days were prominently increased, and the muscle fiber necrosis suggested skeletal muscle-specific toxicity. Same drugs administered mice didn’ t show the increase in serum levels of biomedical parameters. Conclusion In this study, rhabdomyolysis was induced in rats by coadministration of the clinically using statin and fibrate. Wistar rats may beuseful to identify risk factors in the animal study of rhabdomyolysis such as Haff disease.

Establishment and Implementation of the Accreditation System for Laboratory Animal Institutions in China

2020, 37(03): 53.

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The accreditation system for laboratory animal institutions is a national accreditation system with laboratory animal welfare and ethics as the core evaluation content. From the social and economic background of the laboratory animal institutions accreditation system research and establishment, this paper introduces the historical process, the main research result, the content of the evaluation system framework and relevant supporting documents and the situation of the system operation of China accreditation system of laboratory animal institutions. By means of comparability analysis, the characteristics and advantages of China’ s accreditation system for laboratory animal institutions in terms of system design, evaluation content, evaluation result and supporting services are analyzed and emphasized. Up to now, 8 laboratories in China have obtained the accreditation qualification of this system. Although this system has been just starting, it has shown its unique role in strengthening the welfare and ethics of laboratory animals in China and promoting the development of science and technology of laboratory animals.

Experience in the Implementation of Telemetry System Implant Surgery in Rhesus Monkeys

2020, 37(03): 60.

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Objective To investigate the various conditions and treatment measures encountered in the perioperative period of implanted implants by using telemetry technology. Method The problems occurred in the implantation of 8 cases of macaques in this institution were classified, analyzed and summarized. Analyze the causes of preoperative anesthesia and skin preparation, difficulty in administration, intraoperative blood loss, anesthesia degree, postoperative incision and bruise, animal self-destructive suture or implanted lead wire, and propose prevention or solution. To be implemented, the prognosis of no infection or inflammation was confirmed by the result of weight, body temperature, blood routine and biochemical indicators before and after surgery, and by examining the postoperative cardiovascular system indicators and comparing with the basic values of the monkey in daytime, it was confirmed that the postoperative physiological state was good. Result Through the standardized surgical procedure and the perfect plan for coping with abnormalities, the implant implantation was successfully performed. The operation was successful and the postoperative recovery was good. The monkey’ s weight, blood routine, biochemistry, electrocardiogram, respiration, blood pressure, indicators such as body temperature showed no difference before and after surgery. Conclusion The surgical procedure of monkey application telemetry system is successfully established, and the success rate of telemetry animal model modeling is improved. It can be used as a reference for the application of domestic monkey telemetry model in safety pharmacology research.

tudy for Cryopreservation and Orthotopic Transplantation of Genetically Engineering Mice Ovary

2020, 37(03): 65.

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Objective To explore a method of cryopreservation and orthotopic transplantation of genetically engineering mice ovary of C57BL / 6 J background, establish a cryopreservation system. Method The ovaries from 10 d of C57BL / 6 J and three genetically engineering mice were freezed with DAP213 10 d fresh ovaries and thawed ovaries were grafed to the 4 weeks C57BL / 6 J and C57BL / 6 J( Cg) —Tyrc- 2 J / J( B6 albino) recipient, then mated with 10 weeks C57BL / 6 J male mice after 5 weeks breeding, observed offspring. Result The grafted C57BL / 6 J fresh ovaries and freeze-thawed ovaries and genetically engineering mice freeze-thawed ovaries could be restored reproduction, and gotten normal pups when mated with the same strain males. Conclusion Cryopreservation of C57BL / 6 J and genetically engineering mice ovaries with DAP213, were thawed and grafted to recipient, then mated with male, gotten normal pups. Cryopreservation of ovaries is a important method for mice resource bank.

Research advance of rodent animal endometriosis models

2020, 37(03): 72.

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Research advance of rodent animal endometriosis models

Detections Methods of Peripheral Neuropathy in Diabetic Mouse Models

2020, 37(03): 78.

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Diabetic peripheral neuropathy is a peripheral neurological dysfunction caused by sustained hyperglycemia that most commonly presents as distal degenerative neuropathy with abnormal pain, numbness and paresthesia and so on. Clinically, the diagnosis and treatment of DPN is still a great challenge in chronic complications of diabetes, which is related to its unclear pathogenesis. Rats have long been preferred in the rodent diabetes model, but advances in technology have gradually driven mice to become an important tool for the study of diabetic neuropathy. At present, there are many screening method for peripheral neuropathy in diabetic mice but no uniform standard. This paper summarizes the commonly used diabetic mouse models of diabetes and relevant method of detecting peripheral neuropathy including sensory function test, neuroelectrophysiology, pathological examinations and motor function test, aimed to provide a scientific reference for the researchers who conduct the peripheral neuropathy in diabetic mice.

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