Abstract:[Abstract] Objective: To investigate the effects of QDPR on regulating the autophagy of HEK293T cells. Methods：HEK293T cells were transiently transfected with recombinant plasmid DNA rQDPRwt and recombinant plasmid DNA rQDPRmut by calcium phosphate. After 72h, the expression of rat QDPR in 293T cells was detected by Western Blot. Then, the effects of QDPR on autophagy related gene ( including LC3 and Beclin1) were analyzed by RT-PC R, and Western blot was used to monitor the changes in autophagy associated protein level. Results: 1) The recombinant plasmid DNA rQDPRwt and recombinant plasmid DNA rQDPRmut were successfully constructed. 2）The fusion protein can express in HEK293T cell. 3) Compared with control vector group, the mRNA expression of LC3 was significantly up-regulated in rQDPRwt group (p<0.05), and the mRNA expression of Beclin1 showed no significant difference among the 3 groups (p>0.05); 4）The Western blot analysis revealed that LC3-II and Beclin1 increased in rQDPRwt group when compared with control group, and there were no difference of protein levels among the 3 groups. The protein expression of LC3-I，II and Beclin1 showed no significant difference between rQDPRmut group and control group. Conclusion: QDPR may activate the autophagy of HEK293T cells by increasing the expression of autophagy associated genes of HEK293T cells.

Abstract:Objective To explore the histological difference of spleen in BALB/c nude mouse with green fluorescent protein (GFP) at different ages, and compared with the histology of spleen in BALB/c nude mouse. Methods The spleens were gained, respectively, from 32 BALB/c nude mice with GFP and 32 BALB/c nude mice at assigned ages. The weight of spleen and spleen index were measured; the histology changes of the spleen were observed under light microscope. The sum of lymphocyte cells were counted and analyzed. Results The spleen index of 28 days BALB/c nude mouse with GFP are higher significantly than 14 days. Compared with 14 days BALB/c nude mouse with GFP, the sum of lymphocyte cells of 49 days, 70 days are reduced significantly. The sum of lymphocyte cells of 14 days, 28 days, 49 days, 70 days BALB/c nude mouse with GFP are higher significantly than 14 days, 28 days, 49 days, 70 days BALB/c nude mouse. Conclusion The GFP gene had some effect on the development and the immune function of the spleen in the BALB/c nude mouse with green fluorescent protein (GFP) at different ages.

Abstract:Objective To investigate the feasibility and effect of rat epidural anesthesia models by modifying fixation of the epidural catheter. Methods 24 male Wistar rats, used to build epidural anesthesia models, were randomly divided into experimental group (n = 12) and control group (n = 12). After inserted the catheter, the catheter were fixed by some blocks designed with sterile adhesive tape and the catheter connector were replaced by sterile medical tape in the experimental group, while catheter was fixed with wire wound and catheter connector was sutured on the back of the neck in the control group. 0.1% ropivacaine 30ul was injected into epidural space once a day and continued 28 days. Then incidence of postoperative infection, the time to failure of the epidural anesthesia models was recorded and the epidural catheter depth measured on the 28th day. Results No significant difference was noted in the incidence of postoperative infection between two groups (P>0.05). The effective duration of models in the experimental group is significantly longer than that in the control group(26.2±1.7d & 18.5±3.0d，P<0.05). The epidural catheter depth were 1.81±0.07cm in the experimental group, respectively, whereas in the control group, 1.44±0.55cm of the depth were detected, the degree of dispersion is lower than that in the control group(P<0.05). Conclusion Modified catheter fixation can significantly improve the stability and success rate of epidural anesthesia animal models.

Abstract:Objective To develop a standardized ELISA kit for mouse Sendai virus and test it in clinical application. Methods Sendai virus seed batches and BHK-21 cell banks were set firstly; then the crafts for virus production and antigen purification were established; the detection system in ELISA flat was also optimized; the positive and negative serum controls were normalized, a critical concentration positive serum was also set in this system. A totally 672 sera samples were analyzed by the current ELISA kit, and the results were confirmed by IFA and Western blot. Results the Sendai virus seed batches stored at -80℃ were stable for more than 6 months; the virus production craft and antigen purification craft improved the antigen quality; the control systems reduced the effect from environment. The detection sensitivity of ELISA kit is higher than Western blot, much higher than IFA. Conclusion the standardized mouse Sendai virus detection ELISA kit can detect the Sendai virus infection condition, and the detection result is stable and reproducible.

Abstract:Objective To construct pEGFP-C1-BAFF, pEGFP-N1-BAFF, and pIRES2-EGFP-BAFF recombinant plasmids, and to generate establish a SLE disease model of BAFF overexpression zebrafish by microinjection and fluorescence screening. Methods We cloned BAFF full length cDNA encoded by 807bp nuclear acid using spleens RNA of zebrafish by RT-PCR and constructed 3 types of BAFF overexpression plasmids: pEGFP-C1-BAFF,Objective We construct pEGFP-C1-baff, pEGFP-N1-baff, and pIRES2-EGFP-baff recombinant plasmids, and establish a SLE disease model of Baff overexpressed zebrafish by microinjection and fluorescence screening. The aim is to evaluate the meaning of Baff overexpression zebrafish in study of SLE mechanism and drug selection. Methods We cloned baff full length cDNA encoded by 807bp nuclear acid using spleens RNA of zebrafish by RT-PCR and constructed 3 types of baff overexpression plasmids: pEGFP-C1-baff, pEGFP-N1-baff and pIRES2-EGFP-baff. Baff protein expression was estimated by in-vitro cell transfection and Western blotting. Results 3 types of baff overexpression plasmids were examined by agarose gel electrophoresis. Western blotting and in-vitro transfection confirmed the expression of Baff-GFP fusion protein. We also obtained the baff over expression zebrafish by microinjection and fluorescence screening. Conclusion All the plasmid we constructed in our research have been estimated and could be used to generate the disease model of baff overexpression zebrafish which can be manipulated for immune-disease research. pEGFP-N1-BAFF and pIRES2-EGFP-BAFF. BAFF protein expression was estimated by in-vitro cell transfection and Western blotting. Results 3 types of BAFF over expression plasmids were examined by agarose gel electrophoresis. Western blotting and in-vitro transfection confirmed the expression of BAFF-GFP fusion protein. We also obtained the BAFF over expression zebrafish by microinjection and fluorescence screening. Conclusion All the plasmid we constructed in our research have been estimated and could be used to generate the disease model of BAFF overexpression zebrafish which can be manipulated for immune-disease research.

Abstract:Objective To establish a highly efficient method for the extraction, separation and purification of rat tail collagen. Methods The rat tail tendon has been obtained through stripping the rat tail; the rat tail collagen stoste has been developed through processing with Tris-HCl buffer, pepsin; sodium chloride solution has been repeatedly used for graded salting-out; acetic acid has been used for the purification of collagen. To get purified rat tail collagen, inorganic salt has to be removed with ultra-pure water dialysis. Appraisal is to be made through such technical means such as SDS-PAGE protein electrophoresis, amino acid content analysis. Results Rat tail collagen of high purity can be developed with the method established in this study, whose purity can reach electrophoresis level. When compared with those commercialized rat tail collagen products imported from overseas there is no difference. The effects the parameters for the extraction, separation and purification have exerted on yield and purity has been deliberated, established The optimal conditions for the extraction of rat tail collagen has been established, pepsin usage: 1:500, enzyme solution time: 72h, salting-out concentration: 2mol/L, the acid solution used for extraction: 0.05 mol/L acetum. Conclusion For rat tail collagen enlargement production to provide the proper processing parameters have been provided for the enlargement of production of rat tail collagen, for a large gain rat tail collagen and deeper efficacy research provides the theoretical support and practical basis have also been provided for the obtaining of large quantities of rat tail collagen so as to carry out deeper research on its efficacy.

Abstract:objective To generate the heart-specific KCNQ1V180L expression transgenic mice and an animal model for the study of KCNQ1 function and its effects on arrhythmia. Methods The transgenic vector was constructed by inserting the human KCNQ1V180L gene into the down stream of α-MHC promoter. The transgenic mice were created by the method of microinjection. The genotype of transgenic line was identified by PCR and the expression level of the gene was determined by Western Blot. The pathologic changes were analyzed with echocardiography and electrocardiography (ECG). Results Two lines of C57BL/6J transgenic mice with high levels of KCNQ1V180L expression were established. The heart of KCNQ1V180L transgenic mice showed thick ventricular wall, smaller ventricular chamber and increased fractionl shortening (FS%) compared with that of the non transgenic mice. Ventricular repolarization dysfunction was determined by ECG. Conclusions The KCNQ1V180L transgenic mice showed a similar phenotype with human long QT syndrome (LQTS). The transgenic mouse could be an useful animal model for the research of KCNQ1 gene function and the relationship between KCNQ1 mutation and arrhythmia.

Abstract:[Abstract] Objective Visfatin(also known as nicotinamide phosphoribosyltransferase or Nampt)is an adipokine associated with obesity ,but the relationship is unclear. The aim of the present study was to determine whether fat accumulation was related to visfatin level. Methods The expression levels of visfatin were detected with western blot from the adipose tissues of visfatin transgenic mice, wild type mice and knockout visfatin( /-) mice. The female mice were feed a high fat diet from 2 to 9 months of age. The body weight was recorded at 2, 4, 6, 8, 9 months of age. Then the mice were sacrificed at 9 months of age and the total weight of subcutaneous and visceral adipose tissue was scaled. The histopathology of visceral adipose tissue was observated under light microscope. Results The expression of visfatin in transgenic mice increased by 37% compared with that of wild type mice due to the transgenic expression of visfatin, while the expression level of visfatin in visfaitn( /-) mice decreased by 55% compared with that of wild type mice due to the one copy gene knockout of visfatin. After high fat diet for 7 months, the average body weight of visfatin transgenic mice was 27.8?.8g, wild type mice was 33.6?.1g and the konckout visfatin( /-) mice was 37.6g?.9g. The total weight of subcutaneous and visceral adipose tissue in visfatin transgenic mice decreased by 40% compared with that of wild type mice while knockout visfatin( /-) mice increased by 37% compared with that of wild type mice. Histopathology observation showed that visfatin transgenic mice had smallest size of visceral adipose cell and knockout visfatin( /-) mice had biggest size of visceral adipose cell. Our results indicated that the visfatin level in mice exhibited a negative effect on body weight, the total weight of subcutaneous and visceral adipose tissue, the size of visceral adipose cell. Conclusions our resulted suggested that the accumulation of adipose tissue was restrained by visfatin when mice were fed a high fat diet.

Abstract:Objective To establish a mink model of canine distemper virus, and different canine distemper virulent strain virulence was evaluated by the mink animal model for laying the foundation for the study of the canine distemper virus. Methods Different canine distemper viruses were separated from specimens ,cell-cultured, passaged and determined virulence of CDV. And we compared the virulence of different generation virulent strains, stable virulent strains were used for establishing mink canine distemper animal model and evaluated their virulence. Results we screened 3 canine distemper virulent strains and made dog animal experiments, the three viruses infected dogs and they showed a strong clinical symptoms, and mink animal model was established using different generations. Conclusion: we successfully established the canine distemper animal model and evaluated virulence of CDV.

Abstract:α-synuclein, a presynaptic protein, was found to be the major component of the lewy bodies in inherited and sporadic Parkinson’s disease（PD）. α-synuclein point mutations or genetic alteration can cause Parkinson’s disease（PD）. It is thought to be involved in the processes of neurodegeneration. This paper reviewed the toxic mechanisms of α- synuclein in Parkinson disease and α- synuclein transgenic animal models.

Abstract:【Abstract】 Objection To determine the optimal anesthetic method, our study compared the effect of three anesthetic methods in orthopedic animal experiments. Methods 80 experimental animals were randomly divided into three groups, group A was given with sumianxin II by intramuscular injection, group B was administrated with 0.6% pentobarbital sodium by intravenous injection, and group C was given with 0.6% pentobarbital sodium by introvenous injection combined with sumianxin II by intramuscular injection. Anesthesia was performed in orthopedic animal experiments surgery. Induction period of anesthesia (maintenance of anesthesia time), the recovery period, the anesthetic effect, mortality rates were observed after anesthesia. Results Anesthesia induction period is long, the effect is not good in group A; anesthesia induction period was short, but the bad effects of anesthesia and the mortality rate was high in group B; however, induction period of group C was short and safety. Conclusion pentobarbital sodium combined with sumianxin II is a good anesthetic method compared with pentobarbital sodium or sumianxin II respectively. Its induction period is short and safety. Therefore, it is an excellent and safe method to anesthetize animal in orthopaedic using Sumianxin II combined with pentobarbital sodium. 【Keywords】 Sumianxin II; pentobarbital sodium;experimental animal;anesthesia

Abstract:[Abstract] Objective: To explore the factors in duck hepatitis B virus infection model by different kinds of ducklings and observe the antiviral effect in models. Methods: Collecting duck sera , qualitative detecting viral DNA by PCR method and detecting the changes of viral DNA load by the quantitative PCR method in duck sera. Observing the antiviral effects in duck DHBV infection models after antiviral drug treatment. Results: Natural infection rate of duck DHBV is different, Cherry Valley duck for 8.75%, two batches in Hubei duck were 17.80% and 10.68% respectively. Both intravenous and intraperitoneal injection can cause ducklings infected with DHBV, infection rates of two ways were 80% and 65% respectiviely. Ducks infected with DHBV , the viral load is maintained at 106 ~ 108 copies / mL and persist for more than 20 days. Changing trends of antiviral effect in different DHBV models are coincident after treating by the antiviral drugs. Conclusion: Duck species and artificial routes of infection can affect DHBV infection rate, in vivo the viremia of duck is stably persistent after DHBV infection. Therefore, DHBV infection model is an available model in antiviral drug screening.

Abstract:【Abstract】The mouse model of middle cerebral artery occlusion (MCAO) is applied in the research of cerebral ischemia and reperfusion widely. In addition to measuring infarction volume, behavioral evaluation of functional neurological deficit is also an important indicator in estimating the severity of stroke. Existing tests are mostly transplanted from rat evaluation system. There are many different tests being used, and no unified standard. This review describes the advantage and disadvantage of most commonly used behavior tests in mouse model of ischemia.

Abstract:There are many problems in Virological Quality Monitoring of laboratory Animals,for example, monitoring items,test methods and samples, and frequency of monitoring. This article introduces briefly the health monitoring recommendation for virus detection of laboratory animals in China and foreign countries. In order to meet the needs for the development of the laboratory animal science, the author put forward some suggestions for the solution of problems in virus detection of laboratory animals in China.