Abstract:Object With the development of the stem cell, the study of rat stem cell became urgent. The purpose of this study was to generate EGFP transgenic rat models for fluorescence imaging, tracking of stem cells homing and cell transplantation. Methods The enhanced green fluorescent protein (EGFP) transgenic rats were obtained by microinjection and the expression of EGFP was determined under in vivo fluorescence imaging system and laser scanning confocal microscope. The expression of EGFP in blood cells, bone marrow cells and bone marrow stem cells was investigated with flow cytometry. Results The EGFP transgenic rat SD-TgN(CAG-EGFP-1)ZLFILAS was established. EGFP was systematically expressed in tissues of heart, liver, muscle, lung, pancreas, brain, bladder, stomach, kidney, intestines and spleen, and EGFP labeling ratios in blood cells and bone marrow stem cells were 94.4% and 97.8% respectively. Conclusions A systematic EGFP transgenic rat line was generated which showed EGFP labeling ratios higher than 95% in bone marrow stem cells. This transgenic rat may be a useful rat tool for fluorescence imaging and stem cells tracking.

Abstract:OBJECTIVE: To establish production management and the methods of microbiological quality control on breeding SPF chickens raised in the isolator. METHODS: The sera obtained from 10, 20, 40-week-old SPF chickens including introduced and self-breeding ones were used to monitor microbiological quality in order to detect the effectiveness of the current production management and the methods of microbiological quality control on SPF chickens. RESULTS: Because the hatching process was not strickly controlled during the early stage of breeding SPF chickens in the isolator, there were the emergence of microbiological contamination. Microbiological condition of the isolator breeding SPF chickens was effectively controlled after improving the operation management during the hatching process. CONCLUSION: If the disinfection during all breeding stages is strengthened, the microbiological quality of SPF chickens in the isolator could be ensured. The appropriate control on the density of SPF chickens in the isolator is not only beneficial to the breeding of SPF chickens and the recycling of hatching eggs, but also helpful to the control of environmental microbiology.

Abstract:Objective: Evaluate the IVF ( in vitro fertilization) rate of the post-thawed oocytes and the post-thawed punched oocytes of C57BL/6J fertilized with fresh or frozen sperm of 9 different strains. Methods: (1) The changes of nuclear, microfilament and zona pellucida glycoprotein-2 of C57BL/6J oocytes before and after cryopreservation were studied By immunofluorescence technique. (2)The IVF rate of C57BL/6J post-thawed oocytes or laser-punched oocytes fertilized with fresh or frozen sperms of 9 strains were compared.Results：(1) there were no obvious changes of nuclear before or after cryopreservation, microfilament were changed slightly when handled with cryopretectant or after cryopreservation. Zona pellucida glycoprotein-2 were hurted after cryopreservation or handled with cryopreservant ; (2) the IVF rate of fresh sperm of 9 strains with post-thawed oocytes were 17.6～42.9%(averaged 29.6%), which were significantly lower than the IVF rate of fresh sperm with laser-punched oocytes, 29.1～72.3% (average 49.7%); the IVF rate of post-thawed sperms of 9 strains with post-thawed oocytes were 5.4～23% (averaged 15.5%), also significantly lower than the IVF rate of post-thawed sperms fertilized with laser-punched oocytes, 16.7～48.6% (average 28.8%). Conclusions: (1) Cryopreservation would have a remarkable effect on microfilament and zona pullucida glycoprotein-2, but have not obvious effect on nuclear; (2) The IVF rate of oocytes would decreased remarkably after cryopreservation, but if the laser-punched system were used, the IVF rate would increased apparently. It showed that the laser-punched system would be useful for oocytes cryopreservation.

Abstract:Objective: to study and improve percutaneous puncture rabbit cisterna magna success rate method and the treatment for common problems.methods: using 2 ml aseptic syringe needle puncture cisterna magna of 30 New Zealand rabbit 2-2.5kg , to find the best puncture depth, angle, solve common problem. Results: 25 puncture success, the puncture depth in between 1.2-1.6 cm, an average of 1.432 cm, 5 failure. Discuss: rabbit cerebellomedullary cistern puncture don't limited to fixed depth, Angle, should be based on animal size, body posture timely adjust; In addition the right puncture needle can improve the success rate of puncture.

Abstract:【Abstract】 Objective: To establish the experimental ischemic vertigo rat model for the research of the anti-vertigo drugs. Methods: The rat model of ischemic vertigo was induced by the right common carotid artery (CCA) and subclavian artery (SCA) ligation. The escaped latency from electricity of rats was recorded to measure the degree of vertigo, and the blood flow of right vestibular nucleus was determined for 5 min before modeling and 30 min after modeling. The rate change of blood flow was calculated to evaluate the ischemic level. Results: 1.The latency of the model rats significantly prolonged when compared with the sham rats (P<0.01), which indicated that the experimental ischemic vertigo rat model was established. In addition, the latency of rats was remarkably decreased after treated with difenidol hydrochloride tablets and tianbaoning tablets, when compared with the model rats (61.9% and 52.0%, respectively, P<0.01). 2. The blood flow volume of right vestibular nucleus of the model rats reduced obviously after ligating the right CCA and SCA when compared with the sham rats ( P<0.01), which demonstrated the success of the model. The blood flow volume of right vestibular nucleus was significantly improved by treating with tianbaoning tablets after ligation, and the blood flow increased 92.8% at 5min, 83.8% at 10min, 77.3% at 15 min,77.9% at 20 min,65.7% at 25 min,55.8% at 30 min after ligation, when compared with the model group (P<0.01). Conclusion: The experimental ischemic vertigo rat model could be induced by the right common carotid artery and subclavian artery ligation, which can be used for the research on the mechanism of anti-vertigo drugs.

Abstract:【Abstract】 Objective To research the impact of PPARα agonist on liver and kidney function and blood fat index in PPARα transgenic mouse, and to detect these indices in the model if they can be applied to the study of pharmacodynamics. Methods Twenty-seven mice, at the age of 6 weeks, were divided into three group(each n=9), and fed with high-fat forage for a mounth.They were control group one，high dose group(60 mg/kg of fenofibrate),low dose group(30 mg/kg of fenofibrate).At the same time, 9 C57BL/6 mice were set as the control group two. The time of gavage administration lasted one month. The liver function index, kidney function index ,and blood fat index were tested before and after the drug administration. The growth of the mice was also observed. Results ①The comparison after fenofibrate administration :Compared with control group one, in the PPARα transgenic mouse with the fenofibrate dosage of 60 mg/kg、30 mg/kg, the level of triglycerides (TG) was reduced markedly (P＜0.05）and the high-density lipoprotein cholesterol (HDL-C) 、CHO level was increased markedly (P＜0.05). However, there was no obvious statistics difference on weigh among all groups(P＞0.05) . ②The comparison before and after fenofibrate administration :Compared with the value before fenofibrate administration, the ALT、AST、ALP、BUN、TG were obviously reduced after the administration of 60 mg/kg fenofibrate (P＜0.05）,in addition, the CHO、HDL-C were obviously raised (P＜0.01）.However, in the 30 mg/kg fenofibrate administration group, ALP was obviously reduced (P＜0.05）,and the CHO、HDL-C were obviously raised (P＜0.05）.Conclusion Compared with C57BL/6 mouse ,the PPARα transgenic mouse has the sensibility in evaluation of PPARα agonist. It is a new animal model .

Abstract:【Abstract】 Objective To explore the effect on the body weight and the mucosa and muscularis mucosae thickness of the forestomach on Sleeve Gastrectomy（SG）model in Wistar rat, and to provide morphology base for the further study weight loss mechanisms of SG．Methods 20 male Wistar rats were randomly divided into two groups, 12 wistar rats underwent sleeve gastrectomy as SG group．8 male Wistar rat underwent a laparotomy and stomach manipulation as sham group．Preoperative weight of the rats was measured，Preoperative, postoperative every week within postoperative 4 weeks, and then every two weeks for weight measurement．Rats were killed 12 weeks after surgery，then made Paraffin sections, observed the morphological changes of forestomach by histological methods. Results All rats remained alive in sham group，10 rats remained alive in SG group．In sham group，weight gain was achieved obviously after operation，whereas in SG group，weight gain was achieved until the 2th week．From then on , weight gain in SG group was significantly lower than in the sham group （P < 0.05）． Postoperative forestomach in SG group rats compensatory dilatated, Morphometry results under light microscopy suggest that mucosa and muscularis mucosa thickness in SG group thinning than that of sham group ( P < 0.01 ), mucosal shallowing. Conclusion Observation on the forestomach morphology on SG model in Wistar rat may contribute to the further elucidation of weight loss and weight rebound mechanism in the rat model.

Abstract:Objective to compare the three methods of ELISA, IFA and WB in the detection of Sendai virus in rat sera. Methods the SeV antigen proteins separated by SDS-PAGE were used in the WB method; 20 sera from germ free rats, 227 sera from SPF rats and 63 sera from clean rats were analyzed by ELISA, IFA，those candidate positive samples were tested by WB. Results The 20 germ free rats were determined SeV negative by all of the 3 methods; all of the SPF rats were determined SeV negative by IFA, but 1.32% of these SPF rats were determined SeV positive by ELISA, and 2/3 of these ELISA determined positive sera were confirmed by WB; The SeV positive rate in clean rats were 18.12% by ELISA, 11.34% by IFA, and 15.87% by WB. Conclusion the detection sensitivity gradient of the 3 methods from high to low is ELISA, WB, and IFA. And WB should be an alternative SeV detection method for those indeterminate samples by IFA and ELISA.

Abstract:Abstract: lymphocytic choriomeningitis Virus (LCMV) is a significant zoonosis. Recently, human infected with lymphocytic chorimeningtis virus (LCMV) are diagnosed more efficient. LCMV infection rate of laboratory animals is regulated at the low level. In Chinese national standards, LCMV is a required test item for guinea pigs and hamsters, but for mice only test when it necessary. However, LCMV is a routine test item for rats in some other areas. To strengthen the awareness of LCMV test, we will introduce information about laboratory animals LCMV infection status in different countries and areas.

Abstract:Objective To obtain capsid p27 of SIV for detecting . Methods The p27 gene of SIV was synthesized and inserted into pMAL-p5x vector to construct pMAL-p5x-p27 recombinant plasmid.And then the recombinant plasmid was transformed into E.coli BL21 to express protein and the express product was purified by Amylose Resin maltose-binding protein affinity column. Results SDS-PAGE analysis indicated that pMAL-p5x-p27 can be expressed in E.coli BL21 and the protein molecular weight was about 70×103Da; The final p27 protein purification could reach 90%. Conclusion p27 of SIV was successfully expressed in E.coli，which lays a foudation for developing the methods of detecting SIV antibody.

Abstract:Abstract: Objective To explore the effect of Adriamycin with different doses used for the Focal Segmental Glomurular Sclerosis Nephrosis rat model . Methods The Focal Segmental Glomurular Sclerosis Nephrosis rat induced by left nephrectomy plus intravenous injection with different Adriamycin doses,to observe the effect of different Adriamycin doses on rat model of survival rate,24-hour urinary protein excretion, serum creatinine,urea nitrogen, and kidney pathology.Results the survival rate decreased with the increase of injection dosage in different doses group and survival rate of injection dosage of 4.5mg/kg~6mg/kg was reduced to less than 50% at fourth week and eighth week,survival rate had a significant negative correlation with injection dosage at eighth week（r=0.9045，P<0.01）. Each group of injection dosage appeared urinary protein significantly increased at first week（P<0.01）, serum creatinine significantly increased at second week（P<0.01）and urea nitrogen significantly increased at fourth week（P<0.01）, and all showed progressive increase, serum urea nitrogen of each group is higher than the normal except the 3mg/kg group（P<0.01，P<0.05）, 24-hour urinary protein excretion, serum creatinine and urea nitrogen had a significant positive correlation with injection dosage at eighth week（r=0.9429，P<0.01 ；r=0.9384，P<0.01；r=0.9568，P<0.01）. Renal pathology of all groups showed Focal Segmental Glomurular Sclerosis,and the hardening degree aggravation while injection dosage rising.Conclusion Mortality of 3mg/kg group is Lowest and accord with Human Focal Segmental Glomurular Sclerosis，it is the more ideal dose of Adriamycin rat model.