Abstract:Objective To analyse Random Amplified Polymorphic DNA(RAPD) for laboratory Beagle dogs so as to know their genetic background.Method\ 16 RAPD primers were selected and used to amplify 40 samples.Results\ In this study,93 bands were gained in agarose gel.The homology coefficient of all samples ranged from 80.4% to 100%,average at 93.28%.Assisted by DNAman software,the homology trees were produced by cluster analysis with all individuals and group samples.Conclusion\ Those genetic background materials can provide some technologic direction for Beagle dog breeding and applying in bio-medicine research.

Abstract:目的　观察二恶英 (2 ,3,7,8 tetrachlorodibenzo p dioxin ,以下略为TCDD)对斑马鱼 (Zebrafish)初期胚胎的毒性作用。方法　斑马鱼胚从受精后 2 4h直至观察放在 2 4孔培养皿内水浴染毒。采用形态学、组织学观察法及细胞色素P4 5 0 1A(CYP1A)抗体染色法。结果　 0 1μg LTCDD未观察到明显的变化 ,但是 0 3～ 10 μg L浓度的TCDD染毒时 ,首先观察到后主静脉的血流减缓与停滞为共同特征的毒性症状。在染毒群同时还观察到心囊、卵黄囊、头部、躯体等不同程度的水肿以及头部的畸形。在 180h ,摄食对照群 ,0 1μg L、0 3μg L、0 5 μg L、1μg L和 10 μg L各自的死亡率分别是 0、0、5 %、6 0 2 %、6 1 8%、10 0 %。CYP1A抗体染色发现对照群无阳性反应 ,而染毒群在血管上皮看到极强的阳性反应。结论　本试验表明TCDD对斑马鱼胚胎的循环系统有极强的损坏作用 ,而且这种损坏与CYP1A相关联

Abstract:Objective This study was to explore a simple and quick polymerase chain reaction (PCR) method for the genotyping of heat shock factor 1(HSF1) knockout mice. Method\ Two pairs of primers were designed to amplify genomic DNA fragment of wild-type HSF1 gene and the same region on HSF1 targeting vector respectively. A gradient PCR strategy was used to test the best annealing temperature. Results\ A 562 bp band was found in wild-type HSF1 mice, a 377 bp band in homozygous mutated HSF1 mice and both bands in heterozygous mice. The genotyping results were completely coincided with those from typical Southern blot. Conclusion \ PCR is a simple, fast and practical method for the genotyping of HSF1 gene knockout mice.

Abstract:Objective In order to study the effect of the genotype of different strains on mouse embryo cryopreservation in the glycerol vitrification solution.Methods\ The 6 5mol/L glycerol and 6% (w/v) BSA were used as the vitrification solution for the cryopreservation of morulae and blastocysts embryos of CBA,NOD,C 57 BL/6J,ICR and CD1 mouse.Results and Conclusion\ The rates of in vitro development of CBA,NOD,C 57 BL/6J,ICR and CD1 mouse embryos to expanded blastocysts ranged from 31 3% to 88% after vitrification.The rates of CBA,NOD,C 57 BL/6J,ICR and CD1 mouse embryos to normal fetuses was 21%,23 5%,11%,38%,35 5%.The rates of in vitro development of CBA,NOD,C 57 BL/6J6,ICR and CD1 mouse morulae and early blastocysts were significantly higher than expanded blastocystst.The in vitro development and in vivo development were significantly influenced by inbred strain,closed colony and different stages of mouse embryo.The result of mouse embryo cryopreservation in the vitrification solution may be connected with the different genotype of the different strains and different stages of mouse embryo.

Abstract:Objective To creat a new rodent model of Parkinson's disease (PD) for the investigation of its pathological mechanism. Methods At different survival time after injection of lipopolysaccharide(LPS) in the substantia nigra (SN) of rats, the levels of DA and their metabolites were measured by using HPLC, and the tyrosine hydroxylase (TH) positive neurons and microglia were observed by immunohistochemistry. Results The levels of DA and their metabolites were decreased and the lowest level ( p <0 01) appeared at 14th day after injection of LPS. The number of TH positive neurons was also reduced from the second day after injection, and a large amount of microglial cells were activated with their shapes changed. Conclusions The injected LPS could result in a damage of dopaminergic neurons in the SN.

Abstract:目的　以人apoE7基因制备的高脂血症转基因小鼠模型已建成 ,要用于探查这一突变基因致病的分子机制 ,首先揭示人apoE7在小鼠体内诱发了那些基因的表达异常 ,是重要的环节之一。cDNA削减文库是寻找异常高表达的已知和未知基因有效的手段 ,而cDNA文库更是获取全长基因所必须。方法　自正常小鼠与转基因小鼠肾脏同时提取总RNA ,再分离mRNA ,逆转录制备cDNA后 ,用两次分子杂交削除转基因鼠cDNA中与正常小鼠相同的部分 ,再用PCR与择需PCR法扩增转基因鼠特异的基因 ,克隆入pGEM T载体后 ,制备削减文库。未经削减的转基因小鼠cDNA克隆入λgt11载体 ,制备基因表达文库。结果和结论　自高脂血症转基因小鼠肾脏构建的削减文库得到约 4 0 0个削减克隆 ,测定了部分克隆的序列 ,并进行同源性比较。λgt11 cDNA文库滴度 1 7× 10 7pfu ml,随机测定的克隆其插入片段长约 5 0 0bp以上。

Abstract:Objective To study the sensibility of airway of new strain Zmu 1:DHP guinea pig inhaling the chemical mediator from the respiratory tract. Method With increasing concentration, histamine (His) and acetylcholine (Ach) were breathed in respectively by the animals, and the time and the mediator concentration of the asthma onset were recorded in comparison with DHP guinea pig. Results When inhaling the gas of 0 02% His, the respiratory rate and the volume of flow of Zmu 1:DHP guinea pig were higher significantly than DHP guinea pig ( P <0 05). When 0 04% His, both of the strains were not divergent remarkably in asthma ( P >0 05). When 0 06% His, these of Zmu 1:DHP guinea pig were lower notably than DHP guinea pig ( P <0 05). Conclusion The sensibility of Zmu 1:DHP guinea pig is higher significantly to the reaction of asthma than DHP guinea pig when the lower His gas is inhaled.

Abstract:目的　比较不同品种 (系 )小鼠接种E2G8细胞株的某些生物学特性。方法　将E2G8细胞稀释成不同浓度 ,接种KM、DBA 2、6 15、C57BL 6及BALB c小鼠皮下或腹腔 ,观察小鼠出现可见肿块 腹水时间及小鼠生存时间 ;测量肿块直径 (cm)及瘤重 ;荷瘤小鼠腹腔注射环磷酰胺 (CTX) ,观察E2G8细胞肿瘤模型对CTX治疗的敏感性。结果　E2G8接种BALB c、6 15小鼠皮下毒性最大 ,DBA 2、C57BL 6小鼠次之 ,接种KM小鼠皮下毒性最小 ;接种DBA 2、C57BL 6、6 15小鼠腹腔毒性较大 ,致死性高 ,对BALB c和KM小鼠的致死性弱 ;E2G8瘤细胞在KM和 4种纯系小鼠皮下均能很好生长肿瘤 ,其中在KM小鼠皮下肿瘤生长最大 ,在BALB c小鼠皮下肿瘤最小 ;KM、C57BL 6、6 15、DBA 2、BALB c小鼠皮下接种瘤细胞第 12天瘤重分别为 (2 4 0± 1 30 ) ,(0 90± 0 4 8) ,(1 2 0± 0 38) ,(1 10± 0 2 9) ,(0 80± 0 30 )g ;E2G8细胞接种 5个不同品种 (系 )小鼠制备的肿瘤模型 ,用CTX治疗 ,抑瘤率由高到低依次为6 6 7% (6 15 ) ,5 5 0 % (BALB c) ,5 3 3% (C57BL 6 ) ,5 0 % (KM) ,36 4 (DBA 2 ) ,其中 6 15小鼠建立的模型对CTX的治疗最敏感。结论　E2G8细胞株接种 5个不同品种 (系 )小鼠 ,所测生物学特性不完全相同 ,但是从腹腔 皮下接种、CTX

Abstract:Objective To explore the effect of Chinese medicine of tonifying kidney plus strengthening vital energy on production of IL 1,IL 2,IL 12 and their mRNA expression in mice with nephrasthenia. Methods\ Animal model of kidney deficiency was made by corticosterone. The model animal was medicated with tonifying kidney plus strengthening vital energy for 6 weeks, then, the production of IL 1 by peritoneal macrophages, IL 2 and IL 12 by splenetic lymphocytes were assayed with the methods of biological activity analysis and ELISA separately. The mRNA expressions of IL 1,IL 2,IL 12 were examined by RT PCR. Results\ The production of IL 1, IL 2 and IL 12 declined in model animal group, but they were restored in mice treated with Chinese medicine in different degree,and the 3 kinds of mRNA expression were also increased when the mice were given Chinese medicine. Conclusion\ Tonifying kidney plus strengthening vital energy is an ideal method for treating mice with nephrasthenia due to corticosterone, so that it can improve the state of immunosenescence.

Abstract:Objective To compare the immunological features of FMMU albino guinea\|pig and pigment ones, including the serum content of complement and immunoglobulin, lymphocyte transformation rate and immunological function of erythrocyte.Methods\ The content of immunoglobulin(IgG,IgA,IgM), complement 3(C3), complement 4(C4) and the total activity of complement of two groups were measured by Clinical Chemistry Analyzer; the lymphocyte transformation rate of FMMU albino guinea\|pig and pigment ones were measured by lymphocyte transformation test; RBC\|C3bR rosette \|forming rates of two groups were measured by red cell C3b receptor rosette test. Results\ The serum content of immunoglobulin(IgG,IgA,IgM), complement 3(C3), complement 4(C4) and the total activity of complement of the FMMU albino guinea pigs were lower than those in pigment ones. The transformation rate of the FMMU albino guinea pigs was a little lower than that of pigment ones. There was no significant difference in the values of RBC\|C3bR rosette\|forming rate between FMMU albino guinea\|pigs and pigment ones, but RBC\|IC rosette\|forming rate was lower than that in pigment ones. Conclusion\ Closed colony FMMU albino guinea\|pigs have their unique immunological features, and their immunological function is weaker than that of pigment ones as a whole.

Abstract:Objective\ To collect age\|specific embryos, increase the resources of embryos, and establish the bank of frozen embryos in mice. Methods\ The in vivo development and movement of zygotes, the in vitro maturation and fertilization of oocytes , the in vitro culture and rapid\|freezing of embryos were studied. Results\ (1) Zygotes developing to pronuclei occurred at 12-20h after injection of hCG;2\|cell, 4\|cell, and 8\|cell embryos formed at 42-48 h, 48-60 h, and 60-68 h, respectively. The embryos of the above stages were in the oviducts. Morula, early blastocysts, and blastocysts, which were in the uterus horns, formed at 75-70 h, 90-92 h, and 92-96 h, respectively. (2)Adding FSH and hCG to the culture medium could increase significantly the percentage of in vito fertilization of oocytes. Adding FCS and FSH and hCG simultaneously to the culture medium was better than adding hormone alone. FCS could increase significantly the rate of the embryo development. (3)Adding EDTA to the culture medium could effectively overcome the 2\|cell block in mice. The rates of embryos developed to 2\|cell stage and to 8\|cell stage were 100% and over 55%, respectively. Adding sodium lactate and pyruvate sodium to the culture medium could stimulate significantly the embryonic development. (4) After frozen in the freezing solution (D\|PBS+glycerol+sucrose), and thawed in DPBS +sucrose solution, the survival rate of morula and early blastocysts were 69.3% and 60.4%, respectively. Conclusion\ The present results laid the preliminary basis for the establishment of embryo bank in mice.

Abstract:Objective To develop the rat model of ordinary male Wistar Rat induced by filling with L methionine. Method\ The drug doses of group A and B were 230 mg/rat and 450 mg/rat respectively. The blood lipid level and pathology of aortic arch were examined every week. Results\ Taking L methionine resulted in the disturbance of blood lipid, and the local tunica edema of aortic arch in both group A and B at the 8th week after. The pathological change of group B (high dose group) was more severe than group A (low dose group), and occasionally the invasion of theca cells and mononuclear leukocytes could be seen. The control group had no changes mentioned above. Conclusion\ Filling with L methionine may cause disturbace of blood lipid and pathological changes of tunica intima of artery in Wistar Rat.

Abstract:Objective To study the sterilization effect of glutaraldehyde compound disinfectant on some swine viruses, and to determine the optimum disinfectant concentration.Methods\ PPV,PRRSV and PRV,treated with glutaraldehyde compound disinfectant at different concentration,were inoculated on cell monolayers.The optimum disinfectant concentration was determined according to the cytopathologic effects.HCV reacted with the disinfectant at different concentration was inoculated into rabbits through vein injection,the optimum concentration was decided according to the body temperature changes.Results\ PPV, PRRSV and PRV could be disinfected effectively by 1% disinfectant in 20 min.And HCV could be killed effectively by 0\^5% disinfectant in 10 min.Conclusion\ Glutaraldehyde compound disinfectant possesses good sterilization effect with the concentration of 1% and 0\^5% to some swine viruses,and is suitable for disinfecting the stock farm,hospital and epidemic area of infectious diseases.It is a kind of practicable method of in vitro experiment to detect and judge the sterilization effect of disinfectant with the cell infection method.

Abstract:Objective To establish a dog spinal cord injury animal model for both research and clinical practice. Methods Dog spinal cord was injured by intention, and recovered through the stimulation of direct electric field. Results The dog spinal cord injury animal model is established with nervous function recovered. Conclusion In different periods, the injured dog spinal cord and its function can be recovered by the stimulation of direct electrical field.