Abstract:Objective Sheep fetal fibroblasts(SFF) were transfected by enhanced green fluorescent protein(EGFP) gene to investigate the effect of the foreign gene on the biological characteristics of SFF. Methods SFF were dispersed and cultured in vitro.pEGFP-C1 was transfected to the passage 1 SFF by liposomes.The SFF were incubated in medium containing G418 for 10-12 days.Resistant colonies were picked and subcultured until use.The SFF were examined by cell morphology,growth curve or the nuclear type of chromosome.Sex-determining region Y gene(SRY) of the SFF was identified.Results The biological characteristics did not differ significantly between the resistant colonies and the control cells.Expressive amount of foreign gene was revealed by fluorescent intensity.Conclusion The present research can provide a support for the transgenic cell with EGFP as the report gene in vitro.Transgenic donor cells integrated with EGFP gene provide an efficient new strategy for the production of transgenic farm animals.

Abstract:目的通过建立A549人肺腺癌细胞/615-SCID小鼠模型,评价重度联合免疫缺陷615-SCID小鼠在建立人类肺癌转移模型方面的应用价值.方法将1×107A549细胞接种到615-SCID及SCID小鼠右上肢背部皮下,观察成瘤时间、成瘤率、肿瘤生长速度及转移发生.结果两品系小鼠接种后的成瘤率均为100%,615-SCID小鼠移植瘤潜伏期较长、生长较缓慢,更容易发生转移.结论 615-SCID小鼠比SCID小鼠更易于构建人类肺腺癌转移模型,对于肺癌转移特性研究具有较大的意义.

Abstract:Objective To establish goat mammary gland epithelial cell lines for evaluation of mammary gland-specific expression components.Methods A pair of primers were synthesized according to the previously published DNA sequence and used to amplify the SV40 T gene from COS-1 cells by high fidelity PCR.The SV40 T gene was subcloned into eukaryotic expression vector pTarget with its neo gene deleted and the resulting recombinant vector pTarget-LT was transfected into primary goat mammary gland epithelial cells.The transformed cell clones were obtained by limited dilution and repeated passage.Results An expected PCR product of 2.5?kb was obtained,which was 99% identical in sequence to the previously published SV40 T gene.Following transfection and repeated in vitro passage,49 clones were obtained with a clone forming rate of 26.7% and a logarithmic growth period at the forth day post-plating.Dot blotting analysis showed that the SV40 T gene was integrated into the transformed cell genome.The transformed cells had a normal karyotype with no tumorigenicity to nude mice and were not able to grow in soft agar culture.One of the transformed cell clone had been passed for more than 30 generations,which maintained normal morphology of mammary gland epithelial cells and was able to form glandulous structures on collagen matrix.Conclusion These results showed that the transformed mammary gland epithelial cell line shared similar biological characteristics of other transformed cell lines.

Abstract:Objective To establish special pathogen free Kunming mouse of Hubei closed colony.Method The purification mouse was made by excising operation of uterus in the situation of bacteria free. The SPF Kunming mouse strain of Hubei closed colony was developed with the following surveillance on microbes and parasites in first generation.Results 57 mice were got from 67 cases by operation, and 509 baby mice with purification,from which 460 grew into adults and the state of microbiology in those mice was in keeping SPF.Conclusion The method can keep the excellent laboratory animal stain with biological purification.

Abstract:Objective To investigate the existence and expression of porcine endogenous retrovirus(PERV)in Chinese experimental miniature pig(CEMP).Method Three pairs of specific primer(gag,pol,env)and three pairs of type-specific primer(A,B,C)were synthesized,then using the PCR and RT-PCR methods,we detected the PERV of DNA and RNA samples derived from the peripheral blood lymphocytes of CEMP.Results The results showed that the PERV existed and expressed in all samples,and the expression of PERV belonged to PERV-A and PERV-B,but the expression of PERV-C wasn't detected in all samples.Conclusion The PERV exists and can be expressed in Chinese experimental miniature pig.This research may be helpful to development of the Chinese experimental miniature pig and to assessment of the pathogen safety of PERV.

Abstract:目的探索小鼠基因组39个微卫星的PCR条件,评价微卫星在小鼠遗传检测中的运用.方法采用梯度法探索39个微卫星的PCR条件;选择本中心不同来源及引种时间的C57BL/6、BALB/c、DBA/2J、CBA/N、FVB/NJ、ICR共6个品系(8个组)小鼠,每组采用10只个体的鼠尾,提取DNA并混合成DNA池,用39个微卫星扩增后电泳观察、比较种系纯度.结果小鼠微卫星的PCR条件差异较大,Mg2+浓度多数在1.5 mmol/L左右,退火温度多数在59℃左右.在6个品系小鼠的39个微卫星位点中,C57BL/6、BALB/c、DBA/2J都是纯合的; 其余品系有1～3个杂合位点. BALB/c在D5Mitl68、D8Mit320、D13Mit262三个位点,DBA/2J在D14Mit205位点与数据库记录有差异.结论本研究为小鼠39个微卫星提供了候选的PCR条件,并对6个品系小鼠的微卫星概貌及微卫星的运用价值进行了探讨.

Abstract:目的测定6～8月龄Beagle犬正常生理、生化等数据,为药理、毒理试验提供对照参考.方法血液学、血液生化学、血清激素水平、血压、呼吸、心电图、骨髓象、脏器系数等指标均按现临床检测方法进行.结果 48头6～8月龄Beagle犬(雌雄各半)的正常生理、生化参数,血清激素水平,骨髓象及主要脏器的系数均取得平均值及标准差范围.结论 6～8月龄正常Beaele犬生理、生化等参数的个体差异较小,本研究结果可作为新药Beagle犬长毒试验中各项测试的正常值参考.

Abstract:Objective To compare the sensitivity and specificity of outer membrane protein (OMP) and lipopolysaccharide (LPS) of Pasteuralla pneumotropica used as serological diagnostic antigens to detect Pasteuralla pneumotropica.Methods LPS,OMP and whole cell(WC) were used as ELISA and Western Blot antigens to characterize the IgG antibody of mice with natural and experimental infection of Pasteuralla pneumotropica, meanwhile the cross_reaction of three antigens to common laboratory animal pathogens was also evaluated by ELISA and Western Blot methods.Results Three antigens were tested with sera from experimentally and naturally infected mice in different ages via ELISA.LPS showed the greatest antibody ratio,WC antigen showed lower,and OMP lowest.In naturally infected mice colony,the antibody ratio of LPS in 4_week age mice was 80%,while that of whole cell antigen and OMP only 25% and 20% respectively,showing that LPS was the most sensitive one.Three kinds of antigens were also tested with sera from uninfected mice and antisera of common laboratory pathogens via ELISA and Western Blot.WC antigen had a high A value in ELISA,but it was proved to be unspecific using Western Blot.LPS was the most specific antigen,and OMP second.Conclusion The mixed species specific and typical specific OMP and LPS in ELISA to detect latent infection of P.pneumotropica in laboratory animals have more specific and sensitive than WC antigen.

Abstract:Objective To isolate and identify piroplasms from a Japanese blood donor by using NOD/shi-scid mice. Methods The donor's blood specimen was inoculated into NOD/shi-scid mice, whose circulating erythrocytes(RBC) had been replaced with human RBCs(hu-RBC-SCID mice). Results Parasites proliferated within the human RBCs in the mice, resulting in a high level of parasitemia, and the parasites were related closely to %Babesia microti.% Conclusion Our results demonstrated the usefulness of hu-RBC-SCID mice for isolation of parasites from human and for maintenance of the parasite infectivity for human RBCs.

Abstract:Objective To establish a kind of culture system to improve %in vitro% development of Kunming mouse pronuclear embryos. Methods Mouse pronuclear embryos were obtained through superovulation, then the normal morphologic zygotes were selected and cultured in M16, CZB and KSOM respectively, or co_cultured with mice oviduct epithelia. The cell number of blastocysts obtained from in vito/in vitro were counted. Results The ratio of blastocyst formation was improved in KSOM and CZB by addition of fetal bovine serum(14.71% vs 85.71%,6.45% vs 10.81%). Co_culture system increased percentage of cleavage and blastocyst formation, and was advantageous for embryos quality and synchronized development. Conclusion Mouse pronuclear embryos developed in vitro highly in KSOM-{FBS} cocultured with oviduct epithelia.

Abstract:Objective To analyze the genetic type of NJS and its relative mice.Methods Five 10bp random primers were used to amplify DNA of NJS and its relative mice and DNA analyzed among individuals in the population of NJS.Results There were several fragments of genetic DNA shared by two mice but specific fragments were seen.Different individual NJS had various bands,but the frequency of common bands was high.Based on RAPD patterns,similarity index(F) was 0.927(0.880-0.980).Conclusion RAPD could be used to distinguish NJS and its relative mice. There is higher degree of homogeneity and stability in heredity among individuals in the population of NJS and the degree of population differentiation is at a lower level.

Abstract:目的研究犬胃内pH值和胆汁反流的特点.方法应用便携式pH监测仪和便携式光纤分光光度仪(Bilitec 2000),持续监测禁食状态下5只犬胃内的pH值和胆红素吸光值(abs),每只犬同条件下监测2次,共进行10次监测.结果 pH值的波动范围1.2～7.8, 平均5.4±0.5,pH>4的时间百分比为(91.2±7.3)%.10次监测中仅1次胆汁反流阳性,其他9次abs的波动范围为0～0.20,平均0.08±0.02.结论犬胃内pH水平较人高.首次将Bilitec用于犬胃内胆红素的检测,发现胆汁反流发生率较低,建议以abs>0.20为犬胃内胆汁反流的诊断标准.

Abstract:目的探讨原花青素(Procyanidolic,pc)对急性呼吸窘迫综合征(Acute respiratory distess syndrome,ARDS)大鼠疾病模型的治疗作用.方法大鼠用百草枯(Paraquat,PQ)一次性灌胃,剂量250 mg/kg,治疗组分别于染毒6、24、48 h各给予PC治疗剂量500 mg/kg,染毒组与治疗组分别于24、48、72 h,取大鼠血浆和支气管肺泡液(BALF),测试其血浆和BALF中超氧化物歧化酶(SOD)、丙二醛(MDA)谷胱甘肽过氧化物酶(GSH-Px)的活力,各组取肺病理组织做病理组织观察研究.结果治疗组与染毒组比较,血浆和BALF中GSH-P\-X、SOD活力逐渐升高,差异有显著性 P <0.05,MDA含量下降,差异有显著性 P <0.05,染毒与对照组比较,血浆和BALF中MDA含量高于对照组,差异有显著性 P <0.05,SOD和GSH-P\-X活力下降,差异有显著性 P <0.05,肺组织病理学观察,ARDS组肺泡壁充血,炎症细胞浸润,Ⅰ型与Ⅱ型细胞破坏,肺泡内有炎症渗出物,对照组肺泡壁完好,肺泡内没有炎症渗出物,没有炎症细胞浸润;给予PC治疗后,肺泡壁充血、出血减轻,炎症渗出物减少;结论 PC对ARDS模型大鼠有一定的治疗效果.

Abstract:Objective To study the infectivity of Pasteurella pneumotropica (P.pneumotropica) from different sources between laboratory rats and mice. Methods 30 laboratory rats and mice were inoculated with 3 %P.pneumotropica% isolates from rats, mice and wild mice, to compare the biologic characteristics, by SDS_PAGE, Western blot and RAPD_PCR, of the 3 isolates at different days of post_inoculation with that before inoculation. Results Laboratory rats and mice were infected easily by 3 %P.pneumotropica% isolates, which were carried in them until the end of the test (4w). lsolates reobtained from post_inoculated animals had no difference distinctly in biologic characteristic, protein component, antigenic bands and genotype. Conclusion The same isolate of %P.pneumotropica% can infect both laboratory rats and mice.

Abstract:Chronic obstructive pulmonary disease(COPD) is an important chronic respiratory system disease with high morbidity and mortality.It mainly consists of chronic bronchitis and pulmonary emphysema,characterized by slow progressive development.The patients with COPD have weak work ability and poor life quality. Numerous researches both at home and abroad are focused on the potential pathogenesis of COPD in recent years.However,until now the progress in COPD research is still very slow.The main reason for this is not only multiple causes but also complex pathogenesis of COPD.The aim of this review is to sumup the COPD animal models and to objectively evaluate the advantages and disadvantages of each model.