Abstract:Objective TW mouse is a new domestic inbred strain,established by the Tianjin Center for Disease Control and Prevention.The aim of the present study was to carry out genetic characterization and detect rare genes in these inbred mice.Methods 25 alleles,including Akp-1 and others,were assayed in the inbred mice by genetic biochemical labeling technique using a homemade biochemical label detection Kit.Results Rare genes were detected at 8 biochemical loci: Amy1,Es2,Es3,Es10,Gpd1,Hbb,Np1 and Pgm2 in the TW mice.Conclusions Polymorphism of DNA sequences in the mice can be detected by biochemical polymorphism analysis with allele isomorphic enzymes.It may be of great importance to provide abundant animal gene bank to serve the identification of disease genes in humans and as an invaluable resource for experimental animal research as well.

Abstract:Objective To establish a golden hamster model of hyperlipidemia.Methods 20 golden hamsters were randomly divided into control and experimental groups,10 per each group.The hamsters of experimental group were treated with high cholesterol diet for 15 days.During the experiment,the gross health status was observed.Finally,the contents of serum total cholesterol(TC),triglyceride(TG),high-density lipoprotein-cholesterol(HDL-C),low-density lipoprotein-cholesterol(LDL-C) and very low-density lipoprotein-cholesterol(VLDL-C) were assayed,and pathological changes of the liver were examined by light and electron microscopy.Results\ The golden hamsters showed to be well toleranble to high cholesterol diet.After 15 days,the contents of serum TC, TG,HDL-C,LDL-C and VLDL-C were 9.06,4.19,2.43,6.21,18.88 fold-increased,respectively(P<0.01),but the ratio of HDL-C/(LDL-C VLDL-C) decreased significantly compared with that in the normal group(P<0.05).The pathological examination exhibited liver structural alterations.Conclusion It is a quite rapid and reliable way to establish hyperlipidemia model of golden hamster,which can continuously provide enough plasma for hyperlipidemia research.

Abstract:目的探讨新生猪缺氧后心肌连接蛋白Cx43 mRNA和蛋白含量的变化。方法对照组(C组):新生猪6头,未吸入低氧;缺氧组(H组):新生猪6头,吸入低氧(FiO2=0.1)维持1 h。于缺氧结束后5 h,免疫荧光染色检测心肌组织细胞缝隙连接蛋白Cx43蛋白的表达,实时荧光定量逆转录多聚酶链式反应(RT-PCR)检测心肌组织Cx43 mRNA的表达。结果H组心肌组织Cx43蛋白的表达显著低于C组(P<0.01),但两组心肌组织Cx43 mRNA表达差异无统计学意义(P>0.05)。结论缺氧可导致心肌Cx43蛋白的表达或分布发生变化。

Abstract:目的阐明H3亚型鸭流感病毒与其他亚型流感病毒的关系。方法对活禽市场分离的3株H3N8亚型鸭源流感病毒聚合酶PB1基因进行了序列分析。结果3株鸭源H3N8流感病毒聚合酶PB1基因核苷酸同源性为99.9%,与H9N2亚型流感病毒(DK/ST/2143/00)的同源性为96.31%~96.44%,而与H3N8亚型鸭流感病毒(Mal/Alberta/279/98)为88.65%~88.79%。系统进化树分析表明,本实验中的3株病毒属于相同的分支,且与A/duck/Hong Kong/Y439为代表的H9N2亚型禽流感病毒位于一进化分支,说明三株H3N8亚型流感病毒重排了H9N2亚型禽流感病毒的基因片段。结论不同亚型禽流感病毒在贮存宿主体内的重排以及重排病毒的新特点如鸭H3N8亚型流感病毒对禽的致病性,应当引起我们的高度重视。

Abstract:Objective To explore whether the Oct4 transcription factor can improve the mouse-porcine interspecies nuclear transfer development or not.Methods Oct4 cDNA was amplified by RT-PCR.Two recombinant expression plasmids were constructed as the pEGFP-N1-Oct4-EGFP fusion vector and pEGFP-N-1-Oct4-EGFP termination vector,and pEGFP-N1 as the negative control,then transfect the NIH3T3 cell with lipofectamineTM2000.RT-PCR assay showed that oct4 DNA fragment could be amplified,and confirmed by fluorescence microscopy.Then the cell was inserted into the perivitelline space of the enucleated porcine oocyte to investigate the development of interspecies embryo.Results All the non-transgenic NIH3T3 group,transgenic Oct4-EGFP NIH3T3 and pEGFP-N-1 groups could support the early development of cloned interspecies embryo,and the cleavage rate and development rate into the eight-cell stage were not significantly different.Conclusion It is clear that NIH3T3 can support the early embryonic development of mouse-porcine interspecies nuclear transfer.But maybe due to the low development of NIH3T3 derived nuclear transfer embryo and our limited condition of micromanipulation,the Oct4 gene does not have an improving effect on interspecies nuclear transfer.Further studies are still needed to elucidate its concrete mechanism.

Abstract:Objective To investigate the pathophysiologic role of T lymphocytes in renal ischemia-reperfusion injury in mice.Methods BALB/c wild-mice(group A, n = 24) and BALB/c nude-mice(group B,n = 24) were divided into four groups(n = 6),respectively,the groups A-1 to A-4 and groups B-1 to B-4.Bilateral renal pedicles were occluded by microvascular clamps for 45 minutes.Then the kidneys were reperfused.Samples were collected at sham operation and 24 h,48 h and 72 h after IRI.Serum creatinine(Scr),urine protein concentration,renal histology and splenic CD3~ /CD4~ /CD8~ positive cells(group A) were examined,respectively.Results\ Scr and urine protein concentration increased and histological injury was observed in group A-(2-4) and B-(2-4).The injury in group A were more severe than that in group B(P<0.05).FACS analysis showed,that in comparison with group A-1,percentage of splenic CD3~ T cells in group A-2 and A-3 increased significantly(P<0.05),but a similar ratio of CD4~ /CD8~ between group A1 and group A-(2-3)(P>0.05)).Conclusion T lymphocytes play an important pathophysiologic role in renal ischemia-reperfusion injury,and may mediate post-ischemic decline in renal function and lead to histological injury.

Abstract:目的建立卷尾猴正常月经周期卵泡生长发育变化规律及排卵的超声监测(ultrasound examination,US)技术。方法采用阴道细胞涂片及超声监测技术,对5只卷尾猴连续3个周期检查阴道细胞变化,对卵泡生长发育进行超声监测。结果阴道脱落细胞在生殖周期内呈周期性变化,月经周期为(21.13±2)d。阴道细胞学涂片显示卷尾猴的月经周期分为以下各期:月经期(menstrual phase,M)、滤泡期(follicular phase,F)、排卵前期(pre-ovulationphase,Po),黄体期(luteal phase,L)。US技术在滤泡期及排卵前期可观测到直径约3~9 mm的优势卵泡(dominantfollicle,DF)的生长过程。当DF较前一天至少小10%,轮廓不规则,回声较以前强,阴道细胞涂片显示淋巴细胞出现,提示排卵已发生,黄体开始形成。结论B型超声技术能实时监测卷尾猴的卵泡生长发育过程,预知优势卵泡排卵时间。

Abstract:目的利用昆虫细胞/杆状病毒系统表达猪瘟病毒(CSFV)E2蛋白,用于E2蛋白功能、开发CSF新型疫苗以及建立相关血清学诊断方法等研究。方法采用RT-PCR扩增CSFV E2基因,将PCR产物克隆到pGEM-T-Easy载体,将该基因插入到pFast-BacHT A载体中,构建重组转座载体后转化DH10Bac感受态细胞,获得重组Bacmid质粒后转染sf9昆虫细胞,传毒3代,对表达蛋白进行Western-blot及免疫组化鉴定。结果成功克隆CSFVE2基因,其核苷酸序列为1119 bp。SDS-PAGE电泳结果显示表达E2蛋白相对分子质量约为43×103,Western-blot和免疫组化结果证实表达蛋白能够被CSFV标准阳性血清识别。结论在Bac-to-Bac杆状病毒系统中的成功表达了CSFV E2蛋白,与CSFV标准阳性血清具有较好的反应性。

Abstract:目的探索建立化疗损伤性卵巢功能早衰动物模型的适宜方法及最佳时间。方法以70只ICR雌性小鼠为研究对象,腹腔连续注射顺铂7~14 d,观察不同剂量和时间条件下顺铂对小鼠的体重、卵巢功能、肝肾毒性及卵巢组织抗氧化系统各指标的变化。结果顺铂可引起小鼠体重明显下降,卵巢功能减退,出现肝肾毒性,并可引起卵巢抗氧化及氧化损伤指标的异常,与对照组比较均有明显差异。结论3.0~4.0 mg/(kg.bw)的顺铂作用7 d后即可引起小鼠出现明显的卵巢功能衰退、肝肾损伤,氧化损伤是顺铂产生其毒性作用的可能机制之一。该模型的生殖内分泌和病理组织学变化,与人类化疗损伤性卵巢功能早衰病变过程相似。

Abstract:Objective In order to establish a substitute material of small-caliber vascular graft,an experiment with heterogenic graft transplantation in canine model was carried out.The study was designed to evaluate the safety,reliability of the epoxy treated porcine blood vessels as vascular graft.Methods 17 female dogs were used in this experiment: 10 dogs received the epoxy treated porcine graft(PG),while the other 7 dogs received standard Gortex e-PTFE grafts(e-PTFE).The animal model was established by using the graft materials to make arteriovenous fistulas of the right femoral artery and vein.In order to evaluate the patency,all the grafts were assessed by Doppler ultrasonic examination and angiography.The grafts were taken out and evaluated by histopathological examination after 3 months.Results The grafts were assessed by Doppler ultrasonic examination at 1 and 2 weeks after operation and showed that all vessels kept free circulation of blood.The patency rate was 100%.Angiography was performed at 1 month after operation,the PG group kept patency in 6 dogs(60%),and e-PTFE group in 5 dogs(71.4%).At 3 months after operation,the angiography showed patency in 5 dogs of PG group(62.5%) and 4 dogs in P group(66.7%).The difference of the patency bewteen the two groups was statistically not significant(P>0.05).The transplanted vessels were examined by light and scanning electron microscopy at 3 months after operation.No stenosis of the anastomotic stoma was found and the endothelial cells were found to be with nomal structure.There were no signs of calcification in the graft and surrounding tissues.Conclusion\ This preliminary study demonstrated that the epoxy treated porcine blood vessel(PG) as a heterogenous graft is shown to be reliable.However,much more animal research is needed.

Abstract:Objective To establish an experimental animal model of fetal growth restriction(FGR) by temporary interruption of the uterine artery blood flow during late stage of pregnancy,and provide a useful animal model to investigate the FGR mechanism induced by hypoxia.Methods Based on the fact that the rat uterine artery is a branch of ovary artery,the pregnant rats undergone an operation temporarily interrupting the ovary artery blood flow at 15th day of pregnancy and caesarean birth at 21~(st) day of pregnancy.After the caesarean birth,the body weight of fetus,weight of placenta,weight of the brain,heart,liver,lungs and kidneys of the fetus in every group were compared.The prognosis of each group and the effect of different time of artery blood interruption(10 min,20 min,30 min and 40 min) were compared and analyzed.Results The rat FGR model was successfully established by interrupting the ovary artery in pregnant rats for 20 minutes.This method was simple to perform and needs less time than the operation interrupting the artery blood for 30 or 40 minutes,while the fetal mortality was similar between experimental group and control group (P>0.05).In the rat model,the weights of new-laid rats and their placenta,brain,heart,liver,lungs and kidneys were all decreased in comparison with those in control group(P<0.05).Conclusion By temporarily clamping the ovary artery blood flow in pregnant rats,an animal FGR model has been successfully established in rats.The model is simple to perform and well reproducible,with autogenous control,and provides a useful approach in FGR research.

Abstract:目的筛选一种既提高精子转染外源DNA效率,又保持解冻后精子活力的山羊精液冷冻—解冻方法。方法应用正交设计L9(34),因素分别为稀释液种类、稀释比例、降温时间和解冻液,每个因素选择3个水平,检测和比较解冻后精子转染外源DNA效率和精子活力。结果所选冷冻—解冻各因素对精液转染效率影响不显著[F(8,18)=1.032,P=0.449];平衡时间对冷冻—解冻活力影响极显著[F(2,24)=9.972,P=0.001],平衡1h极显著小于平衡2 h和4 h的精液活力(P=0.003,P=0.000),以平衡4 h最好。用筛选的冷冻—解冻方法处理精液,解冻后精子的活力极明显降低(P=0.002);生存指数降低,GOT释放量增加,菌落数减少,与鲜精相比差异显著(P=0.018;P=0.016;P=0.018);精子畸形率增加,顶体完整率降低,与鲜精相比差异不显著(P=0.494;P=0.084)。结论优化了提高精子转染外源DNA效率的山羊精液冷冻—解冻方法。

Abstract:目的探明小鼠两种促性腺激素受体(FSHr、LHr)在卵巢的位置分布,揭示促性腺激素(GTH)调节卵巢机制及与卵泡发育分化的关系。方法运用免疫组化ABC法对小鼠卵巢FSHr、LHr分别进行定位染色,结合图像分析系统处理分析阳性切片。结果①FSHr阳性物质主要见于GC、TC、卵母细胞及间质细胞。随卵泡的发育,FSHr、LHr阳性细胞数量呈增长趋势,卵泡早期与中期之间阳性细胞数量差异显著,平均吸光度变化差异不显著。②LHr阳性物质主要见于卵泡TC、间质细胞、GC、卵母细胞,阳性物质着色以卵泡中、晚期较强,阳性细胞数量以卵泡中期与晚期之间差异显著。平均吸光度变化差异不显著。结论卵泡颗粒细胞、膜细胞上受体是接受促性腺激素的主要调节部位,受体数量与卵泡大小和发育程度有一定的正相关。

Abstract:Objective To evaluate the beneficial effects of a Chinese medicine,Goutengsan,on intelligence of mice.Methods Mice were randomly divided into 6 groups: 3 experimental groups,of which were daily administrated(i.g.) with Goutengsan extract at a high,middle and low dose,respectively.The model and control groups were administrated with distilled water,whereas mice receiving administration of aniracetam at 0.2 g/kg body weight served as a positive control.After continuous administration for 3 weeks,scopolamine,sodium nitrite,40% ethanol were used to establish models of acquirement disorder,stability disorder and reappearance disorder of memory,respectively.The intelligence-promoting effect of Goutengsan extract was examined by Y-labyrinth method.Results The Goutengsan extract significantly improved the above-mentioned three memory hindrance models,especially in the group of reappearance disorder of memory.Conclusion The Chinese medicine Goutengsan can considerably improve the learning and memory capacities and show apparent intelligencepromoting effect in animal learning and memory disorder models.

Abstract:目的探讨西藏小型猪的生理生化指标特性。方法采用全自动血球计数仪和全自动生化仪,对1.5~5月龄西藏小型猪(♂21,♀24)血液的部分生理和生化指标进行测定和分析。结果西藏小型猪血液生理指标的性别差异不显著,部分生化指标性别差异显著;但多数指标在不同月龄间差异极显著(P<0.01)或显著(P<0.05),并有近半数项目随月龄的增长呈减少或增加的趋势,其余多无明显的变化规律。所测生理生化指标中部分与巴马小型猪相近,与普通猪差异较大,与人类水平接近。结论西藏小型猪、其他小型猪和人类的生理生化特性有一定的相似性。

Abstract:Objective The differences of growth and reproductive perfomance between F1 and F0 generations of Tibet mini-pigs raised in Guangzhou were investigated.Methods\ Litter size,number of births alive,number of piglets weaned and body weight at birth,30 days and 60 days of F1 generation after birth were measured and compared with those of F0.Results\ The findings indicated that litter size,number of births alive and number of piglets weaned,the average body weight at birth,30 days and 60 days of F1 and F0 after birth were 3.83,2.30;3.29,2.20;3.14,3.11;0.66 kg,0.53 kg;3.20kg,2.30 kg;4.50 kg,4.01 kg,respectively.Litter size and Survival rate after weaning of F1 are higher than that of F0.Conclusions\ The Tibet mini-pigs have adapted to the environment in Guangzhou and are able to normally grow and reproduce.

Abstract:目的选择HMG-CoA还原酶抑制剂洛伐他汀作为模型药物,在巴马香猪体内进行生物利用度及药代动力学研究,探讨巴马香猪对洛伐他汀的处置规律。方法12头6~8月龄健康雄性巴马香猪随机分为灌胃组、静脉组和空白对照组,其中灌胃组和静脉组通过胃管和耳静脉分别给与洛伐他汀8、1.2 mg/kg。给药后前腔静脉采血,用RP-HPLC-UV法检测血浆洛伐他汀浓度;用3P97计算药代动力学参数,用统计矩法计算血药浓度-时间曲线下面积(AUC0-∝)。结果灌胃组t1/2和Ke分别为3.43 h和0.20 h;静脉组C0、Ke、Vdt、1/2和CL分别为180.27 ng/mL、0.17h、6.7L/kg3、.97 h和66.97 mL/min。统计矩法计算得到灌胃组和静脉组的AUC0-∝分别为每小时409.41,1627.78ng/mL,绝对生物利用度F为3.8%。结论洛伐他汀在巴马香猪体内的绝对生物利用度人和犬基本一致,提示巴马香猪与人和犬对药物的处置具有一定的相似性。

Abstract:Miniature pigs have been becoming important laboratory animal for life science researches.Many excellent miniature pig breeds exist in China.Miniature pig breeds or resources in China belong to resource breed,outbred colony or inbred strain.Chinese miniature pig breeds and resources were stated and analyzed in th paper.

Abstract:In the research of transgenic mice models,the controlled expression of target gene is very important to the application of the transgenic mice models.The implement of the expression of the transgene in time and in space has got great progress.This article introduces the principles,application,progresses in research of Tet system and the Cre/LoxP system,and presents the strong and weak points of those two control systems.The focal point is the outcome of the combined application of those two systems.Furthermore,to introduce the application of Cre/Loxp system in directional expression of HPV11 complete gene in mice.Accordingly,put forward an assumption to solve the problem of orientation expression of the multi-gene.