Abstract:ObjectiveTo assay the coagulation factor IX -knockout mice. MethodsThe DNA of mice was analysed by PCR and the plasma of the mice was measured by one stage method. ResultsPCR analysis of FIX mice was positive and FIX activity was below 5%. ConclusionThe result showed that the coagulation FIX -knockout mice could be generated steadily and exhibited a phenotype similar to human hemophilia B.

Abstract:目的　对雌性树的麻醉、生殖器官解剖与卵母细胞形态特征进行观察 ,为制备转基因树奠定基础。方法　用 1%的戊巴比妥钠 (10 - 3ml g体重 )肌内注射对树麻醉 ,比较在不同环境温度下对麻醉效果的影响。对雌性树生殖器官解剖结构、卵母细胞形态特征等进行观察。结果　①在 2 5～ 2 8℃、18～ 2 0℃下麻醉的持续时间分别为 80min、130min ;②雌性树的子宫为双角子宫 ,卵巢外有包膜 ;③卵母细胞富含色素 ,卵母细胞的透明带与质膜的韧性较山羊强。结论　在 2 5～ 2 8℃下 ,用 1%的戊巴比妥钠 (10 - 3ml g体重 )对树麻醉时间比较适中 ,便于实验操作而且苏醒快。雌性树生殖器官解剖结构、卵母细胞形态特征观察表明 ,树的胚胎移植、转基因等实验操作与小鼠类似 ,但树的受精卵须进行离心处理。

Abstract:ObjectiveTo investigate the function of a novel mouse imprinting gene which was cloned by DD -PCR and RACE, a knockout strategy was applied to target this gene, so in this study the targeting vector was constructed and the recombinant cell clones were primarily screened by PCR. Methods Two large fragments flanking the coding region of this gene were cloned into the pKO Scrambler NTKV -1906 vector after the genomic sequence of this gene was analyzed by computer, and then the linearized replacement vector was transfected into the mouse ES cells and drug -resistant cell clones were picked out for screening by a PCR -based strategy. Results The targeting vector and a control vector were constructed successfully and three recombinants were obtained by PCR screening for further use. Conclusion This targeting vector and the recombinants provided a support for identification of the function of the mouse novel imprinting gene . At the same time, a PCR -based screening was established and successfully applied to obtain three positive clones for further use, which would reduce the work of screening in the knockout process.

Abstract:ObjectiveTo clarify the G -banding pattern and spontaneous aberration frequencies of an inbred line of Chinese hamster of shanyi colony. Methods The bone marrow chromosome preparation and G -banding pattern for Chinese hamster were applied. Results Of the 30 G -banding cells from 14 Chinese hamsters 7 cells were selected for banding analysis. The chromosomes were distinguished by characteristic banding patterns. In addition,800 metaphase cells were analysed by conventional Giemsa stain. The results showed that break was 0.25%, acentric aberration and translocation were 0.05%. The spontaneous aberration frequencies were very low. Conclusion The discernment of G -banding pattern of an inbred line of Chinese hamster is for providing scientific basis to study on abnormal structure and gene map.

Abstract:ObjectiveThe Lats1 gene knockout mice were selected as an example to study the technology on germplasm storage of the genetic mice in Resource Bank of National Genetic Mice. Methods Techniques such as embryonic transplantation, genotype and embryo cryopreservation were applied. Result 24 Lats +/- mice were got. After reproduction, 110 embryos were frozen in resource bank. Conclusion The above -mentioned approaches can insure the successful germplasm storage of Lats1 gene knockout mice in resource bank.

Abstract:目的建立东方田鼠生化与分子遗传学标记检测法.方法参考近交系小鼠、大鼠生化遗传标记检测方法,对东方田鼠某些同功酶进行活性测定.根据东方田鼠特异DNA序列,合成引物,扩增东方田鼠基因组DNA,将PCR产物回收、序列分析并进行生物信息学分析.结果东方田鼠Trf、Es-3和Ce-2三个生化位点呈现遗传多态性,而Id1、Mod-1、Car-2、Gpd-1、Gpi-1、Hbb、Es-1七个生化位点无遗传多态性.用合成的特异引物扩增东方田鼠基因组DNA,得到了丰富的多态性资料.对其中的20只东方田鼠的扩增产物进行测序并进行多序列比较,发现10个等位基因以及16个单核苷酸多态性(SNP)位点.结论初步建立了东方田鼠生化及分子遗传学标记.分子遗传学标记与生化遗传标记相比,具有杂合率高、重复性好、易于操作等优点.这些结果为深入研究东方田鼠的遗传背景、生物进化规律积累了基础资料.

Abstract:ObjectiveCompared with Schmidt Criterion and the others, the authors studied the characteristics of acute necrotizing pancreatitis (ANP) in the rat model triggered with sodium taurocholate (biliary pancreatitis) in order to develop more precise and reasonable method of pathological evaluation in practice. Methods Forty -eight Sprague -Dawley (SD) rats were assigned into 3 different groups, i.e., treatment (16), ANP (21) and sham -operation (11). Acute necrotizing pancreatitis was triggered with injection of sodium taurocholate via the biliopancreatic duct. Schmidt Criterion was employed after modification for histopathological scoring to make sure the accuracy of different methods in evaluation, combined with transmission electronmicroscope. Results Autopsy of the rats showed large amounts of bloody ascites, the largest, 6% of the body weight. Severe hemorrhage and acinar necrosis, destroyed pancreatic lobules with lots of red blood cells between them were demonstrated under microscope. Inflammation, congestion and hemorrhage were also found in liver, heart, lungs and kidneys. Of the 4 parameters of the Schmidt Criterion, the difference of hemorrhage was not revealed significantly between groups, while the other 3, inflammation, edema and acinar necrosis, too elaborate and troublesome. We used the mean count of the red blood cells between pancreatic lobules or grade scoring and found out the significant difference of pancreatic hemorrhage. Conclusions 1. The bile salt model of acute necrotizing pancreatitis triggered with sodium taurocholate is characterized by large amounts of bloody ascites, severe hemorrhage and acinar necrosis, microthrombi in capillaries in pancreatic tissues, and injuries in multiple extra -pancreas organs; 2. On the basis of modification of 3 parameters in Schmidt criterion, i.e., edema, acinar necrosis and inflammation, and the mean count of red blood cells, we suggested a new criterion for more precise histopathological assessment.

Abstract:目的　对比观察滋补肾阴方与温补肾阳方对卵巢切除所致骨质疏松大鼠的治疗作用 ,以探讨它们之间的疗效是否存在差异。方法　将雌性Wistar大鼠 4 9只随机分为正常对照组、假手术组、卵巢切除组、滋阴组 (灌服生药含量为 1 0 5g ml的滋补肾阴方 ,按每 10 0g体重 0 7ml剂量 )、补阳组 (灌服生药含量为 0 78g ml的温补肾阳方 ,按每 10 0g体重 0 7ml剂量 )。于造模 3个月后开始给药 ,1次 日 ,连续 6d ,休息一天后 ,再连续灌服 6d。给药 2个月后 ,将大鼠处死 ,取胫骨行不脱钙骨切片 ,甲苯胺蓝染色 ,对其进行骨组织形态计量。结果　切除卵巢 5个月后 ,大鼠胫骨TBV %显著降低 ,TRS %以及TFS %、AFS %、MAR、BFR、OSW和mAR皆显著增高 ,给大鼠灌服滋阴补肾方和温补肾阳方 2个月后 ,均能使上述指标发生逆转 ,但程度有所不同 ,温补肾阳方的效果要明显优于滋补肾阴方。结论　滋补肾阴方与温补肾阳方对卵巢切除所致的骨质疏松均有明显的治疗作用 ,但温补肾阳方的疗效要优于滋阴补肾方。

Abstract:ObjectiveThe follicular fluid (oFF) and hypoxanthine were added in maturation medium to prevent the GVBD occur,prolong the transcription of the oocyte genome, improve the maturation level of oocyte cytoplasm, enhance the IVF embryo developmental potential. Method The oocytes recovered from the ovaries collected in abattoir were cultured in vitro maturation medium containing follicular fluid(oFF) and hypoxanthine in different concentration. The nuclear phases were checked after 24hrs. Results Oocytes were incubated in culture medium with 50% and 100% follicular fluid for 24h, and 19% and 33 .3% of them were maintained in GV stage, respectively. At a concentration of 4mM hypoxanthine in culture medium, the meiosis resumption of ovine oocytes was inhibited significantly. 21 .6% of oocytes were maintained in GV stage, and 44 .6% were in PI stage. But the inhibitory effect could not be reversed by the induction of FSH. Conclusion The concentration of inhibitory factors had liner relationship with meiosis arrest. There were dose -dependent effects in the control of meiosis.

Abstract:ObjectiveTo study the reaction of Guizhou miniature pig to the DNA damage and repair caused by chemical substances or drugs. MethodsSingle cell gel electrophoresis was used to study the DNA damage and repair caused by 5 -azacytidine(5 -azaC) in phytohemagglutinin (PHA) -stimulated and unstimulated lymphocytes. Results5 -azaC induced significant DNA migration(comet tail) in unstimulated lymphocytes, and the DNA migration after 2h repair incubation was of no significance compared with pre -incubation. In contract, the comet tail induced by 5 -azaC significantly reduced after 2h repair incubation compared with pre -incubation. ConclusionThe DNA damage caused by 5 -azaC in unstimulated lymphocytes of Guizhou miniature pig was not significantly repaired within 2h post -incubation, but significantly repaired in PHA -stimulated cells.

Abstract:ObjectiveTo study the effect of self ilium periosteum free state transplantation on curing femoral neck fracture. Methods 7 ordinary adult beagles, 14 hip joints in all, were for animal models of femoral neck fracture. After fixation, self ilium periosteum was stuck to the fracture surface with protein adhersive. Observation was with naked eyes and by X -rays and pathological section observed under microscope. ResultsOne month later, X -ray showed that the fracture line blured, transplanted periosteum grew with femoral neck. Under microscope, blood capillary in the periosteum increased sharply. Cartilage and bone like matter were seen. Three months after operation, X -ray showed that fractures was united, a lot of bone matter were seen where the periosteum was transplanted. Under microscope, developed blood vessel net in the periosteum and lots of bone cells were seen, new bone trabecula growing into the original ones in the femoral neck and united with them. ConclusionsFree state transplanted self ilium periosteum can survive and help bone reunit with the blood vessel net reconstructed. It is a feasible and easy way to cure femoral neck fracture.

Abstract:目的对雌性树鼩的麻醉、生殖器官解剖与卵母细胞形态特征进行观察,为制备转基因树鼩奠定基础.方法用1%的戊巴比妥钠(10-3ml/g体重)肌内注射对树鼩麻醉,比较在不同环境温度下对麻醉效果的影响.对雌性树鼩生殖器官解剖结构、卵母细胞形态特征等进行观察.结果①在25～28℃、18～20℃下麻醉的持续时间分别为80 min、130 min;②雌性树鼩的子宫为双角子宫,卵巢外有包膜;③卵母细胞富含色素,卵母细胞的透明带与质膜的韧性较山羊强.结论在25～28℃下,用1%的戊巴比妥钠(10-3ml/g体重)对树鼩麻醉时间比较适中,便于实验操作而且苏醒快.雌性树鼩生殖器官解剖结构、卵母细胞形态特征观察表明,树鼩的胚胎移植、转基因等实验操作与小鼠类似,但树鼩的受精卵须进行离心处理.

Abstract:目的鉴定凝血因子Ⅸ基因剔除小鼠.方法采用PCR扩增检测小鼠的DNA样品以及采用一期法检定小鼠血浆FIX活性和血浆凝血酶原时间(plasma prothrombin time,PT)、白陶土部分凝血活酶时间(Kaolin partialthromboplastin time,KPTT)值.结果小鼠PCR检测为阳性,FIX活性＜5%.结论凝血因子Ⅸ基因剔除小鼠能稳定遗传,鉴定结果提示该小鼠符合人血友病B相应临床症状.

Abstract:ObjectiveTo study the methods of establishing mesangial proliferative glomerulonephritis rapidly. Methods30 SD rats were divided into three groups: the control groupA, the groupB receiving SEB by intravenous injection, bovine serum protein orally,immunological adjuvants injected subcutaneously and the groupC being the same as group B in addition to left hepatic lobels removed.Urinary albumin,IgA immunofluorescence and pathohistological assays were made 14 weeks later. ResultsUrinary albumin was found in group C more earlier than the other groups, but with no statistic significance 14 weeks later. The pathological examination showed mild to moderate mesangial expansion with a few cells in kidneys of groups B and C, the IgA immunofluovescence was more stronger than the control group. Conclusion Both group B and gruop C can induce the mesangial proliferative glomerulonephritis, and the urine protein occurred in group C earlier, with no statistic significance 14 weeks later.

Abstract:ObjectiveTo find out the influences of ages and sexes on normal blood biochemical parameters in the Lagurus lagurus closed colony. Methods 23 blood biochemical parameters of male group, female group, 1-2 monthes group and 24 monthes group in Lagurus lagurus closed colony were determined by using AEROSET auto blood biochemical analyser. Results The same parameters in two age groups had no difference, so did in two sex groups. Conclusion In this experiment, ages(1-2 monthes and 24 monthes) and sexes showed no influences on the 23 normal blood biochemical parameters in the Lagurus lagurus closed colony.

Abstract:ObjectiveTo establish the ARDS animal model in rats. MethodKM mice and SD rats were divided into control group and paraquat -exposed groups (with a single dose of 100mg/kg in mouse and 120mg/kg in rat). Each group was subdivided into three smaller groups (24 -hour group, 48 -hour group, and 72 -hour group). Blood was collected from the femoral artery of mouse for blood gases analysis. Tissues from lung of rats were embedded in paraffin for histological analysis. Result1.After the paraquat was given, respiration movements of the animals were getting more and more difficult with cough, pink excretions in nasal cavity, progressively decreased appetite, and weight loss. 2.Lung changed from pink color to hepatization in gross view. 3.By blood gases analysis, the blood PH value and PaO 2 gradually decreased, while PaCO 2 and pulmonary coefficient went in the other way. Statistical analysis (t -test) showed an obvious difference between experimental groups and control group ( P <0 .01). 4.Histological section showed a typical inflammation. Transparent membrane formed in the pulmonary alveoli. ConclusionARDS animal model can be successfully produced by means of exposure to paraquat.

Abstract:采用PCR法对不同样品进行检测 ,其目的是寻求在小鼠肝炎病毒 (MHV)感染早期 ,可以尽快检测出MHV的一种最佳检测材料。将被感染小鼠的新鲜粪便、干燥粪便、笼具过滤网上的尘埃以及病毒液作为基因检测样品 ,进行检测并比较检出的灵敏度。将灵敏度量化 ,以稀释成不同浓度的cDNA用巢式PCR法能够检出的稀释界限的倍数表示。其结果 :第一 ,从过滤网中的尘埃所得到的cDNA是新鲜粪便的 10 0倍 ;第二 ,从粪便检出的灵敏度不以粪便的干燥而发生变化。第三 ,如果将病毒液放在室温下 ,会随室温干燥 ,检出的灵敏度会大幅度的降低。从以上结果看 ,在过滤网上的尘埃中有大量的MHV存在 ,尘埃中的一部分病毒 ,即使放在室温也能够以稳定状态存在。作者认为 ,将饲育装置的排气过滤器中粘着的尘埃收集 ,作为检测的样品 ,结合巢式PCR法进行测定 ,对于频繁地检测单个饲育笼具或单元式饲育室内的MHV比较简便易行 ,是一种较好的早期监测感染鼠肝炎病毒的方法。

Abstract:嵌合体动物 (Ｃｈｉｍｅｒａｓ)是指由两种或两种以上具有不同遗传性的细胞系组成的聚合胚发育成的个体。早期嵌合体动物主要作为发育生物学中的一种十分有用的工具 ,用于研究胚胎发育过程中组织器官的发生等 ,1986年建立“胚胎干细胞 (ｅｍｂｒｙｏｎｉｃＳｔｅｍＣｅｌｌ,ＥＳｃｅｌｌ)介导的转基因的途径”[1-2 ] ,可制作带有定点突变基因的动物品系。用基因打靶策略可以精确构建疾病的动物模型 ,在各个水平研究基因功能 ;用基因诱捕策略可以鉴定新基因和分析它们在生物现象中重要性 ,这是其他转基因方法所不能比拟的。嵌合体动物的获得…