Abstract:Objective In order to enrich the present Tet mice animal bank and provide an useful tool for studies on pathogenesis of HCV-C gene,a study on establishment of a double transgenic mouse model containing both ApoE-rtTA and TRE-HCV-C elements was carried out.Method\ Two constructed fragments carrying ApoE-rtTA and TRE-HCVC were individually introduced into fertilized eggs by microinjection.The injected eggs were implanted into the oviducts of female mice,from them offsprings were obtained.Grossed the two strains double transgenic mice were got and identified with RT-PCR and western blot.Results\ Three mice carrying ApoE-rtTA gene and five mice carrying HCV-C gene were identified by PCR and Southern blotting.Double transgenic mice were got by crossing the two transgenic mice.It turned out 3 transgenic mice carrying ApoE-rtTA and 125 positive offsprings,and also 5 transgenic mice carrying TRE-HCV-C and 16 positive offsprings.Five double transgenic mice were generated.Conclusions\ Transgenic mice carrying tet-on systems have been developed,which enriched the world "Tet-mouse" bank,and provides an animal model for research on HCV core protein.Moreover,an ideal model is established for studies on some genes related to hepatic disease in vivo and provides a technical platform for research on gene expression with temporal,quantitative and site-speciality regulation.

Abstract:目的探讨静脉注射博莱霉素(BLM)和博莱霉素A5(BLM-A5)对小鼠肺组织损伤的不同作用。方法8周龄雌性C57BL/6小鼠49只,随机分为BLM组19只、BLM-A5组20只和对照组10只,分别经尾静脉一次性注射BLM 150 mg/kg、BLM-A5 150 mg/kg和生理盐水。观察每组小鼠体重、生存率,肺组织病理改变及肺组织羟脯胺酸的含量。结果①BLM组和BLM-A5组小鼠最低体重分别为静脉注射处置前的65.46%和66.87%,均较对照组明显降低(P值均小于0.001)。②BLM组、BLM-A5组和对照组小鼠的生存率分别为:43%、55%和100%。③BLM组小鼠注射处置后28 d,在胸膜下及血管周围形成广泛、稳定的间质纤维化病理改变,而BLM-A5组无明显纤维化形成。BLM组与BLM-A5组、正常组比较,肺纤维化病理评分均明显增高(P值均小于0.001)。④BLM-A5组和对照组注射处置后28 d,右肺羟脯氨酸含量分别为471.6±49.4 nmol和405.0±74.6 nmol,两组间无统计学差异。BLM组为978.4±106.1 nmol,与BLM-A5组、对照组比较分别存在显著性差异(P值均小于0.01)。结论静脉注射BLM与BLM-A5,对小鼠肺组织损伤的作用存在着明显的差异,BLM可导致肺间质形成广泛的纤维化,而BLM-A5仅引起轻微的肺组织损伤,不形成明显的间质纤维化。

Abstract:Objective In order to explore the mechanism of stress-induced immune dysfunction,two kinds of animal models with depressed immune response under stresses were established.Methods\ The mice were divided into 3 groups: the restraint-stressed group,heat-stressed group and control group.The experimental mice were stressed by restraint and heat,respectively.After being stressed,the mice of each group were subdivided into two parts,one part was injected by Escherichia coli.The death rate of each group was calculated.In other of them,spleen indexes of the mice of each group were compared,and the histopathological changes were examined.Results\ The death rates of the restrained stress group and heat-stressed group were all higher than that of control group(P<0.005).The death rates were not significantly different between the restrained stress group and heat-stressed group(P>0.05).The spleen indexes of both the restraint stress group and heat-stress group were lower than that of the control group(P<0.05).The histopathological examination revealed white pulp decreased with disarrangement in spleen of stressed mice.Conclusion\ The mice models of stress-induced immune dysfunction have been successfully established and there may be a correlation between the spleen structural alterations and immune dysfunction.The changes are not significantly related to types of stresses.

Abstract:Objective To develop a diabetic mouse model to investigate metabolic changes of blood glucose level and body weight in the MKR mouse,a model in which expression of a dominant-negative IGF-I receptor in the skeletal muscle leads to systemic insulin resistance and diabetes.Methods\ MKR mice were received from the National Institutes of Health,USA,and genotyped by a specific PCR reaction using tail DNA.These mice were followed up for blood glucose level and body weight once a week from 1 to 16 weeks of age.The serum insulin level and glucose tolerance were determined at two months,using wild-type Kunming mice or C57 mice as controls.Results\ The offspring of MKR mice exhibited stable transgenic hereditary.These mice were hyperglycaemic from 3 weeks of age onward.The body weight increase slowed down after 4 month.There was 20-fold hyperinsulinemia in MKR vs.wild-type littermates,and very significant glucose intolerance in 2month-old MKR mice.The hyperglycemia is more severe in male especially older male MKR mice,than their female counterparts.Conclusion\ We have estabolished the MKR model,set up a PCR reaction to genotype and revealed sexual dimorphic changes in the type 2 diabetes.

Abstract:目的使用屋尘螨(HDM)提取液致敏和激发C57BL/6小鼠构建气道变态反应性炎症模型的方法。方法模型组和对照组,各8只。模型组以HDM致敏和激发小鼠,分别于第0、7、14、21天腹腔注射HDM,10 d后,连续雾化吸入HDM 3 d。对照组以PBS代替HDM。进行支气管肺泡灌洗液(BALF)细胞总数计数和分类计数,肺组织病理检查,ELISA测定BALF上清中IL-4、IL-5和IFNγ-水平。结果模型组可见明显炎性细胞浸润,以嗜酸性粒细胞为主。而对照组未见明显炎性细胞浸润。BALF中细胞总数计数和嗜酸性粒细胞比例较对照组明显升高(P<0.01);BALF上清中IL-4、IL-5水平较对照组明显升高(P<0.001),IFNγ-水平较对照组降低(P<0.01)。结论使用HDM致敏和激发C57BL/6小鼠成功地建立了小鼠气道变应性炎症模型。

Abstract:Objective The purpose of this study was to determine the restoring-capacity of a Chinese herbal medicine decoction to the ultrastructure of hepatocytes in eel with liver injury induced by Cu-exposure.Methods Eels Anguilla anguilla were injured with cupric sulfate and treated with a Chinese herbal medicine decoction mainly composed of Salvia miltiorrhiza,Schisandra chinensis(Turcz.)Baill and Radix Glycyrrhizae.At the end of the trial period,fish were killed,dissected and the liver was examined by electron microscopy.Results Compared with the control fish,the hepatocytes of Cupoisoned control fish showed mitochondria swelled with disintegration or disappearance of cristae,dilation,degranulation and disintegration of rough endoplasmic reticulum.Intracisternal sequestration,autophagosomes and vacuoles were often observed.Some nuclei of hepatocytes presented nuclear crenation.Compared with the Cu-poisoned control fish,the ultrastructure of hepatocytes of drug-treated fish showed recovery notably.Conclusion Our findings demonstrated that this Chinese herbal medicine decoction may promote the recovery of ultrastructure of hepatocytes in the copper-damaged eel liver,indicating an effective protective effect against those damages.

Abstract:目的比较CCl4和脂多糖诱导的两种急性肝损伤模型中肝细胞凋亡的病理特点。方法雄性BABL/c小鼠随机分为正常组、CCl4模型组与脂多糖模型组。CCl4和脂多糖模型小鼠分别腹腔注射0.03 mL/kg 50?l4/橄榄油混合液和10μg/kg脂多糖与900μg/kg半乳糖胺。9 h后采集标本,检测肝细胞凋亡,血清ALT、AST活性,肝组织SOD活性、MDA含量。结果与正常组比较,CCl4和脂多糖模型小鼠血清ALT与AST活性均明显升高(P<0.01),且脂多糖模型高于CCl4模型(P<0.01);模型小鼠均有明显肝细胞凋亡(P<0.01),且两组间无显著差异,但CCl4模型呈局灶性,以中央静脉周围为主;脂多糖模型呈弥漫性,肝细胞坏死与肝组织炎症更显著(P<0.01)。CCl4和脂多糖模型肝组织均SOD活性升高、而MDA含量下降(P<0.01),但CCl4模型更显著(P<0.05)。结论CCl4与脂多糖诱导的急性肝损伤小鼠模型均有明显的肝细胞凋亡,但CCl4模型以中央静脉区为主,脂质过氧化损伤更为明显;脂多糖模型呈弥漫性,肝细胞坏死与肝组织炎性反应较重。

Abstract:Objective To explore the characteristics and risk factors of ventricular fibrillation(Vf) in open-chest acute myocardial infarction models of swine.Methods Coronary artery was surgically occluded at different percentage point of LAD varying from 25% to 75% in fifty-seven miniswines.Cardiac fraction(LVEF) using echocardiography was assessed before and 1 hour after myocardial infarction(MI).Coronary angiography was used to confirm the position of ligation after MI.The weight,gender,heart rate(HR) before and after MI,LVEF,thoracotomy route(intercostal or parasternal),operation duration,percentage point of LAD ligation,arhythmia after MI were recorded and compared between Vf group and Vf-free group.The risk factors of Vf were determined by logistic regression analysis.Results Eighteen of the 57 pigs suffered from ventricular fibrillation within 35 min after LAD ligation and 11 cases died.Compared with Vf-free group,the Vf group featured a higher position of LAD ligation,a higher incidence of ventricular tachycardia and maximal HR>160 bpm post MI.Logistic regression analysis showed the position of LAD ligation was a sole risk factors of Vf after MI with a statistical significance.Pigs with HR>160 bpm or <60 bpm post MI was susceptible to Vf.Conclusion The position of LAD ligation is a vital determinant of Vf in establishing acute MI model of swine.Attentions should be paid to the first 30 min after LAD ligation,HR,and ventricular tachycardia.

Abstract:Objective To explore the genetic relationship and genetic diversity of plateau Juema pigs,and whether it is affected by foreign genealogical factors or not.Methods To analyze the alleles frequency,genetic distance,the phylogenetic tree and principal component etc.by using 9 microsatellite DNA markers in 5 pig populations.Results In 125 individuals from the 5 pig populations,assessed by 9 microsatellites,148 alleles were detected altogether,in which Juema pig is the least(55).The genetic distance D_A value and the standard genetic distance D_S value between Juema and Lanzhou pig populations were the largest,0.6781 and 1.3312,respectively.D_A and D_S formed 4 types of phylogenetic tress by using unweighted pair-group method using the average approach and neighbour-joining.It indicated that Juema pig population is a sort and the other 4 populations are another.As the representative of Juema pig,principal constituent(PC1) was 62.174%,which had major difference from other 4 pig populations.Conclusion Comparing Juema pig with Lanzhou,Wuwei,Lintao and Qinhai pigs,the genetic distance and principal constituent discrepancy of Juema pigs is great,and its genetic relationship is far,therefore Juema pig is purer,not influenced genealogically by the other four pig populations.

Abstract:目的探讨用非放射性标记的寡聚核苷酸探针(GTG)5进行近交系小鼠的DNA指纹分析。方法用非放射性标记的寡聚核苷酸探针(GTG)5制作BALB/c、C57BL/6J、DBA/2、C3H近交系小鼠的DNA指纹图,对这四个品系的小鼠进行遗传检测,分析各品系内和品系间的遗传变异性。结果(GTG)5探针可产生具有良好多态性的DNA指纹图,平均图带数为8~12条。各品系内的DNA指纹图平均相似系数(-x)在0.96~1.00的范围内,具有相同指纹图的概率(P)均在3.1×10-1以上,极显著地高于品系间的相似系数(0.22~0.39)和相同指纹图的概率(P<1.07×10-4)。结论(GTG)5可用于制作近交系小鼠的DNA指纹图以对其进行遗传检测。

Abstract:目的 通过化学遗传学方法建立视黄酸缺乏的斑马鱼模型,探讨视黄酸缺乏对斑马鱼胚胎心脏前后轴发育即房室分化的影响.方法 在斑马鱼胚胎孵育的5 hpf,用不同浓度梯度的视黄醛脱氢酶2抑制剂DEAB(1×10-6、5×10-6、10×10-6、25×10-6 mol/L)处理斑马鱼胚胎,实时观察斑马鱼胚胎发育的全过程.通过给予斑马鱼胚胎外源性视黄酸,观察其对DEAB的拮抗作用.应用胚胎整体原位杂交观察视黄酸缺乏对心脏特异基因vmhc和amhc表达的影响.结果 斑马鱼胚胎的生存率随着DEAB处理浓度的增加而降低,当DEAB浓度≥5×10-6 mol/L时,斑马鱼的畸胎率达100%.5×10-6 mol/L DEAB的致畸作用能够被1×10-9mol/L外源性视黄酸所拮抗.整体原位杂交结果显示视黄酸缺乏会导致斑马鱼胚胎心脏房室分化异常,表现为vmhc表达细胞的范围增大,amhc表达细胞的范围缩小.结论 通过外源性DEAB处理能有效地建立视黄酸缺乏的斑马鱼模型,DEAB影响胚胎发育存在剂量依赖性.视黄酸在斑马鱼心脏前后轴发育过程中起重要调控作用,心脏发育早期视黄酸缺乏会抑制心房的发育而支持心室的发育,出现房室分化异常.

Abstract:Objective To compare two types of orthotopic implant models of human gastric cancer in nude mice using glue paste technique and to provide an ideal animal model for research in the field of pathogenesis and experimental treatment of gastric cancer.Methods The orthotopic implant models of human gastric cancer in nude mice were established using OB and FS organic glue paste technique,respectively.the tumor growth characteristics,metastatic and morphological characteristics were analyzed and compared.Results In the model using OB glue no serious tumor necrosis was observed,the rate of ascite formation was 85.7%,the rate of pyloric obstruction was 57.1%.In the model using FS organic glue the rate of extensive tumor necrosis was 100%,the rate of ascites formation was 14.3%,no pyloric obstruction was observed,metastases in the lung and brain were observed in three cases.Conclusions The orthotopic implant model of human gastric cancer in nude mice using FS organic glue paste technique,simulating the clinic process of patients,has been successfully established.It provides an ideal animal model for studies on mechanism of metastasis and treatment of human gastric cancer.

Abstract:Objective In order to provide valuable information for learning and memory basic study,a series of behavioral experiments were carried out to explore the difference of learning and memory ability of different strain mice.Method 80 BALB/c and 80 ICR mice were randomly divided into Morris water maze test group, jumping estrade test group,shuttle box test group and rota-rod test group.The learning and memory ability of the mice were assessed.Results Morris water maze test: Escape times of BALB/c mice in Morris maze had no significant change in their space learning and memory ability during the 9 runs water maze training.However,escape time of ICR mice was gradually decreased.From the 4~th run to the 9~th run,the escape time of ICR mice was significantly decreased in comparison with the preceding 3 runs(P<0.001).Jumping estrade test: wrong times in 5 minutes and jumping latent time of ICR mice before and after jumping estrade experiment had significant differences,respectively.Shuttle box test: The positive escape times,passive escape times,electro-stimulus time of ICR mice pro-shuttle box test training were higher than those of post-shuttle box test training,respectively.But those of BALB/c mice had no significant difference between pro-shuttle and post-shuttle box test training.ROTA-ROD Test: the keep-running time of ICR mice in ROTA-ROD test was significantly higher than that of BALB/c mice.Conclusion These results suggest that ICR mice are more suitable for some kind of learning and memory experiments than BALB/c mice.

Abstract:Objective To study the mechanisms of action of fasudil at different doses on the cardiomyocyte apoptosis in the rats with pressure overloaded heart failure following coarctation of the ascending aorta.Methods A rat model of heart failure was created by coarctation of ascending aorta.40 female Wistar rats with heart failure were divided randomly into four groups,including natrium chloride (0.1 mL),fasudil(1 mg/kg),fasudil(5 mg/kg),fasudil(20 mg/kg)(n=10 per group),which were treated by i.p injection b.i.d for 4 weeks.Another 10 age-matched sham-operated rats were used as control.The rats were observed for another 20 weeks after coarctation of the ascending aorta operation.Hemodynamic parameters,ratio of LV weight to body weight,cardiomyocyte apoptosis,protein expression levels of bcl-2 and c-myc were investigated in all the groups.Results Fasudil intervention reduced cardiomyocyte apoptosis,decreased protein expression level of c-myc and improved protein expression level of bcl-2 in rats with chronic cardiac dysfunction,in a dose-dependent manner.The number of cardiomyocyte apoptosis and the protein expression level of cmyc were significantly lower and the level of bcl-2 was markedly higher in the group of high dose fasudil than those in the groups of low dose fasudil.Conclusions Fasudil can effectively decrease cardiomyocyte apoptosis,reduce the protein level of c-myc and increase bcl-2 protein level dose-dependently,which may be one of mechanisms of prevention and treatment of heart failure.

Abstract:Objective To study the localization and distribution of somatostatin in rat mammary gland.Method Immunohistochemical methods were used to detect somatostatin in mammary gland of virgin rats,on days 6,12,18 of pregnancy,and days 6,12,18 of lactation.Results Intense staining for somatostatin in cytoplasm of epithelial cells and in the secretory material at all stages mammary gland.Conclusion Somatostatin is located in the whole epithelial cells cytoplasm and the secretory material of rat mammary gland.

Abstract:目的在兔急性心梗再灌注模型中通过硬膜外给予吗啡,探讨其对兔心肌急性缺血再灌注损伤的保护作用。方法采取开胸结扎LAD(left anterior descending,左前降支)的急性心肌梗死模型,结扎冠状动脉LAD 30min后,松开结扎线,再灌注4 h。新西兰兔22只随机分成2组:①硬膜外吗啡组:经硬膜外注入吗啡(0.25 mg/kg,配成0.5 mL溶液);②对照组:经硬膜外注入生理盐水0.5 mL。缺血再灌注后逐层关胸,4周后行超声心动图检查。结果缺血再灌注后4周,两组LVEDD和LVESD均明显增大,但以对照组为甚,组间比较差异有显著性(P<0.05);而LVEF和LVFS,在缺血再灌注后4周硬膜外吗啡组分别降低了9.36%和8.24%,对照组分别降低了13.53%和12.18%,对照组比硬膜外组更显著(P<0.05),左室心功能硬膜外吗啡组好于对照组(LVEF:69.73%vs.62.99%;LVFS:31.39%vs.26.59%;LVEDD:14.35 vs.16.96;LVESD:9.52 vs.12.01),差异有统计学意义(P<0.05)。结论硬膜外注射吗啡能改善缺血再灌注后左心功能,并对缺血再灌注心肌有明显的保护作用。

Abstract:核移植来源的胚胎干细胞 (ES) 细胞在动物疾病模型上的成功治疗作用以及核移植来源的患者特异的ES细胞研究进展有望将这一技术即治疗性克隆应用于人类疾病治疗.本文对治疗性克隆的研究进展、治疗性克隆研究中存在的问题以及治疗性克隆的应用前景作一综述和讨论.

Abstract:GK大鼠是1975年由Goto等在日本仙台从Wistar大鼠中反复选择形成的非肥胖性2型糖尿病鼠种。该鼠具有葡萄糖刺激的胰岛素分泌受损,β细胞分泌受损,空腹高血糖,肝糖原生成增多,肝脏、肌肉和脂肪组织中度胰岛素抵抗等特点,晚期合并各种并发症,与人类2型糖尿病进展极为相似。