Abstract:Objective To observe the pathological changes in the liver of p21~(HBsAg/HBsAg) transgenic mice.Methods\ The p21~(HBsAg/HBsAg) transgenic mice(2,6,12, 18,24 months) of SPF grade and corresponding p21~(+/+) mice were dissected and the livers and hepatic tumor tissues were examined by histopathology and electron microscopy.Results \ Distinct pathological changes appeared in the liver of p21~(HBsAg/HBsAg) transgenic mice both macroscopically and microscopically.With the increase of age,the liver of the transgenic mice turned to be dark and sclerotic with formation of scattered nodules and tumors.Under the light microscope,the hepatocytes were swollen and inflammatory cell infiltration occurred in the liver.Fatty degeneration appeared with punctuate,focal and crumblike necrosis.Atypical hyperplasia and hepatocellular carcinoma(HCC) developed.Resembling normal hepatocytes,the hepatpocarcinoma cells were well differentiated,forming cord-like and acinus-like structures.Under the electron microscope,the nuclei of carcinoma cells were dark stained with invagination of nuclear membrane.Mitotic figures were observed.Mitochondria were swollen with less cristae.HCC were found in four cases of 18-month-old p21~(HBsAg/HBsAg) transgenic mice(4/10,40%),and six cases of 24 months old(6/10,60%),respectively,of which metastasis occurred in two mice.Conclusions\ Prominent pathological changes occurred in the liver of p21~(HBsAg/HBsAg) transgenic mice.Well differentiated hepatocarcinoma can be developed in the mice at the age of 18 months.Metastasis of HCCs can be seen in elderly transgenic mice.

Abstract:目的观察TGF-β1基因过表达诱发大鼠肺间质纤维化动物模型的组织病理学变化.方法成年健康SD大鼠30只,随机分为空白对照组10只,模型组10只和空载体对照组10只,其中对模型组大鼠气管内一次性注射携带TGF-β1基因腺病毒载体(AdTGF-β1 223/225,1×109 pfu,0.4 mL),空白对照组大鼠气管内注射等量的生理盐水,空载体对照组大鼠气管内注射等量的腺病毒空载体.结果一次性肺气管内注射 AdTGF-β1 223/225可造成肺急性炎症和肺间质纤维化,而对照(腺病毒空载体)并不引起相应的变化.结论肺组织局部分泌的细胞因子TGF-β1可以单独诱发肺间质纤维化.

Abstract:Objective To establish a tetracycline-dependent regulation of SV40 Tag transgenic mice.Methods\ p205-sv40Ta p205-rtTA-C3 and pTRE-Tag dual transgenic mice was generated by pro-nuclear microinjection and embryo transplantation.Positive founder mice were identified through both PCR and Southern blot.Results\ A rtTA and Tag dual positive transgenic mice,two rtTA single and one Tag single transgenic mice were confirmed by PCR and Southern Blot.The expression of Tag mRNA was detected by RT-PCR in the ovary of the dual transgenic mice which were administered with tetracycline in the drinking water.Conclusion\ The results of this study indicate that it is possible to establish a tetracycline-dependent regulation of SV40 Tag transgenic mice by administeration of tetracycline in the drinking water.

Abstract:目的探讨低频超声终止妊娠的可能性及其对胎盘形态学的影响。方法利用低频聚焦声束超声(353 kHZ“超声终止早孕治疗机”样机)6种声强各60 s体内直接照射妊娠第10天兔胚胎,妊娠20 d时观察各组胚胎死亡率和胎盘、子宫的形态学变化。结果对照组胚胎总死亡率为12.65%(32/253);照射组中声强30 W/cm2、35W/cm2及≥40 W/cm2组的胚胎死亡率分别为25.00%(12/48)、72.88%(43/59)和100%(64/64、43/43、40/40、35/35)。超声辐照后的即刻改变:光镜下主要为胎盘(绒毛及子宫蜕膜)出血、坏死;电镜下胎盘细胞正常结构消失,子宫平滑肌、内膜上皮细胞中线粒体肿胀。变化随剂量的增加而明显。照射后10 d观察:光镜下见绒毛、蜕膜细胞仍有变性或坏死;电镜下胎盘组织细胞坏死呈致密团块状,子宫内膜上皮、平滑肌及血管内皮细胞均未见异常。结论胎盘较子宫对低频超声更敏感,更易受损,子宫照射后变化轻微且可恢复,低频超声抗早孕是有效而可行的。

Abstract:Objective To creat a pancreatitis-associated acute lung injury(PALI) model in rats by retrograde infusion of 5% sodium tauroculam(TAC) into the biliopancreatic duct.Methods\ A modified procedure of retrograde infusion of TAC into the biliopancreatic duct was used to induce acute hemorrhagic necrotizing pancreatitis(AHNP) in rats.Rats were divided into AHNP,control (laparotomy only,no TAC injection) and dexamethasone(DXM) treatment(after successfully TAC injection,supramaximal dose of DXM was injected) group.The rats were killed at 3,6,12 hours after operation.Serum amylase and lipase were assayed.The middle lobe of right lungs were removed and used for lung edema histopathological evaluation.The ratio of protein content in bronchialo-alveolar lavage fluids (BALFs) to serum proteins was used as lung permeability index.Results\ In the AHNP group,lung permeability index,wet to dry weight ratio and histopathological alterations were increasing with time,higher at 6,12 hours than at 3 hours determined by one way ANOVA analysis(P<0.05).No significant difference was found in the 6,12 hours groups in regard to the former two injury indexes.Compared with AHNP group,injury was less severe in DXM treatment group at 6,12 hours after operation. Conclusions\ Apparent lung injury occurs at 6 hours after pancreatitis induced by retrograde infusion of TAC into the biliopancreatic duct in rats.The pulmonary morphological and functional changes in the model rats are similar to those in AHNP patients with acute lung injury,suggesting its value in PALI studies.

Abstract:目的建立检测兔病毒性出血症(RHD)血清抗体的间接ELISA方法并组装成试剂盒,进行初步应用。方法通过差速离心和蔗糖密度梯度离心提纯RHDV病毒粒子作为包被抗原,优化反应条件和筛选试剂,建立间接ELISA检测方法,组装试剂盒并对其总体性能进行评价。结果试剂盒敏感性为血凝抑制试验的4~32倍,重复性和特异性良好,保质期为4℃3个月。对105份兔血清进行随机检测,与血凝抑制试验的复核率为95%。结论本试剂盒可以在检测和科研中初步应用。

Abstract:Objective To study the application of ovary transplantation for preserving pancreatic cancer transgenic mice.Methods Ovary of SV40T positive transgenic mice were transplanted into ovarectornized FVB female mice through micro-surgery,then were crossed with normal FVB male.The positive offsprings were confirmed by PCR.Ovary tissue slides of foster mother were examined.Results Ovaries of 19 donors were transplanted into 32 female mice,among which 21 were pregnant.96 in 159 offsprings survived.35 with a positive transgenic mice were identified by PCR,with a positive rate of 25.4%.Conclusion Ovary transplantation may be used for transgenic mice preservation.

Abstract:Objective In order to assess and verify the identity of beagle dogs bred in Guangzhou and from USA on molecular level,polymorphism of microsatellite loci of the beagle dog genome were analysed.Methods\ Using the short tandem repeats(STR) technique and sequencing of microsatellite loci to assess the DNA of the two dog populations.Results\ Six alleles were found in the samples.Among them,the populations of dogs from USA and dogs bred in Guangzhou had 6 and 5 alleles,respectively.Rates of alleles of microsatellite loci were calculated according to genotypes.Heterozygosity,polymorphism information content(PIC),rate of homozygosity,coefficient of gene differentiation of microsatellite loci were calculated by means of correlated formulas.It showed that individuals bred in Guangzhou had the heterozygosity 0.7010 and PIC 0.6747.In comparison,the individuals from USA had the heterozygosity 0.7876 and PIC 0.7515.Coefficient of gene differentiation is small(0.021).Conclusion\ Our findings indicate that the two populations did not form different independent groups.

Abstract:目的建立检测SV5的PCR方法并加以初步应用。方法根据GenBank中报道的SV5序列,针对其中的SH基因设计引物进行PCR反应,扩增产物进行测序并用BLAST软件进行同源性比对,同时利用限制性内切酶的酶切反应以证实此PCR反应的特异性。在此基础上设计巢式PCR提高此方法的灵敏度。利用此方法对20份猴肾源细胞培养物和40份血清标本进行检测。结果利用设计的引物扩增出的序列测序结果证实与报道的SV5SH基因相对位置的序列一致。AccⅢ限制性内切酶可对PCR产物进行特异性酶切。巢式PCR比一次PCR的敏感度有所提高。用此方法检测的20份猴肾源细胞培养物和40份血清标本结果为阴性。结论初步建立了检测SV5病毒的PCR方法,排除实验室用20份猴肾源细胞培养物和40份血清标本SV5的污染。

Abstract:目的用脑内注射胶原酶方法建立小鼠脑出血模型。方法在动物脑立体定位仪下将肝素化Ⅶ型胶原酶0.15μL(0.5 U/μL)注射到昆明小鼠右侧尾状核内;分别在术后8、12、24、72 h对小鼠运动功能进行检测,通过在各时间点测量血肿直径来观察出血高峰的时间,并对出血部位脑组织进行光镜观察来验证模型制作是否成功。结果术后小鼠即表现左侧肢体偏瘫症状,至12~24 h症状最重。形态学观察证实右侧尾状核区血肿形成,且出血高峰的时间与肢体偏瘫症状发展过程基本一致。不同个体间血肿体积和部位基本一致。结论应用Ⅶ型胶原酶脑内注射的方法建立的小鼠脑出血模型,是一种方法简单,效果可靠,易于大批量制作的动物模型。

Abstract:目的建立大鼠睡眠呼吸暂停综合征(SAS)动物模型。方法选取Wistar雄性青年大鼠与老年大鼠各30只。各组分为常氧对照组及间歇性缺氧组,每组6只。实验中抽取气体用于气体成分分析,测氧仪监测吸入气氧浓度,在缺氧终点测定右室肥厚指数、右室压力以了解缺氧对右心的影响;取肺、肾组织石腊包埋,病理切片,光镜观察。结果①随缺氧时间延长,出现右室肥大、肺动脉高压。②肺组织学观察:肺血管壁增厚、肺泡间隔增宽;肾小管上皮水肿变性。结论通过模拟间歇性缺氧机制成功建立大鼠睡眠呼吸暂停综合征动物模型。

Abstract:Objective To construct human bone morphogenetic protein-2(hBMP-2 ) expressing eukaryotic vector(pcDNA3-hBMP2) and examine its expression in rabbit bone marrow stromal cells(MSCs).Methods pcDNA3-hBMP2 vector was constructed using gene clone and recombination technique.After being confirmed by enzyme cut,pcDNA3-hBMP2 was transfected into cultured rabbit MSCs in vitro with the help of liposome reagent FuGENE6.The expression of hBMP-2 was determined by in situ hybridization and immunohistochemical analysis.Results The pcDNA3-hBMP2 vector was constructed successfully.The rabbit MSCs transfected by pcDNA3-hBMP2 showed transient and stable expression of BMP2.Conclusion pcDNA3-hBMP2 can express hBMP2 in bone marrow stromal cells.

Abstract:Objective To establish a canine model of acute cardiac insufficiency (ACI) induced by coronary artery occulsion and tachycardiac stress.Methods Studies were performed in 12 dogs using rapid ventricular pacing after the left anterior decending(LAD) coronary artery was ligated.Cardiac output(CCO) reduction was expected to 50% of the baseline.Hemodynamic measurements were performed in the dogs under different CCO conditions.Echocardiography was used to examine the cardiac size and left ventricular ejection fraction(LVEF).Blood sample was taken for examination of plasma renin (PRA) and endothelin(ET) activity.Urine output(UO),serum creatinine(Scr),creatinine clearance(Ccr)were also assayed.Results CCO was decreased by 50% steadily after right ventricular pacing following ligation of LAD.Arterial pressure and arterial oxygen saturation decreased while right atrial pressure,pulmonary capillary wedge pressure,systemic vascular resistance increased significantly.The cardiac cavity became larger while LVEF decreased.PRA,ET,Ccr increased while UO and Ccr decreased.Conclusion A canine model of ACI can successfully be created by right ventricular pacing following ligation of LAD.

Abstract:目的研究阿霉素肾病大鼠肾组织中表皮生长因子(EGF)及其受体EGFR的表达分布以及表达量与尿蛋白之间关系。方法选择第5天、14天、28天作为动态观察的时点,同期设立正常对照。采用荧光定量RT-PCR、免疫组织化学及计算机图像定量分析EGF mRNA以及EGF、EGFR蛋白在肾组织的表达,同时测定24 h尿蛋白定量。WT1和EGFR双重免疫组化确定EGFR在肾小球内确切细胞定位。结果阿霉素注射后第5天,EGFmRNA即较正常增高,28 d明显增高并高于5 d和14 d。正常对照组EGF阳性细胞主要分布于远曲小管和髓袢,阿霉素组EGF还在集合管和近曲小管上表达;EGF阳性表达范围和强度随尿蛋白增加而增加;EGFmRNA表达量以及EGF在肾小管中的表达强度与24 h尿蛋白量呈正相关。肾小管上皮细胞广泛表达EGFR,阿霉素组EGFR在小管表达均高于正常,但组间各时点差异无显著性;随尿蛋白增加EGFR在肾小球内表达逐渐增多。EGFR在肾小球和肾小管中的表达强度均与24 h尿蛋白量呈正相关。WT1和EGFR双重免疫组化显示阿霉素肾病组EGFR可在足突细胞上表达,正常组则无。结论阿霉素肾病大鼠的肾小球脏层上皮有EGFR的表达。EGF/EGFR可能参与了阿霉素肾病的发病过程以及蛋白尿的形成。

Abstract:Objective To improve the techniques for establishment of a stable and reproducible animal model of heart transplantation in rat for further research on acute rejection.Methods To expose the recipient's abdominal arteries at first.Then to obtain the donor's heart and upper right lung.The donor's ascending aorta was anatomized with recipient's abdominal aorta and then blood flowed through donor's organ.Results 80 heterotopic heart transplantations were performed,4 rats died of anastomotic bleeding,2 died of the stenosis of anastomosis and 1 died of the failure of heart rebound, and the total successful rate was above 80%.Conclusion The modified surgical techniques and careful operative management can contribute to better success in heart transplantation in rats.

Abstract:Objective To develop a mouse model of myasthenia gravis.Methods C57BL/6J mice were immunized with the mixture of acetylcholine receptor extracted from Torpedo Californica and complete Freund's adjuvant.After twice strengthened immunization,several indexes including myodynamia,electromyogram,diaphragma end-plate electron microscopy and serum anti-nAChR level were tested.Results Compared with the control,the myasthenic mice displayed myodynamia-attenuating,and the electromyogram showed an apparently declined amplitude of gastrocnemius combined action potential.The electron microscopic examination demonstrated flattened postsynaptic membrane,its folds diminished and showed vacuolar degeneration.The serum antibody level in the myasthenic mice increased significantly.Conclusion We have successfully established a mouse model of myasthenia gravis,which may be helpful in investigation of mechanisms of MG and pursue new ways to treat autoimmune diseases.

Abstract:Objective Tuberculosis infection in the live monkeys was usually diagnosed by tuberculin test.It was difficult in tuberculous monkeys' management that quarantine method was single,and the sensitivity and specificity of tuberculin test was low.The purpose of this study is to establish an ELISA detection method of mycobacterium antibody in monkeys.Methods Three antigens(cytoplasm protein,PPD-bovis and TDM) were applied in the ELISA test.The application value was evaluated by detecting the sera of 28 monkeys following a natural outbreak of tuberculosis and 121 healthy monkeys.Results On the basis of the sensitivity and specificity of detecting results,they were analyzed by receiver operating characteristic(ROC) curve.The sensitivity of the cytoplasm protein,PPD-bovis,TDM was 89.3%,75% and 64.3%,respectively.The specificity was 93.4%,95.9%,88.4%,respectively.The areas under ROC curve were 0.964,0.858 and 0.844,respectively.All the areas exceeded 0.5,P<0.01.Conclusion\ These results indicate that all the three antigens can be used in ELISA detection test of serum of tuberculous monkeys.They have good serum diagnostic accuracy.Among them,the cytoplasm protein is a good antigen to detect tuberculous monkeys' antibody,showing the highest sensitivity and specificity in the ELISA tests.The result of ELISA test using TDM in serum-diagnosis of tuberculous monkeys is not satisfactory.

Abstract:Diabetic nephropathy(DN) represents the most frequent cause of end-stage renal disease(ESRD) and threatens the life of patients with diabetes mellitus(DM).However,the pathomechanisms which cause renal damage in diabetes have not been completely clarified.An appropriate animal model may provide an opportunity for investigation of diabetic nephropathy.The animal models of diabetic nephropathy include induced,spontaneous and transgenic rodent or non-rodent.Some of the characteristics of these animal models are similar to those of human diabetic nephropathy in the development,pathogenesis,and functional and structural lesions of renal disease.The animal models of diabetic nephropathy can be applied in research of preventive medicine,etiology and mechanism of human diabetic nephropathy,and also to explore new medicines and therapeutic strategies to prevent progressive renal disease in diabetes.

Abstract:Objective Gout is a disease caused by hyperuricemia as a result of disturbance in purin metabolism and/or reduction of excreation of uric acid in the urine.Consequently,uric acid deposits in tissues.Presently,there has not been animal model of urarthritis similar to human gout.In this article the methodology,mechanisms and applications of current animal models of hyperuricemia and urarthritis were reviewed,their characteristics and drawbacks were analyzed,and prospective directions of animal models of hyperuricemia and urarthritis were discussed.

Abstract:Molecular markers have become main tools for genetic diversity studies of macaca mulatta.Detection methods and applications of molecular markers(RFLPs,RAPD,mtDNA,microsatellite DNA,SNPs) were reviewed in this article,which may offer references for genetic diversity studies of macaca mulatta in China.