Abstract:Objective To investigate the changes of gene expression in the cardiac outflow tract (OFT) in Cx43 knockout mouse embryo, and to elucidate the genes involved in the pathogenesis of pulmonary outflow obstruction. Methods The cDNA was retrotranscripted from RNA extracted from OFT tissues of both Cx43 knockout and Cx43 wild type mouse embryos on embryonic day (ED) 14.5. The biotin-labeled cRNA derived from the transcription of cDNA was fragmented as probes. The probes were hybridized with Affymetrix Mouse Genome 430 2.0 Array. Gene Array Scanner was used to screen the signals of hybridization and the expression of genes was detected. Results Among the differentially expressed genes, there were 287 upregulated and 199 downregulated in Cx43 knockout OFT tissue as compared with those in Cx43 wild type heart. Functions of proteins encoded by the altered genes encompassed all functional categories, with largest percentage in genes involved in regulation of transcription, cell cycle, etc. Among the differentially expressed genes in the Cx43 knockout heart were those related to Galpha13 signaling pathway. Conclusions Genes related with Galpha13 signaling pathway differently expressed in ED14.5 Cx43 knockout OFT tissue may be involved in the pathogenesis of pulmonary outflow obstruction.

Abstract:目的观察单纯腹主动脉缩窄造成的心肌肥厚能否转变成为心力衰竭。方法实验选用8周龄的Wistar大鼠,使用7-0号尼龙线对其肾上腹主动脉进行缩窄手术,造成后负荷性心肌肥厚模型(LVH,n=10),同时设置假手术组(Sham,n=10)和正常组(Con,n=10)作为对照。术后第20周和第38周使用超声多普勒和多导生理仪对大鼠血流动力学进行检测。解剖后取出心脏,计算心脏/体重比,并通过HE染色和天狼猩红染色观察心脏形态和纤维化程度。结果腹主动脉结扎后第20周,LVH组大鼠心室壁肥厚,舒张功能下降(E/Aratio:LVH组:1.0±0.25,Con组:1.6±0.12)。术后38周,左心室壁肥厚程度有所下降,但是心室腔扩大,心脏收缩和舒张功能明显下降(EF:LVH组:44.8±8.42,Con组:70.9±5.19;MaxdP/dt:LVH组:4916±1267.3,Con组:14225±932.1;MindP/dt:LVH组:-3246±1217.3,Con组:-12138±725.2)。腹主动脉缩窄术后的动物心脏重量明显增加(3.58±0.32vs.2.34±0.15),HE染色和天狼猩红染色显示LVH组大鼠在术后38周心脏纤维化明显。结论腹主动脉缩窄造成的后负荷增高动物模型首先出现向心性心肌肥厚,伴以舒张功能下降,进而收缩功能下降,发展为心力衰竭。

Abstract:目的探讨旨在降低死亡率的二甲基亚硝胺(DMN)肝纤维化改良造模方法及其病理特点。方法大鼠分为常规方法造模组(常规组)与改良方法造模组(改良组),每组再分为模型组与正常对照组,均设染毒后4周和8周共2个时间观察点。常规组造模以DMN10μL/kg大鼠体重的剂量,腹腔注射,每周连续3d,连续4周,共12次;改良组同上剂量的DMN腹腔注射,每2d一次,连续4周,共14次。观察大鼠体重、肝体比与脾体比,试剂盒测定血清肝功能,HE染色与天狼猩红染色分别观察肝组织炎症与胶原沉积,盐酸水解法测定肝组织羟脯氨酸含量。结果2种造模方法均在造模4周时形成典型的肝纤维化,模型大鼠的体重、肝体比、脾体比、血清肝功能等均有相应的变化趋势。与常规组比较,改良组模型大鼠造模8周后死亡率显著降低,肝细胞炎症坏死稍轻,而肝窦毛细血管化明显。结论隔日1次腹腔注射较之连续3d腹腔注射DMN是一种较为稳定的肝纤维化改良造模方法,具有肝窦毛细血管化与小叶内纤维化的肝纤维化病理特征。

Abstract:目的观察杂种犬心肌缺血再灌注损伤过程心电图(ECG)的动态变化规律,为研究干细胞移植对犬心肌缺血再灌注损伤后的治疗作用提供基础。方法选用杂种犬24只,结扎冠状动脉左前降支中远1/3处,分3组分别于30min、60min、90min后松开。应用MPA-2000生物信号分析系统,在结扎前后及松开后连续动态记录ECGII导联QRS波群的变化。结果在开始结扎阻断冠状动脉血流时,QRS主波向上Rs型;重新开放血流血管再通之初,83.3%(20/24)的犬QRS波表现为主波向下rS型或QS型,伴QRS波增宽,然后r波波幅逐渐增大,逐渐演变成主波向上的Rs型,QRS时程恢复正常。15min、30min和大于30min的演变率分别为25.00%(6/24)、41.7%(10/24)和16.7%(4/24);且演变率与冠脉阻断时间相关。结论犬心肌缺血再灌注后ECG变化有一定规律,QRS波群变化趋势有可能作为心肌细胞功能恢复程度以及心肌保护效果较为直观的判断指标。

Abstract:Objective To establish an animal model of subintimal angioplasty(SIA),and observe the pathologic changes of the components in the treated arteries.Methods 16 Bama mini-pigs were used in this study.Atherosclerotic occlusion model was created in 7 swine.SIA was performed in normal common carotid artery(9 swine)and atherosclerotic common carotid artery(7 swine).The patency rate and vascular pathologic changes were surveyed and examined.Results Technical success rate of SIA performed on swine common carotid arteries was 93.8%(15/16).The total patency rate after SIA was 53.3%(8/15).There was no significant difference in the patency rates after SIA between the groups of normal artery and ASO artery(P=0.43).Comparing to the normal artery,the media of the artery wall was thicker after SIA,and more collagen fibers as well as elastic fibers were present.Smooth muscle cells(SMC)in the media were observed in both contractile and synthesizing appearance.Conclusions(1)An animal model of subintimal angioplasty in Bama swine has been successfully designed and established.The following factors are of great importance to ensure the success:choosing swine as the experimental animal,performing SIA on the common carotid artery,closing the true luminal of the artery with suture,and using anticoagulation drugs after the operation.(2)The pathologic changes after SIA was similar with that after PTA,such as recovered endothelium,SMCs phenotype changed from contracted to synthesis,and increased amount of some extracellular matrix(collagen and elastic fibers).

Abstract:Objective To establish an experimental animal model with insulin resistance and/or type 2 diabetes mellitus.Methods High energy fructose diet was given to mesocricetus to produce insulin resistance and hyperlipidemic models.Diabetes was induced by a single intraperitoneal injection of streptozotocin(30 mg/kg)on these insulin resistant animals.Normal animals were used as control.Serum glucose,lipid profiles and insulin levels were measured to verify the diabetic and hyperlipidemic models.The liver,pancreas and aortic arch were examined to distinguish the histological difference among the animals of three groups.Results The mesocricetus fed with high energy fructose diet for 6 weeks developed insulin resistance,obesity and dislipidemia,80% of which became type 2 diabetic after single intraperitoneal injection of streptozotocin.Conclusion Insulin resistant and diabetic mesocricetus models can be produced successfully with high energy fructose diet and single intraperitoneal injection of streptozotocin.

Abstract:Objective To explore the correlation between the exudation amount of Evans blue in mouse model of blood stasis induced by macromolecular dextran and the degree of blood stasis.Method Preparing 10%,5% and 2.5% dextran solution with 0.5% Evens blue in normal saline.Mice were injected the preparation into tail vein,0.1 mL/10 g.The skin colour of ear and tongue was examined.Evens blue was extracted from the ears by anhydrous acetone and Na2SO4 solution and determined at 590 nm by ultraviolet spectrophotometer at 0.5,1 and 2 hours after dextran injection.Results Compared with the Evens blue-group mice,the whole skin of tongue,in 10%,5% and 2.5% dextran groups appeared blue and the blue color became deeper along with the increasing content of dextran(P<0.05 or P<0.01).At 0.5 hour after dextran injection,the skin of tongue,mice appeared plaque blue.One hour later,the whole skin appeared obvious blue,while its blue faded away about two hours later.Conclusion The degree of blood stasis induced by macromolecular dextran is apparently correlated to the exudation amount of Evens blue,and the degree of blood stasis is regularly related with the content and time of exudation amount of Evens blue.

Abstract:目的 探讨125I标记的Flt4多抗(125I-Flt4PcAb)对荷瘤小鼠前哨淋巴结(sentinel lymph node,SLN)的检测,为应用Flt4PcAb进行SLN特异性定位提供实验依据.方法 建立BALB/c裸小鼠后肢荷瘤模型,7周后应用125IFh4PcAb检测肿瘤的SLN,健侧作为对照,切取(月园)窝淋巴结探测γ射线的每分钟计数率(count per minute,Cpm),并进行HE及角蛋白免疫组化染色;分析不同状态淋巴结摄取125I-Flt4PcAb的特点.结果 BALB/c裸小鼠后肢皮下注射Tca-8113细胞悬液,移植瘤成功率达100%;患侧70枚(月园)窝淋巴结中,HE染色3枚(4.3%)出现肿瘤转移;角蛋白免疫组化染色为5枚(7.1%);125I-Flt4PcAb摄取方面,肿瘤转移淋巴结、反应性增生淋巴结和正常淋巴结之间均有显著差异(P＜0.05).结论 BAIB/c裸小鼠后肢皮下注射Tca-8113细胞悬液可成功致瘤,但淋巴结转移率低;免疫组化染色较HE染色检测肿瘤转移更敏感;125I-Flt4PcAb能够检测到荷瘤鼠的SLN,肿瘤转移淋巴结对125I-Flt4PcAb的摄取下降.

Abstract:目的自发性肿瘤的研究是实验动物学的研究内容之一,对其流行病学和病理学的研究可为动物模型的开发、肿瘤发病机制的探讨提供有价值的线索。方法对SPF级大鼠生产群进行流行病学调查和临床病理学观察。收集大鼠自发乳腺肿瘤病例,通过巨检与镜下观察对肿瘤进行定性并分类。结果SD大鼠自发乳腺肿瘤发生率为10.5%,其中良性肿瘤为8.5%,恶性肿瘤为2.0%;Wistar大鼠乳腺自发肿瘤发生率为2.0%,其中良性肿瘤为1.5%,恶性肿瘤为0.5%。SD大鼠、Wistar大鼠乳腺良性肿瘤自发率均高于恶性肿瘤,并以纤维腺瘤和乳腺腺瘤最为多见。结论丰富了封闭群SPF级大鼠生物学背景资料,并为人类相应疾病动物模型的建立提供了一种途径。

Abstract:Objective To obtain venous endothelial cells to be used in the research of blood vessel tissue engineering.Methods The external jugular vein of canine was resected under aseptic conditions.The primary endothelial cells were harvested,cultured,identified and multiplicated,and observed by optical microscopy.Result The endothelial cells of the canine external jugular vein,cultured and multiplicated in vitro,not only had good bioactivity,but also met the need of experiments in amount.Conclution Endothelial cells of canine external jugular vein cultured in vitro may fit the need of blood vessel tissue engineering.

Abstract:Objective To investigate the protective effect of carvedilol on renal toxicity induced by cisplatin(CP).Method Male Wistar rats were randomly divided into four groups:saline control group,carvedilol control group,CP group and carvedilol protection group.The rats were killed at 3 to 6 days after drug administration.The kidney tissues were examined by light(HE and PAS staining)and electron microscopy.Results The renal histopathological and ultrastructural damages were markedly alleviated in the carvedilol protection group in comparison with those in the CP group.Conclusion By intragastric administration of carvedilol,obvious protective effect can be achieved on the cisplatin-induced nephrotoxicity.Antioxidation,scanvaging of free radicals,and enhancement of the activities of some antioxidant enzymes may be involved in the mechanisms of action in this process.

Abstract:目的观察摘除泪腺后,兔泪液S1T值、角膜荧光素染色和虎红染色的变化及其角膜上皮超微结构的变化,评估兔干眼症模型的建立。方法摘除兔泪腺和Harder氏腺,比较泪腺摘除前后S1T、角膜荧光素染色和虎红染色分值的变化,并在透射电镜下观察角膜上皮形态学的变化。结果泪腺摘除后平均S1T值(15.88±6.29mm/5min)低于摘除前的平均S1T值(20.25±5.52mm/5min),差别有统计学意义(P=0.0062)。平均角膜荧光素染色和虎红评分(分别为8.22±1.99和7.67±0.87)明显高于摘除前(分别为0.22±0.44和0.67±0.5),差别有统计学意义(两组均P<0.0001)。以上3个检查指标的变化说明摘除泪腺后出现水液性泪液分泌缺乏,导致角膜上皮点状剥脱,干燥及坏死。在形态学上泪腺摘除前后的角膜上皮也发生了明显的改变,透射电镜检查发现液化的表层上皮细胞,表层细胞膜破裂。以上的改变均符合干眼症的特征。结论摘除兔泪腺和Harder氏腺后兔泪液分泌减少,角膜上皮坏死,能有效形成泪液缺乏型干眼模型。

Abstract:目的探讨雌/雄激素比例对去势大鼠前列腺体积及其脏器系数的影响。方法选用雄性SD大鼠随机分为37组,分别为:35个不同雌/雄激素比例组、正常和去势对照组。给药组分别注射不同配伍剂量的雌/雄激素、对照组均注射生理盐水各一个月。最后一次注药后24h取血、处死、取材,称取前列腺重量,测量体积,并计算脏器系数。结果当丙酸睾丸酮给药剂量分别为每只鼠0.02、0.5、2.5和12.5mg时,均未见器官增生程度与苯甲酸雌二醇注射剂量呈负相关。丙酸睾丸酮给药剂量每只鼠为0.1mg时,随着苯甲酸雌二醇注射量增加,前列腺体积及其系数均增大,而当每只鼠苯甲酸雌二醇注射量达到50μg/kg时,器官增生程度不再随苯甲酸雌二醇注射剂量增加而增加。结论雌激素发挥作用需以雄激素存在为前提,雌激素并非总是呈现对抗雄激素促进前列腺增生的作用,而是在一定范围内协同雄激素促进前列腺增生。

Abstract:目的测定SPF级金黄地鼠的生物学特性数据,为生物制品的生产、检定及科学研究提供标准化的SPF级金黄地鼠。方法在5周龄、10周龄和30周龄群体中随机抽样20只地鼠,雌雄各半。测定生长发育、繁殖性能、淘汰种鼠干物质及粗脂肪含量、解剖学特性以及血液生理生化指标等生物学特性指标。结果从SPF级金黄地鼠的生长和繁殖特性来看,通过严格的选择和淘汰,SPF级金黄地鼠的生长、繁殖等特性已保留下来,同比原种群得到进一步提高。其体重和主要脏器重量、脏器系数的数据在5、10和30这三个日龄段基本稳定一致,呈现正常的生长规律;该鼠群的血常规、血液生化指标和免疫指标也比较稳定一致,个体间的差异很小。结论本实验测定的SPF级金黄地鼠的主要生物学特性,将为SPF级金黄地鼠进一步标准化提供极其重要的依据。

Abstract:Objective To explore the feasibility of local implantation of endothelial progenitor cells(EPC)from peripheral bloods to promote neoangiogenesis and restore contractility in ischemic myocardium.Methods 5 mL of peripheral blood was obtained from the femoral artery of Sprague-Dawley rats.Myocardial infarction was induced by ligating the left anterior descending coronary artery(LAD).Four weeks after ligation,autologous EPCs isolated from the peripheral blood of each rat were locally injected into the ischemic myocardium.In the control groups,cell culture medium was injected.Results Contractility of ischemic myocardium in the rats of EPC groups was significantly better than that in the control group.Simultaneously,much higher microvessel density(MVD)was observed in the EPC group.Conclusion Local implantation of autologous EPC improves post-ischemic myocardial contractility and resultantly cardiac function improves.Together with higher MVD,these favorable outcomes encourage future clinical trials of intramyocardial implantation of autologous EPCs in the treatment of chronic myocardial ischemia.

Abstract:Objective Two pairs of E.suis specific primers were designed to establish a new nested-PCR method in order to assess the E.suis DNA in the blood of E.suis infected rats.The practicability and operability of the nested-PCR techniques used in E.suis rat model were verified.Methods Rats were infected intraperitoneally,subcutaneously or intramuscularly with positive blood from infected swine.Blood samples were colleted and normal blood smear staining was done.The E.suis was purified and its DNA was extracted.Nested-PCR was done and the PCR products were assessed by electrophoresis in 0.85% agarose gel.Results A 498 bp specific fragment was amplified with nested-PCR.It was verified by sequencing.During the experiment no obvious clinical symptom was found in infected rats.Conclusion A rat model of infection with eperythrozoon has been established.In the transcription latent period,nested-PCR is more sensitive and it may become an important and necessary technique in E.suis models assessment.

Abstract:Objective To observe the behavior and content of dopamine(DA)in striatum of mice treated with different doses of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP),and to explore the optimal conditions of establishment of a mouse model of parkinson's disease.Methods C57BL mice were administered at different doses of MPTP.Pole test was conducted to determine the motor coordination and the content of DA in striatum in different groups of mice was determined by HPLC.Results All MPTP treated mice showed different disability in pole test and remarked decrease in content of TH and DA.Conclusion Symptoms of Parkinsonism in mice can be induced by MPTP administration.In this model,different dose of MPTP and different drug administration route should be used according to the goal of actual study.

Abstract:目的建立一套比较理想的小鼠精子冷冻复苏的方法。方法采用R18S3和FERTIUPTM-CPA两种保护剂,对DBA/2、C57BL/6J、KM和B6.129S7-Ldlrtm1Her/J四个品系的小鼠精子进行冷冻,冷冻精子用三种方法复苏,以体外受精率来评价精子冷冻的效果。结果以R18S3作为冷冻保护剂,DBA/2(73.3%,88.4%,55.6%)和KM(64.9%,60.2%,39.6%)品系小鼠冷冻精子复苏后体外受精率差异显著(P<0.05),C57BL/6J(3.0%,10.3%,3.7%)和B6.129S7-Ldlrtm1Her/J(0%,5.0%,0%)结果差异不显著(P>0.05);以FERTIUPTM-CPA作为冷冻保护剂,DBA/2(33.6%,14.1%,91.6%)、C57BL/6J(8.4%,21.0%,4.9%)和B6.129S7-Ldlrtm1Her/J(8.2%,10.0%,28.9%)品系小鼠冷冻精子复苏后体外受精率差异显著(P<0.05),而KM(48.1%,48.0%,48.1%)小鼠冷冻精子复苏后体外受精率差异不显著(P>0.05)。结论对于DBA/2和KM品系小鼠来说,用R18S3或FERTIUPTM-CPA冷冻精子,选择一种恰当的复苏方法,均可以得到较理想的体外受精率,而C57BL/6J和B6.129S7-Ldlrtm1Her/J品系小鼠无论采用哪种冷冻保护剂,选择何种方法复苏精子,得到的体外受精率都较低。

Abstract:The traditional polymerase chain reaction(PCR)technique played a very important role in detection of nuclear acids.Recently,based on PCR,the real-time PCR that can quantitate the products of PCR have been developed and applied in various fields,especially in detection of viral pathogens.In this review,the real-time PCR technique and its application will be introduced.