• Issue 3,2003 Table of Contents
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    • Breeding and Characterization of a New Severe Combined Immunodefecient 615-SCID Mouse

      2003(3):129-133.

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      Abstract:Objective To breed a new severe combined immunodefecient 615-SCID mouse with 615 genetic background. Methods The scid mutant gene of SCID mouse was transferred into 615 inbred strain by cross-backcross method. Results The new established 615-SCID mouse with brown hair like 615 mouse is homozygous for the scid mutant gene lacking functional T and B lymphocytes. The conspicuous histological features of the 615-SCID mouse was observed. The total leukocyte count of the peritoneal cavity of 615-SCID varied from 1500 to 4500/mm 3. The proportion of lymphocytes is below 30%. B(CD19) and T(CD3, CD4, CD8) lymphocytes are both similar to SCID. But the proportion of T cell is less than SCID mouse. ELISA-test found that the serum Ig is less than 0.02mg/ml. The thymus glands were very small, usually comprising one or two lobules and occasionally undetectable grossly. No lymphocytic cortex is evident in thymus. The spleen varied in size, smaller than SCID mouse. The splenic folliculi are virtually absent. And lymph nods have only cortex. Conclusion 615-SCID mouse expresses an obvious sensitivity to radiation, lacks detectable B and T cells.

    • Antitumor Effect of Recombinant Adenovirus Expressing hIL-2 and hTNF-α

      2003(3):134-137.

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      Abstract:目的 研究腺病毒介导的细胞因子基因hIL 2、hTNF a对人肺腺癌细胞系的生物学特性的影响 ,探讨重组腺病毒的体内抗肿瘤作用。方法 将分别携带有hIL 2基因、hTNF a基因的重组腺病毒 (rAd hIL 2、rAd hTNF a)感染人肺腺癌Anip973细胞系 ,通过细胞生长实验、克隆形成实验等观察其对肿瘤细胞的作用 ;利用PCR技术对转染后的细胞进行检测并应用ELISA试剂盒检测转染后的细胞IL 2、TNF a的分泌量 ;通过肿瘤局部注射rAd hIL 2、rAd hTNF a的方法观察其在小鼠体内的抗肿瘤作用。结果 rAd hIL 2、rAd hTNF a经纯化后 ,滴度可达 10 1 0 PFU ml,当病毒为 30MOI时 ,对Anip973细胞的转染率达 90 %以上。体外转染的肿瘤细胞生长能力、克隆形成率等无明显变化。 2 4h细胞培养上清IL 2的分泌量为 5 0pg 2× 10 5细胞 ,TNF a的分泌量为 2 0pg 2× 10 5细胞。体内实验表明注射rAd LacZ、rAd hIL 2、rAd hTNF a的小鼠肿瘤生长缓慢 ,体积明显小于对照组 (P <0 0 5 ) ,生存期显著延长。结论 腺病毒介导的细胞因子基因hIL 2、hTNF a转染的肿瘤细胞形态学未见明显变化。转染hIL 2、hTNF a基因的肿瘤细胞可表达IL 2及TNF a。rAd hIL 2、rAd hTNF a重组腺病毒具有抗肿瘤作用

    • Establishing a Rat Model Similar to the Adult Patient of the General Type 2 Diabetes by Long-Term Fat-Enriched Fed and Lower Dose of Streptozocin-Treated Rats

      2003(3):138-141.

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      Abstract:目的 建立类似人类普通 2型糖尿病大鼠模型。方法 采用长期高脂高能量饮食结合小剂量链脲佐霉素 (STZ)腹腔注射的方法。结果 模型大鼠有高血糖 (18 78± 1 97)mmol L、高甘油三酯血症 (10 95 4± 5 5 4 )mmol L、高胆固醇血症 (2 4 4 2± 0 74 )mmol L、脂肪肝、高胰岛素血症 (0 937± 0 2 6 )mmol L、胰岛素敏感性下降及不低于正常对照组的体重。结论 这是一种类似人普通 2型糖尿病大鼠模型

    • Preliminary Study of Mice Mutation Induced by ENU

      2003(3):142-146.

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      Abstract:Objective To study the efficiency of mice mutation induced by ENU. To screen and acquire the mutant mice with some new traits showing dominant heredity. Methods 33 C 57BL/6 and 18 DBA/2 male mice about 8-10 weeks old were injected intraperitoneally with a dose of ENU 100 mg/kg once a week and total three times. Mated with the same strain female mice, their progeny were screened for mutations. Results Treated males became transient sterility and the average length of sterile period was 9-13 weeks. In 1241 mice screened, 61 mice were produced with mutations, such as abnormal eyes, dwarf, white belly spot,and so on. The mutation percentage was about 5%. There were two mutation strains showing dominant heredity. Conclusion The mouse models for human diseases could be obtained by ENU-induced mutation. ENU mutagenesis on a large scale had important value for functional genome research.

    • Study on the Model of Systemic Inflammatory Response Syndrome-Acute Lung Injury in Rats

      2003(3):147-150.

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      Abstract:目的 研究大鼠经“一次打击”和“二次打击”后导致全身炎性反应综合征 肺损伤 (systemicinflammato ryresponsesyndrome acutelunginjury ,SIRS ALI)的动物模型 ,并探讨其意义。方法 用梯级剂量的大肠埃希菌脂多糖(lipopolysaccharide ,LPS)致伤大鼠 ,复制“一次打击”模型 ;用小剂量油酸致伤大鼠后 ,再予小剂量LPS ,复制“二次打击”模型。结合动脉血气分析、外周血白细胞计数、肺湿重 干重比值 (W D)及肺组织病理观察 ,评估两种SIRS ALI动物模型。结果 在“一次打击”模型中 ,当LPS≥ 8mg kg时 ,动物出现顽固的低氧血症和明显的病理组织学改变 ,类似SIRS失控 ;小剂量油酸加LPS两次打击 ,亦可导致SIRS失控。结论 较大剂量LPS一次性致伤 ,类似临床严重感染导致单相速发型SIRS ALI的发生过程 ;而油酸加LPS两次致伤 ,可模拟创伤后继发感染导致双相迟发型SIRS ALI的发生过程

    • Establishment of Method of Extracting Genomic DNA from Tyzzer Organism

      2003(3):151-155.

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      Abstract:Objective To construct DNA library ,genomic DNA was extracted from Tyzzer organism. Methods Three methods (density gradient centrifugation combining enzymatic digestion method, enzymatic digestion method, and filtration salting-out combining centrifugation method) were used to purify Tyzzer organism from infectious livers, and results of purification were compared. Genomic DNA was extracted from Tyzzer organism by three methods (benzyl chloride-method, kit and phenol method), character of which was analyzed. Results Tyzzer organism was purified effectively by filtration salting-out combining centrifugation method. Genomic DNA extracted by phenol method was high quality and specificity.Conclusion Genomic DNA was first extracted from Tyzzer organism successfully, which can apply to many experiments of molecular biology.

    • Establishment of Male Wistar Rats Model with Ulcerative Colitis(UC) and the Activity Change of Peripheral Blood NK Cell

      2003(3):156-160.

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      Abstract:目的 通过建立Wistar大鼠溃疡性结肠炎 (UC)模型 ,探讨外周血NK细胞活性的变化和发病机理的关系。方法 采用饮用Dextransulfatesodium (DSS)水溶液 ,复制UC动物模型 ,随机分为 4组 ,即模型组 :每天饮用生理盐水 6ml(简称M) ;锌治疗组 (Zn组 ) :每天 2次灌肠给予硫酸锌 180mg kg ;硒治疗组 (Se组 ) :每天给予硒力口服液 (含硒 6 0 μg ) 6ml;锌 +硒治疗组 :(ZS组 )每天给予上述剂量硫酸锌 硒力口服液 6ml;均给予 15d ,另设正常对照组 (C组 ) ,每天饮用生理盐水 6ml。用微量乳酸脱氢酶 (LDH)释放法测定各组外周血NK细胞活性。采用火焰原子吸收法测定结肠粘膜锌含量。采用荧光分光光度计比色法测定结肠粘膜硒含量。结果 DSS所致大鼠UC模型简单易行 ,与人类病变相类似。模型组外周血NK细胞活性较正常组降低 (P <0 0 5 ) ,经硒治疗后则有明显回升 (P<0 0 5 )。应用锌、硒治疗后结肠粘膜局部相应微量元素的含量明显提高 (P <0 0 1)。结论 经硒治疗后NK细胞活性明显升高 ,微量元素 (锌和硒 )对UC动物模型有一定的疗效

    • Establishment and Application of ELISA for Detection of Antibody to Simian B Virus

      2003(3):161-164.

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      Abstract:目的 建立猴B病毒抗体ELISA检测方法。方法 采用方阵滴定法 ,比较不同血清稀释度、3种酶结合物、2种抗原对检测结果的影响 ,与国外同类参比实验室进行检测结果的比较 ,确定ELISA法的最佳实验条件。结果 HSV抗原包被浓度 10 μg ml,抗人IgG HRP为 1∶2 0 0 0时 ;或B病毒抗原包被浓度 10 μg ml,抗人IgG HRP为1∶4 0 0 0时 ,阴性血清和阳性血清的A值差距最大。与国外B病毒检测专业实验室的检测结果符合率分别为98 0 2 %和 97 5 2 %。结论 建立了猴B病毒抗体的ELISA检测方法 ,提高了检测方法的准确性。

    • Study of an Acute Rejection Model for Pancreas Transplantation in Miniature Swine

      2003(3):165-169.

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      Abstract:目的 建立小型猪胰腺移植动物模型 ,探索早期诊断急性排斥反应的方法。方法  4 0只猪随机配对行胰腺移植 2 0次 ,将供胰所带的血管与受体髂血管吻合 ,所带小段十二指肠与空肠吻合 ,术中监测平均动脉压、中心静脉压及血气。术后测定受体的血淀粉酶、血糖 ,监测外周血免疫指标 ,彩色多普勒检测供胰血流及超声引导下活检、组织病理检查。结果 移植手术成功率为 90 % ,受体平均动脉压在吻合血管开放后有明显下降 ,与血流开放前差异有显著性 (P <0 0 5 ) ,输血有助于手术成功 ,受体术后平均存活 (12 6± 2 3)d。外周血免疫学监测指标早于供胰的组织病理改变 ,两者的改变均早于急性排斥反应的临床表现。结论 小型猪适用于胰腺移植模型的建立 ,加强术中循环功能的管理、及时输血有利于受体成活 ;超声引导下穿刺活检供胰的组织病理学检查与监测外周血免疫指标均适于早期诊断急性排斥反应

    • Establishment of a Model of Explosive Open Abdominal Injury and Study of Risky Factors of Early Death

      2003(3):170-172.

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      Abstract:Objective To establish a model of explosive abdominal injury and seawater immersion and to study the risky factors of early death. Methods Thirty healthy dogs were divided into two groups: experimental group(explosive abdominal injury and seawater immersion)and control group(explosive open abdominal injury only). Blood samples were taken at eight different time intervals for assessing blood gas, plasma osmotic pressure and serum electrolytes. The hemodynamic and respiratory changes were recorded. At the end of study lung was harvested for pathologic examination. Results Post_trauma condition in experimental group was more serious than that in control group. Progressive dysfunction of respiratory and circulatory system, acute lung injury, severe hypernatremia and higher osmotic pressure were found only in experimental group. The mean survival time in experimental group was 209 minutes. Conclusion The results of this study indicates that the explosive model of abdominal injury is practicable and can be repeated well. Seawater immersion after open pneumothorax could result in severe physiopathologic changes. These risky factors appear to be associated with early death in experimental models.

    • The Effect of AngiotensinⅡ and Endothelin on the Development of Broiler Pulmonary Hypertension

      2003(3):173-176.

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      Abstract:目的 揭示低温处理后肉鸡血管紧张素Ⅱ、内皮素变化与早期肺动脉高压发生发展间的关系。方法 采用SMUP_2A型生物信号系统监测肉鸡肺动脉压 ,利用放射免疫法动态测定肉鸡血浆和组织中血管紧张素Ⅱ、内皮素的变化 ,比较肉鸡肺动脉压变化与血管紧张素、内皮素间的关系。结果 低温处理后肉鸡肺动脉压呈现升高 ,差异有显著性 (P <0 0 1) ;血浆、右心室组织的血管紧张素Ⅱ含量显著性升高 ,肺动脉低压组、肺动脉高压组血管紧张素浓度也极显著性升高 (P <0 0 1) ;血浆内皮素含量显著高于对照组 ,与对照组相比 ,肺动脉低压组、肺动脉高压组、腹水组血浆内皮素浓度呈显著性升高 (P <0 0 5 ) ,尤其是与肺动脉高压组相比 ,差异有显著性 (P <0 0 1)。试验发现血管紧张素Ⅱ、内皮素的升高早于肺动脉高压的形成。结论 低温环境下血管紧张素、内皮素释放增加可能会造成肺血管收缩性阻力增加和右心室功能增强 ,从而导致早期肺动脉高压的形成

    • Establishment and Identification of Porcine Endogenous Retrovirus-infected HEK293 Cell Model

      2003(3):177-180.

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      Abstract:目的 建立猪内源性反转录病毒感染HEK2 93细胞的模型 ,验证PERV在体外对人源细胞的感染性 ,并进一步研究PERV的生物学特性。方法 将G4 18抗性的HEK2 93细胞与猪源PK 15细胞共培养 ,6周后 ,用含有6 0 0 μg mlG4 18的选择性培养基对共培养细胞进行加压筛选 ,用以除去共培养体系中的PK 15细胞 ;然后应用PCR及RT PCR的方法对所制备的感染细胞模型进行系统鉴定。结果 经 6周共培养及 3周加压筛选后 ,细胞的形态、生长速度及折光性均未见明显变化 ;PCR及RT PCR鉴定表明 ,筛选后的共培养体系中已没有PK 15细胞的存在 ;PERV特异性检测方法检测显示 ,该细胞模型的DNA中已有PERV的整合且有PERV特异性mRNA的表达。结论 本实验成功建立了PERV感染HEK2 93细胞的模型 ,证实了PERV在体外对人源细胞具有感染性 ;为PERV生物学特性的研究奠定了基础

    • Preliminary Study about the Active Ingredients of Chinese Herb M Influencing Proliferation of Endothelial Cells in Rabbit

      2003(3):181-183.

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      Abstract:Objective To study the Chinese herb M on influencing proliferation of endothelial cells and to elucidate its biological mechanism of vascularization. Methods The model of rabbit chest aorta endothelial cells was established in vitro, and rabbit sera with different concentration of the drug were prepared. The serum was added into 96 wells with endothelial cells in culture plate, and the proliferation of endothelial cells was measured with MTT colorimetry. Result the Chinese herb M can promote proliferation of endothelial cells.Conclusion The Chinese herb M may improve vascularization by promoting proliferation of endothelial cells.

    • Studying RAPD Method for Genetic Monitoring in Inbred Strain Mice

      2003(3):184-186.

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      Abstract:Objective To study a new method for inbred strain mice genetic monitoring.Method 6 random primers were used to amplify DNA in 6 inbred strains of mice (TA1, TA2, T739, C57BL/6,615 and Scid). Results The amplified band patterns were obviously different in the products of 4 primers(p2,p3,p5 and p6).Conclusion The results indicated that different strains of mice could be easily distinguished by RAPD method.

    • Analysis of the Correspondence between SV40 Antibody and SV40 DNA Fragments in Rhesus Monkeys'' Blood

      2003(3):187-190.

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      Abstract:目的 检测 5 7份恒河猴血清及对应的猴血中抗SV4 0抗体、SV4 0DNA的携带情况 ,找出抗体滴度与DNA携带的相关关系。方法 用间接免疫荧光法检测猴血中SV4 0中和抗体 ,聚合酶链反应 (PCR)法检测SV4 0st抗原DNA的携带情况。结果  5 7份猴血清中 ,SV4 0抗体阳性率为 93% (5 3 5 7) ,抗体滴度最高为 1∶12 80 ,最低为 1∶10 ,GMT为 15 9.5 6 ,PCR检测猴血淋巴细胞SV4 0DNA阳性率为 2 4 .5 6 % (14 5 7) ,当抗体为 1 80以下时 ,病毒整合于血细胞 ,1∶80以上时 ,对st抗原基因的整合有抑制作用。结论 恒河猴SV4 0DNA的携带情况与抗体阳性和滴度呈反相关

    • The Anesthetic Effect of Sumianxinzhusheye and Ketamine Hydrochloride Injection on Rhesus Monkeys

      2003(3):191-192.

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      Abstract:目的 比较速眠新与盐酸氯胺酮和速眠新复合麻醉对猕猴的麻醉效果。方法 成年健康猕猴 10只 ,动物分成单纯麻醉组 (速眠新组 ,5只 )和复合麻醉组 (速眠新和盐酸氯胺酮组 ,5只 ) ,均采用肌内注射麻醉。结果 速眠新麻醉维持时间仅 0~ 1h ,麻醉效果低于复合麻醉效果。速眠新有较好的镇静、镇痛和肌肉松弛作用 ,复合麻醉使速眠新效果更佳 ,麻醉时间更长、更稳定 (1~ 2h) ,复苏期较快。结论 速眠新和盐酸氯胺酮复合麻醉适合动物实验时间较长的手术 ,是一种较理想的实验动物麻醉方法

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