• Volume 0,Issue 3,2008 Table of Contents
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    • Induction of Apoptosis of Human Ovarian Cancer CoCl Cells by 5-Allyl-7-Gem-Difluoromethylenechrysin

      2008(3).

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      Abstract:

    • >研究报告
    • Construction and Subcellular Localization of SDF-lα-incorporating GFP Fusion Protein Vector

      2008(3):1-4,83.

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      Abstract:目的构建SDF-1α基因与绿色荧光蛋白的融合蛋白表达载体,进而观察SDF-1α基因编码蛋白在细胞内的定位情况。方法用EcoRI内切酶从pMD-T18一SDF-1α重组载体中酶切分离SDF-1α基因的完整ORF,构建pEGFP-C1-SDF-1α的融合表达载体,脂质体转染COS-7细胞,并在荧光显微镜下观察表达的融合蛋白。结果SDF-1α基因在COS-7细胞中高效表达,激光共聚焦的结果显示,SDF-1α基因定位在细胞质内。结论成功构建了pEGFP-C1-SDF-1α的融合表达载体,SDF-1α基因主要在细胞质中表达。

    • Induction of Apoptosis of Human Ovarian Cancer CoCl Cells by 5-Allyl-7-Gem-Difluoromethylenechrysin

      2008(3):5-9.

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      Abstract:目的 观察5-烯丙基-7-二氟亚甲基白杨素(ADFMChR)诱导人卵巢癌(CoCl)细胞凋亡的作用。方法 以体外培养的人卵巢癌CoCl为研究对象。采用软琼脂克隆测定ADFMChR对细胞集落的影响;流式细胞术(FCM)检测ADFMChR诱导细胞凋亡率;凝胶电泳观察ADFMChR诱导基因DNA梯形条带。Westernblot分析ADFMChR对CoCl细胞PPARγ,NF-κB,Bcl-2,Bax蛋白表达的影响。结果软琼脂克隆显示ADFMChR呈剂量依赖性抑制细胞集落形成;FCM分析发现ADFMChR呈剂量依赖性诱导细胞凋亡;ADFMChR(30μmol/L)孵育CoCl细胞48h后,DNA琼脂糖凝胶电泳呈现典型梯形条带。Westernblot分析结果表明ADFMChR以剂量依赖方式上调CoCl细胞PPARγ和Bax蛋白表达,下调NF-κB和Bcl-2蛋白表达。结论 ADFMChR诱导人卵巢癌CoCl细胞凋亡与其活化PPARγ,抑制NF-κB表达和提高Bax/Bcl-2比值有关。

    • Analysis of the Platelet Activated Function of Hyperlipemia Rats

      2008(3):10-13.

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      Abstract:目的分析高脂血症大鼠血小板功能的变化。方法24只Wistar大鼠随机分为正常对照组、高脂血症模型组,4周后检测两组大鼠的血脂:总胆固醇(CHO)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平。应用流式细胞术检测血小板表面a-颗粒膜糖蛋白(CD62P)的表达、酶联免疫法测定血浆中α颗粒膜蛋白.140(GMP140)浓度。所有数据采用Excel软件建库,SPSS11.5统计软件进行分析。结果正常对照组与高脂血症组CHO、TG、LDL-C、HDL-C,CHO分别是1.846mmol/L VS 13.779mmol/L、0.999mmol/L vs 0.746mmol/L、0.164mmol/L vs 10.680mmol/L、0.583mmol/L vs 0.195mmol/L。CD62P、GMP140水平分别为10.089% vs 25.169%、10.665ng/mL vs 27.045.g/mL。结论高脂血症能够促使血小板活化。

    • Expression of P-ERK1/2 and ERK2 mRNA in Lung Tissues of Asthmatic Rats

      2008(3):14-18,84.

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      Abstract:目的探讨细胞外信号调节激酶在哮喘大鼠气道中表达变化及其对气道平滑肌细胞增殖的影响。观察细胞外信号调节激酶是否参与了哮喘气道重构这一病理过程。方法18只6周龄雄性wistar大鼠随机分为对照组、哮喘组、地塞米松干预组各6只。以腹腔注射10%卵蛋白和1%卵蛋白雾化吸入复制慢性哮喘模型。干预组在每次激发前给予地塞米松干预。用免疫组化与原位杂交法检测p-ERK1/2及ERK2mRNA在不同大鼠肺组织的表达程度,采用图像分析系统进行图象分析。结果(1)哮喘模型组气道壁面积和平滑肌厚度较对照组和干预组显著增加(P〈0.05)。(2)哮喘组p-ERK1/2及ERK2mRNA在大鼠肺组织的表达程度较对照组和干预组显著增加(P〈0.05)。(3)直线相关性分析显示,哮喘组气道壁面积和平滑肌厚度与大鼠肺组织中p-ERK1/2表达水平呈正相关(分别为r=0.858,r=0.848,P均〈0.05),哮喘组气道壁面积和平滑肌厚度与大鼠肺组织中ERK2mRNA表达水平呈正相关,(分别为r=0.918,r=0.860,P均〈0.05)。结论哮喘大鼠肺组织p-ERK1/2及ERK2mRNA表达上调,并与气道重构密切相关,该结果提示细胞外信号调节激酶可能参与了气道重构中平滑肌的增殖过程。

    • Expression of P-ERK 1/2 and ERK2 mRNA in Lung Tissues of Asthmatic Rats

      2008(3):14-18.

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    • >研究报告
    • Relationship of the Value of CT Liver and Atherosclerosis

      2008(3):19-22.

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      Abstract:目的探讨肝脏CT值与动脉粥样硬化(AS)程度的关系,寻找AS活体量化观测指标,提高AS干预实验的质量。方法40只日本大耳白兔随机分为A(绞股蓝)组、B(辛伐他汀)组、C(高脂模型)组、E(对照组)组,分别喂饲高脂饲料+绞股蓝5g,kg、高脂饲料+辛伐他汀5mg/kg、高脂饲料、标准饲料。实验前和实验后3周、6周、9周检测三酰甘油(TG)、胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)。第7周和第9周进行CT检查。进行动脉和肝脏病理学检测。结果①TG、TC、LDL-C、HDL-C:A组、B组、C组饲养后TG、TC、LDL-C明显升高(P〈0.05),A组和B组TG、TC、LDL-C低于C组(P〈0.05),A组TG、HDL-C、LDL-C优于B组(P〈0.05)。②CT检查:饲养后7周、9周,A、B、C组CT值明显下降,比较有显著性差异(P〈0.05),A组高于B组、C组(P〈0.01),B组高于C组(P〈0.01)。③主动脉病理学:C组严重的大面积AS,A组和B组AS面积和程度显著低于C组(P〈0.01)。④肝脏病理学:A组、B组、C组肝脏弥漫性肿大,边缘钝而厚,灰黄色,A组较B组、C组肝脏色泽红润、肿大程度轻。A组2/3以上肝细胞发生脂肪样变。B、C组肝脏脂肪样变几乎为100%。结论肝脏CT值与AS的程度有一定的关系,可作为AS程度的量化指标,对指导实验过程有一定的价值。

    • Comparison of Normal Electrocardiogram among Three Different Strain of Minipigs

      2008(3):23-26.

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      Abstract:目的建立清醒状态记录小型猪心电图的方法,并对3个品系小型猪6个导联心电图进行分析比较。方法选用4月龄广西巴马猪7头、5月龄五指山小型猪6头、中国农大小型猪10头,清醒状态下吊床保定,测定Ⅰ、Ⅱ、Ⅲ、aVR、aVL和aVF6个导联的心电图。结果3个品系小型猪心电图的波形与人相似,心率较快,在3个品系之间差别不明显,存在偶发性窦性心率不齐。广西巴马猪、五指山小型猪和中国农大小型猪的P-R间期分别介于0.08—0.10s、0.08—0.11s和0.08—0.11s;QRS波群分别介于0.04—0.06s,0.04—0.07s和0.04~0.05s。3个品系小型猪均有清晰的S-T段,偏移小于1mm。结论本研究获得了3个品系小型猪清醒状态时的心电图基本数据,经比较,3个品系之间无明显差异。

    • Effect of Different Grade Environment on Mice and Rats

      2008(3):27-31.

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      Abstract:目的了解不同等级环境(屏障环境和普通环境)对实验大小鼠的一般生理表现和抗应激能力以及对药物反应的影响,初步判定严格的微生物控制是否影响实验动物的人类模型作用。方法将40只SPF级Wistar大鼠和200只SPF级BALB/c小鼠分别饲养于屏障环境和普通环境内,通过以下实验对比观察不同等级环境对实验大、小鼠及其应用的影响:每周测量动物的体重增长情况;饲养35d后,测定大鼠15项血液学指标、14项血液生化指标和9个脏器的相对重量;以抗缺氧实验和游泳实验判断对小鼠体质和抗应激能力的影响;以5.氟尿嘧啶急性毒性实验和戊巴比妥钠麻醉实验测试其对药物作用的异同。结果在5周观察期内,屏障环境饲养小鼠和大鼠的体重增长均明显快于普通环境的对照动物(P〈0.05);普通环境饲养大鼠的血液RBC、WBC、PCT、PLT、GOT、GGT、GPT、CK、TB值高于屏障环境大鼠(P〈0.05),而CHO、LDL-C和HDL-C值低于屏障环境大鼠(P〈0.05);不同微生物环境条件饲养小鼠的游泳耐力和抗缺氧实验结果接近,但是屏障环境组动物所获数据的变异较小;不同等级环境条件饲养小鼠的戊巴比妥钠麻醉反应和5-氟尿嘧啶中毒反应相似,而屏障环境组动物的麻醉维持时间较短,LD50较低,95%可信限狭窄。结论饲养于屏障环境和普通环境的实验大、小鼠在一般生理表现上出现部分差异,动物的抗应激能力和对药物的反应未发生显著变化;在屏障环境条件中,动物的药物反应更敏感、所获数据更稳定,因此,初步认为严格微生物控制条件下的SPF级动物可以作为生活在自然环境中的人类的模型动物。

    • Superovulation Effect in Specific Pathogen-free Mice

      2008(3):32-39,83.

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      Abstract:Objective To Investigat the superovulation effect in specific pathogen-free mice influenced by age,stock and season. Method We gave superovulation treatment to BALB /c and Km mice aged from 4 to 10 weeks in spring, summer and winter respectively. Statistics were done based on the rate of vaginal plug and ovulation, the number and the type of embryo acquired in day 3.5. Result None of the 3 factors we investigated have an effect on the rate of vaginal plug in specific pathogen- free BALB/c and Km mice. Otherwise these factors have a certain effect on the rate of embryo produced, the number and the type of embryo in specific pathogen-free BALB/c and KM mice. Conclusion Using specific pathogen-free BALB/c mice aged from 9 to 10 weeks in the superovulation experiments may give a satisfied number embryo in day 3.5,while the proper age of Km mice ranges from 4 to 10 weeks. Also, superovulation treatment in specific pathogen-free mice should be done in spring or summer, as it could give you a fine result.

    • Effects of Salinity on Multiplication and Growth of Bareneck goby

      2008(3):40-43.

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      Abstract:目的探索裸项栉鰕虎鱼繁殖和育苗的适宜盐度。方法比较不同盐度梯度条件下裸项栉鰕虎鱼的产卵率、孵化率和生长存活情况。结果裸项栉鰕虎鱼性腺成熟、产卵和孵化的适宜盐度为10‰-20‰,过低或过高盐度该鱼产卵量少,孵化率极低;适宜的盐度有利于裸项栉鰕虎鱼的生长。结论裸项栉鰕虎鱼适盐范围广,适宜的繁殖、生长盐度较低。

    • Discussion of Paralysis Symptom for the Rhesus Monkey Raised in Cage

      2008(3):44-47.

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      Abstract:Objective To use artificial nursing and medication in cage rearing rhesus monkeys which suffered from crural paralysis symptom. Method The paralysis monkeys were cared by artificial nursing, such as adopted scientific and reasonable nutrition ingredients, expanse activity space, suitlcient sunlight, furnished assistant facility, massage and functionality exercise and so on. At the same time they were treated by medication, involved heteropathy of Vitamin A, Vitamin B, Vitamin C, Vitamin D, and assorted with assistant treatment of Transfer Factor, Interferon, Prostaglandins, and Cortin medicine etc. Results Eighteen paralysis symptom monkeys were cured, the return to health rate was 60%, these monkeys recuperated well and could get round in the course of nature. Conclusion The artificial nursing and medication treatment that we adopted might prevent and cure to evacuate obstruction of urine or faces, crural atrophy, gore, dropsy, putrescence and so forth, these steps were available for crural paralysis symptom in Rhesus monkey, there would be beneficial not only to protection the resource, but also to the reduction of pain or death caused for the paralysis monkeys.

    • Detection of the Blood Biochemistry of Conventional and SPF Wistar Rats in Liaoning Area

      2008(3):48-50.

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      Abstract:目的建立本省、本地区实验动物生理指标数据背景资料,为科学研究和新药安全性评价提供准确、可靠、科学的参考依据。方法普通级、SPF级Wistar大白鼠(6~8周龄)各60只,雌雄各半,腹主动脉采血,用全自动生化分析仪及配套试剂盒检测血液生化指标。结果普通级Wistar大白鼠的ALT、ALP、CHO、CK、CRE雌雄之间差异显著(P〈0.05或P〈0.01);SPF级Wistar大白鼠的ALB、ALT、ALP、CHO、BUN、CK、AST、GLU、TBIL雌雄之间差异显著(P〈0.05或P〈0.01);普通级与SPF级Wistar大白鼠的ALT、ALP、TP、BUN、TBIL、CRE、CHO、CK、AST、ALB、GLU差异显著(P〈0.01)。结论本研究的结果证明普通级与SPF级Wistar大白鼠的血液生化指标差异显著,多数生化指标雌雄之间存在显著差异,为应用提供了有价值的参数。

    • >技术方法
    • Establishment of Myocardial Infarction Models by Freezing Regional Dominance of the Middle-upper 1/3 Segment of the LAD in Rats with Liquid Nitrogen

      2008(3):51-54,85.

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      Abstract:目的探讨大鼠左冠状动脉前降支中上1/3所支配的区域液氮冷冻处理后对心肌形态学及心功能的影响,以建立适合移植干细胞再生修复心肌梗死研究的一种新的大鼠心肌梗死模型制作方法。方法80只雄性SD大鼠,随机分为3组即:冷冻组、结扎组、对照组。大鼠麻醉后,行气管插管连通动物呼吸机,打开胸廓暴露心脏,用特制的直径为0.6cm冷冻头置入液氮中冷冻降温后迅速冷冻大鼠左冠状动脉前降支中上1/3所支配的区域,或结扎左冠状动脉前降支中上1/3处。分别于处理后1d、3d、7d、14d、28d观察心脏病理组织学变化,并于处理28d后检测心功能的变化。结果在液氮冷冻大鼠心脏后,大鼠心肌组织出现凝固性坏死,继而有肉芽组织长人梗死灶内,最后形成疤痕。用液氮冷冻法可成功复制心肌梗死大鼠动物模型。与冠状动脉结扎法相比较,操作简单,手术时间短,死亡率低.心肌梗死面积变异小。结论液氮冷冻法作为一种复制心肌梗死模型的方法,有其自身的优势.可用于心肌梗死发生机制和干细胞治疗等方面的研究。

    • Preparation of Monoclonal Antibodies of Mycoplasma Hyorhinis and Establishment of Antibody-sandwich ELISA Method

      2008(3):55-58.

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      Abstract:目的 制备多种抗猪鼻支原体的单克隆抗体,建立双抗体夹心ELISA方法用于该病原体的检测.方法 用猪鼻支原体CVCC361免疫BALB/c小鼠,采用杂交瘤技术和酶联免疫吸附实验筛选出抗该病原体的单克隆抗体;运用免疫双向扩散试验、Westem blotting确定IgG亚类及针对抗原的相对分子质量;筛选出配对抗体,建立双抗体夹心ELISA的检测方法,并评价其灵敏度和特异性.结果 共筛选出17株单克隆抗体,抗体亚类分别为ISG1、IsG2、 IgG2b、IgG3,免疫印迹结果表明单抗ZBI、ZB2及ZB16与相对分子质量为35×103的抗原有特异性结合,而ZB3和ZBl0与相对分子质量为70 × 103的抗原有特异性结合.确定了2个配对抗体(ZBI-ZBI-HRP和ZB1-ZB2-HRP),可检出最小抗原量为30ng/mL,检出猪鼻支原体活菌8.34×102CFU/mL,与人呼吸道常见的致病菌及支原体均无非特异性反应.结论 筛选的单克隆抗体具有较高的特异性和敏感性,应用双抗体夹心ELISA方法可用于猪鼻支原体的检测.

    • Method Foundation of Detection of SV40 T Antigen Genome in Wild and Caged Macaca mulatta

      2008(3):59-63.

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      Abstract:目的建立猴外周血单核细胞SV40DNA的PCR检测方法,对猕猴SV40T抗原基因进行检测。方法使用PCR方法对分别来自野外(云南宁蒗71只、景东60只)及笼养猕猴(64只)的SV40大T抗原基因进行检测,同时对扩增出的阳性结果进行测序。结果在195份样本中,有4只来自野生猴样本扩增出SV40大T抗原基因(3.1%,4/131),1只来自笼养猴样本扩增出SV40大T抗原基因(1.6%,1/64)。测序结果显示:猴血样本扩增片段序列与GenBank中的SV40大T抗原C-末端的基因序列片段有15个核苷酸不同(3.3%差异),与SV40.776标准株的序列基本一致,但SV40—776在nt3020处有一缺口。结论云南野生及笼养猕猴猴群均能检出SV40T抗原基因。因此建立SV40病毒的DNA检测技术,对用于科研及疫苗生产的实验猕猴的病毒学质量控制具有重要意义。

    • Development of Double-antigen Sandwich ELISA Kit of Simian T-lymphotropic Virus Type-I

      2008(3):64-67.

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      Abstract:目的研制灵敏度和特异性高的检测实验猴血清中T淋巴细胞趋向性病毒-1型(STLV-1型)/E体的双抗原夹心ELISA(dsELISA)检测试剂盒。方法采用经原核表达系统表达并纯化的人T淋巴细胞白血病病毒-1型(HTLV-1型)的Env蛋白作为包被用抗原,建立了检测STLV-1的dsELISA诊断方法。通过优化反应条件和筛选试剂,确定了dsELISA诊断试剂盒的相关条件,并经敏感性、特异性和重复性试验考查该试剂盒质量。结果试剂盒特异性好,批内重复试验变异系数(CV)〈7%,批间重复试验CV〈10%。对200份猴血清进行随机检测,与国际公认的诊断试剂盒(美国BioReliance公司)的符合率为97%。结论本试剂盒可初步应用于临床上实验猴STLV-1型抗体的检测。

    • Improved Technique of Cervical Heterotopic Heart Transplantation in Mice by Self-made Cuff

      2008(3):68-71.

      Abstract (1881) HTML (0) PDF 0.00 Byte (0) Comment (0) Favorites

      Abstract:目的探讨小鼠心脏移植模型的建立与改良。方法供体采用BLAB/c小鼠,受体采用C57小鼠。用24GACuff管法套扎供心肺动脉与颈外静脉,用自制Cuff管套扎供心升主动脉与颈总动脉。结果供心冷缺血时间少于15min,手术时间大大缩短,成功率明显提高。结论改进后的模型显著降低了手术难度,可用于移植免疫的各种研究。

    • One Method of Lumbar Puncture and to Place a Soft Plastic Tub of Beagle

      2008(3):72-74.

      Abstract (1709) HTML (0) PDF 0.00 Byte (0) Comment (0) Favorites

      Abstract:Objective To establish a new test-determinited the cerebrospinal fluid of Beagle. Methods Using ketamine to anesthetize, collect the cerebrospinal fluid afer lumbar puncture and to place a soft Plastic tube. Results It could collect the cerebrospinal fluid more than 48 hours if we successed make lumbar puncture and to place a soft plastic tube. Conelusion Lumbar puncture and to place a soft Plastic tube is a simple and utility method to continiun collect the cerebrospinal fluid.

    • A Simple Device for Holding Rats

      2008(3):75-76,83.

      Abstract (1539) HTML (0) PDF 0.00 Byte (0) Comment (0) Favorites

      Abstract:Objective To provide a practical device and skills for injection of caudal veins, knee joints and belly cavity of rats. Method Using plastic bottle for injection and haemostatie forceps to make a simple device for holding rats. With this device, the rats were treated by injection of caudal veins, knee joints or belly cavity. Result The rats could be hold well in the device. With the device, the injection of caudal veins, knee joints and belly cavity could be finished smoothly, quickly and safely. Conclusion This device is economical, practical and easy to be made. It is a good device for holding rats.

    • Comparison of the Model Establishment Method of Syndrome of Deficiency of Kidney YANG between Adenine and Hydrocortisone

      2008(3):77-80.

      Abstract (2157) HTML (0) PDF 0.00 Byte (0) Comment (0) Favorites

      Abstract:目的比较腺嘌呤诱导与氢化可的松诱导的SD大鼠肾阳虚模型的优劣,从而得到临床症状、生化指标更符合中医“肾阳虚”型的动物模型,以供科研、教学使用。方法雄性SD大鼠100只,随机分成正常组、腺嘌呤造模组及氢化可的松低剂量组、中剂量组、高剂量组各20只,造模成功后检测血肌酐(Scr)、尿素氮(BUN)、尿液中17-羟皮质类固醇(17-OH)和血浆中甲状腺激素(13、T4)、睾丸酮(T)、雌二醇(E2)及模型大鼠骨密度。结果无论从症状主证方面,还是客观指标方面(除骨密度外),腺嘌呤诱导的SD大鼠肾阳虚型动物模型在生化指标研究方面均优于氢化可的松诱导组。结论腺嘌呤诱导的肾阳虚证SD大鼠模型在症状主证方面及其相关的生化研究方面优于氢化可的松组。

    • >简讯
    • Observation on Skin Histopathology of SD Rat

      2008(3):81-82,86.

      Abstract (2357) HTML (0) PDF 0.00 Byte (0) Comment (0) Favorites

      Abstract:Objective To observe the histological structure of skin in SD rat at different stages. Method By using 10 percent formaldehyde paraffin section, and staining with hematoxylin and eosin. Result The skin thickness was thin in SD rat which lacked sratum lucidum at day 1. They had a few shallower and irregularly positioned hair follicles and well-developed sebaceous glands. The skin hair follicle size increased significantly, An increase in thickness of the epidermis was observed in SD rat as a result of an increase in the number of cell layers in basal layers and stratum corneum, Hair follicles had moved down to the subdermal fat tissue layer at month 6. Analoisis in sebaceous glands and sweat gland were observed in SD rat with very thin elastic fibers at month 24. Conclusion Histological structure through the skin of SD rat at different stages showed difference.

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