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The morphological and cytopathological characters of the viruses were observed by electron microscopy The types of virus were identified with I-ELISA, DAS-ELISA, TAS-ELISA, RT-PCR and IC-PCR. The CP genes of the main viruses identified in this research were amplified by PCR, cloned into pGEM-T and sequenced. The pumpkin germplasm from Heilongjiang and Yunnan provinces was screened for resistance to the main viruses using friction and roots immersed inoculation.

应用电子显微镜负染法和超薄切片法观察病原粒体形态及细胞病理学特征；利用酶联免疫吸附测定法中的双抗体夹心ELISA、间接ELISA、三抗体夹心ELISA以及反转录聚合酶链式反应、免疫捕捉PCR方法鉴定了采集样品中的病毒种类；对被确定为南瓜主要病毒病原的外壳蛋白基因进行了PCR扩增，克隆到pGEM-T载体并进行测序；应用摩擦接种法和浸根接种法对云南、黑龙江省部分南瓜品系、品种进行了抗性筛选。

Total mRNA was extracted from a venom gland of snake Daboia russellii siamensis following the manufacturers protocol of the PolyATtract System 1000 kit purchased from Promega Biotech. cDNAs encoding C-type lectins were amplified by RT-PCR and subcloned into a pMD18-T vector.

按照Promega公司的mRNA提取试剂盒操作手册，从圆斑蝰蛇的毒腺中提取mRNA；利用RT-PCR的方法进行体外扩增，获得C-型凝集素蛋白的基因，克隆到pMD18-T载体中。

The RT-PCR product was inserted into pTG19-T vector and transformed into E. coli successfully. By blastn, the sequence results of Kunming mus musculus were in complete accordance with the conservative sequence of Genbank NR_003278 (791bp-1153bp). By Blastn in NCBI, the sequence with little difference among animals was confirmed to be conservative. After Blastn, fourteen complete CDS coding for different animals were chosen. According to VECTOR NIT 9.0 software, the similarities between Kunming mus musculus and bos taurus, homo sapiens, erinaceus europaeus, cricetulus griseus, sus scrofa, dasypus novemcinctus, rattus norvegicus, rabbit, equus caballus, macaca fascicularis, didelphis virginiana, monodelphis domestica and vombatus ursinus was 67%, 100%, 100%, 36%, 100%, 100%, 67%, 100%, 100%, 92%, 99%, 99% and 99%. In the phylogenetic tress constructed with the forteen 18S rRNA by Treeview, the Kunming mus musculus clustered with cricetulus griseus, sus scrofa and rabbit, which was nearer to cricetulus griseus and was most far away from macaca fascicularis.(3) After sencodary structure analyses of 18S rRNA of mus musculus, an oligonucleotide fragment for RNAi was designed and synthesized, which was transformed into plasmid, and restriction enzyme analyses and sequencing results should the expression plasmid pGPH1/ GFP/Neo-mouse-sh 18S rRNA were constructed for RNAi successfully.

结果①通过RT-PCR检测显示18S rRNA基因在小鼠卵巢组织和单个GV期、MⅠ期卵母细胞中均有表达，且在未成熟卵母细胞中，MⅠ期的表达明显强于GV期的表达；②RT-PCR产物克隆测序结果显示：昆明小鼠18S rRNA基因保守区序列与基因库序列[NR_003278保守区部分(791bp～1153bp)]完全一致；Blastn比对结果发现：在不同物种中差异较小，选出14种生物18S rRNA全序列经VECTOR NIT 9.0软件分析，提示昆明小鼠18SrRNA与牛、人类、刺猬、中国仓鼠、猪、犰狳、褐鼠、兔子、马、食蟹猴、负鼠、短尾猊、袋熊的18S rRNA的相似率依次为67%，100%，100%，36%，100%，100%，67%，100%，100%，92%，99%，99%，99%；Clustal 1.81和Treeview构建出的分子进化树表明：在上述14种生物中昆明小鼠与中国仓鼠进化关系最近，与兔子、猪聚成一簇，与食蟹猴进化关系最远；③根据18S rRNA二级结构设计并合成RNA干扰寡核苷酸片段，重组质粒经过限制性内切酶及测序表明成功构建了pGPH1/GFP/Neo-mouse-sh 18S rRNA干扰表达质粒。

Methods:TNF-α and TGF-β_1 mRNA expression on placenta decidual tissue from 15 early spontaneous abortion women and 20 normal early pregnant women were examined by real-time fluorescence quantitative RT-PCR.

方法采用实时荧光定量RT-PCR法检测了15例早期自然流产及20例同期正常早孕要求行人工流产者蜕膜组织的TNF-α和TGF-β1mRNA表达水平。

The remaining 9 rats in control group, 16 rats in treated grouop and 15 rats in model group were decollated and obtained brains to detect the relative mRNA expression of nuclear receptor-related factor 1 (Nurr1) and muscarinic acetylcholine receptor 5(M5) byusing RT-PCR, which was the consequence of those comparing with reference geneβ-actin mRNA expression.

将对照组、治疗组、模型组其余9、16、15只大鼠，断头取脑，采用RT-PCR方法检测大鼠脑组织孤儿核受体及乙酰胆碱毒蕈碱型受体5(M5)mRNA相对于内参基因β-actin mRNA的相对表达值。

In sham-operation group,animals were decollated and their brains were taken out at 24 hours postoperatively. In all the groups, hippocampal brain injury was detected using histopathological method combined with semi-quantified RT-PCR methods for detecting caspase-3 mRNA. Adult rats aged 90 days were used in modified y maze to examine their learning and memory functions. All these three experiments were independent.

各组海马区脑损伤行组织病理学检测，半胱氨酸天冬氨酸蛋白酶-3 mRNA的表达采用半定量反转录-聚合酶反应方法进行检查，成年90 d龄的大鼠进行三等分迷宫测试其学习记忆能力，3个实验各自独立。

We collected normal brain tissues from 10 patients who underwent decompressive surgery because of brain injury. The expression of CENP-F was detected by RT-PCR and immunohistochemical staining respectively in normal brain tissues, and these were used as control for the CENP-F expression.

3收集10例颅脑外伤后行手术减压的正常脑组织，分别采用免疫组化和RT-PCR两种方法分别检测正常脑组织中CENP-F的表达，并作为对照组。

FCM analysis showed that the apoptotic rate of MDA-MB-231 cells and intracellular Ca(superscript 2+) concentration increased gradually with the increase of deguelin concentration. RT-PCR and Western blotting analysis showed that the expression of Bcl-2 mRNA and protein was clown-regulated and that of Sax was op-regulated after deguelin treatment.

流式细胞术检测显示细胞凋亡率和细胞内钙离于浓度随鱼藤素浓度的增加而增多；RT-PCR和蛋白印迹检测结果发现鱼藤素能使Bcl-2 mRNA和蛋白表达减少，而Bax表达增加。

AIM: To investigate the change of cell cycle checkpoint kinases Chk1/2 in human gastric cancer MGC803 cells induced by diallyl disulfide. METHODS: mRNA expression was assayed by means of RT-PCR in MGC803 cells treated with DADS.

目的：观察二烯丙基二硫化物对人胃癌MGC803细胞的细胞周期检查点激酶1(Chk1)和细胞周期检查点激酶2(Chk2)的影响。

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As of Tuesday, Google's results were still censored in China.

截至周二，谷歌的搜索结果仍受中国审查。

In order to make the positive action increase and negative one decrease, the sub-forces of the social factors must be adjusted to form a centripetal force.

在这一过程中，人的主体性发挥是社会有机体健康发展的灵魂。