快速检测猪瘟兔化弱毒疫苗株的TaqMan荧光定量RT-PCR方法的建立

在猪瘟病毒兔化弱毒疫苗株（HCLV）的5′端非编码区设计一对引物和一条TaqMan探针,通过优化反应条件,成功建立了特异性检测HCLV的荧光定量RT-PCR方法．结果表明：该方法检测的最低拷贝数为45拷贝／μL,灵敏度比普通PCR方法高10^4倍,在较广的范围内（4．5×10^1-4．5×10^6拷贝／μL）有良好的线性关系（r=0．994）;分别以乙型脑炎病毒、猪繁殖与呼吸综合征病毒、副猪嗜血杆菌、牛病毒性腹泻,黏膜病病毒作为模板进行TaqMan RT-PCR扩增,未出现阳性信号：4个不同浓度标准品组内试验变异系数为1．90％～5．82％,组间试验变异系数为4．02％～5．69％：HCLV3个cDNA样本组内试验变异系数为3．72％～4．93％;组间试验变异系数为2．99％～4．02％．该方法具有很好的灵敏性、特异性及稳定性,能够快速准确定量检测HCLV,为HCLV疫苗的研制、猪瘟病毒分子生物学等方面研究提供了一种快捷有效的工具．     

玉米幼苗热激诱导抗冷性过程中钙的效应

 玉米幼苗在冷胁迫前经过热激处理或CaCl₂浸种后再热激处理,其存活率,抗氧化酶GR的活性、可溶性蛋白质中热稳定蛋白质和游离脯氨酸的含量在的冷胁迫中均发生了变化,发现热激处理幼苗的这些参数高于对照,而最高的是CaCl₂浸种后再热激处理的,表明热激能提高玉米幼苗的抗冷性,而Ca²⁺对上述热激处理有加强作用.     

细胞凋亡过程中热休克蛋白的调节

热休克蛋白(heatshockproteins,HSPs)是一类进化上高度保守,广泛存在于自然界原核、真核细胞中的蛋白质。HSPs在正常细胞中呈基础性表达,维持细胞的基本形态和功能;它在应激环境下的细胞中,可参与细胞的损伤和修复,发挥应激保护作用。新近的研究发现,HSPs在细胞凋亡中发挥重要作用。     

轴对称零件高温热冲击动态热应力分析

采用泛函分析法，导出非定常温度场热弹性轴对称问题的线性时空有限元基本方程，并开发了预测轴对称零件承受高温热冲击载荷时热弹性动态应力分布的分析软件。在传热方程中不考虑热耦合项的情况下，建立了轴对称零件承受高温热冲击载荷时的有限元分析理论和离散模型。     

Expression profile analysis of 9 heat shock protein genes throughout the life cycle and under abiotic stress in rice

Plant heat shock proteins (Hsps) facilitate protein folding or assembly under diverse developmental and adverse environmental conditions. Nine OsHsps were identified in our previous study from a proteomic analysis of rice cv. IRAT 109 leaf samples at the seedling stage under drought stress. To obtain additional information on the 9 OsHsp genes, this study focused on an expression profile analysis at different development stages throughout the life cycle of rice, and under different abiotic stresses and phyto-hormone treatments. The 9 genes exhibited distinctive expression patterns in different organs or development stages. Five of the genes (OsHsp72.90, OsHsp72.57, OsHsp71.18, OsHsp24.15 and OsHsp18.03) showed high expression in the endosperm, indicating that OsHsp genes may play important roles in rice seed development. All 9 OsHsps were up-regulated under heat, polyethylene glycol, and abscisic acid treatment, whereas salt stress caused up-regulation of 6 genes (OsHsp93.04, OsHsp71.10, OsHsp71.18, OsHsp72.57, OsHsp24.15 and OsHsp18.03) and cold stress resulted in down-regulation of OsHsp93.04 and OsHsp72.57. These diverse expression profiles imply potential functional diversity of the Hsp gene family in rice.     

Quantitative detection of ING1 mRNA under different gene regulation based on molecular beacon

As an important tumor suppressor, normal expres- sion of ING1 is associated with the maintenance of cell normal physiological functions. Its low expression of- ten causes aberrant events of cell survive, growth and differentiation[1] which results in loss…     

A preliminary study on functional domains of small heat shock protein Hsp16.3

Hsp16.3, the small heat shock protein (sHSP) from Mycobacterium tuberculosis, was originally identified as an immunodominant antigen,which possesses three functional domains typical of sHSP family, namely the N-terminal hydrophobic region, α-crystallin domain and a short non-conserved C-terminal extension.To further understand the functional assignment of these independent regions, the three functional domains of Hsp16.3 were defined and the two N- or C-terminal truncated Hsp16.3 remnants were successfully cloned, expressed and purified.In the far and near circular dichroism analysis, the results showed that these remnants expressed similar secondary and tertiary structures to that of wild-type protein.During the reassembly of wild-type nonamer, the C-terminal truncated remnant could interact with the wild-type protein to form hetero-oligomers.When trypsin is used to digest the wild-type Hsp16.3, its α-crystallin domain could resist such degradation.Taken together, these results indicate that the stable secondary and tertiary structures of Hsp16.3 are mainly kept by its α-crystallin domain.     

热激和热胁迫过程中玉米幼苗谷胱甘肽还原酶活性和同工酶的变化

 以玉米品种晴3和大黄为材料,研究结果表明热激幼苗的存活率比未热激幼苗高,这说明热激提高了玉米幼苗的抗热性.热激幼苗的谷胱甘肽还原酶活性明显高于未经热激的幼苗,同时热激处理后,GR的同工酶谱在整个热胁迫期间与对照相比也存在差异,说明热激过程中谷胱甘肽还原酶活性的升高和同工酶的变化与玉米幼苗的抗热性相关.     

氯胺酮相关性膀胱炎大鼠逼尿肌热休克蛋白27的表达及其意义

研究热休克蛋白27(HSP27)在氯胺酮诱导膀胱炎大鼠逼尿肌中的表达及临床意义.基于氯胺酮腹腔内注射诱导大鼠膀胱炎模型,选择不同时间通过尿流动力学检测膀胱收缩功能,收集大鼠膀胱组织,常规病理染色观察膀胱粘膜及逼尿肌结构,免疫组化法检测HSP27在逼尿肌中的表达水平.实验后观察结果表明,同对照组相比,实验组尿动力学表现为膀胱收缩频率显著增加及初感膀胱容量明显减小;对照组膀胱组织无明显病理及免疫表达变化,而实验组可观察到不同程度膀胱粘膜充血,炎症细胞浸润,HSP27表达水平的上升,且HSP27表达水平与诱导时间呈正相关.实验结果表明HSP27的表达水平可能与氯胺酮膀胱炎的逼尿肌收缩活动程度相关.     

睾丸组织高表达新基因THE的克隆及表达特征分析

以Homo.sapiens brain为库,选取编号为CA423810的EST序列,联网到NCBI调用Blast服务器分析,该EST序列是一个代表新基因的未知序列.以该序列作为电子探针,通过Internet采用Blast软件进行GenBank的EST数据库检索,获得了该序列的电子延伸产物EST重叠群.经与人类基因组草图进行序列校正,获得了全长为2 232bp的cDNA序列.利用NCBI的ORFfinder服务器,分析发现该序列具有完整的阅读框架,从而确定了基因的全长cDNA序列.该基因定位于染色体上的5q22,编码由388个氨基酸组成,分子量为44859的蛋白质.运用RTP-CR技术,以新基因电子克隆全长cDNA序列设计基因特异性引物,以11例癌组织及相应的正常组织的cDNA、人胎脑cDNA文库和人睾丸cDNA文库为模板扩增目的片段,并以看家基因GAPDH为内对照,检测目的基因的mRNA表达水平.研究结果表明,新基因只在睾丸组织中高表达,在直肠癌、结肠癌、宫颈癌、胃癌等的癌组织及相应的正常组织中都无明显的表达.因此将该新基因命名为睾丸组织高表达基因(testis high expression,THE).以上结果提示,THE基因可能是在人脑中表达的同一基因的不同剪接本.关键词检索UniGene数据.     

Effects of metal ions and heat shock on the expression of a metallothionein-like gene htMT2 in Helianthus tuberosus

The expression of htMT2 in tissues of Helianthus tuberosus treated with metal ions and heat shock is studied by Northern blot hybridization. The results indicate that htMT2 is not expressed in roots. The expression in leaves and stems can be reduced by Cu2+ and induced by low concentration of Zn2+. However, the expression of htMT2 can be suppressed by Zn2+ when its concentrations are above 500 μmol/L. The effects of Ca2+ on the expression of htMT2 in leaves are similar to that of Zn2+. But Ca2+ induces the expression of htMT2 in stems. The expression levels of htMT2 in various tissues are not significantly affected by heat shock. The results above demonstrate that the expression of htMT2 can be regulated by metal ions and confirm that htMT2 is a new metallothionein-like gene.     

一种马铃薯病毒RT-PCR诊断新方法

对传统的植物病毒RNA苯酚提取法进行改进;同时设计并优化单管RT-PCR(O ne-tube RT-PCR)方法,使传统的RT-PCR两步反应在一个反应体系中同时进行,依据马铃薯Y病毒和马铃薯S病毒的基因组RNA保守序列设计2套特异性引物,采用上述新建立的检测方法对染病的马铃薯叶片组织进行病毒检测,结果与G eneB ank中报道的这两种马铃薯病毒核苷酸序列进行B last比较,同源性达到96%以上,用这种新型诊断方法对马铃薯病毒核酸进行了浓度检测,实验结果证明,这种新型马铃薯病毒诊断方法具有可靠、快速、简便等特点.     

实时定量PCR检测B细胞恶性肿瘤中BCL11A基因的表达水平

目的:建立B细胞淋巴瘤/白血病BCL11A基因的定量检测方法,并分析其在B细胞恶性肿瘤中的表达水平。方法:利用实时定量RT-PCR分析B细胞淋巴瘤(18例)、B细胞性慢性淋巴细胞白血病(B-CLL,8例)、T细胞性急性淋巴细胞白血病(T-ALL,8例)和正常对照(15例)外周血单个核细胞(PBMC)中BCL11A基因的表达水平,以甘油醛-3-磷酸脱氢酶(GAPDH)基因作为内对照。结果:B-CLL组和B细胞淋巴瘤组患者PBMC中BCL11A表达水平均明显高于正常对照(P=0.000)和T-ALL组(P=0.000);T-ALL组和正常对照组BCL11A表达水平无显著差别(P=0.084);B-CLL组和B细胞淋巴瘤组BCL11A表达水平无显著差别(P=0.776)。在B细胞淋巴瘤不同的病例中,BCL11A表达水平有较大差异,其中最小值为0.04,最大值为9.70,中位值为1.00。结论:成功建立BCL11A基因的定量检测方法。     

三热源内可逆热变换器比供热率优化

针对经典热力学分析方法的不足,根据热变换器的功能,投资等方面的特点,按照内可逆联合诺循环模型,利用有限时间热力学理论方法,导出了吸收式热变换器比供热率与操作参数,比供热率与热学性能系数的优化关系,从而使优化结果兼顾了系统有能的合理性和与总传热面积相关的反映设备投资回报的经济性指标,对实际热变换器的优化和设计有更全面的指导意义。     

Development and application of flexible substrate sensors in instantaneous heat flux measurement

A new type of sensor with the flexible substrate is introduced. It is applicable in measuring instantaneous heat flux on the model surface in a hypersonic shock tunnel. The working principle, structure and manufacture process of the sensor are presented. The substrate thickness and the dynamic response parameter of the sensor are calculated. Because this sensor was successfully used in measuring the instantaneous heat flux on the surface of a flat plate in a detonation-driven shock tunnel, it may be effective in measuring instantaneous heat flux on the model surface.     

mRNA差异显示技术研究低温条件下深黄被孢霉基因的表达差异

mRNA差异显示技术已经广泛应用于研究植物、动物和微生物在各种环境条件下基因的差异表达研究.研究以15℃和30℃培养的深黄被孢霉M6-22菌体为材料,利用mRNA差异显示技术研究两种培养条件下基因的表达差异.经实时荧光定量PCR验证,共获得7条差异片段,相似性搜索结果表明它们是6-磷酸葡萄糖异构酶、单糖核苷酸转运蛋白、Ras1鸟苷酸转移因子、依赖于NAD的苹果酸脱氢酶、Δ12-脂肪酸脱氢酶、CLK4关联的丝氨酸/精氨酸丰富蛋白和假定蛋白,涉及糖酵解、蛋白质修饰、信号传导、脂肪酸合成和mRNA加工等生命过程,表明深黄被孢霉M6-22低温适应性是多种途径协同调控的结果.     

纳米金免疫层析法定量检测尿RBP

应用纳米金免疫层析技术(双抗夹心法)和柠檬酸三钠还原法建立一种对尿RBP的快速定量检测方法,并在此基础上研制出快速检测试纸条.该试纸条用纳米金免疫层析定量分析仪可在15 min内实现RBP定量检测,检测限为150μg/L(相当于150 ng/mL),检测范围为150～5000μg/L.与尿微量白蛋白(ALB)、转铁蛋白(TRF)、β2-微球蛋白(β2-MG)、尿纤维连接蛋白(FN)、溶菌酶(LZM)等肾脏疾病标志物无交叉反应.该方法特异性强、检测灵敏度高、范围广、简便快捷,在近端肾小管的损伤、糖尿病肾病等肾脏疾病的早期诊断及过程监控中具有较为广泛的临床应用.