基于新型分子距边矢量ν与多元统计方法对于烷烃13C NMR化学位移和(CSS)估计与预测的深入研究

基于本实验室提出一种新型以势能形式表达的分子距边矢量, 深入地系统研究了核磁共振碳-13谱化学位移和(CSS)规律以及分子拓扑指数矢量在定量结构波谱关系(QSSR)中的应用. 借助多种计量化学方法包括多元线性回归、逐步多元回归、主成分回归、主筛选回归等进行分子拟模和定量相关研究, 发现烷烃¹³C NMR 化学位移和(CSS)与其分子距边矢量及路径长度指数有良好线性相关性, 回归方程及其统计参数为:CSS=bν+cp₃=∑_^mj=0b_jν_j+b₁₁p₃=b₀ν+b₁ν₁+b₂ν₂+b₃ν₃+b₄ν₄+b₅ν₅+b₆ν₆+b₇ν₇+b₈ν₈+b₉ν₉+b₁₀ν₁₀+b₁₁p₃=-13.576+22.179ν₁+28.407ν₂+25 .950ν₃+26.690ν₄+14.498ν₅+5.726ν₆-5.379ν₇-3.214ν₈-15.021ν₉ -25.710ν₁₀+12.278p₃ n=63, R=0.997, EV=99.68%, RMS=3.7348, SD=4.1 18, F= 773.116, U=144228.844, Q=864.938; CV: R²_CV=0.980, EV=98.83%, RMS=7.126 1, SD_CV=7.634, F_CV=221.720, U_CV=142121.891, Q_CV=2971 .896.结果良好.     

Mechanism of the interaction between Au nanoparticles and polymerase in nanoparticle PCR

Nanoparticle PCR is a novel method to optimize DNA amplification. It performs well in improving specificity, enhancing sensitivity and speed. Several mechanisms were proposed in previous studies: one was based on the interaction between gold nanoparticles (AuNPs) and DNA while the other was attributed to the heat transfer property of AuNPs. In this paper, we propose that the interaction between AuNPs and DNA polymerase can significantly influence PCR. First, the addition of DNA polymerase can eliminate the inhibitory effects of excess AuNPs. Second, the addition of AuNPs will increase yield of the desired PCR product and make the optimum concentration of DNA polymerase move to higher value. Third, while excess polymerase might inhibit amplification efficiency, AuNPs can reverse this process and the yield of PCR amplification. Based on these results we propose a possible mechanism that AuNPs might modulate the activity of polymerase and improve PCR amplification.     

An ultra‐high temperature flow‐through capillary device for bacterial spore lysis

Rapid and specific characterization of bacterial endospores is dependent on the ability to rupture the cell wall to enable analysis of the intracellular components. In particular, bacterial spores from the bacillus genus are inherently robust and very difficult to lyze or solubilize. Standard protocols for spore inactivation include chemical treatment, sonication, pressure, and thermal lysis. Although these protocols are effective for the inactivation of these agents, they are less well suited for sample preparation for analysis using proteomic and genomic approaches. To overcome this difficulty, we have designed a simple capillary device to perform thermal lysis of bacterial spores. Using this device, we were able to super heat (195°C) an ethylene glycol lysis buffer to perform rapid flow‐through rupture and solubilization of bacterial endospores. We demonstrated that the lysates from this preparation method are compatible with CGE as well as DNA amplification analysis. We further demonstrated the flow‐through lysing device could be directly coupled to a miniaturized electrophoresis instrument for integrated sample preparation and analysis. In this arrangement, we were enabled to perform sample lysis, fluorescent dye labeling, and protein electrophoresis analysis of bacterial spores in less than 10 min. The described sample preparation device is rapid, simple, inexpensive, and easily integratable with various microfluidic devices.     

HCV基因分型在丙肝诊断和治疗中的应用

目的 建立ＨＣＶ基因分型方法，研究ＨＣＶ基因型与肝病程度的关系，探讨ＨＣＶ基因型与ＩＦＮ－ａ２ｂ抗病毒疗效的关系。方法 ＨＣＶ基因组ＮＳ５区ＰＣＲ扩增产物酶切分型。结果 ７３例ＨＣＶ感染者中有ＨＣＶⅡ型４５例，ＨＣＶⅢ型２８例。Ⅱ型ＨＣＶ感性肝炎３１例，肝硬化１４例；Ⅲ型ＨＣＶ感染者中有慢性肝炎２６例，肝硬化２例。Ⅱ型ＨＣＶ感染者的肝硬化发生率显著高于Ⅲ型ＨＣＶ感染者（Ｐ〈０．０５）。所有病人均应     

地高辛掺入法检测HCV RNA含量在肝病诊断中的应用

目的：研究血清HCVRNA含量与HCV感染者肝病程度的关系。方法：应用地高辛掺入法检测血清HCVRNA含量。在PCR过程中将DIG－dUTP掺入到PCR产物中去，再用PCRELISA法检测PCR产物，结合标准参考样本而达定量目的。结果：丙肝后肝硬化组血清HCVRNA含量显著高于丙型肝炎组。结论：地高辛掺入法检测血清HCVRNA含量可以为HCV感染者的病情判断提供科学依据。     

一种快速检测植物病毒RNA蓄积量的竞争PCR方法

用Oligo6.6软件在pUC18上设计1对比目的基因多100～200 bp的内参引物,摸索了目的基因和内参共同扩增的竞争PCR反应条件,测定了嘧肽霉素处理对烟草组织中烟草花叶病毒RNA相对含量的影响,同时用间接ELISA方法验证了该方法.试验结果表明:竞争PCR的方法能够快速、相对准确的测定植物组织中病毒RNA蓄积量的变化情况.     

Physical location of terminal markers belonging to five linkage groups in maize RFLP map using in situ hybridization

Ten terminal or subterminal RFLP markers belonging to linkage groups 1, 3, 5, 6, and 10 in maize RFLP map were physically locted onto maize mitotic chromosomes with in situ hybridization. All biotinylated probes from 600 to 2 250 bp were detected by DAB staining. The markers belonging to linkage groups 1, 3, 5, 6, and 10 correspondingly located at the chromosomes 1, 3, 5, 6, and 10. All of the tested markers except bnl6.25 and umc44 were duplicated sequences. Each of them was also labeled on another chromosome besides on the chromosome corresponding to its linkage group. The marker bnl3. 04 was triplicated sequences and the signals were detected on three nonhomologous chromosomes. In the tested ten markers, there were only four located at the ends of corresponding chromosomes. Others were located at sites midway along the chromosome arms or near the centromeres. The region covered by two terminal or subterminal markers in each of linkage groups 1, 3, 5, and 6 occupied 80.02%, 38.25%, 82.30% and 51.16% of the region of both short and long arms in chromosomes 1, 3, 5, and 6 respectively. Only two terminal markers of linkage group 10 covered the whole chromosome 10. In some linkage groups, two terminal or subterminal markers covered a short genetic distance but were physically distant, while two covering a longer genetic distance were physically closer. Supported by The National Natural Science Foundation of China and the Doctorate Vesting Point Foundation of the Education Committec of the People's Republic of China Mao Ninghui: born in 1986, used to be an MS student of Wuhan University in 1992–1995 and now is working in Fudan University, Shanghai 200433     

稻瘟菌毒素胁迫下水稻根部生理变化及相关基因表达研究

以丽江新团黑谷和谷梅二号为研究对象,利用稻瘟菌粗毒素分别对两者的幼苗根部处理0、6、12、24、48h,并进行地上部和地下部生理指标和相关基因表达分析研究.结果表明,毒素处理后,除SOD外,POD和CAT的活性均升高,MDA含量也升高.与水杨酸和茉莉酸途径相关的8个基因与本底水平相比都表现出上调或下调的趋势,可见这两个途径都得到了表达.特别是48h时,PAL、JIOsPR10、PR1b和PR4在稻瘟病抗性品种谷梅二号的根或叶中的表达量远高于敏感品种丽江新团黑谷,说明这4个基因的表达在调节谷梅二号抗稻瘟菌毒素中具有重要作用.     

多氯联苯胁迫下红树蚬荧光定量PCR内参基因的筛选

【目的】筛选出多氯联苯(PCBs)胁迫下红树蚬(Polymesoda erosa)的实时荧光定量PCR(qRT-PCR)最适内参基因,为进一步开展分子毒理学研究奠定基础。【方法】以β?actin、18S rRNA、GAPDH和#?tubulin为候选内参基因,qRT-PCR测定PCBs胁迫下4个候选内参基因在红树蚬外套膜、鳃、闭壳肌和肝胰腺组织中的表达水平,应用geNorm、NormFinder和BestKeeper软件分析其表达稳定性。【结果】geNorm软件分析表明β?actin表达最稳定;NormFinder软件分析发现,外套膜、鳃和肝胰腺组织中β?actin的稳定性最好,闭壳肌组织中β?actin(0.454)表达稳定性略微低于#?tubulin(0.425),排第2位;BestKeeper软件分析表明,外套膜和鳃组织中β?actin最稳定,肝胰腺和闭壳肌组织中GAPDH最稳定,β?actin排位第2位。【结论】筛选β?actin为红树蚬在PCBs胁迫下的qRT-PCR最适内参基因。