DNA barcoding of Gaultheria L.in China (Ericaceae: Vaccinioideae)

Four DNA barcoding loci,chloroplast loci rbcL,matK,trnH-psbA,and nuclear locus internal transcribed spacer (ITS),were tested for the accurate discrimination of the Chinese species of Gaultheria by using intraspecific and interspecific pairwise P-distance,Wilcoxon signed rank test,and tree-based analyses.This study included 186 individuals from 89 populations representing 30 species.For all individuals,single locus markers showed high levels of sequencing universality but were ineffective for species resolvability.Polymerase chain reaction amplification and sequencing were successful for all four loci.Both ITS and matK showed significantly higher levels of interspecific species delimitation than rbcL and trnH-psbA.A combination ofmatK and ITS was the most efficient DNA barcode among all studied regions,however,they do not represent an appropriate candidate barcode for Chinese Gaultheria,by which only 11 out of 30 species can be separated.Loci rbcL,matK,and trnH-psbA,which were recently proposed as universal plant barcodes,have a very poor capacity for species separation for Chinese Gaultheria.DNA barcodes may be reliable tools to identify the evolutionary units of this group,so further studies are needed to develop more efficient DNA barcodes for Gaultheria and other genera with complicated evolutionary histories.     

Homology modeling threedimensional structure of AnxB1 and reducing its immunogenicity by sequence-deleted mutagenesis

AnxB1,a novel annexin previously isolated from Cysticercus cellulose,shows high thrombi affinity and anticoagulant activity in vivo.In order to investigate the relationship between structure and biological function,a predicted three-dimensional(3D)model of AnxB1 was generated by homology modeling.This model contains four homologous internal-domains and the Cα trace of domain Ⅰ,Ⅱ and IV shows high similarity.Based on the structure characterization,four sequence-deleted mutants were constructed and expressed as GST fusion proteins in E.coli.Two of the mutants,GST-M3 and GST-M4 reserved high anticoagulant activity(p＜0.01 vs.GST).Furthermore,compared with the wild type GST-AnxB1,the immunogenicity of GST-M3 and GST-M4 was reduced significantly(p＜0.01)and the molecular weight was lowered to 27 kD and 34 kD,respectively.These observations laid a solid foundation for further study on developing new thrombolytic agents with higher efficiency and lower side effect.     

刺五加皂苷合成关键酶基因表达的半定量RT-PCR分析

根据已克隆的刺五加鲨烯合酶(squalene synthase,SS)、鲨烯环氧酶(squalene epoxidase,SE)和β-香树酯醇合成酶(β-amyrin synthase,bAS)基因序列信息设计引物,通过半定量RT-PCR分析了SS、SE和bAS基因在刺五加不同生长发育时期和不同器官中表达量的变化.结果表明,SS、SE和bAS基因在各生长发育时期和各器官中均有表达,但表达量差异显著(P<0.05),三者均在盛花期表达量最高,之后降低,进入果实成熟期后SS和bAS的表达量迅速回升,SE无显著变化.SS和bAS在叶片和根中的表达量较高,SE表达量的最大值出现在叶片和幼茎中.刺五加SS、SE和bAS基因的表达间存在显著的正相关关系(P<0.05).研究结果为进一步分析关键酶基因对刺五加三萜皂苷生物合成的影响奠定了基础.     

微水相超声波协同固定化脂肪酶催化酯交换过程优化

超声波协同固定化脂肪酶催化制备生物柴油的最佳工艺条件为：超声波功率70W、叔丁醇为反应介质、叔丁醇用量3％（v／v）、醇油比3：1且甲醇分三批加入、反应温度40℃、水含量为2％（v／v）。副产物甘油对固定化脂肪酶使用寿命影响最大,使用后的固定化脂肪酶用丙酮洗去表面的甘油,进行酯交换反应,酶的稳定性大为提高,可连续使用16批次。     

武夷山九曲溪水体营养水平与原生动物群落变化相互关系研究

2000年研究了武夷山九曲溪水体的营养状态与原生动物群落结构关系。发现并非水越清洁PFU原生动物种类就越多,这要决定于水体的营养状况。当水体是贫营养时,水越清洁,PFU原生动物种类反而会越少。研究结果表明,即使在低营养水平的水体中,PFU法一样能准确反映人类活动对当地水环境的影响。     

周围神经损伤后外源性GKNF对神经元的保护作用

采用硅管套接大鼠切断的坐骨神经模型,局部给予胶质细胞源性神经营养因子（ＧＤＮＦ）,应用尼氏染色、酶组织化学染色方法,观察到外源性ＧＤＮＦ能减少脊髓修复侧前角运动神经元死亡的数目,降低脊髓前角运动神经元及脊神经节感觉神经元中胆碱酯酶（ＣＨＥ）及酸性磷酸酶（ＡＣＰ）变化的幅度。这表明外源性ＧＤＮＦ能保护周围神经切断后引起的神经元损伤．     

促细胞黏附生长的自组装肽水凝胶

随着细胞与组织工程的迅猛发展,能够促进细胞黏附、生长和分化的生物材料基质支架的研究日益重要。具有生物相容性且含水量超过99％的自组装肽水凝胶因其很好地符合理想的生物材料基质支架标准而备受重视。这类自我互补的两亲寡肽含50％的带电残基,并且以交替的离子亲水性和不带电的氨基酸残基周期性重复为特征;在其寡肽的氨基末端可用直接固相合成法修饰几个短序列生物活性模体进行功能化,用以促进不同细胞的黏附生长和靶向定位。现对自组装肽水凝胶的结构特征、自组装机制、对细胞黏附生长的影响以及未来自组装肽生物材料设计的目标进行综述．     

A sensitive signal-on assay for MTase activity based on methylation-responsive hairpin-capture DNA probe

This work develops a simple, sensitive and signal-on electrochemical sensor for methyltransferase (MTase) activity analysis. The sensor is composed of a methylene blue-modi?ed "signaling DNA probe" and a "capture DNA probe" tethered methylation-responsive hairpin DNA (hairpin-capture DNA probe). The thiol- modified hairpin-capture DNA probe at 5' end was firstly self-assembled on gold electrode via Au-S bonding. Methylation-induced scission of hairpin-capture DNA probe would displace the hairpin section and remain the "capture DNA probe" section on the gold electrode. Subsequently, the remained "capture DNA probe" on the gold electrode can hybridize with the methylene blue-modi?ed "signaling DNA probe", mediating methylene blue onto the gold electrode surface to generate redox current. It was eT on state. The developed facile signal-on electrochemical sensing system showed a linear response to concentration of Dam MTase range from 0.1 to 1.0 U/mL. The detection limit of Dam MTase activity was determined to be 0.07 U/mL and the total detection time is 7h. The sensor also has the ability to provide information about the dynamics of methylation process. Furthermore, we demonstrated that this sensor could be utilized to screen inhibitors or drugs for Dam MTase.     

Choline Transporters in Human Lung Adenocarcinoma： Expression and Functional Implications

Choline is an essential nutrient for cell survival and proliferation, however, the expression and function of choline transporters have not been well identified in cancer. In this study, we detected the mRNA and protein expression of organic cation transporter OCT3, carnitine/cation transporters OCTN 1 and OCTN2, and choline transporter-like protein CTL1 in human lung adenocarcinoma cell lines A549, H 1299 and SPC-A-1. Their expression pattern was further confirmed in 25 human primary adenocarcinoma tissues. The choline uptake in these cell lines was significantly blocked by CTL1 inhibitor, but only partially inhibited by OCT or OCTN inhibitors. The efficacy of these inhibitors on cell proliferation is closely correlated with their abilities to block choline transport. Under the native expression of these transporters, the total choline uptake was notably blocked by specific PI3K/AKT inhibitors. These results describe the expression of choline transporters and their relevant function in cell proliferation of human lung adenocarcinoma, thus providing a potential ＂choline-starvation＂ strategy of cancer interference through targeting choline transporters, especially CTL1.     

不同加热方式对嗜肺军团菌环介导等温扩增检测法的影响

目的:研究不同的加热方式对嗜肺军团菌环介导等温扩增检测法的影响方法:用已知的13株嗜肺军团菌样本,采用空气浴、水浴和PCR仪同时进行环介导等温扩增,观察沉淀反应、荧光反应以及产物电泳结果。结果:水浴和PCR仪加热LAMP反应的沉淀产物较多,荧光反应较强,电泳检测结果较为明显。空气浴的3种检测结果均较弱。结论:采用水浴和PCR仪进行环介导等温扩增反应的效果较好,从仪器设备的成本及实验条件考虑,采用水浴是环介导等温扩增反应首选的加热方式。