用指纹图谱评价环境样本DNA的提取效果

研究了DNA指纹图谱技术在评价环境样本DNA提取效果方面的应用。采用 3种DNA提取方法提取垃圾渗滤水和活性污泥中的微生物DNA ,并用ARDRA和RISA图谱加以评价。实验结果表明 ,RISA图谱分析法可以作为评估DNA提取效果的有效手段。     

Bullera hoabinhensis sp. nov., a new ballistoconidiogenous yeast isolated from a plant leaf collected in Vietnam

VY-68, a ballistoconidiogenous yeast strain, isolated from a plant leaf at Cuc Phuong National Park of Ninh Binh Province, Vietnam, was assigned to the genus Bullera based on morphological and chemotaxonomical characteristics. Based on the sequence analyses of 18S rDNA, D1/D2 region of 26S rDNA, and internal transcribed spacer regions (ITS), VY-68 was phylogenetically closely related to Bullera pseudoalba and Cryptococcus cellulolyticus. DNA-DNA reassociation experiments among VY-68, B. pseudoalba and C. cellulolyticus revealed that strain VY-68 is a distinct species, and the latter two are conspecific. Bullera hoabinhensis is proposed for VY-68.     

褶纹冠蚌精子的超微结构研究

利用电镜褶纹冠蚌精子的形态和结构作了研究,结果表明：精子全长约40-43μm,由头部、中段和鞭毛组成。头部呈子弹头形,长约2.6μm,直径约1.5μm,内含细胞核,核属浓缩型,外被核膜,5个球形的线粒体构成了精子的中段,中段长约0.6μm,最大直径约1.8μm。近端中心粒位于核基部的凹陷处,并通过致密的无定形的基质与远端中心粒相连,远端中心粒与鞭毛领之间通过硬功夫个围中心粒器紧密相连,鞭毛长约37-40μm。精子顶体退化,仅由几个顶体囊泡组成。     

Optimization of trans-Resveratrol bioavailability for human therapy

We have developed an innovative soluble galenic form to overcome the low absorption of trans-Resveratrol (t-Res) as a dry powder. We present here data on pharmacokinetics, bioavailability, and toxicity of t-Res in human volunteers treated with this soluble form, plus additional data on biological effects in rodents. Fifteen healthy volunteers of both sexes received 40 mg of t-Res in two forms, the soluble formulation (caplets) and the original powder (capsules), in a crossover design. Blood samples were collected at 15 min, 30 min, and every hour for 5 h. Plasma concentrations of t-Res and its metabolites were analyzed by liquid chromatography and mass spectrometry. The single dose (40 mg) of the soluble t-Res was well absorbed and elicited biologically efficient blood levels (0.1–6 μM) for several hours, despite metabolization into glucuronide and sulfate conjugates coupled to renal elimination. In contrast, t-Res administered as a dry powder barely elicited efficient blood levels for a short duration. The new formulation led to 8.8-fold higher t-Res levels in plasma versus the powder. t-Res metabolism was not modified and neither intolerance nor toxicity were observed during the study and the following week. The soluble formulation elicited a robust anti-inflammatory effect in various tissues of mice fed a high-fat diet, while dry powder t-Res was almost inactive. Our data suggest that significant improvements in t-Res bioavailability and efficiency can be obtained by this soluble galenic form, also allowing lower doses. The use of t-Res in human therapy is thus greatly facilitated and the toxicity risk is reduced.     

果蝇中Ecp蛋白的表达、纯化与抗体制备

获得特异性抗Ecp多克隆抗体,为进一步研究ecp的功能奠定基础。利用特异引物,通过RT-PCR扩增出编码Ecp蛋白的全长cDNA,克隆至谷胱甘肽S转移酶融合蛋白表达载体pGEX-4T-1(His)6C中,转染大肠杆菌DH 5α,经诱导表达后,利用谷胱甘肽琼脂糖珠从细胞裂解物中特异吸附融合蛋白,经凝血酶裂解,释放出Ecp蛋白,再经Ni-NTA亲和层析,最终获得高纯度的Ecp蛋白。用纯化的Ecp蛋白免疫新西兰家兔,亲和层析纯化抗Ecp抗体。利用该抗体进行的Western Blot结果表明：Ecp蛋白在野生型黑腹果蝇胚胎、三龄幼虫神经系统、成虫、成虫头部组织中均有明显表达,提示ecp可能是一个重要的管家基因。     

Inhibitory effect on NO production of triterpenes from the fruiting bodies of Ganoderma lucidum

Four new lanostane triterpenes, butyl lucidenate P (1), butyl lucidenate D₂ (2), butyl lucidenate E₂ (3) and butyl lucidenate Q (4) along with 11 known compounds (5–15) were isolated from the fruiting bodies of Ganoderma lucidum. Their chemical structures were established mainly by 1D and 2D NMR techniques and mass spectrometry. Their anti-inflammatory activity was evaluated against LPS-induced NO production in macrophage RAW 264.7 cells. Compounds 1, 3, 4, 9, 10 and 15 showed inhibitory potency with IC₅₀ values of 7.4, 6.4, 4.3, 9.4, 9.2 and 4.5 μM, respectively. Compounds 1, 3 and 15 dose-dependently reduced the LPS-induced iNOS expressions. Preincubation of cell with 1, 3 and 15 significantly suppressed LPS-induced expression of COX-2 protein.     

糖类在植物组织培养中的效应

糖类是影响植物组织培养成功与否的关键之一。迄今为止,已用于植物组织培养的糖类有５０多种。在植物体细胞组织培养中,蔗糖一直作为标准碳源,然而其他糖类物质如：葡萄糖、麦芽糖和山梨醇对植物体细胞培养也产生了一定的影响。在植物花药培养中,蔗糖较好,但应注意麦芽糖对花药培养的促进作用。     

超临界CO2流体对纤维素酶催化反应的影响

超临界二氧化碳流体预处理对纤维素超分子结构及纤维素酶催化反应有重要影响。一定含水量的微晶纤维素用SC-CO2在10MPa,50℃处理30min,其结构发生了有利于进一步被酶解的变化。上述超临界条件单独作用于纤维素酶时,并未造成酶催化活力的降低;但与纤维素共同进行SC—CO2处理时,纤维素酶则失去催化活性,但这种处理却能提高纤维素进一步被酶解的效率。一定范围内处理时的酶用量与酶解效率的增加正相关。纤维素的含水量对SC-CO2处理后的酶解效率有显影响。     

High-level expression of a truncated wall-associated protein A from the dental cariogenic Streptococcus mutans

Streptococcus mutans plays a primary role in the formation of dental caries. Previously, in our laboratory, an S. mutans genomic library was prepared, and the wapA gene was cloned into the shuttle vector, pSA4/4B2. To generate overexpression of wapA and to facilitate efficient purification of the WapA protein for use as an immunogen, an expression vector with the strong tac promoter was used. In order to answer questions regarding the optimization of solubility and expression based on gene size or the hydrophobicity of the protein product, 12 truncated constructs of the wapA gene were prepared using PCR. The truncated products were subcloned into the pGEX-6P-1 glutathione S-transferase (GST) fusion vector and expressed in E. coli BL21. The fusion proteins were analyzed by SDS-PAGE and confirmed by analysis with anti-GST and anti-WapA antibodies. Our study suggests that abrogation of the wapA promoter is necessary for expression of this gene in this expression system. Deletion of the signal peptide and the hydrophobic C terminus of WapA increased expression compared with the full-length construct, and truncation at the protease cleavage site of the C-terminal region greatly increased the stability of the protein without a loss in reactivity with the anti-WapA antibody. Western immunoblot analysis with anti-WapA antiserum clearly showed that the majority of the epitopes of the GST-WapA fusions are located in the N-terminal region of WapA. The immunogenicity of the various WapA fusion products is being examined in mice and rats to further map the immunologically dominant regions of the protein. This method effectively increased the expression of WapA and should contribute to the further understanding of gene expression of E. coli, as well as aid in the characterization of this protein for future immunologic evaluation.