Antioxidant flavonoids from the rhizomes of Helminthostachys zeylanica

Eight flavonoids, ugonins E-L (1-8), were isolated from the rhizomes of Helminthostachys zeylanica, and their structures were established on the basis of spectral evidence. Compounds 3-8 were evaluated for their antioxidative activity in the 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assay. Compounds 6, 7 and 8 were more active than Trolox, with IC20 values of 5.29 +/- 0.32, 7.23 +/- 0.22, and 7.93 +/- 0.31, respectively.     

Stereoselective esterase from Pseudomonas putida IFO12996 reveals alpha/beta hydrolase folds for D-beta-acetylthioisobutyric acid synthesis

Esterase (EST) from Pseudomonas putida IFO12996 catalyzes the stereoselective hydrolysis of methyl dl-beta-acetylthioisobutyrate (dl-MATI) to produce d-beta-acetylthioisobutyric acid (DAT), serving as a key intermediate for the synthesis of angiotensin-converting enzyme inhibitors. The EST gene was cloned and expressed in Escherichia coli; the recombinant protein is a non-disulfide-linked homotrimer with a monomer molecular weight of 33,000 in both solution and crystalline states, indicating that these ESTs function as trimers. EST hydrolyzed dl-MATI to produce DAT with a degree of conversion of 49.5% and an enantiomeric excess value of 97.2% at an optimum pH of about 8 to 10 and an optimum temperature of about 57 to 67 degrees C. The crystal structure of EST has been determined by X-ray diffraction to a resolution of 1.6 A, confirming that EST is a member of the alpha/beta hydrolase fold superfamily of enzymes and includes a catalytic triad of Ser97, Asp227, and His256. The active site is located approximately in the middle of the molecule at the end of a pocket approximately 12 A deep. EST can hydrolyze the methyl ester group without affecting the acetylthiol ester moiety in dl-MATI. The examination of substrate specificity of EST toward other linear esters revealed that the enzyme showed specific activity toward methyl esters and that it recognized the configuration at C-2.     

濒危植物秦岭冷杉地理分布和生物生态学特性研究

秦岭冷杉(Abies chensiensis)为中国特有三级保护植物,仅岛屿化分布于陕西、甘肃、四川、河南的秦巴山地和湖北神农架中高山地区,分布海拔1300—2300m。为了对秦岭冷杉种群进行深入研究和有效保护,通过样地调查和相关资料分析,对其地理分布、群落特性、种群特性进行了系统研究。秦岭冷杉群落包含种子植物40科63属102种,可划分成11个地理区系成分,其中温带分布型属占84．21％,热带分布型属占15．79％,温带分布是群落的基本特性。秦岭山区的秦岭冷杉群落可分为5个群丛,基本特点为：生境阴湿;群落郁闭度大,结构复杂;种群个体年龄差异大;天然更新主要发生在保护较好的阴坡林窗地段。秦岭冷杉苗期生长缓慢,在30—70年龄阶段,高生长旺盛;在40—100年龄阶段,胸径生长较快。秦岭冷杉开始结实年龄在30—40年龄阶段,结实间隔期为3—5年。秦岭山区成熟个体平均产球果8个／株,饱满种子56粒／球果。种群更新的关键问题是种子产量低,林下幼苗稀疏,更新缺乏后继资源。种子向幼苗转化率低是生活史中的脆弱环节。在未来经营保护中,应以就地保护为主,通过抚育管理,促进天然更新;丰年应适时采种,就地育苗,扩大人工种群。     

两份太空诱变玉米雄性不育突变体的遗传研究

从搭载神舟4号飞船的4份玉米自交系后代中选育出两份雄性不育突变体, 对其进行育性鉴定, 并分析不育性状的稳定性及遗传特点。以不育材料的不育株为母本, 同群体的可育株和其他自交系为父本进行杂交, 结合自交、回交分析其后代的育性表现; 同时, 以具有正常细胞质的自交系为母本, 育性完全恢复的测交F1植株为父本进行反交, 对其反交的F1及F2进行育性观察分析。结果表明：这两份不育材料不育株的花药内无花粉或含少量畸形花粉, 败育彻底, 花粉败育表现为典败型。不育性状在不同年份、不同季节、不同地点下稳定遗传, 属可遗传的单基因控制的隐性核不育类型。     

免疫荧光法检测原核和真核细胞中表达的轮状病毒外壳蛋白VP4

目的：用免疫荧光法快速检测原核和真核细胞中表达的轮状病毒（RV）外壳蛋白VP4。方法：以抗VP4的抗体为一抗、FITC标记的羊抗豚鼠IgG为二抗,用免疫荧光方法检测在大肠杆菌BL21（DE3）中重组表达的同源RVVP4;检测SA11或Wa株RV感染MA104细胞后不同时间段病毒VP4的合成及其在感染细胞中的分布情况。结果：用免疫荧光法可直接检测到原核细胞中表达的外源蛋白,也可检测到病毒蛋白在真核细胞中的分布情况。结论：免疫荧光法可特异、方便、快速地检测RV VP4在原核和真核细胞中的表达;来源于RV TB—Chen株的VP4抗体可特异性识别同源病毒VP4,交叉识别SA11或Wa株的VP4。     

A组人轮状病毒VP6基因克隆及在大肠杆菌中的高效表达

轮状病毒(rotavirus RV) 结构蛋白VP6位于病毒三层衣壳结构的中间层,在病毒粒子的形成过程中起重要的作用。从临床样品中分离的人轮状病毒TB-Chen株VP6基因通过RT-PCR得到扩增产物。以pET作为表达载体,将VP6蛋白编码基因序列插入到质粒pET中成功构建原核表达质粒pET-VP6。实验表明,带有pET-VP6质粒的大肠杆菌BL21(DE3)可以高效表达目的蛋白VP6,重组表达产物VP6占菌体总蛋白的27.4%, 其分子量约为45 kDa,并且能被豚鼠抗SA11血清抗体识别（Western blot）。这一结果为进一步研究VP6的结构和功能奠定了重要的物质基础。     

蚕豆保卫细胞中钙调素的免疫电镜定位

以蚕豆横切和平切气孔为材料,对钙调素进行了免疫胶体金电镜定位的结果表明:在蚕豆保卫细胞的细胞核、细胞质、细胞膜、叶绿体、液泡、高尔基体、细胞壁中都有金颗粒分布,在线粒体上的分布较少.     

植物脱落酸合成缺陷与反应敏感型突变体

介绍了高等植物体脱落酸生物合成缺陷型突变体,生物合成途径,以及对脱落酸反应超敏感和不敏感的反应型突变体的研究进展。     

Docosahexaenoic acid inhibition of inflammation is partially via cross-talk between Nrf2/heme oxygenase 1 and IKK/NF-κB pathways

We examined the underlying mechanisms involved in n-3 docosahexaenoic acid (DHA) inhibition of inflammation in EA.hy926 cells. The present results demonstrated that pretreatment with DHA (50 and 100 μM) inhibited tumor necrosis factor-alpha (TNF-α)-induced intercellular adhesion molecule 1 (ICAM-1) protein, mRNA expression and promoter activity. In addition, TNF-α-stimulated inhibitory kappa B (IκB) kinase (IKK) phosphorylation, IκB phosphorylation and degradation, p65 nuclear translocation, and nuclear factor-κB (NF-κB) and DNA binding activity were attenuated by pretreatment with DHA. DHA triggered early-stage and transient reactive oxygen species (ROS) generation and significantly increased the protein expression of heme oxygenase 1 (HO-1), induced nuclear factor erythroid 2-related factor 2 (Nrf2) translocation to the nucleus and up-regulated antioxidant response element (ARE)-luciferase reporter activity. Moreover, DHA inhibited Nrf2 ubiquitination and proteasome activity. DHA activated Akt, p38 and ERK1/2 phosphorylation, and specific inhibitors of respective pathways attenuated DHA-induced Nrf2 nuclear translocation and HO-1 expression. Transfection with HO-1 siRNA knocked down HO-1 expression and partially reversed the DHA-mediated inhibition of TNF-α-induced p65 nuclear translocation and ICAM-1 expression. Importantly, we show for the first time that HO-1 plays a down-regulatory role in NF-κB nuclear translocation, and inhibition of Nrf2 ubiquitination and proteasome activity are involved in increased cellular Nrf2 level by DHA. In this study, we show that HO-1 plays a down-regulatory role in NF-κB nuclear translocation and that the protective effect of DHA against inflammation is partially via up-regulation of Nrf2-mediated HO-1 expression and inhibition of IKK/NF-κB signaling pathway.     

First record of Eretmophyllum (Ginkgoales) with well-preserved cuticle from the Middle Jurassic of the Ordos Basin,Inner Mongolia,China

The genus Eretmophyllum, a characteristic and widespread taxon of the order Ginkgoales, is restricted in the Mesozoic floras of Eurasia. In China, although some specimens were assigned to Eretmophyllum based on gross leaf morphology, none have epidermal structure, which is essential for species delimitation within this genus. In this paper, we describe a new species, Eretmophyllum neimengguensis n. sp., from the Middle Jurassic Yan’an Formation of the Ordos Basin, China. This is the first record of the genus Eretmophyllum (Ginkgoales) from the Ordos Basin as well as from China based upon the gross leaf morphology and the epidermal structure. The new record significantly extends the stratigraphic and geographic distribution of Eretmophyllum in China and Eurasia. It also improves our understanding of its leaf morphology, epidermal structure, and past diversity. The taxonomic position and the stratigraphic and geographical range of the genus Eretmophyllum are discussed. Briefly, leaf morphological and epidermal characters of the new species, associated plant assemblages, and sedimentology indicate that the regional climate of the Ordos Basin was warm and humid with seasonal temperature and precipitation fluctuations in warm-temperate zone during the Middle Jurassic. Furthermore, given the relationship between the distribution/diversity of the genus and the climate, the genus Eretmophyllum may be confined to the warm-temperature climate, is intolerant of heat and frost, and can be considered as an indicator of humid and warm climatic conditions.