Interaction with magnesium and ADP stabilizes both components of nitrogenase from Klebsiella pneumoniae against urea denaturation

The nitrogenase enzyme of Klebsiella pneumoniae consists of two separable proteins, each with multiple subunits and one or more oxygen sensitive metallocenters. The wild-type nitrogenase proteins are stable to electrophoresis in high concentrations of urea under anaerobic conditions. Addition of Mg+2 and ADP greatly increases the stability of the smaller Fe protein (from <4 to >6 M for full unfolding), an effect directly analogous to stabilization in p21ras induced by Mg+2 and GDP. Stabilization by Mg+2 is slight for the holo MoFe protein (from approximately 1.5 to approximately 2.4 M) but more dramatic for the apo protein form of the MoFe protein accumulated by certain Fe protein (nifH gene) mutants. The potent product inhibitor of nitrogenase function, MgADP, increases stability of the MoFe protein more than Mg+2 alone, to approximately 3.6 M, showing that nucleotides interact with the MoFe protein. Mutations of the nifM gene result in slower accumulation of less stable Fe protein, indicating that NifM is involved in correct folding of the Fe protein. Mutationally altered proteins are often difficult to purify for study because of their inherent instability, low expression level, or oxygen lability. Crude extracts of 11 different mutants of Fe protein (nifH gene) were examined by transverse urea gradient gels to rapidly screen for stabilizing interactions in the presence or absence of substrate or inhibitor analogs. Amino acid alterations D44N and R188C, at the interface of the dimer, in the vicinity of the nucleotide binding site(s), have significantly lower stability than the wild-type enzyme in the absence of Mg+2 but comparable stability in its presence, showing the importance of Mg+2 in the subunit interactions. Mutations N163S and E266K, in which residues normally involved in hydrogen bonding far from the active site were altered, are more labile than the wild-type even with Mg+2 added. Seven other mutants, though nonfunctional, did not appear altered in stability compared to the wild-type.     

MicroRNA-30a sensitizes tumor cells to cis-platinum via suppressing beclin 1-mediated autophagy

Autophagy is activated in cancer cells during chemotherapy and often contributes to tumor chemotherapy resistance. In this study, we characterized the role of microRNA-30a (miR-30a) in the coordination of cancer cell apoptosis and autophagy, which determines the sensitivity of cancer cells to chemotherapy. First, the autophagy activity in cancer cells increased after cis-dichloro-diamine platinum (cis-DDP) or Taxol treatment, as indicated by the enhanced expression of beclin 1, a key regulator of autophagy, and increased number of LC3-positive autophagosomes. Second, miRNA screening using a TaqMan probe-based quantitative RT-PCR assay identified that miR-30a, a miRNA that targets beclin 1, was significantly reduced in tumor cells by cis-DDP treatment. Forced expression of miR-30a significantly reduced beclin 1 and the autophagy activity of tumor cells induced by cis-DDP. Third, the blockade of tumor cell autophagy activity by miR-30a expression or 3-methyladenine significantly increased tumor cell apoptosis induced by cis-DDP treatment. Finally, an in vivo tumor implantation mouse model clearly showed that elevation of miR-30a in implanted tumor cells by administration of the recombinant lentivirus expressing miR-30a strongly enhanced cis-DDP-induced apoptosis of tumor cells. In conclusion, our results demonstrate for the first time that miR-30a can sensitize tumor cells to cis-DDP via reducing beclin 1-mediated autophagy and that increasing miR-30a level in tumor cells represents a novel approach to enhance the efficacy of chemotherapy during cancer treatment.     

Research progress on the effects of soil erosion on vegetation

The relationship between vegetation and soil erosion deserves attention due to its scientific importance and practical applications. A great deal of information is available about the mechanisms and benefits of vegetation in the control of soil erosion, but the effects of soil erosion on vegetation development and succession is poorly documented. Research shows that soil erosion is the most important driving force for the degradation of upland and mountain ecosystems. Soil erosion interferes with the process of plant community development and vegetation succession, commencing with seed formation and impacting throughout the whole growth phase and affecting seed availability, dispersal, germination and establishment, plant community structure and spatial distribution. There have been almost no studies on the effects of soil erosion on seed development and availability, of surface flows on seed movement and redistribution, and their influences on soil seed bank and on vegetation establishment and distribution. However, these effects may be the main cause of low vegetation cover in regions of high soil erosion activity and these issues need to be investigated. Moreover, soil erosion is not only a negative influence on vegetation succession and restoration, but also a driving force of plant adaptation and evolution. Consequently, we need to study the effects of soil erosion on ecological processes and on development and regulation of vegetation succession from the points of view of pedology and vegetation, plant and seed ecology, and to establish an integrated theory and technology for deriving practical solutions to soil erosion problems.     

大豆异黄酮对大鼠乳腺癌细胞内cAMP/PKA信号途径的影响

本实验研究了大豆异黄酮对SHZ-88大鼠乳腺癌细胞内cAMP／PKA信号途径的影响。实验设3组：空白对照组、50μg／ml大豆黄酮及15μg／ml染料木素组。采用放射免疫测定法（RIA）检测了胞内cAMP的浓度、腺苷酸环化酶（adenylate cyclase,AC）和磷酸二酯酶（phosphodiesterase,PDE）的活性,用（γ-^32P）ATP掺入法测定cAMP依赖性PKA的活性,半定量RT-PCR法分析cAMP反应元件结合蛋白（cAMP response element binding protein,CREB）mRNA表达的变化。结果表明：在处理后5min,大豆黄酮组和染料木素组细胞的cAMP浓度分别比对照组升高了9．5％和11．0％（P〈0．05）：10min时,分别比对照组升高31．0％和40.3％（P〈0．01）。3组细胞的AC活性在处理时间内没有明显变化。但在处理后5min,大豆黄酮组和染料木素组细胞的PDE活性分别降至对照组的71．8％和71．6％（P〈0．05）。处理后20min,大豆黄酮组和染料木素组细胞PKA活性分别上升到对照组的125．8％和122．3％（P〈0．05）;到40min时仍维持在高水平。大豆黄酮组和染料木素组细胞CREB mRNA的表达量在处理后3h分别比对照组增加31．6％和51．1％（P〈0．05）;6h后开始下降。这些结果提示,大豆异黄酮能够激活大鼠乳腺癌细胞内cAMP／PKA信号途径;而且是通过抑制磷酸二酯酶的活性,导致胞内cAMP浓度升高而实现的。     

Ecophysiological response and morphological adjustment of two Central Asian desert shrubs towards variation in summer precipitation

As part of global climate change, variation in precipitation in arid ecosystems is leading to plant adaptation in water-use strategies; significant interspecific differences in response will change the plant composition of desert communities. This integrated study on the ecophysiological and individual morphological scale investigated the response, acclimation and adaptation of two desert shrubs, with different water-use strategies, to variations in water conditions. The experiments were carried out on two native dominant desert shrubs, Tamarix ramosissima and Haloxylon ammodendron, under three precipitation treatments (natural, double and no precipitation, respectively), in their original habitats on the southern periphery of Gurbantonggut Desert, Central Asia, during the growing season in 2005. Changes in photosynthesis, transpiration, leaf water potential, water-use efficiency, above-ground biomass accumulation and root distribution of the two species were examined and compared under the contrasting precipitation treatments. There were significant interspecific differences in water-use strategy and maintenance of photosynthesis under variation in precipitation. For the phreatophyte T. ramosissima, physiological activity and biomass accumulation rely on the stable groundwater, which shields it from fluctuation in the water status of the upper soil layers caused by precipitation. For the non-phreatophyte H. ammodendron, efficient morphological adjustment, combined with strong stomatal control, contributes to its acclimation to variation in precipitation. On account of its positive responses to increased precipitation, H. ammodendron is predicted to succeed in interspecific competition in a future, moister habitat.     

云南省香格里拉大峡谷藏族神山在自然保护中的意义

位于云南省西北部迪庆藏族自治州境内的香格里拉大峡谷拥有独特的自然人文景观和丰富的生物多样性。藏族作为当地最重要的原住民族,他们的文化对当地的自然环境具有重要的影响。为了解藏族传统文化中的神山崇拜对香格里拉大峡谷地区自然环境的影响,本文应用民族生态学、植物生态学和文化人类学的研究方法,结合文献资料的分析,对香格里拉大峡谷的藏族神山进行了调查和研究,并对处于不同保护状态下（神山与非神山）的两个典型植物群落即高山松(Pinus densata)群落和云南黄果冷杉(Abies ernestii var. salouenensis)群落进行了比较。结果显示,在相同的样地面积内（20 m×20 m）,神山植物群落在物种数量（20,14）和群落盖度（100%,85%）方面都明显高于非神山植物群落（11,2;70%,60%）。结合入户调查资料,发现广泛分布于香格里拉大峡谷的藏族神山构成了一个“自下而上”的乡土保护体系,它不仅在保护生物多样性、维护当地脆弱的生态环境等方面发挥着重要的作用,而且还可以提供多种非林产品,实现多种生态功能。文章探讨了发挥乡土保护体系的保护功效及应用传统文化知识以促进自然保护的重要性,建议应当尊重和保护原住民的传统文化,发掘其中优秀的自然保护知识和经验,引导其发挥更加有效的自然保护功能,使民族传统保护体系成为现代自然保护体系的有力补充。     

红花油体提取条件优化及稳定性研究

油体是储藏脂肪的亚细胞单位,其表面包裹一层磷脂和油体蛋白。这种稳定的结构可以保护油体面对环境的压力,使油体可以应用在食品、化妆品及制药工业中。研究红花油体的提取方法及红花油体乳液的基本性质,旨在为以油体为基质的载体体系研究奠定基础。以pBS为介质,采用梯度离心法,比较了不同提取条件对红花油体的提取效率的影响;对其在不同pH值、NaCl浓度条件下红花油体的平均粒径和稳定性进行测定。结果表明,红花油体在pH值≥6条件下,平均粒径为1.75-2.05μm和p H值≤6条件下,平均粒径1.50-1.75μm;NaCl浓度0.2和0.4 mg/m L时,红花油体分散较为均匀,NaCl浓度1.2和2.0 mg/m L时,红花油体出现聚集现象。蔗糖浓度0.1和0.2 mg/m L时,红花油体分散较为均匀,蔗糖浓度为0.4-1.0 mg/m L时,红花油体比较密集,随着蔗糖浓度的增加,红花油体的粒径逐渐开始不均一。红花油体的最佳提取条件是pH7,NaCl浓度0.2 mg/m L,蔗糖浓度0.1 mg/m L,稀释后的红花油体溶液在不加入保护剂或者不经过物理方法处理下,保存起来不稳定。     

胶原／壳聚糖复合膜的制备及止血效果的研究

目的 以胶原和壳聚糖制备复合膜,检验其止血效果,并探讨其止血原因。材料与方法：以酸解法从牛腱中提取胶原,用甲壳素制得壳聚糖,以胶原和壳聚制成复合膜,通过动物实验测不同配比的复合膜对出血创面的止血时间,并与其它止血材料做对比。结果：各种配比的复合膜的止血效果均比明胶等一般止血材料好。结论：胶原/壳聚糖复合膜有良好的止血作用,可望在外科手术上得到广泛应用。     

热玫瑰小双孢菌来源的丙酮酸磷酸双激酶的表达及应用

以热玫瑰小双孢菌基因组DNA为模板, 通过PCR扩增得到了编码PPDK的基因, 将此基因片段插入到表达载体pET28a(+)中构建得到了重组表达质粒pET28a(+)-PPDK, 将重组表达质粒pET28a(+)-PPDK转化到大肠杆菌BL21(DE3)中, 经过IPTG诱导, 重组菌成功表达了N端带有6-His Tag的重组PPDK。经SDS-PAGE分析, 重组PPDK单体分子量为101 kD。经过镍亲和层析和超滤后, 重组PPDK蛋白基本达到电泳纯, 并被成功应用于焦测序中。