海南红树林淡紫拟青霉胞外多糖提取条件的优化

为了探讨影响红树林淡紫拟青霉胞外多糖提取的因素,确定最佳提取方案,设置不同的发酵液浓缩倍数、三氯乙酸用量等因素,设计单因素实验测定多糖提取最佳条件。然后设计正交试验,检测多糖在不同条件下的提取率,以获得最佳提取工艺。结果发现,发酵液浓缩3~5倍时多糖提取率最高,10%的三氯乙酸对蛋白质脱除效果最好;正交试验表明影响多糖提取的因素依次为乙醇用量、沉淀时间和温度,最优方案为3倍95%乙醇、4 ℃沉淀24 h。该条件下,多糖提取率可达57.835%±1.206%。研究结果为红树林淡紫拟青霉胞外多糖的提取研究提供了参考。     

A Scalable and Accurate Targeted Gene Assembly Tool (SAT-Assembler) for Next-Generation Sequencing Data

Gene assembly, which recovers gene segments from short reads, is an important step in functional analysis of next-generation sequencing data. Lacking quality reference genomes, de novo assembly is commonly used for RNA-Seq data of non-model organisms and metagenomic data. However, heterogeneous sequence coverage caused by heterogeneous expression or species abundance, similarity between isoforms or homologous genes, and large data size all pose challenges to de novo assembly. As a result, existing assembly tools tend to output fragmented contigs or chimeric contigs, or have high memory footprint. In this work, we introduce a targeted gene assembly program SAT-Assembler, which aims to recover gene families of particular interest to biologists. It addresses the above challenges by conducting family-specific homology search, homology-guided overlap graph construction, and careful graph traversal. It can be applied to both RNA-Seq and metagenomic data. Our experimental results on an Arabidopsis RNA-Seq data set and two metagenomic data sets show that SAT-Assembler has smaller memory usage, comparable or better gene coverage, and lower chimera rate for assembling a set of genes from one or multiple pathways compared with other assembly tools. Moreover, the family-specific design and rapid homology search allow SAT-Assembler to be naturally compatible with parallel computing platforms. The source code of SAT-Assembler is available at https://sourceforge.net/projects/sat-assembler/. The data sets and experimental settings can be found in supplementary material.     

Rutaecarpine suppresses atherosclerosis in ApoE−/− mice through upregulating ABCA1 and SR-BI within RCT

ABCA1 and scavenger receptor class B type I (SR-BI)/CD36 and lysosomal integral membrane protein II analogous 1 (CLA-1) are the key transporter and receptor in reverse cholesterol transport (RCT). Increasing the expression level of ABCA1 and SR-BI/CLA-1 is antiatherogenic. The aim of the study was to find novel antiatherosclerotic agents upregulating expression of ABCA1 and SR-BI/CLA-1 from natural compounds. Using the ABCA1p-LUC and CLA-1p-LUC HepG2 cell lines, we found that rutaecarpine (RUT) triggered promoters of ABCA1 and CLA-1 genes. RUT increased ABCA1 and SR-BI/CLA-1 expression in vitro related to liver X receptor alpha and liver X receptor beta. RUT induced cholesterol efflux in RAW264.7 cells. ApoE-deficient (ApoE^−/−) mice treated with RUT for 8 weeks showed ∼68.43, 70.23, and 85.56% less en face lesions for RUT (L), RUT (M), and RUT (H) groups, respectively, compared with the model group. Mouse macrophage-specific antibody and filipin staining indicated that RUT attenuated macrophages and cholesterol accumulations in atherosclerotic lesions, respectively. Additionally, ABCA1 and SR-BI expression was highly induced by RUT in livers of ApoE^−/− mice. Meanwhile, RUT treatment significantly increased the fecal ³H-cholesterol excretion, which demonstrated that RUT could promote RCT in vivo. RUT was identified to be a candidate that protected ApoE^−/− mice from developing atherosclerosis through preferentially promoting activities of ABCA1 and SR-BI within RCT.     

Ca2+ entry via TRPC channels is necessary for thrombin-induced NF-kappaB activation in endothelial cells through AMP-activated protein kinase and protein kinase Cdelta

The transient receptor potential canonical (TRPC) family channels are proposed to be essential for store-operated Ca2+ entry in endothelial cells. Ca2+ signaling is involved in NF-kappaB activation, but the role of store-operated Ca2+ entry is unclear. Here we show that thrombin-induced Ca2+ entry and the resultant AMP-activated protein kinase (AMPK) activation targets the Ca2+-independent protein kinase Cdelta (PKCdelta) to mediate NF-kappaB activation in endothelial cells. We observed that thrombin-induced p65/RelA, AMPK, and PKCdelta activation were markedly reduced by knockdown of the TRPC isoform TRPC1 expressed in human endothelial cells and in endothelial cells obtained from Trpc4 knock-out mice. Inhibition of Ca2+/calmodulin-dependent protein kinase kinase beta downstream of the Ca2+ influx or knockdown of the downstream Ca2+/calmodulin-dependent protein kinase kinase beta target kinase, AMPK, also prevented NF-kappaB activation. Further, we observed that AMPK interacted with PKCdelta and phosphorylated Thr505 in the activation loop of PKCdelta in thrombin-stimulated endothelial cells. Expression of a PKCdelta-T505A mutant suppressed the thrombin-induced but not the TNF-alpha-induced NF-kappaB activation. These findings demonstrate a novel mechanism for TRPC channels to mediate NF-kappaB activation in endothelial cells that involves the convergence of the TRPC-regulated signaling at AMPK and PKCdelta and that may be a target of interference of the inappropriate activation of NF-kappaB associated with thrombosis.     

Effects of Predator and Prey Dispersal on Success or Failure of Biological Control

Biological control, defined as the reduction of pest populations by natural enemies, is often a component of integrated pest management strategies. Augmentation of natural enemy numbers by planned releases is a common biological control method, the successes and failures of which have been extensively reviewed. The effectiveness of biological control is influenced by how populations of predators and prey (or hosts and parasitoids) disperse in patchy environments. Here, we address the question of whether such dispersal leads to beneficial or detrimental pest control outcomes by developing a simple predator-prey model with constant releases of natural enemies in a two-patch environment. Theoretical and numerical results for all possible cases indicate that population dispersal has significant effects on the persistence of pests. For some ranges of dispersal rates or parameter space, dispersal is beneficial for pest control measures but this is not so for other ranges when it is detrimental. Therefore, knowledge of pest and natural enemy dispersal is crucial for understanding the effectiveness of biological control in a patchy environment. Finally, the model is generalised for multi-patch systems.     

NASBA荧光分子信标技术定量检测丙型肝炎病毒

建立NASBA荧光分子信标探针检测技术,并对国家HCV标准品、人工构建HCVRNA野生株及HCV抗体阳性不同人群进行检测。实验结果:该方法检测HCV的灵敏度为103拷贝ml血清,阴性参比品的符合率为100%;检测的线性范围为103拷贝～109拷贝ml血清;精密性(CV值)小于6%,在HCV抗体阳性人群中HCVRNA的检出率在45%～65%之间。结论:该方法在HCVRNA临床定量检测中具有良好的灵敏度、特异性、重复性与实用性。     

喀喇昆仑山-喜马拉雅山脉生态区类型与保护地空间分布格局

喀喇昆仑山-喜马拉雅山脉是泛第三极区域的重要组成部分。对中国与阿富汗、巴基斯坦、印度、尼泊尔、不丹和缅甸等6国交界的喀喇昆仑山-喜马拉雅山脉地区生态区和保护地分布开展研究。喀喇昆仑山-喜马拉雅山脉地区总面积902843.76 km²,跨古北界和东洋界两大全球重要的生物地理区域,分布有14个生态区,其中有55处保护地。保护地由国家公园和自然保护区组成,主要分布在东喜马拉雅亚高山针叶林、喜马拉雅亚热带阔叶林和东喜马拉雅阔叶林3个生态区内,面积为159063.30 km²,占喀喇昆仑山-喜马拉雅山脉地区总面积的17.62%。保护地中有41处国家公园,占保护地总数的74.5%;有25处为多国毗邻,占总数的45.45%。保护地的地理集中指数都大于完全平均分布值（37.796）,空间分布呈集聚状态;其中自然保护区分布的不均衡度高于国家公园,分布在中国,尼泊尔和印度三国境内。核密度分析显示中尼边境与中印边境区域的保护地分布集中度高。20世纪30年代开始,特别是80年代以来各国建立保护地,目前已经形成全球著名的保护地集群带。对于进一步推进喀喇昆仑山-喜马拉雅山脉地区国家公园跨境合作具有重要意义。