稳定表达猪APOBEC3F的PK15细胞单克隆株的建立

目的:利用慢病毒载体建立稳定表达猪载脂蛋白B mRNA编辑酶催化多肽样蛋白3F(pAPOBEC3F,简称pA3F)的PK15细胞系。方法:以实验室前期构建的pBPLV-flag-pA3F质粒为模板,PCR扩增pA3F基因,克隆到pLenti-Puro-3Flag载体构建成pLenti-Puro-pA3F-3Flag质粒,Western印迹鉴定其在HEK293T细胞中的表达;将pLenti-Puro-pA3F-3Flag质粒与包装载体共转染HEK293T细胞,包装成Lenti-pA3F慢病毒并测定病毒滴度;将Lenti-pA3F慢病毒感染PK15细胞,通过嘌呤霉素压力筛选并结合有限稀释法,筛选稳定表达pA3F的细胞单克隆株,最终Western印迹检测细胞单克隆株中pA3F的表达。结果:菌落PCR和序列测定表明pLenti-Puro-pA3F-3Flag质粒构建正确,Western印迹显示该质粒可在HEK293T细胞中表达pA3F蛋白;包装了Lenti-pA3F慢病毒,滴度为1.32×10~8TU/mL,慢病毒感染PK15细胞后可检测到pA3F蛋白的表达;经Western印迹鉴定,筛选得到的单克隆细胞可稳定表达pA3F蛋白。结论:建立了稳定表达pA3F的PK15细胞单克隆株,为进一步深入研究猪内源性反转录病毒在pA3F作用下的免疫逃逸机制奠定了基础。     

Association of genetic variants with response to iron supplements in pregnancy

The incidence of iron deficiency anemia in pregnancy is high in India where iron supplementation is a regular practice. The response to oral iron is influenced by several factors such as age, body mass index, gravida, socioeconomic status, food, vitamin deficiency and compliance to supplements. The major challenge is to understand the various modulators of iron status in this high-risk group so that we can improve the diagnosis and the management of these patients. The current study was designed to evaluate the iron status during pregnancy and to identify factors which might be influencing their response to oral iron. We investigated a total of 181 pregnant women with anemia (Hb < 11 g/dl) and evaluated the impact of probable factors on anemia and their iron status. Assessment of the response was based on hemoglobin and serum ferritin or transferrin saturation level after 8 and 20 weeks of iron supplementation. Socioeconomic, clinical, hematological, biochemical and genetic factors were all evaluated. Molecular analysis revealed that HFE variant allele (G) (rs1799945) was significantly associated with an adequate response to iron supplementation. We identified five subjects with a sustained poor response, and targeted re-sequencing of eleven iron-related genes was performed in them. We have identified seven novel variants in them, and in silico analysis suggested that these variants may have an iron regulatory effect. Taken together, our findings underscore the association of genetic variants with response to supplements in pregnancy, and they can be extended to other diseases where anemia and iron deficiency coexist.

Electronic supplementary material

The online version of this article (doi:10.1007/s12263-015-0474-2) contains supplementary material, which is available to authorized users.     

丹江口水库马口鱼肠道寄生蠕虫群落结构

从2004年8月到2005年11月在丹江口水库调查的847尾马口鱼(Opsariichthys bidens)肠道内共获得寄生蠕虫14种,其中线虫7种,复殖吸虫4种,棘头虫2种,绦虫1种。该寄生蠕虫群落以广谱性寄生虫为主。杜父鱼驼形线虫(Camallanus cotti)、鱊头槽绦虫(Bothriocephalus acheilognathi)、卡斯杆咽线虫(Rhabdochona cascadilla)和木村小棘吻虫(Micracanthorhynchina motomurai)为群落的核心种。研究结果表明,寄生蠕虫群落随着马口鱼的体长和食性的改变而发生显著的变化。群落核心种的感染强度与马口鱼体长呈显著相关关系。种间相关性分析表明群落结构呈非随机组合,种间感染强度存在显著的相关性。同时群落种间关系受到马口鱼的体长和食性以及季节变化的显著影响。马口鱼肠道寄生蠕虫群落在宿主较大个体或夏秋季节中更容易形成显著的种间相关性。     

滇西北鼠疫自然疫源地大绒鼠体表寄生虫研究(英文)

大绒鼠是中国滇西北鼠疫疫源地的主要储存宿主,它也是横断山脉地区小型哺乳动物的典型代表.用U 检验和相关分析的统计方法对2003～2004年云南洱海(中国滇西北著名的淡水湖泊)周边916只大绒鼠体表寄生虫进行了调查.调查点位于我国十一大鼠疫自然疫源地之一,此地也是我国恙虫病和流行性出血热的流行地区.在此,将大绒鼠的体表寄生虫群落和体表寄生虫医学和兽医学的重要性进行了描述.有756只大绒鼠寄生有体表寄生虫,侵染率为83%.采集到的体表寄生虫有86种,包括51种恙螨, 23种革螨, 7种蚤和5种吸虱.其中17种以前已经被证明是人类疾病的主要媒介.因此大绒鼠很可能成为鼠疫、恙虫病和流行性出血热等病原体的贮存宿主.     

中国云南洱海周边小兽体表吸虱多样性

在2003—2004年对云南大理洱海周边小兽体表寄生吸虱进行了调查,诱捕小兽3303只,隶属4目7科、15属、21种,共收集14635只吸虱,共计5科、6属、21种。调查点位于洱海周边3个不同方位,恰好处于无量山（洱海东部）、哀牢山（洱海南部）和苍山（洱海西部）3座山的连接地段,由于洱海的天然隔离使这3个方位形成了同地域（经度、纬度、海拔和动物地理区划相同）异生境的地理景观。此项研究的目的是调查同地域异生境里小兽体表吸虱在物种多样性、丰富度、群落结构、相似性和分布。结果表明吸虱的物种多样性低,群落结构简单。吸虱的丰富度、分布和物种多样性随着宿主和生境的不同而存在着显著的差异,这可能暗示着生态环境影响着吸虱和它相对应的小兽宿主的构成和分布。某种宿主经常有某种固定的吸虱,在动物分类上近缘关系较近的小兽宿主,其体表昆虫群落相似性较高。洱海周边不同方位同种小兽宿主上的吸虱物种相同,这个结果强烈暗示着吸虱与其对应宿主之间有明显的协同进化关系。     

Apoptotic effect of cleaved high molecular weight kininogen is regulated by extracellular matrix proteins

We previously reported that cleaved high molecular weight kininogen (HKa) and its domain 5 (D5) inhibit critical steps required for angiogenesis and in vivo neovascularization (Colman et al. 2000: Blood 95:543-550). We have further shown that D5 is able to induce apoptosis of endothelial cells, which may represent a critical part of the anti-angiogenic activity of HKa and D5 (Guo et al. 2001: Arterioscler Thromb Vasc Biol 21:1427-1433). In this study, we demonstrate that HKa- and D5-induced apoptosis is closely correlated with their anti-adhesive effect. An important new finding is that the apoptotic activity of HKa and D5 is highly regulated by their interactions with different extracellular matrix (ECM) proteins. HKa inhibited cell adhesion to vitronectin (Vn, 90%) and gelatin (Gel) (40%), but it had no apparent effect on cell adhesion to fibronectin (Fn). D5 showed a similar pattern on cell adhesion but was less potent than HKa. HKa induced apoptosis of endothelial cells grown on Vn and Gel but not cells grown on Fn which closely parallels with its anti-adhesive potency. Further results revealed that the anti-adhesive effect and the apoptotic effect of HKa are associated with its ability to inhibit phosphorylation of focal adhesion kinase (FAK) and paxillin, two important signal molecules required for cell adhesion and cell viability. We conclude that the anti-adhesive activity of HKa and D5 is responsible for their apoptotic effect and that Vn is likely an ECM component that mediates the effect of HKa and D5.     

CLOSTERIUM MONILIFERUM-EHRENBERGII (CHAROPHYCEAE, CHLOROPHYTA) SPECIES COMPLEX VIEWED FROM THE 1506 GROUP I INTRON AND ITS2 OF NUCLEAR rDNA

To assess phylogenetic relationships and speciation modes in Closterium , we sequenced two noncoding regions of the nuclear ribosomal cistron, the 1506 group I intron in small subunit and the internal transcribed spacer 2, for a total of 58 strains of the Closterium moniliferum-ehrenbergii species complex. These include both homothallic and heterothallic C. moniliferum Erenberg ex Ralfs v. moniliferum , heterothallic C. moniliferum v. submoniliferum (Woronichin) Krieger, and heterothallic C. ehrenbergii Meneghini ex Ralfs that can be divided into several mating groups. We found no or very little sequence divergence within single mating groups of C. ehrenbergii and among all heterothallic strains of C. moniliferum v. moniliferum or C. moniliferum v. submoniliferum. Nevertheless, sequence divergence was much greater between those mating groups of C. ehrenbergii and also among the three traditional taxa . Maximum parsimony and maximum likelihood analyses showed that the taxon C. ehrenbergii was not monophyletic. The two varieties of C. moniliferum appeared as a sister clade to certain mating groups of C. ehrenbergii . Among the clades that were recovered in different trees by maximum parsimony and maximum likelihood analyses, we consistently found two large conspicuous clades: clade I consisted of mating groups A, B, C, H, K, and L of C. ehrenbergii whose zygospores have smooth-walls, and clade II contained the mating groups D, E, I, J, and S whose zygospores are scrobiculate. Phylogenetic incongruences observed are discussed from the viewpoints of the different molecular nature of the group I intron and internal transcribed spacer 2, as well as putative rapid diversification of the mating groups and probable ancient ancestral hybridization.