山丹黄参的分布及微量元素含量研究

山丹黄参是伞形科迷果芹属的迷果芹（Sphallerocarpus gracilis)经测定,该植物含有大量人体必需的各种微量元素,其中P,Mg、Ｋ、Ｃu、Ｃr、Ｃo含量比党参（Codonopsis pilosula)和当归（Ａngelica sinensis)还高,Ｚn,Mn的水平也不低,是一种营养丰富的绿色食品,具有广泛的应用与开发前景。     

幽门螺杆菌HspA融合蛋白口服疫苗的构建

构建表达幽门螺杆菌的保护性抗原分热休克蛋白A亚单位(HspA）和霍乱毒素B亚单位（CtxB）的重组融合蛋白的生物工程菌株,以此制备幽门螺杆菌的口服疫苗。用PCR方法扩增hspA和ctxB两个目的的基因片段,将它们分别克隆至pSK（＋）质粒上,然后插入含T7启动子ET－22b（＋）的表达载体中,构建嗓基因的表达质量pET－hct,转化E.coliBL21(DE3）,经IPTG诱导表达融合蛋白HCT。经测序,hspA-ctxB(hct)融合基因片段由726bp组成,可以编码242个氨基酸残基的多肽。经SDS－PAGE和免疫印迹分析检测发现,融合基因表达的蛋白质相对分子质量约为30kD。融合蛋白经镍离子柱纯化、复性后,和HspA共同标记同位素^125I,然后给小鼠灌胃,结果观察到HCT组小鼠血清中的^125I的放射量要明显高于HspA组（P＜0．001）,且吸收峰值时间明显提前。融合蛋白中的CtxB可明显促进小鼠对HspA的吸收,HCT融合蛋白可以作为预防和治疗幽门螺杆菌感染的侯选口服疫苗。     

中国普通野生稻遗传分化的RAPD研究

多数学者已认定亚洲栽培稻（ＯｒｙｚａｓａｔｉｖａＬ．）的祖先是普通野生稻（Ｏ．ｒｕｆｉｐｏｇｏｎ）。然而栽培稻的籼、粳分化是发生在驯化之前还是在驯化之后,也即普通野生稻是否存在籼、粳分化的问题,是十几年来稻作起源研究中争论的热点之一。Ｓｅｃｏｎｄ［１］用多个同工酶位点的分析结果得出结论,普通野生稻在驯化为栽培稻之前就已经发生了籼、粳分化,即有籼型普通野生稻和粳型普通野生稻之分。Ｍｏｒｉｓｈｉｍａ和Ｇａｄｒｉｎａｂ［２］用２４个形态和生理性状及１２个同工酶位点和杂交亲合力等方法证明普通野生稻没有发…     

In vitro study of DNA interaction with melamine and its related compounds

In vitro studies on the interactions between native herring sperm DNA (HS-DNA) and melamine as well as its related compounds (MARCs), that is, ammeline, ammelide, and cyanuric acid, have been investigated by spectrophotometric, spectrofluorometric, melting temperature, and viscosimetric techniques. It was found that any of the MARCs might interact with HS-DNA by a groove mode of binding via hydrogen bonds. The interaction constants between any of the MARCs and HS-DNA were at 10?-10? L mol?1, determined by both spectrophotometric and spectrofluorometric methods. The thermodynamic studies suggested that the interaction processes were exothermic favored (ΔH?相似文献   

Human platelet lysate supports ex vivo expansion and enhances osteogenic differentiation of human bone marrow-derived mesenchymal stem cells

MSCs (mesenchymal stem cells) with their versatile growth and differentiation potential are ideal candidates for use in regenerative medicine and are currently making their way into clinical trials, which requires the development of xeno-free protocols for their culture. In this study, MSCs were cultured in 10% FCS or 7.5% HPL (human platelet lysate)-supplemented media. We found that both groups of MSCs showed a comparable morphology, phenotype and proliferation. The percentage of cells in the S- and G2-/M-phases, however, was slightly up-regulated (P<0.01) in HPL group. HPL contains PDGF (platelet derived growth factor)-AB and IGF (insulin-like growth factor)-1. In addition, compared with FCS group, MSCs in HPL group showed an increase in osteogenic differentiation and a decrease in adipogenic differentiation. In conclusion, MSCs in HPL-supplemented media maintained similar growing potential and phenotype, while osteogenic potential was enhanced. HPL offers a promising alternative to FCS for MSC expansion for clinical application, especially in bone injury diseases.     

WNK4 kinase inhibits Maxi K channel activity by a kinase-dependent mechanism

WNK [with no lysine (k)] kinase is a serine/threonine kinase subfamily. Mutations in two of the WNK kinases result in pseudohypoaldosteronism type II (PHA II) characterized by hypertension, hyperkalemia, and metabolic acidosis. Recent studies showed that both WNK1 and WNK4 inhibit ROMK activity. However, little is known about the effect of WNK kinases on Maxi K, a large-conductance Ca(2+) and voltage-activated potassium (K) channel. Here, we report that WNK4 wild-type (WT) significantly inhibits Maxi K channel activity in HEK αBK stable cell lines compared with the control group. However, a WNK4 dead-kinase mutant, D321A, has no inhibitory effect on Maxi K activity. We further found that WNK4 inhibits total and cell surface protein expression of Maxi K equally compared with control groups. A dominant-negative dynamin mutant, K44A, did not alter the WNK4-mediated inhibitory effect on Maxi K surface expression. Treatment with bafilomycin A1 (a proton pump inhibitor) and leupeptin (a lysosomal inhibitor) reversed WNK4 WT-mediated inhibition of Maxi K total protein expression. These findings suggest that WNK4 WT inhibits Maxi K activity by reducing Maxi K protein at the membrane, but that the inhibition is not due to an increase in clathrin-mediated endocytosis of Maxi K, but likely due to enhancing its lysosomal degradation. Also, WNK4's inhibitory effect on Maxi K activity is dependent on its kinase activity.     

InlB-mediated Listeria monocytogenes internalization requires a balanced phospholipase D activity maintained through phospho-cofilin

Internalization of Listeria monocytogenes into non-phagocytic cells is tightly controlled by host cell actin dynamics and cell membrane alterations. However, knowledge about the impact of phosphatidylcholine cleavage driven by host cell phospholipase D (PLD) on Listeria internalization into epithelial cells is limited. Here, we report that L. monocytogenes activates PLD in Vero cells during the internalization. With immunostaining it was shown that both PLD1 and PLD2 surrounded partially or completely the phagocytic cup of most L. monocytogenes. Either up- or down-regulation of PLD expression (activity) diminished Listeria internalization. Both PLD1 and PLD2 in Vero cells were required for efficient Listeria internalization, and could substitute for each other in the regulation of Listeria internalization. Further, exogenous InlB activated host cell PLD1 and PLD2 via the Met receptor, and restored host PLD activation by InlB-deficient L. monocytogenes. InlB-induced PLD activation and Listeria internalization were tightly controlled by phospho-cycling of cofilin. PLD1, but not PLD2, was involved in cofilin-mediated PLD activation and Listeria internalization. These data indicate that cofilin-dependent PLD activation induced by InlB may represent a novel regulation mechanism for efficient Listeria internalization into epithelial cells.     

水稻条纹叶枯病抗性位点的检测和效应分析

利用111个家系组成的热研2号（Oryza sativa subsp．japonica‘Reyan2’）／Milyang23（Oryza sativa subsp．indica‘Milyang23’）重组自交系（recombinant inbred lines,RIL）群体（F7）,采用重病区田间自然接种方法,以病情指数作为条纹叶枯病的表型值,鉴定了2个亲本及111个RIL家系对条纹叶枯病的抗性。使用QTL Cartographer软件复合区间作图法,对水稻（Oryza sativa）条纹叶枯病抗性基因进行了QTL分析。结果检测到2个抗水稻条纹叶枯病的QTL,分别位于第2和第11染色体上,其中第11染色体上的QTL贡献率为19．58％,表明这是一个主效的QTL,该QTL及其附近的分子标记,可以用于水稻条纹叶枯病抗性分子标记辅助育种。     

人SUMO-3基因原核表达及多克隆抗体制备

构建人SUMO-3基因的原核表达载体pET41a(+)-SUMO-3,表达重组GST-SUMO-3融合蛋白,制备人SUMO-3多克隆抗体。试验结果显示,通过PCR方法从重组质粒pEYFP-SUMO-3中克隆到的SUMO-3 N端93个氨基酸的基因序列与NCBI上提供的序列一致,重组质粒pET41a(+)-SUMO-3构建成功;重组pET41a(+)-SUMO-3在E.coli.BL21 (DE3) pLysS中表达GST-SUMO-3融合蛋白,分子量为44.0 kDa,与预期分子量一致;采用亲和层析纯化融合蛋白GST-SUMO-3并免疫家兔,获得人SUMO-3抗体;Western blot 检测显示该抗体可以特异性识别SUMO-3,ELISA检测结果成阳性,抗体效价约为1: 20000。实验结果为进一步研究人SUMO-3及SUMO第二类家族的功能提供了有用工具。