High expression level of TAL1 gene in newly diagnosed acute myeloid leukemia

AIM: To investigate the characteristic of T-cell acute lymphocytic leukemia 1 (TAL1) gene expression in acute myeloid leukemia (AML) cell lines and in primary AML cells from de novo AML patients with different subtypes. METHODS: Real-time PCR was used to determine the expression of TAL1 mRNA in acute leukemia cell lines (Jurkat, CCRF-CEM, HL-60 and NB4 cell lines) and peripheral blood mononuclear cells from 47 newly diagnosed AML patients. Twelve healthy individuals were served as healthy control group. RESULTS: A significantly increased level in TAL1 mRNA was found in AML cell lines (HL-60 and NB4), T-cell acute lymphacytic leukemia (T-ALL) cell lines (Jurkat, CCRF-CEM) and primary AML cells compared with the healthy controls. Over-expression of TAL1 was found in all detected AML subtypes, the highest level of TAL-1 mRNA was found in AML-M1 and AML-M5 subtype (P<0.05). CONCLUSION: High expression of TAL1 in AML might influence the differentiation and proliferation of myeloid cells, further investigation needs to confirm whether it would be as a biomarker for pathogenesis of AML.     

降钙素基因相关肽对猪小肠上皮细胞钠离子依赖型Ⅱb磷转运蛋白表达及磷吸收的影响

本试验旨在研究不同浓度降钙素基因相关肽(CGRP)对猪小肠上皮细胞钠离子依赖型Ⅱb磷转运蛋白(NaPi-Ⅱb)表达和磷吸收的影响.选用第3～8代猪小肠上皮细胞进行试验,按单因素试验设计,将试验细胞分为1个对照组和5个CGRP(1×10-11～1×10-7 mmol/L)试验组,每组设6个重复.用4-甲基偶氮唑盐(MTT)法测定细胞生长速度,采用比色法测定CGRP干预24 h后细胞上清液磷含量,用RT-PCR法检测NaPi-Ⅱb mRNA相对表达量,以及蛋白质印迹(Western blot)法检测NaPi-Ⅱb蛋白表达.结果表明:与对照组相比,不同浓度CGRP对细胞生长没有显著影响(P＞0.05);各试验组均极显著或显著地促进了磷的吸收和NaPi-ⅡbmRNA的相对表达量(P＜0.01或P＜0.05);除1×10-10 mmol/L组外均显著提高了NaPi-Ⅱb蛋白的表达(P＜0.05).研究结果表明,CGRP通过诱导小肠上皮细胞中NaPi-Ⅱb的表达促进细胞磷的吸收,其中以1×10-9、1×10-8 mmol/L浓度的CGRP促进效果较为显著.     

桑树中的1-脱氧野尻霉素含量变化规律研究

综合桑品种、桑树生长季节、桑树不同药用部位以及桑叶的不同发育阶段等因素探讨桑树(Morus L.)体内1-脱氧野尻霉素(1-deoxynojimycin,1-DNJ)的积累规律,期望发掘富含1-DNJ的生物资源。利用反相高效液相色谱法测定了18个代表性桑品种的摘芯芽和2个桑品种全年不同季节、不同发育时期的桑叶以及不同季节桑枝皮中1-DNJ的含量,并测定了与桑树亲缘关系相近的构树(Broussonetia papyrifera L.)等7种植物中的1-DNJ含量。在18个供试桑品种的摘芯芽中,1-DNJ含量以秋雨和沙2×伦教109最高,质量分数均达0.328 0%左右,黄鲁的1-DNJ含量最低。农桑14号和陕桑305两个桑品种冬、春季不同发育时期桑叶中的1-DNJ含量以冬芽最高,质量分数分别达到0.345 3%、0.255 7%,在开叶期和叶片成熟期较高,在缓慢生长期较低;不同季节桑叶中1-DNJ的含量均是嫩芽>嫩叶>成熟叶>老叶,其中嫩芽和嫩叶中1-DNJ含量以晚秋最高,成熟叶中以夏季最高,而老叶中的1-DNJ含量以春季最高;桑枝皮中1-DNJ的含量以中秋和晚秋期较高,夏季最低。测定的7种非桑属植物均不含1-DNJ。结果说明桑树是富含1-DNJ的主要植物资源,其中在桑园管理过程可以大量获得的桑树摘芯芽以及桑树伐条剪下的桑枝(桑皮)中均富含1-DNJ,具有重要的开发利用价值。     

钱塘江粗唇鱼危细胞色素b基因片段序列分析及系统发生研究

为研究钱塘江粗唇(鮠)与其他鲿科鱼类的亲缘关系,扩增并测定了粗唇(鮠)的细胞色素b基因片段序列.用Blastn分析所获序列与其他鲿科鱼类细胞色素b基因的同源性,用Mega4软件构建系统发生树.     

过表达MyBPC1对牛骨骼肌卫星细胞增殖分化的影响

旨在探究肌球蛋白结合蛋白C1（myosin binding protein C1,MyBPC1）对牛骨骼肌卫星细胞增殖与成肌分化的影响,为进一步研究MyBPC1在细胞分化和肌肉发育过程中的调控作用提供依据。本研究利用西门塔尔胎牛原代牛骨骼肌卫星细胞体外诱导成肌分化模型模拟牛骨骼肌的生长发育过程。采用qRT-PCR和Western blot检测MyBPC1的细胞时序表达谱。试验分为两组。在RNA水平每组4个重复,每个重复20 μL;在蛋白水平每组3个重复,每个重复15 μg。采用qRT-PCR和Western blot检测牛骨骼肌卫星细胞转染MyBPC1的过表达效果,并进一步检测细胞增殖期标志因子Pax7、Ki67以及细胞分化期标志因子MyHC、MyOG的表达变化情况,观察牛骨骼肌卫星细胞肌管形成状态。结果,MyBPC1在牛骨骼肌卫星细胞分化前后表达水平存在极显著差异,牛骨骼肌卫星细胞诱导分化后MyBPC1的mRNA和蛋白表达量均极显著高于增殖期（P<0.01）。过表达MyBPC1后,细胞分化形成的肌管数量明显多于对照组,增殖标志因子Pax7的mRNA水平和蛋白表达水平无显著差异,分化标志因子MyHC的mRNA水平和蛋白表达水平极显著高于对照组（P<0.01）。过表达MyBPC1可以促进牛骨骼肌卫星细胞体外成肌分化,为进一步开展MyBPC1对牛骨骼肌卫星细胞的调控机制奠定基础。     

利用环介导等温扩增(LAMP)技术快速检测辣椒疫霉菌

<正>辣椒疫霉(Phytophthora capsici)是一种重要植物病原菌,能造成植株坏死、果实腐烂,严重影响产量[1]。辣椒疫霉侵染植物的早期病症并不明显,易被忽视。因此,对辣椒疫霉早期快速、准确检测显得尤为重要。聚合酶链式反应(PCR)为动植物病原物检测的重要方法,但需要较昂贵的仪器、试剂与耗材,后期的电泳检测也费时费力,致使这一技术很难在生产一线普及推广。Notomi等2000年研发了环介     

Insights into the genetic diversity and evolution of Little cherry virus 1

Little cherry virus 1 (LChV‐1), a member of the recently proposed genus Velarivirus, is a sweet cherry pathogen that has been recently reported to infect other Prunus species and is associated with various plant disorders. In this work the incidence of the virus on its putative hosts and possible mechanisms driving its evolution were investigated. Due to problems encountered with LChV‐1 detection, a new nested RT‐PCR assay was developed and applied. The virus was found to be prevalent in cherry plantations in Greece and only occasionally detected in other Prunus species. Sequences corresponding to the partial RNA‐dependent RNA polymerase (RdRp), heat‐shock protein homologue (HSP70h) and coat protein (CP) genes were determined from Greek LChV‐1 isolates originating from different hosts; these were analysed, along with published homologous genomic regions from other isolates. Phylogenetic analysis of the three genes revealed the segregation of four evolutionary distinct groups showing no host or geography‐based clustering. Mean genetic distances among the four groups were high with the CP region showing the highest divergence, although intragroup variability levels were low. Nevertheless, estimations of the mean ratio of nonsynonymous substitutions per synonymous site to synonymous substitutions per synonymous site (dN/dS) for the partial RdRp, HSP70h and CP indicated that these genomic regions are under negative selection pressure. Interestingly, a recombination event was identified at the 3′ end of RdRp on a Greek virus isolate, thus highlighting the role of this mechanism in the evolutionary history of LChV‐1.     

Variability in infection and reproduction of Meloidogyne javanica on tomato rootstocks with the Mi resistance gene

The response of 10 commercial or experimental tomato rootstocks with the Mi resistance gene to an initial inoculum of a Mi‐avirulent population of Meloidogyne javanica was determined in pot tests conducted in spring and summer. In a field test, the rootstocks were subjected to continuous exposure to high initial population densities (2050 ± 900 second‐stage juveniles (J2) per 250 cm³ soil) of the nematode. The presence of the Mi locus in the resistant rootstocks and cultivars was confirmed using the PCR co‐dominant markers REX‐1 and Mi23. Nematode infectivity (egg masses) and reproduction (eggs g^?1 root) were highly variable in the spring tests. Rootstocks PG76, Gladiator and MKT‐410 consistently responded as highly resistant, with nematode multiplication rate (Pf/Pi) < 1 and reproduction index (RI) < 10%, and they were as efficient as standard resistant tomato cultivars at nematode suppression. The relative resistance levels of rootstocks Brigeor, 42851, 43965, Big Power and He‐Man varied depending on the susceptible standard used for reference or the duration of the test. Rootstocks Beaufort and Maxifort were susceptible to M. javanica (Pf/Pi > 50 and RI > 50%). Rootstocks PG76 and He‐Man, and the resistant tomato cv. Caramba showed high levels of resistance in the test conducted in summer, whereas MKT‐410 and 42851 and the resistant tomato cv. Monika were moderately resistant. In the field, seven rootstocks showed high levels of resistance and one (He‐Man) showed an intermediate level, whereas Beaufort and Maxifort were susceptible.     

转基因玉米表达的Cry1Ab、PAT和EPSPS蛋白对日本通草蛉幼虫的安全风险评估

为评价我国自主研发的转基因玉米表达的Cry1Ab、PAT和EPSPS蛋白对日本通草蛉Chrysoperla nipponensis幼虫生长发育的安全风险,通过将外源蛋白混入日本通草蛉人工饲料中的方法,以加入砷酸二氢钾(KH_2AsO_4)的饲料为阳性对照,研究了日本通草蛉幼虫取食后的生长发育状况。结果表明:日本通草蛉取食含Cry1Ab和EPSPS蛋白饲料的幼虫发育历期、茧期、结茧率、羽化率及成虫体重等生物学参数与取食正常饲料处理相比均没有显著差异;而取食含PAT蛋白饲料的幼虫发育历期和结茧率分别为10.7 d和96.6%,与取食正常饲料的对照11.1 d和89.9%差异显著,即饲料中添加PAT蛋白显著提高了幼虫的存活率和发育速率;取食含KH_2AsO_4饲料的日本通草蛉幼虫不能存活到茧期,说明KH_2AsO_4具有显著的杀虫活性。ELISA检测结果表明,取食了分别添加有3种外源蛋白饲料的日本通草蛉幼虫体内可以检测到相应的Cry1Ab、EPSPS和PAT蛋白,含量分别为2 758.8~5 210.7、35 018.0~54 426.6、16.8~149.8 ng/g。表明转基因玉米所表达的Cry1Ab、EPSPS和PAT蛋白对日本通草蛉幼虫没有显著的不利影响。     

Cry1Ac毒素对小菜蛾幼虫中肠蛋白酶和羧酸酯酶活性的影响

为了明确Cry1Ac毒素对小菜蛾Plutella xylostella幼虫中肠蛋白酶和羧酸酯酶活性的影响,利用蛋白酶专性底物测定其中肠各蛋白酶活性在不同缓冲液中的最适pH,并用不同浓度Cry1Ac毒素饲喂小菜蛾幼虫后测定该毒素的致死中浓度LC_(50),观察中肠蛋白酶和羧酸酯酶活性的变化,分析Cry1Ac毒素与昆虫中肠酶活性间的关系。结果表明,小菜蛾幼虫中肠总蛋白酶在Tris-HCl、KH_2PO_4/NaOH和Glycine/NaOH三种缓冲液中最适pH均为11.0;强碱性胰蛋白酶的最适pH分别为10.0、10.5和10.5;弱碱性胰蛋白酶的最适pH均为8.5;胰凝乳蛋白酶的最适pH分别为8.5、9.0和9.0。Cry1Ac毒素对小菜蛾的致死中浓度LC_(50)为0.343μg/mL。用不同浓度Cry1Ac毒素饲喂小菜蛾幼虫后,其中肠各蛋白酶活性均随时间的变化而变化。弱碱性胰蛋白酶和胰凝乳蛋白酶的活性明显升高,且随浓度升高呈上升趋势;在取食36~72 h期间低浓度处理组的羧酸酯酶活性与对照组间无显著差异,而高浓度处理组羧酸酯酶活性显著高于对照组。表明Cry1Ac毒素能够显著影响小菜蛾的中肠蛋白酶和羧酸酯酶活性。