ANLL中转录因子GATA—2基因点突变

了解转录因子ＧＡＴＡ－２基因在ＡＮＬＬ中的表达和突变情况,方法：采用ＲＴ－ＰＣＲ检测８５例ＡＮＬＬ病人外周血单个核细胞中ＧＡＴＡ－２基因的表达情况,ＰＣＲ产物进一步经单链构象多态性（ＳＳＣＰ）分析以了解基因突变情况。结果：绝大多数ＡＮＬＬ都表达了ＧＡＴＡ－２基因（８９．４％）,ＳＳＣＰ分析发现一例Ｍ２型的ＰＣＲ产物出现异常迁移带,核苷序列分析显示在ＧＡＴＡ－２基因第８９２位的核苷酸出现点突变,即第     

QIAquick Gel Extraction Kit提取和纯化PCR产物用于AML中GATA-2基因突变的分析

转录因子ＧＡＴＡ家族是１９８９年首次发现的锌指结构家族转录因子。其中ＧＡＴＡ２在造血干细胞的增殖和分化中起重要作用［１］。绝大多数急性髓细胞白血病（ＡＭＬ）细胞表达ＧＡＴＡ－２基因［２］。在分析ＧＡＴＡ－２基因表达中，发现ＧＡＴＡ－２突变的现象［...     

急性髓细胞白血病中转录因子GATA—2基因的表达和？…

目的：分析急性髓细胞白血病（ＡＭＬ）中转录因子ＧＡＴＡ－２基因的表达和突变情况。方法：应用反转录－多聚酶链反应（ＲＴ－ＰＣＲ）检测８５例ＡＭＬ的ＧＡＴＡ－２基因表达情况。２５例正常人的外周血或骨髓作为对照。结果：８５例ＡＭＬ中７６例表达了ＧＡＴＡ－２基因。各ＡＭＬ亚型均见ＡＧＴＡ－２表达。在８５例ＡＭＬ发现２例出现至今未报道的ＧＡＴＡ－２基因的突变。在一例ＡＭＬ－Ｍ２病人发现ＧＡＴＡ－２ｃＤＮＡ     

各种布尔矩阵最大广义逆

设Ａ是布尔矩阵，依据４个性质、ＡＧＡ＝Ａ，ＧＡＧ＝Ｇ、（ＧＡ）￣Ｔ＝ＧＡ、（ＡＧ）￣Ｔ＝ＡＧ的不同组合，定义了五种广义逆Ａ￣－、Ａｒ￣－、Ａ＿ｍ￣－、Ａ＿ｌ￣－、Ａ￣＋，这里Ｇ是布尔矩阵．本文中，我们证明了，如果Ａ￣－、Ａｒ￣－、Ａｍ￣－、Ａ＿ｌ￣－、Ａ￣＋，存在，那么它们一定有最大广义逆，其表示分别为（Ａ￣ＴＡ￣ＣＡ￣Ｔ）￣Ｃ、（Ａ￣ＴＡ￣ＣＡ￣Ｔ）￣ＣＡ（Ａ￣ＴＡ￣ＣＡ￣Ｔ）￣Ｃ、（Ａ￣（ＴＣ）ＡＡ￣Ｔ）￣Ｃ、（Ａ￣ＴＡＡ￣（ＴＣ））￣Ｃ、Ａ￣Ｔ．     

泥螺BULLACTA EXARATA的室内人工育苗

泥螺ＢＵＬＬＡＣＴＡＥＸＡＲＡＴＡ的室内人工育苗王一农，岑利权，韩建婵（浙江水产学院水产养殖系，宁波３１５０１０）ＩＮＤＯＯＲＡＲＴＩＦＩＣＡＬＲＥＡＲＩＮＧＯＦＭＵＤＳＮＡＩＬ，ＢＵＬＬＡＣＴＡＥＸＡＲＡＴＡＷａｎｇＹｉｎｏｎｇＣｅｎＬｉｑｕａｎｈ...     

野蚕DHPBAN基因启动子的克隆及序列分析

为研究昆虫滞育激素性信息素合成激活肽基因在进化过程中的变化以及基因表达调控的异同,用ＰＣＲ方法克隆了野蚕的ＤＨＰＢＡＮ基因的启动子并测序,这是第二个被克隆的昆虫ＤＨＰＢＡＮ基因启动子．与家蚕相比,在核苷酸水平上同源性达９４％．野蚕ＤＨＰＢＡＮ基因启动子的ＴＡＴＡ框的保守序列为ＴＡＴＡＴＡＡＡ,位于－４７—－４０处;ＣＡＡＴ框是ＧＧＣＣＣＡＴＣＴ,位于－８３—－７５处;野蚕与家蚕一样具有ＴＡＡＴ保守序列,这段序列是一类调控蛋白的结合核心位点,家蚕的位于－１８５—－１７６,野蚕的位于－１７６—－１６７．在－６４—－５６部位,核苷酸序列表现出较大的不同．     

POLYCYCLE－COMPOUNDING STRUCTURES AND REGIONAL LAYERSLIP－DIPSLIP SYSTEMS：IMPLICATIONS FOR THE TARIM BASIN，XINJIAN

ＰＯＬＹＣＹＣＬＥ－ＣＯＭＰＯＵＮＤＩＮＧＳＴＲＵＣＴＵＲＥＳＡＮＤＲＥＧＩＯＮＡＬＬＡＹＥＲＳＬＩＰ－ＤＩＰＳＬＩＰＳＹＳＴＥＭＳ：ＩＭＰＬＩＣＡＴＩＯＮＳＦＯＲＴＨＥＴＡＲＩＭＢＡＳＩＮ，ＸＩＮＪＩＡＮ￥ＳｕｎＹａｎ；ＪｉａＣｈｅｎｇｚａｏ（Ｄｅ...     

检测丽蚜小蜂取食寄主银叶粉虱的多重聚合酶链式反应

结合Ｄ２引物和ＢＥＦ引物,优化了一个可以用来检测丽蚜小峰（Ｅｎｃａｒｓｉａ ｆｏｒｍｏｓａ Ｇａｈａｎ）取食寄主银叶粉虱（Ｂｅｍｉｓｉａ ａｒｇｅｎｔｉｆｏｌｉｉ Ｂｅｌｌｏｗｓ＆Ｐｅｒｒｉｎｇ）的重多聚合酶链式反应。特异于银叶粉虱的ＢＥＦ引物（正向５′－ＡＡＴ ＴＣＡ ＣＡＣ ＡＴＡ ＣＧＴ ＴＡＧ ＣＣＣ ＣＴ－３′和反向５′－ＴＡＡ ＣＣＧ ＡＡＣ ＣＡＴ ＣＡＡ ＣＡＧ ＡＴＴ ＡＴＴ－３     

GHRP—scu—PA—32K突变体的构建,表达及纯化产物的部分性 …

在ｓｃｕ－ＰＡ３２Ｋ的ｃＤＮＡ分子基础上经定点突变,在Ｎ端紧接Ｌｅｕ＾１以前引入编码ＧＨＲＰ四肽的寡核苷酸序列（ＧＧＴＣＡＴＡＧＧＣＣＴ）,构建了ＧＨＲＰ－ｓｃｕ－ＰＡ－３２Ｋ的突变体ｃＤＮＡ。将它克隆到表达载体ｐＣＭ－β－ｄｈｆｒ共转染ＣＨＯ／ＤＨＦＲ＾－细胞。筛选到的稳定表达株在无 清培养基的表达量为５８０ＩＲ／（１０＾６细胞．２４ｈ）。经锌离子螯合亲和柱纯化的产物,ＳＤＳ－ＰＡＧＥ显示为一蛋     

纺锤菌素－DNA相互作用的分子力学分析

文章介绍了纺锤菌素－ＤＮＡ相互作用分子力学研究的最新成果，对纺锤菌素－ＤＮＡ相互作用的性质、相互作用能作了详细的叙述，并与ｘ射线晶体衍射结果作了比较．在此基础上，我们对纺锤菌素ＤＮＡ相互作用进行了分子力学分析、考察了纺锤菌素与ｄ（Ａ６）、ｄ（Ｔ６），ｄ（ＴＡＴＡＴＡ）２、ｄ（ＣＧＣＧＣＧ）２和ｄ（ＣＧＣＧＡＡＴＴＣＧＣＧ）２的相互作用，得到了一些有益的结论．     

Elf-1基因在急性髓细胞性白血病中的表达情况

目的:建立实时定量PCR检测Elf-1基因表达水平的方法,了解急性髓细胞性白血病(AML)患者外周血Elf-1基因表达水平.方法:采用SYBR Green Ⅰ荧光定量PCR和相对定量分析法检测33例AML患者:M2型17例、M3型6例、M5型10例和20例健康人外周血的单核细胞的Elf-1表达情况,以β2微球蛋白基因(β2M)作为内参,采用公式2~(-△Ct)×100%计算Elf-1基因表达水平.结果:AML组Elf-1表达水平(13.518±19.197)%明显高于健康对照组(2.044±1.321)%(P<0.01),不同AML亚型之间Elf-1的表达水平没有显著性差异(P=0.52),但各亚型AML的Elf-1的表达水平均与健康对照组比较均有显著性差异(P<0.01).结论:建立SYBR Green Ⅰ荧光实时定量PCR分析外周血Elf-1表达水平的方法,检测急性髓细胞性白血病Elf-1基因高表达情况.     

Induction of embryonic stem cells to hematopoietic cells in vitro

In order to get hematopoietic cells from embryonic stem (ES) cells and to study development mechanisms of hematopoietic cells, the method of inducing embryonic stem cells to hematopoietic cells was explored by differenciating mouse ES cells and human embryonic cells in three stages. The differentiated cells were identified by flow cytometry, immunohistochemistry and Wright's staining. The results showed that embryoid bodies (EBs) could form when ES cells were cultured in the medium with 2-mercaptoethanol (2-ME). However, cytokines, such as stem cell factor (SCF), thrombopoietin (TPO), interleukin-3 (IL-3), interleukin-6 (IL-6), erythropoietin (EPO) and granular colony stimulating factor (G-CSF), were not helpful for forming EBs. SCF, TPO and embryonic cell conditional medium were useful for the differentiation of mouse EBs to hematopoietic progenitors. Eighty-six percent of these cells were CD34+ after 6-d culture. Hematopoietic progenitors differentiated to B lymphocytes when they were cocultured with primary bone marrow stroma cells in the DMEM medium with SCF and IL-6. 14 d later, most of the cells were CD34-CD38+. Wright's staining and immunohistochemistry showed that 80% of these cells were plasma-like morphologically and immunoglubolin positive. The study of hematopoietic cells from human embryonic cells showed that human embryonic cell differentiation was very similar to that of mouse ES cells. They could form EBs in the first stage and the CD34 positive cells account for about 48.5% in the second stage.     

Induction of embryonic stem cells to hematopoietic cellsin vitro

In order to get hematopoietic cells from embryonic stem (ES) cells and to study development mechanisms of hematopoietic cells, the method of inducing embryonic stem cells to hematopoietic cells was explored by differenciating mouse ES cells and human embryonic cells in three stages. The differentiated cells were identified by flow cytometry, immunohistochemistry and Wright’s staining. The results showed that embryoid bodies (EBs) could form when ES cells were cultured in the medium with 2-mercaptoethanol (2-ME). However, cytokines, such as stem cell factor (SCF), thrombopoietin (TPO), interleukin-3 (IL-3), interleukin-6 (IL-6), erythropoietin (EPO) and granular colony stimulating factor (G-CSF), were not helpful for forming EBs. SCF, TPO and embryonic cell conditional medium were useful for the differentiation of mouse EBs to hematopoietic progenitors. Eighty-six percent of these cells were CD34⁺ after 6-d culture. Hematopoietic progenitors differentiated to B lymphocytes when they were cocultured with primary bone marrow stroma cells in the DMEM medium with SCF and IL-6. 14 d later, most of the cells were CD34^－CD38⁺. Wright’s staining and immunohistochemistry showed that 80% of these cells were plasma-like morphologically and immunoglubolin positive. The study of hematopoietic cells from human embryonic cells showed that human embryonic cell differentiation was very similar to that of mouse ES cells. They could form EBs in the first stage and the CD34 positive cells account for about 48.5% in the second stage.     

Knockdown of <Emphasis Type="Italic">MRP4</Emphasis> by lentivirus-mediated siRNA improves sensitivity to adriamycin in adriamycin-resistant acute myeloid leukemia cells

Chemotherapy remains the standard treatment for acute myeloid leukemia;however,the emergence of drug resistance is a major hurdle in the successful treatment of leukemia.The expression of multidrug resistance-associated protein 4(MRP4)induces re- sistance in the adriamycin-resistant acute myeloid leukemia cell line,K562/ADR.The aim of this study was to investigate whether knockdown of MRP4 by lentivirus-mediated siRNA could improve the sensitivity of K562/ADR cells to adriamycin.Five lenti- virus-mediated short hairpin RNAs(lv-shRNAs-MRP4)were designed to trigger the gene silencing RNA interference(RNAi) pathway.The efficiency of lentivirus-mediated siRNA infection into K562/ADR cells was determined using fluorescence mi- croscopy to observe lentivirus-mediated GFP expression.MRP4 expression in infected K562/ADR cells was evaluated by real- time PCR and Western blot analysis.The MTS assay was used to measure cell viability and flow cytometry was used to measure apoptosis.The transfection efficiency of K562/ADR cells was over 80 percent.The gene silencing efficacy of lv-shRNA1-MRP4 was superior to the other constructs.Infection of K562/ADR cells with lv-shRNA1-MRP4 led to strong inhibition of MRP4 mRNA and protein expression.Combined treatment with lv-shRNA1-MRP4 and adriamycin decreased cell growth and increased apoptosis compared to treatment with lv-shRNA1-MRP4 or adriamycin alone.These data indicate that in K562/ADR cells MRP4 is involved in drug resistance mechanisms and that lentivirus-mediated knockdown of MRP4 may enhance sensitivity to adriamycin.     

白血病间接免疫荧光法免疫表型分析

目的:研究白血病免疫表型的不同,明确诊断。方法:回顾性分析本院56例白血病患者免疫表型资料。结果:30例急性髓系白血病(AML)均表达髓系抗原,部分伴有淋巴抗原,但其阳性率明显低于急性淋巴细胞白血病(ALL)。7例M3的CD9抗原阳性率为100%,明显高于AML的其它亚型白血病。形态学诊断为ALL的12例白血病中,2例免疫表型为T-ALL,10例B-ALL。4例为杂合性急性白血病,它的诊断标准主要靠免疫分型。在10例慢性粒细胞性白血病(CML)中,4例发生急粒变,且急变期CD34抗原阳性率高于慢性期。结论:免疫表型分析与形态学、细胞化学三者结合为白血病的诊断提供了更可靠、更有价值的依据。     

nm23-H1基因与人早幼粒白血病细胞HL-60增殖的相关性分析

目的探讨nm23-H1基因的表达与白血病细胞HL-60增殖之间的关系。方法：以25ng／mL阿糖胞苷处理HL-60细胞，MTT法测定细胞生长抑制率，NBT还原比色法判断细胞分化状况，RT-PCR检测nm23-H1基因表达的变化；构建nm23-H1基因的真核表达质粒pEGFP-N1-nm23-H1，转染HL-60细胞，通过细胞生长曲线和血清依赖性实验检测nm23-H1基因的过表达对HL-60细胞生长的影响。结果：小剂量Ara-C对HL-60细胞的生长呈时间依赖性抑制，作用4d后细胞NBT还原能力增强且nm23-H1基因的表达下调；转染nm23-H1基因的HL-60细胞生长加快、血清依赖性下降。结论：Ara-C对HL-60细胞增殖的抑制作用与下调nm23-H1基因的表达有-定关系；nm23-H1基因在HL-60细胞中的过表达有促细胞增殖的作用，即增高了HL-60细胞的恶性程度。     

Morphological transformation of human keratinocytes expressing the LMP gene of Epstein-Barr virus.

The association of Epstein-Barr virus (EBV) with nasopharyngeal carcinoma (NPC) has been known for some time, but the precise role of EBV in this cancer is poorly understood, due partly to the lack of an in vitro system for studying NPC cells and the effect of EBV on epithelial cells. Biopsies of NPC tumours have revealed expression of the EBV latent membrane protein (LMP) in 65% of cases, suggesting that in at least some NPC tumours LMP may contribute to cell transformation. Here we address the question of the effect of LMP expression on epithelial cells. Transfection of an immortalized, non-tumorigenic keratinocyte cell line (RHEK-1) with the LMP gene causes a striking morphological transformation: the originally flat, polygonal colonies change to bundles of spindle-shaped cells that form multilayer foci, and cytokeratin expression is down-regulated. Our results suggest that LMP expression may be an important causal factor in the development of NPC.     

T—ALL／淋巴瘤中BCL11B基因转录本的分析

目的:分析T-ALL/淋巴瘤中BCL11B基因转录本的特点。方法:利用逆转录聚合酶链式反应(RT-PCR)和实时定量聚合酶链式反应(RQ-PCR)方法分析12例T-ALL/淋巴瘤外周血单个核细胞(PBMC)中BCL11B基因的表达情况;限制性内切酶消化确定PCR产物的特异性。结果:3例T-ALL/淋巴瘤患者中出现2种BCL11B转录本:野生型和第3外显子缺失型,其余9例均表达第3外显子缺失型的转录本。两组病人PBMC中BCL11B表达水平无显著差别(P=0.301)。结论:3例T-ALL/淋巴瘤病人中存在2种BCL11B基因转录本,但在表达水平上无明显差别。