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Results The amaranthine nuclei had a distinct outline and distributed evenly on the slide. The area of nuclei in liquidbased preparation was much smaller and chromatin was more condensed in comparison with those in conventional smear. The nuclei in conventional smears had the largest value of area and the value of AOD, and CV of IOD, the IOD ratio among diploid, tetraploid and octaploid DNA was most close to 2 and 4, and the CV of IOD exceeded 6%.

结果 三种涂片内肝细胞核分布均匀,轮廓清晰,呈紫红色,与传统涂片法相比,液基法内肝细胞核面积明显减小,染色质高度浓缩;相同倍体的肝细胞核在传统涂片中面积最大,平均光密度最低,各倍体间的比值最接近2和4,积分光密度CV值最低(。5)。

Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastocysts followed by fusion of cytoplasm and nucleus and cleavage in vitro. During the fusion of cytoplasm, the DNA methylation levels of the fused embryos are as high as these of two-cell diploid embryos in vivo Then the embryos are rapidly demethylated when the nucleus begin to fuse, resulting in the lowest DNA methylation levels when the nucleus are fused completely. After that, the DNA methylation levels of the fused embryos are gradually increased until the morula stage. However, whereas an asymmetric distribution of DNA methylation is established in vivo-derived blastocysts with a higher methylation level in the inner cell mass than that in the trophectoderm, we can not detect the asymmetric distribution in most in vitro-derived tetraploid blastocysts.

结果表明:利用电融合方法制备的小鼠四倍体胚胎在体外培养体系中经历细胞质融合、细胞核融合及细胞继续分裂发育直到囊胚期的过程,在细胞质融合的时候胚胎卵裂球同体内体外培养二倍体胚胎一样,呈现高度甲基化状态;在细胞核开始融合的时候,甲基化水平急速下降,在细胞核完全融合的时候甲基化水平达到最低点;随着胚胎继续分裂,胚胎甲基化水平逐渐增加,在桑葚胚期甲基化水平最高;但是囊胚期四倍体胚胎内细胞团同滋养层细胞甲基化荧光信号没有差别,这与体内体外培养二倍体囊胚内细胞团细胞甲基化荧光强度高于滋养层细胞甲基化荧光强度不同。

Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastocysts followed by fusion of cyto-plasm and nucleus and cleavage in vitro. During the fusion of cytoplasm, the DNA methylation levels of the fused embryos are as high as these of two-cell diploid embryos in vivo Then the embryos are rapidly demethylated when the nucleus begin to fuse, resulting in the lowest DNA methylation levels when the nucleus are fused completely. After that, the DNA methy-lation levels of the fused embryos are gradually increased until the morula stage. However, whereas an asymmetric distribu-tion of DNA methylation is established in vivo-derived blastocysts with a higher methylation level in the inner cell mass than that in the trophectoderm, we can not detect the asymmetric distribution in most in vitro-derived tetraploid blastocysts.

结果表明:利用电融合方法制备的小鼠四倍体胚胎在体外培养体系中经历细胞质融合、细胞核融合及细胞继续分裂发育直到囊胚期的过程,在细胞质融合的时候胚胎卵裂球同体内体外培养二倍体胚胎一样,呈现高度甲基化状态;在细胞核开始融合的时候,甲基化水平急速下降,在细胞核完全融合的时候甲基化水平达到最低点;随着胚胎继续分裂,胚胎甲基化水平逐渐增加,在桑葚胚期甲基化水平最高;但是囊胚期四倍体胚胎内细胞团同滋养层细胞甲基化荧光信号没有差别,这与体内体外培养二倍体囊胚内细胞团细胞甲基化荧光强度高于滋养层细胞甲基化荧光强度不同。

Results (1)The permillage of myocyte apoptosis in infarct area in elder patients with sudden death caused by AMI was significantly higher than that in control group(663.00‰±117.12‰ vs 34.30‰±20.68‰,P<0 .01).However the permillage of myocyte apoptosis in 1-vessel,2-vessel,and 3 - vessel disease were 514.28±165.12‰,564.38‰±102.33‰ and 668.25‰±127. 19‰,respectiverly,significantly higher as compared to control group(All P< 0.01).But no significance was found among the three groups.(2)The size of DNA f ragment about 180~200 bp was found only in those patients with two and three ve ssels involoved.(3)The electron microscope showed the characteristics of myocyte apoptosis episodes,the others showed the characteristics of necrosis.

结果 TUNEL法发现,猝死者梗死区的心肌细胞凋亡千分数老年组(663.00‰±117.12‰)明显高于正常对照(34.30‰±20.68‰)(P<0.001);心肌细胞凋亡千分数在冠脉1支病变者(514.28‰±165.12‰)<2支病变者(564.38‰±102.12‰)<3支病变者(668.25‰±127.19‰),虽然均明显高于正常对照(均P<0.01),但三组之间比较则无统计学上的差别(均P>0.05);冠脉2~3支病变者梗死区的心肌细胞DNA电泳可见相差约180~220 bp的阶梯片段;电镜发现猝死者梗死区内的心肌细胞核膜完整、染色质浓集、电子密度增加的凋亡特征,有的则出现核膜破裂、染色质溶解成碎屑的坏死现象。

The display exhibits the molecular structure of the capacity for heredity existing in the nucleus. It is composed of chain-trend ribose and molecule of phosphoric acid root, and basic pairs of molecules linked between the double chains.

本展品所展示的是存在于细胞核内具有遗传特性的分子结构模型,主要由组成链走向的核糖和磷酸根分子以及双链之间互连的碱基分子组成。

This displays the molecular mechanism heredity in the nucleus. DNA is composed of chain-trend ribose and root molecules of phosphoric acid, and basic pairs of molecules linked in double chains.

本展品所展示的是存在于细胞核内具有遗传特性的分子结构模型,主要由组成链走向的核糖和磷酸根分子以及双链之间互连的碱基分子组成。

To investigate the expression, location and function, long distance PCR was used to expand the targets containing open reading frame. The expressive vectors of prokaryotic and eukaryotic cells were constructed after product purification and connecting with vectors. The prokayotic vectors were induced to express the protein by IPTG and the protein was seen by denaturalization PAG electrophoresis and dyeing. The eukayotic expressive vectors were transfected into culture cells and the protein was found in the nulceolus under the observation of the confocal microscope. The transfected cells were chosen by the geneticin (G418). The cell cyele was examined by flow cytometer and the cell numbers were reduced obviously in the stage of G〓-G〓 and G〓-M, but increased distinguishably in the S stage.

为研究磷酸化应激诱导蛋白的表达、定位和功能,采用了长距离聚合酶链反应扩增含有开放阅读框架的靶片段,经产物纯化、与载体连接,分别构建了STIP1基因的原核表达载体和真核表达载体;IPTG诱导原核表达载体,变性聚丙酰胺凝胶电泳与染色后,可见相应大小的蛋白质表达;STIP1基因的真核表达载体转染细胞系后,共聚焦显微镜下观察STIP1蛋白主要分布于细胞核内;用药物筛选STIP1基因转染的细胞后,流式细胞仪检测细胞周期,可见G〓-G〓期和G〓-M期的细胞明显降低,S期细胞明显增多。

Using the intact G1-nuclei prepared from an isogenic yeast strain lacking mitochondrial DNA , we demonstrated that the DNA synthesis observed within the first 10-20 min was largely due to mitochondrial DNA synthesis, whereas nuclear DNA synthesis did not begin until after a 10-20 min lag period.

利用从缺少线粒体DNA的菌株中制备的完整G1期细胞核,证明了最初10-20min的滞后期内的DNA合成主要是由线粒体DNA合成造成的,而核DNA合成是在经过10-20 min后才开始。

Result 1 Magnetic nanoparticles, magnetic nanoparticles modified with antisense oligodeoxynucleotide of human telomerase reverse transcriptase induced HL-60 tumor cells to apoptosis, we could see typical morphologic change of apoptosis cells: karyopyknosis, chromation"s condensing and aggregation in nuclear, forming crescent-shaped or annulus structures to lean on edge of cell nucleus"s membrane and posing apoptosis body by Atomic Force Microscope, Fluorescence microscope, transmission electron Microscope 2 There was a significant difference compared with control group(p.01), inhibition ratio had significant positive correlation with medication dosage and time ;during 0.8-8μM dosage amplitude, inhibition ratio accrescenced by dosages increasing. However, the inhibition ratio would decrease when dosage over 8-80μM.

结果 1 磁性纳米粒子、修饰有端粒酶反义寡核苷酸的磁性纳米粒子诱导HL-60细胞发生凋亡,原子力显微镜、光学显微镜、荧光显微镜和透射电镜下均观察到HL-60细胞呈现典型的凋亡细胞的形态变化:细胞核固缩,核内染色质浓缩、凝聚、形成新月形或环状结构紧靠在细胞核膜边缘,并形成凋亡小体。2 磁性纳米粒子、修饰有端粒酶反义寡聚脱氧核苷酸的磁性纳米粒子对HL-60肿瘤细胞的生长和增殖有明显的抑制作用,与对照组相比有显著性差异(p<0.01),在剂量为0.8-8μmol/L范围内,抑制率随剂量的增加而增加,当剂量超过8μmol/L时,抑制率反而下降;3 磁性纳米粒子、修饰有端粒酶反义寡聚脱氧核苷酸的磁性纳米粒子可增强p53基因的表达活性,引起DNA降解损伤,反向调节细胞周期活动,促使细胞从G0期进入G1期,抑制肿瘤细胞的生长。4 修饰有端粒酶反义寡聚脱氧核苷酸的量子点能通过内吞作用进入HL-60肿瘤细胞的细胞核,可以在细胞内进行定位和促进HL-60肿瘤细胞的凋亡。

Erve growth factor combined with its receptor, activated Ras - MAP kinase , mitogen - activated protein kinase kinase is the important regulating factor that make IkB kinase, IKK, phosphated, IkB kinase make the IκBα:(the subunit of NF -κB ) phosphated, the phosphated IkBα degraded, and p65 - p50 heterodimer can be formed, then the heterodimer translocated to nucleus and combined with the promoter domain or other consensus sequence.

GF与其受体相结合,最后可以激活Ras-有丝分裂激活的蛋白激酶途径,有丝分裂激活的蛋白酶激酶(MEKK1)是IγB激酶发生磷酸化的重要调节因子,IKK使NF-κB的亚单位IKBα发生磷酸化,磷酸化的IKBα发生降解,而最后留下p65-p50二聚体,该二聚体然后转位到细胞核内与κB基因的增强子区域或与他的顺式作用序列结合。

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