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Methods Fourteen mutated vpr fragments were selected from patients with HIV. Both eukaryotic expression vector pcDNA3.1 and PCR products were purified, double-cut by Hind Ⅲ and BamH Ⅰ, and the cut products were legated and transformed into competent cells JM109. The 14 reconstructed plasmids were transfected into Hela cells. Cells with pcDNA vpr-wt, pcDNA vpr-Fs and pcDNA3.1 blank cells, and without pcDNA3.1 cell were established.

以14个带有HIV-1vpr基因片段的PcD-NA3.1真核表达载体构建重组质粒,将其转染Hela细胞,并设立保守株vpr基因转染细胞、突变株vpr-FS基因转染细胞、空载体转染细胞和未转染细胞作为对照,经逆转录多聚酶链式反应检测目的基因转染成功后,Pi染色,用流式细胞仪检测被转染细胞的细胞周期分布和细胞凋亡率。

Methods 14 mutanted vpr fragments selected from Chines patients with HIV. Both eukaryotic expression vector pcDNA3.1 and PCR products purified , double-cut by HindⅢ and BamH and the cut products legated and were into competent cells JM109. The 14 reconstructed plasmids electronically transfected into hela cells, and established cells with pcDNA vprwt 、pcDNA vpr-Fs and pcDNA3.1 blank cells, and without pcDNA3.1 cell.

将14个带有HIV-1 vpr基因片段的pcDNA3.1真核表达载体构建重组质粒,将其转染Hela细胞,并设立保守株vp r基因转染细胞、突变株 vpr-FS基因转染细胞、空载体转染细胞和未转染细胞作为对照,经RT-PCR检测目的基因转染成功后,经Pi染色用流式细胞仪检测被转染细胞的细胞周期分布和细胞凋亡率。

The inhibition of SPG-Rg3 on the growth of MCF-7 cells was detected by MTT assay,IC50 was calculated to obtain the effective concentration; flow cytometry was used to observe the cell cycle of MCF-7 cells; AO/EB fluorescence double-dye technology was performed to observe the apoptosis of cells in morphology,immunocellularchemistry and RT-PCR were utilized to investigate the apoptosis of MCF-7 cells and its relationship with caspase-8 from protein level and molecule level.

采用MTT法观察SPG-Rg3对MCF-7细胞生长的抑制作用,并计算出IC50,依据IC50值确定SPG-Rg3的有效浓度;流式细胞术检测SPG-Rg3作用后MCF-7细胞周期的变化;AO/EB双染从形态学上观察SPG-Rg3对MCF-7细胞凋亡的作用;免疫细胞化学染色和RT-PCR技术分别从蛋白水平和分子水平上检测SPG-Rg3对MCF-7细胞的诱导凋亡作用及其与caspase-8基因的关系。

METHODS: Spleen cells of 1, 2, 4, 6, 8, 10, 12, 16, 20 and 24 h in the coculture system were observed and the expression of RNA editase was detected by semiquantitative PCR and was contrasted with the mouse spleen cells in a singleculture system. The mouse model with the immune system restrained by FK506 was made, its spleen cells were cocultured with stimulated cells and then the effect of FK506 on RNA expression of editase was detected by semiquantitative PCR.

观察共培养1,2,4,6,8,10,12,16,20及24 h的小鼠脾细胞,用半定量PCR的方法检测RNA编辑酶表达的变化,并以相同时段单独培养的小鼠脾细胞RNA编辑酶的表达情况进行对照;制备免疫功能受FK506抑制的小鼠模型,取其脾细胞与刺激细胞进行相同时段的共培养,用半定量PCR的方法检测FK506对脾细胞RNA编辑酶的影响。

A portion of the ce cells derived from cocultures of transgenic tobacco cells and untransformed carrot cells can survive on the medium containing Km (200mg·l〓). Some of the carrot plants regenerated from ce cells also express Km〓 and GUS activity.

以烟草转化细胞为材料,与胡萝卜细胞共培养后,选出的胡萝卜细胞中有部分细胞具有Km〓,ce再生植株中也表现一定的Km〓,部分植株中检测到GUS活性。

The cells of the third stratum were typical mast cells without the tidal line in rat cartilage, cells which arranged into pillar with the distinct tidal line in rabbit cartilage, large irregular cells and coenocytes with the obvious tidal line in human cartilage.

在软骨损伤和骨关节炎的实验研究中,多数研究是以兔和大鼠为实验动物,而且大部分教科书中的软骨组织学结构是以兔的软骨进行描述的。

The adult stem cells in intestinal crypts, i.e., intestinal stem cells, may relate with the pathogenesis of colorectal cancer closely. This paper reviews the recent advances about intestinal stem cells and cancer stem cells of colorectal cancer.

近年较多的文献报道结直肠癌组织中存在肿瘤干细胞,而这些具有干细胞特性的瘤细胞可能来自突变的肠干细胞,本文就肠干细胞和结直肠癌肿瘤干细胞的研究进展作一综述。

Results of the experimental groups: Comparison with the control groups, in 6h and 8h groups eminences of ependymal cells with cobblestone-like appearance were seen, their microvilli were further richer, and a number of microholes could be found on their base. In body part of SFO cell membrane of some ependymal cells were broken completely, their cytoplasm was no longer seen, and cell nucleus located at one side of the cell, 5-7μm in diameter and dividing lobe-like in shape. Beside those cells some ependymal cells appeared many folds on the top of surface. In 2h and 16h groups the observation results seemed no significant differences from the corresponding control groups.

实验组结果:与对照组相比,在6小时和8小时实验组中,可见成团的鹅卵石状室管细胞隆起于表面,微绒毛更为丰富,微绒毛根部可见到若干微细小孔,在SFO体部的一些室管膜细胞的胞膜完全破裂,胞质几乎荡然无存,仅剩胞核居于细胞的一侧,直径有5.7μm,呈分裂叶状;与这些细胞邻近的室管膜细胞表面最高处多呈现皱襞。2小时与16小时的实验组的观察结果与对照组相比未发现明显差异。

Methods Cells were maintained in DMEM medium with 10% fetal bovine serum. Five days before the beginning of experiments, the cells were seeded in phenol red-free DMEM medium containing 5% charcoal dextran-treated FBS. The cells were harvested, and seeded in 6-well culture plates or in 75ml flacks. After NP, BisA and DBP treatments for 72h, the cells were harvested, and mRNA and protein expression of PCNA, bcl-2 and bax were detected by reverse transcription-polymerase chain reaction and immunohistochemistry, respectively.

方法]T47D细胞在含10%胎牛血清的DMEM培养液中进行常规传代培养,实验前将细胞转移至无酚红DMEM(含5%活性碳葡聚糖苷处理过的FBS)中继续培养5d,收集细胞,PBS洗涤后接种於内置盖玻片的6孔板或75ml培养瓶中,用32×10^(-7)mol/L NP、32×10^(-7)mol/L BisA及32×10^(-6)mol/L DBP对细胞分别处理72h,用半定量RT-PCR技术观察这3种化合物对核增殖抗原PCNA、bcl-2及bax mRNA表达的影响,并用免疫组化方法对结果进行验证。

ES cells were maintained on gelatin-coated dishes in Iscove's modified Dulbecco medium containing 0.1 mmol/L of nonessential amino acids, 2 mmol/L of L-glutamine, 0.1 mmol/L of monothioglycerol, 15% fetal bovine serum, and 1000 U/mL leukemia inhibitory factor. Embryonic bodies were firstly formed by ES cells with hang drip method in IMDM without LIF. Six-day-old EBs were trypsinized and the cells were resuspended at a concentration of (3-4)×109 L-1 cells per 1 mL of a 1:1 mixture of culture medium and Matrigel?

实验方法:将鼠胚胎干细胞株接种于被覆明胶的培养瓶内,加入含0.1 mmol/L非必需氨基酸、2 mmol/L L-谷酰胺、0.1 mmol/L二羟基丙硫醇、15%胎牛血清、1 000 U/mL白血病抑制因子的IMDM培养液。3 d后采用&悬滴法&使细胞在不含白血病抑制因子的上述IMDM培养液中形成胚胎体,6 d后胰酶消化得到胚胎干细胞,浓度调整为(3~4)×109 L-1,与Matrigel?

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