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The freezing technology of papanicolaou test is one meliorative technique based on papanicolaou test, the technique emphasizes on every process of sampling,smearing and staining, the most important step is the fixed smears should be soaked into the restoring-liquid of cells morphology based on the theory of penetration and expansion principle,thus achieve the purpose to cells restoring their normal appearance,to promoting cells staining and to improving cells transparency and clarity, it will be favorable to improve the cells recognition rate,the positive rate,the specification,sensitivity and decrease leaking rate and misdiagnosis rate.

冷冻式巴氏涂片技术是建立在传统巴氏涂片基础上的一种改良技术,该法在制片的过程中强调注重取材制片的全过程,其核心步骤是把固定好后的细胞玻片置入细胞形态修复液中,利用渗透膨胀的原理对已干枯萎的细胞进行修复,从而达到恢复细胞形态的目的,促进细胞着色,提高细胞透明度和清晰度,有利于对病变细胞的识别和提高对宫颈病变细胞的阳性检出率、特异度和灵敏度,降低漏诊误诊率。

The results were as follows:The pituitary of Long-snout catfish included adenohypophysis and neurohypophysis. The adenohypophysis have six-cell types in common,i.e adrenocorticotro-phs,lactot-rophs,somatotrophs,gonadotrophs, thyrotrophs, and Melanotroph cells. In neurohypophysis, there were three types of neurosecretory fibres, types A1,A2 and B.There were different typesgranules in type A neurosecretory fibres axone.In type B neurosecretory fibres axone, there were many small transparent vesicles.

结果表明:长吻魷脑垂体包括腺垂体和神经垂体两个部分,腺垂体组织中有六种分泌细胞即促肾上腺皮质激素分泌细胞、催乳激素分泌细胞、生长激素分泌细胞、促性腺激素分泌细胞、促甲状腺激素分泌细胞、促黑色素细胞刺激激素分泌细胞;神经垂体组织中存在A型(A_1、A_2)和B型神经分泌纤维,A型分泌纤维轴突中具有不同类型的分泌颗粒,B型分泌纤维轴突中含有许多透明小囊泡。

The results were as follows: The pituitary of Long-snout catfish included adenohypophysis and neurohypophysis. The adenohypophysis have six-cell types in common,i.e adrenocorticotro-phs,lactot-rophs,somatotrophs,gonadotrophs, thyrotrophs, and Melanotroph cells. In neurohypophysis, there were three types of neurosecretory fibres, types A1,A2 and B.There were different types'granules in type A neurosecretory fibres axone.In type B neurosecretory fibres axone, there were many small transparent vesicles.

结果表明:长吻魷脑垂体包括腺垂体和神经垂体两个部分,腺垂体组织中有六种分泌细胞即促肾上腺皮质激素分泌细胞、催乳激素分泌细胞、生长激素分泌细胞、促性腺激素分泌细胞、促甲状腺激素分泌细胞、促黑色素细胞刺激激素分泌细胞;神经垂体组织中存在A型(A_1、A_2)和B型神经分泌纤维,A型分泌纤维轴突中具有不同类型的分泌颗粒,B型分泌纤维轴突中含有许多透明小囊泡。

Results (1)ATP could inhibit the proliferation of U937 cells with inhibition rate over 43% after 48hour ATP treatment;(2)ATP treated U937 cells numbers increased obviously in G1 phase,and apoptosis peak appeared before G1 phase,apoptotic cell number was 3.4% for 24hour,and was 22.7% for 48hour of ATP treated U937 cells;(3) Nuclear chromatin condensed into sperical masses bound cytoplasma membrane formed apoptotic bodies which is shed from membrane surface into intercellular medium;(4) Apoptotic bodies were nigrosine staining negtive.

结果 (1)ATP对U937细胞的增殖有按摩明显的阻抑作用,加药48h后增殖的抑制率可达43%以上;(2)ATP处理的U937细胞周期发生改变,G1期细胞数按摩明显增多,ATP作用24h时G1期前出现亚二倍体峰——凋亡峰,凋亡细胞数为3.4%,作用48h时凋亡细胞数增多为22.7%;(3)ATP处理的U937细胞首先在核内染色质浓缩成半月形贴近核膜,逐渐向核膜外移动,进入胞浆内再移向质膜内外面,紧贴质膜外面再逐步脱离细胞体,进入细胞基质中,成为游离的凋亡小体。

Methods Cell culture,flow cytometry,HE staining,Nigrosine staining and electron microscopy were used. Results (1)ATP could inhibit the proliferation of U937 cells with inhibition rate over 43% after 48hour ATP treatment;(2)ATP treated U937 cells numbers increased obviously in G1 phase,and apoptosis peak appeared before G1 phase,apoptotic cell number was 3.4% for 24hour,and was 22.7% for 48hour of ATP treated U937 cells;(3) Nuclear chromatin condensed into sperical masses bound cytoplasma membrane formed apoptotic bodies which is shed from membrane surface into intercellular medium;(4) Apoptotic bodies were nigrosine staining negtive.

结果 (1)ATP对U937细胞的增殖有明显的阻抑作用,加药48h后增殖的抑制率可达43%以上;(2)ATP处理的U937细胞周期发生改变,G1期细胞数明显增多,ATP作用24h时G1期前出现亚二倍体峰——凋亡峰,凋亡细胞数为3.4%,作用48h时凋亡细胞数增多为22.7%;(3)ATP处理的U937细胞首先在核内染色质浓缩成半月形贴近核膜,逐渐向核膜外移动,进入胞浆内再移向质膜内外面,紧贴质膜外面再逐步脱离细胞体,进入细胞基质中,成为游离的凋亡小体。

Methods Peritoneal macrophages of guinea pigs were infected in vitro by three different Leptospira strains, the virulent Leptospira interrogans serovar Lai type strain Lai, the avirulent L. interrogans serovar Lai type strain IPAV, and the nonpathogenic L. biflexa serovar Patoc type strain PatocⅠ, respectively, and heat inactivated Staphylococcus epidermidis was added 0.5, 1.5, 3 and 6 h after infection and incubated for 30 min. The effect of Leptospira on the phagocytosis of macrophage was evaluated by the inactivated Staphylococcus epidermidis phagocytosis rate and phagocytosis index. Phagocytosis and ultrastructure of peritoneal macrophages were observed by transmission electron microscopy 3 h after infection, and changes of cytoskeleton of the macrophages were observed by laser scanning confocal microscopy.

用三种不同毒力的钩体(致病性问号钩端螺旋体赖型有毒株Lai株、赖型无毒株IPAV株以及非致病性双曲钩端螺旋体Patoc型PatocⅠ株)分别感染体外培养的豚鼠腹腔巨噬细胞,并分别于感染后0.5、1.5、3和6 h加入热灭活表皮葡萄球菌孵育30 min,通过计算巨噬细胞对灭活表皮葡萄球菌的吞噬率和吞噬指数,检测钩体对巨噬细胞吞噬功能的影响;感染后3 h透射电镜观察巨噬细胞对钩体的吞噬、降解和细胞超微结构的变化;用激光共聚焦显微镜观察细胞对钩体的吞噬和巨噬细胞细胞骨架的变化。

The present study is to culture otocyst cells in fetal, cochlear sensory epithelial cells of neonatal and mature respectively in vitro with refinement of culture media and techniques. Immunocytochemistry and Bromodeoxyuridine labeling are used to detect properties and mitotic status of cultured cells. With the observations about proliferation and differentiation in cultured cells, we want to find the origin of regenerative cells and to understand how these progenitors proliferate and differentiate.

本研究分别对胎鼠听囊、新生鼠听觉上皮和成年鼠听觉上皮用自然培养方法进行体外培养,并应用免疫细胞化学方法,对培养细胞进行染色标记和观察,系统了解哺乳动物听觉细胞发生、发育过程,以及这些细胞在体外的增殖和分化过程,旨在了解哺乳动物听觉细胞听觉上皮感觉细胞的来源,发现细胞生长的前体细胞

RESULTS A eukaryotic expression system for high expression humanmutantCD59 were successfully set up : The recombinant PALTER-MAX plasmid containing human mutantCD59 cDNA and PCDNA plasmid were co-transfected into CHO cell by cation lipoid mediating method ;and the cells were grown in F12 medium containing 400ug/ml G418 for 14 days, positive clones were grown in RPMI1640 medium to get stable expressing cell lines . Highly expressing clones were selected by flow cytometry ,and were named PALTER-CD59-CHO1PALTER-CD59-CHO2 . Flow cytometry indicated that expression rates of PALTER-CD59-CHO1 and PALTER-CD59-CHO2 were 53.7%and 54.5%. Further more, Stable highly expressing CHO cell lines were more detected by immunocytochemistry and immunofluorescence technology . PALTER-CD59 -CHO1 and PALTER-CD59-CHO2 were grown in RPMI1640 to get a large of cells . CD59 protein were obtained by spalling PALTER- CD59- CHO1 and PALTER - CD59 - CHO2 cells . Stable highly expressing cells were further validated by SDS-PAGE, immunoblot analysis and solid enzyme immunoassay . PALTER - CD59 - CHO1 and PALTER - CD59 - CHO2 were glycated in RPMI1640 of 50mM ribose for 72 hours .BCECF releasing test indicted that the releasing rate of PALTER-CD59-CHO1 or PALTER-CD59-CHO2 was less high than PALTER-CHO ,and the releasing rate of glycated PALTER-CD59 -CHOI or PALTER-CD59-CHO2 was higher than unglycated ones . PALTER -CD59-CHO1 and PALTER -CD59 -CHO2 were glycated in RPMI1640 of 50mM ribose for 72 hours .BCECF releasing test indicted that the releasing rate of PALTER - CD59 - CHOI or PALTER - CD59 - CHO2 was less high than PALTER-CHO ,and the releasing rate of glycated PALTER-CD59-CHO1 or PALTER - CD5 9-CHO2 was higher than unglycated ones .

结果 成功构建突变人CD59的真核细胞表达系统:运用阳离子脂质体介导法将含有突变人CD59的PALTER—MAX重组质粒与PCDNA共转染入CHO细胞:用含有400ug/mlG418的F12培养基培养14天,筛选出稳定阳性表达克隆,RPMI1640培养基扩增获得稳定表达细胞株,并用流式细胞术进一步筛选出高效表达细胞株分别命名为PALTER—CD59—CH01、PALTER—CD59—CH02,表达率分别为53.7%、54.5%;应用免疫组化方法、免疫荧光技术进一步鉴定阳性细胞株;RPMI1640培养基大量扩增PALTER—CD59—CH01、PALTER—CD59—CH02细胞株,裂解细胞得到CD59蛋白质;通过SDS—PAGE凝胶电泳技术、免疫印迹技术、固相酶联免疫吸附试验验证了这两中文摘要个阳性细胞株CO59蛋白的高效表达;50mM核糖培养72小时,获得突变人CD59糖化细胞株,BCECF染料释放试验结果显示,PALTER一CD59一CHOI、pALTER一CD59一CHOZ细胞较PALTER一CHO细胞染料释放率低,未糖化PALTER一CD59一CHOI、PALTER一CD59一CHOZ细胞比较糖化后细胞染料释放率低。

Results: The coelomocyte could be divided into 7 types: large grained cell, small granular cell, large hyaline leucocyte, small hyaline leucocyte, lanceolate cell, facultative cell and bacillary flutter. The concentration range of the coelomocyte was 5.62~10.00×10^6/ml. In the observation of phagocytize, pseudopod were obviously found and the phagocytic power was strong in the small granular cell.

结果:观察发现罗氏海盘车的体腔细胞可分成:大颗粒细胞、小颗粒细胞、大透明细胞、小透明细胞、柳叶形细胞、兼性大细胞和杆状颤动体;体腔细胞的浓度范围是:5.62~10.00×10^6个/mL。

The parameter of sphericity can be used to describe the similarity between centroblastic-centrocyte-centroblastic lymphoma cells and elips degree and regular form factor can describe the similarity between centroblastic-mantle-centroblastic lympoma cells.

圆球度可以用来描述中心母细胞--中心细胞--中心母细胞性淋巴瘤细胞之间的相似性;椭圆度、规化形状因子可以描述中心母细胞--套细胞--中心母细胞性淋巴瘤细胞之间的相似性。

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