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An additional micro-structure filter design is adopted in this chip to function as a single cell micro-channel for the purpose of centralizing the cells in a line. DNA or other nano-injectors could be pumped to the electroporation region and be inserted into the target cell via the castellated electrodes with several short electrical pulses.

以交流电动式微帮浦传送DNA或是染剂至On-Chip的电穿透实验区域,给予交错城堡状电极数个脉波的电压讯号,使得已被操控至定位的细胞其细胞膜产生暂时性的微通透孔隙,便能够将DNA或是染剂送入细胞中,观察其生物反应,此一技术将可提供生医医药研发及癌症(drug screening and cancer study)方面的研究应用。

To solve the problem and improve the safety of gene therapy, we used microencapsule, a insulation tool of cell transplantation immunity, to encapsule human fibroblastlike bone marrow stroma cells transfected by part of CEA gene and produced microencapsulated CEA transgeneic cell vaccines; we also studied the expression of vaccine in mouse body as well as its immunologic characteristics.

为解决上述问题、提高基因治疗的安全性,本研究采用细胞移植免疫用隔离工具-微胶囊包裹转染部分CEA基因的人成纤维样骨髓基质细胞HFCL,制备出CEA微囊化转基因细胞疫苗,并进行了疫苗在小鼠体内的表达及其免疫学性质的研究,探讨该疫苗应用于CEA阳性肿瘤治疗的可能性,并为进一步的临床应用研究奠定了基础。

CaN Aβ gene silencing can reduce myocardial hypertrophy in cultured cells, si1280 (21bp) of CaN Aβ gene is the most effective target site for siRNA. The method of intrapericardial injection of plasmid, microbubbles and erzymes can improve transfection efficiency of non-viral plasmid with satisfying targeted transfection. But the scope of transfected myocytes is still limited. CaN Aβ shRNA expressing plasmid transfection in vivo by pericardial injection results in decreased CaN Aβ protein expression of small part of myocytes, and CaN Aβ mRNA only shows decreased trend. The dosage of non-viral vector and the parameters of ultrasound energy should be optimized in further study.

结论RNAi技术抑制CaN Aβ基因表达可有效减轻Ald诱导的心肌细胞肥大程度;CaN Aβ基因中si1280(21bp)片段为实现RNAi的更有效片段;微泡+酶类心包腔内注射超声导入的方法可有效改善非病毒载体在体心肌的转染效率,同时具有一定的靶向性,但总的转染范围仍然有限;采用这一方法进一步进行&一肾一夹&心肌肥大模型大鼠在体心肌细胞的CaNAβ的RNAi干预,发现心肌肥大大鼠心外膜下局部心肌细胞CaN Aβ蛋白水平降低,CaN AβmRNA水平虽有下降趋势,但无统计学差异,提示质粒的用量及超声导入的参数有待进一步研究使其优化。

Adult rats are divided into control group, intrapericardial injection group and negative group, sublingual vein injection group and negative group. 6ds after transfection, issues of heart, liver, lung and kidney are stained with X-gal to observe transfection to myocardium and non-target transfection. 3. CaN Aβ gene silencing in vivo. Adult rats are divided into Control group, hypertrophy model group, intrapericardial injection group and negative group, sublingual vein injection group and negative group.

二整体动物水平上探讨心肌有效转染途径:以PLacZ为报告基因,将成年大鼠分为空白对照纽(生理盐水心包腔内注射后超声导入);转染质粒心包腔内注射组及其阴性对照组:质粒+微泡+酶的混合物心包腔内注射,超声导入;转染质粒舌下静脉注射组及其阴性对照组:质粒+微泡混合液经舌下静脉注射,超声导入。

The micronuclear frequencies of fetal liver blood were 2.67, 3.33 and 4‰ in treatment group, given intraperitoneally combined NET-OEN at doses of 10, 50 and 100mg/kg, respectively. However, the micronuclear frequencies of bone marrow erythroblasts and fetal liver blood in control group were 1.8~3‰.

作者采用微核试验观察了复方庚炔诺酮对小鼠活体骨髓多染性红细胞及胎肝血微核率的影响,检测其有无潜在的致突变作用,从而为评价复方庚炔诺酮的安全性提供依据。

Nosema disease is a kind of zoonoses, of all ages can be affected by pollution, for human carinii infection and host immune function of severely inhibited closely related to people before that Nosema without sexual reproduction period .

微孢子虫病是一种人兽共患病,各年龄组均可受染,人类为孢子虫感染与宿主的免疫功能严重地受到抑制有密切关系,以前人们认为微孢子虫无有性生殖期。

Fig.1 SHEE cultured on coverslide, the living cells were growing in single layer with rich cytoplasm, the nuclei were uniform in size with a nucleolus ph ×400 Fig.2 SHEE had a nucleus with ellipse shape, large nucleolus and the cytoplasm contained mitochondria and tonofibrilEM ×10 000 Fig.3 SHEE was spherical in shape, with pseudopods attached on petri dish and abundant villi on cell surface SEM ×5 000 Fig.4 Same as in Fig.3, cell attached on petri dish, appeared stellate or polygonal in shape, with abundant pseudopods and cytoplasmic processes. Protrusive nuclear region in central part of the cell had more micro-villi SEM ×5 000 Fig.5 Chromosomes of SHEE cells belonged to diploidy type Giemsa ×1 000 Fig.6 The SHEE cells of stained in dark brown by Ki67 immunohistochemistry were the proliferative cells Immunohistochemistry ×400 Fig.7 In SHEE cell culture, the nucleus stained red or pink by PI was dead cell, the green nucleus was living cell Fluorescent ×400 Fig.8 The cell labeled by TdT was apoptotic cell in which the chromatin of nucleus condensed in block, a pyknotic nucleus in the upper right conner was seen TdT labeled ×400

图1 SHEE培养在盖坡片上,活细胞单层生长,胞浆较丰富,细胞核大小一致,有核仁×400 图2 SHEE培养细胞细胞核椭圆形,核仁较大,胞浆有较丰富的线粒体和张力原纤维EM ×10 000 图3 SHEE细胞呈球状,有伪足贴壁,表面有密集微绒毛SEM ×5 000 图4 同上细胞贴壁,呈星状或多角形,有丰富伪足和胞浆突,核区隆起有较多微绒毛SEM ×5 000 图5 SHEE细胞染色体仍属二倍体Giemsa染色×1 000 图6 SHEE细胞Ki67免疫组织化学染棕黄色为增殖细胞×400 图7 SHEE培养细胞出现死细胞,胞核和胞浆PI染色呈红色或淡红色,蓝色细胞核为活细胞荧光显微镜×400 图8 细胞TdT标记阳性为凋亡细胞,染色质凝集呈块状,右上角有一固缩细胞核TdT标记×400

The mutagenicity of seed powder and seed oil of transgenic CryIA cotton to mammals and fish was evaluated in this study. The raw cotton seed powder or seed oil was mixed into mice and fish's diet. Cyclophosphamide and NaF were used as positive controls respectively. Regular cotton seed powder and seed oil were used as negative controls. The micronucleus frequency in mice bone marrow polychromatic erythrocytes and in eripheral erythrocytes of Zebrafish were assayed in this study. The sperm malformation rate in mice was also investigated.

摘 要:分别以环磷酰胺和氟化钠作为阳性诱变剂,以普通棉的棉籽粉及棉籽油作为对照物,同时设阴性对照组,以成年封闭群昆明种雄性小鼠和斑马鱼为实验材料,将棉籽粉或棉籽油掺入饲料中饲喂小鼠或斑马鱼,检测小鼠骨髓嗜多染红细胞(Polychromatic erythrocytes, PCE)微核率(Micronucleus frequency, MN‰)和精子畸变率,以及斑马鱼外周血红细胞微核率。

The mutagenicity of seed powder and seed oil of transgenic CryIA cotton to mammals and fish was evaluated in this study. The raw cotton seed powder or seed oil was mixed into mice and fish's diet. Cyclophosphamide and NaF were used as positive controls respectively. Regular cotton seed powder and seed oil were used as negative controls. The micronucleus frequency in mice bone marrow polychromatic erythrocytes and in eripheral erythrocytes of Zebrafishwere assayed in this study. The sperm malformation rate in mice was also investigated.

分别以环磷酰胺和氟化钠作为阳性诱变剂,以普通棉的棉籽粉及棉籽油作为对照物,同时设阴性对照组,以成年封闭群昆明种雄性小鼠和斑马鱼为实验材料,将棉籽粉或棉籽油掺入饲料中饲喂小鼠或斑马鱼,检测小鼠骨髓嗜多染红细胞(Polychromatic erythrocytes, PCE)微核率(Micronucleus frequency,MN‰)和精子畸变率,以及斑马鱼外周血红细胞微核率。

Results The extracted tubulin could assemble microtubules and form complex with tau in vitro.

结果电子显微镜负染显示纯化的微管蛋白在一定的实验条件下可聚集形成直径为25 nm的微管结构。

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