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These results suggested that many organella could all develop into amyloplast, variation of amyloplast envelope resulted in amylopast proliferation, RER played an important role in synthesis and deposition of protein, ER showed many functions during endosperm development, intercellular space and plasmalemma invagination played an important role in transportation of nutrients, degeneration of starchy endosperm nucleus was a special PCD, Ca〓 and phosphatases played an important role during endosperm development.

上述结果表明:多种细胞器可以趋同发育成淀粉质体,淀粉质体增殖来源于其被膜的活动,粗面内质网在蛋白质的合成与积累中具有重要作用,内质网在胚乳发育中具有多种功能,胞间隙和质膜内陷在营养物质的运输中具有重要作用,淀粉胚乳细胞核的衰退是一种特殊形式的编程性死亡,Ca〓和磷酸水解酶在胚乳发育中起重要作用。

It is concluded that (1) there are no relationships between fusion rate and relativeness of the recipient cytoplasm to nucleus donor cells,(2) cytoplast of the goat MII oocyte can support the preimplantation development of SCNT embryos reconstructed with nucleus from other species,(3) the blastocyst rate of close relative inter-species SCNT embryos is higher than that of distant relative inter-species SCNT embryos.

由此结果得出以下结论:(1)山羊M II期卵母细胞胞质与供核细胞之间的亲缘性不影响两者的融合率;(2)山羊M II期卵母细胞的胞质能支持异种间体细胞核移植胚的着床前发育;(3)亲缘关系近的种间核移植胚的囊胚发育率高于亲缘关系远的种间核移植胚的。

The results showed that 1 there were no significant differences in the rates of cytoplast protrusion and enucleation between oocytes that were incubated in colchicine (0.4 μg/mL) for 0.5 h and oocytes that were incubated in colchicine (0.4 μg/mL) for 1 h, and the rate of cytoplast protusion can be 85.4% while the rate of cytoplast enucleation is 100%. 2 There was no significant difference in oocyte enucleation between oocytes treated with medium containing 0.2 μg/mL colchicine for 0.5 h and oocytes treated with medium containing 0.4 μg/mL colchicine for 0.5 h. 3 A maturation time of 18–23 h did not affect the rates of cytoplast protusion and enucleation by chemically assisted enucleation, whereas the rate of enucleation of oocytes by blind enucleation was found to decrease with a prolonged incubation time. 4 The development rates of reconstructed embryos could not be influenced by these two enucleation methods, increased from oocytes matured for 21–23 h.

结果表明: 1 卵母细胞在0.4 mg/mL的秋水仙素溶液中分别孵育0.5 h和1 h,胞质突起率和去核率没有显著的差异,突起率可高达85.4%,去核率达到100%; 2 0.2 mg/mL或0.4 mg/mL秋水仙素溶液将卵母细胞处理0.5 h,对去核效果没有显著影响; 3 对于体外成熟18~23 h的卵母细胞,随着成熟时间的延长,盲吸法的去核率降低,但没有影响秋水仙素诱导胞质突起的比率和去核率; 4 两种去核方法对重构胚的发育没有产生显著影响,但成熟21~23 h卵母细胞重构胚囊胚的发育率显著高于成熟18~20 h卵母细胞重构胚囊胚的发育率。

The three types of reconstructed embryos(reconstructed embryo of goat-rabbit, goat-bovine and goat-goat) were produced respectively by nuclear transfer using goat ear firbroblast cells as donors and rabbit, bovine and goat ooctyes as recipients, and the factors of cytoplast environments influencing cloned embryo development in vitro, cytoplast effect in nuclear transfer, the methods of transferring cloned embryo and the effects of pregnancy factor on implantation of cloned embryo were studied in this experiment, in order to supply basis for solving the difficulty of implantation of inter-species cloned embryo in recipient and improving the efficiency of cloning.

本试验以波尔山羊耳成纤维细胞为供体,分别以牛、羊、兔的卵母细胞为受体进行体细胞核移植,构建了山羊-兔、山羊-牛和山羊-山羊三种重构胚。研究了不同的胞质环境对重构胚体外发育的影响与核移植中的胞质效应,并对异种克隆胚胎采用了去透明带移植和配种后再移植的方法,探讨移植方法和妊娠因子对异种克隆胚胎在受体动物体内发育的影响,旨在为解决异种克隆胚移植不易着床发育的难题和为提高动物克隆效率提供理论依据。

The experiment results were as following:All cytoplast recipients derived from oocytes of rabbit, bovine and goat can support early development of cloned embryo, the fusion rate of reconstructed embryos of goat-bovine and goat-rabbit were 59.91% and 74.10%, their cleaved rate were 57.48% and 72.57%, blastocyst rate were 9.59% and18.06% respectively, and the fusion of reconstructed embryos of goat-rabbit and goat-goat were 60.01% and 74.10%, their cleaved rate were 56.68% and 72.57%, blastocyst rate were 11.76% and18.06% respectively.

试验获得了如下结果:1。以山羊耳成纤维细胞为供体,以牛、羊、兔卵母细胞做为胞质受体构建重构胚,并经体外培养,发现不同的胞质受体均能支持重构胚的早期发育,但受体胞质对重构卵的重构胚早期发育速度和卵裂率均有显著影响。

Seed vertical, brown, of same shape as utricle; embryo semi-annular; radicle inferior; perisperm scant.

垂直的种子,棕色,相同形状于胞果;胚半环形;胚根下位。外胚乳不够。

This result indicates that WT1 gene plays an important role in differentiation and development of fetal kidney and may be the factor that promotes metanephric blastemal cell to differentiate into epithelial cell.

结果显示小胎龄肾组织中WT1蛋白在胚基细胞和幼稚肾小球细胞核表达而大胎龄组肾组织中WT1在肾小管细胞胞浆表达,阳性率分别为57.1%(8/14)和46.2%(6/13),提示WT1基因在胚胎肾分化发育的过程中起着重要作用,WT1蛋白可能是促进后肾胚基细胞向上皮细胞分化的调控因子,其表达在时间上和空间上都受到严格的调控,WT1的表达异常可能导致胚基细胞分化停滞。17例肾母细胞瘤WT1蛋白表达阳性率为41.2%(7/17),阳性部位在胚基型和上皮型肿瘤细胞核,表达部位和阳性率与早期胚胎肾相似,其中间质型肾母细胞瘤均为阴性,胚基型和上皮型肾母细胞瘤阳性率70%(7/10),两组间阳性率有显著差异。

The vascular bundle of stem is the type of double phloem. The anther is tetrasporangiate. Simultaneous cytokinesis in the microspore mother cell follows meriosis.

茎的维管束是双韧类型;雄蕊的花药四室,小孢子的形成为同时型胞质分裂,悬垂胎座,倒生胚珠,核型胚乳,果实成熟时胚仍处于球形胚阶段。

On the termination date, the cultured explants were all examined by Western blot, HE and transmission electron microscope. Our results showed that after 12-days in culture, the cultivation treated with AS-ODN reduced the synthesis of AMBN and had a deformed dental cusp with thinner enamel matrix. Ultrastructure analyses showed that there was hardly any cisternae of the rough endoplasmic reticulum in the ameloblasts at the tip of the cusp of AS-ODN treatedexplants. However, on average the enamel matrix was thinner compared with that in the control group. Furthermore, the collagen fibers in extracellular matrix were found disorganized. These findings seemed to provide a direct experimental evidence that tended to indicate that the arrested AMBN translation in cultured tooth germs might result in the delay of the tooth development.

经用Western蛋白印迹检测表明,所设计的反义核酸对AMBN InRNA具有良好的封闭效果并成功阻断了牙胚对AMBN的表达;在缺乏AMBN情况下,与对照组相比,实验组牙胚在体外可以继续生长发育至钟状晚期,出现成釉细胞和成牙本质细胞的分化,成釉细胞可以分化成为分泌期型成釉细胞,胞浆中缺少合成蛋白质所必需的粗面内质网和高尔基氏体,缺乏溶酶体,表明对蛋白合成和脚的能力降低;实验组牙胚有牙尖形成和基质分泌,但牙尖形态异常,基质形成减少,牙尖周围基质最厚处为O.6卜m,明显薄于对照组的5.spin,基质中胶原纤维粗细不等,排列稀疏, 3 第四军医大学硕士学位论文未见钙化现象,充分证明了AMBN在牙胚发育中参与釉质基质形成和矿化过程,影响胶原纤维和牙本质基质的合成,促进成釉细胞对蛋白质的合成和釉质基质蛋白降解。

The histological observation showed that the larvae of S .japonicum in superinfected snails were sluggish in development, irregular, shrunken, deformed, relatively small in number as compared with the control. The sporocysts contained few germ balls or immature cercariae and most of the digestive glands of superinfected snails were atrophiej.

组织学观察发现各双重感染组钉螺消化腺萎缩,消化腺盲囊间隙只有少量血吸虫子胞蚴,血吸虫子胞蚴皱缩、变形及不规则,且子胞蚴中只含稀疏的尾蚴胚球,有的子胞蚴中已无胚球;而单独感染血吸虫的对照组中血吸虫均发育到成熟的子胞蚴或尾蚴。

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