卵裂细胞

Fluorescence in situ hybridization was performed to detect the existence of HBV DNA in the blastomere by using the biotinylated probe.

废弃的胚胎卵裂球经激光打孔，活检2～4个卵裂球细胞，利用生物素标记的探针，应用荧光原位杂交技术检测HBV感染情况。

We also confirmed in this paper that it is injected somatic cells but not broken cytoplasm formed multi blastomere through mitosis in the reconstructed embryos.

在本实验中还通过用DNA染料（Hoechst-33342）对核移植体进行染色的方法，对融合后成纤维细胞在核移植体内不同时间的运行及发育模式进行了观察，确认了是成纤维细胞经有丝分裂形成多个卵裂球而不是由于细胞质碎裂形成的两个或多个碎块。

The mitotic error rate in cleavage stage embryos was proven to be higher than the meiotic aneuploidy rate and as a consequence, the genome of a single blastomere is not representative for the genome of the other cells of the embryo.

在卵裂期胚胎这种有丝分裂的错误率被证明是高于减数分裂的非整倍体错误率，因此，单个卵裂球的基因不能代表其他胚胎细胞的基因组。

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。

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。

The cleavation rates and blastocyst development rates of cloned embryos were used to assess the efficiency of different operational procedure. Finally, the best combination of operational procedure, that the spindle-viewer system was used for oocytes enucleating, and donor cell was electrofused into ooplasm by electrical pulse (1.9 kV/cm, 10 ms, two) to reconstruct bovine cloned embryos.

以核移植胚胎的卵裂率、囊胚发育率作为检测指标，对不同的方法所获得的克隆胚胎的卵分裂率与囊胚发育率进行比较，最后筛选获得1个优化的牛体细胞核移植操作程序，即采用Spindle view系统对牛卵母细胞进行去核操作，将供核体细胞注射到卵周隙，然后通过电融合法将供体核引入去核卵细胞质(电融合参数为1.9 kV/cm，脉冲时程10 ms，方波2次间隔2 s)。

Ovine immatured oocytes, matured oocytes and embryos from 2-cell stage to blastocyst produced in vitro were recovered, and RT-PCR was used to semi-quantify mRNA expression. Proteins of Cx43 and Cx45 were detected by using a combination of immunocytochemistry and Laser Scanning Confocal Microscope.

收集绵羊未成熟和成熟卵母细胞以及体外受精早期胚胎各发育阶段的卵裂球细胞，通过RT-PCR半定量检测Cx43和Cx45基因的mRNA表达量；通过免疫组织化学方法结合激光扫描共聚焦显微镜检测Cx43和Cx45蛋白在绵羊体外受精的早期胚胎发育过程中的表达情况及表达区域。

Embryo cells display a synchron cleavage rhythm during the cleavage stage, there is almost no gene transactivation, which is regulate and operated by the mRNAs and protein of oocyte.

卵裂阶段的胚胎细胞呈现同步化的分裂节律，鲜有基因转录活动，其发育由源自卵母细胞发生期间合成的蛋白质和mRNAs调控。

We biopsied 1-2 single blastomere from 6-8 cell cleavage stage intracytoplasmic sperm injection embryo, and using nested PCR amplified the high frequently mutation region G380R of FGFR3 gene of the single blastomere. The products of PCR were digested by restriction enzyme Bfm Ⅰ, then the digested products were detected by 10% PAGE to see whether the embryo inherted the mutation of the patient and to screen out normal embryo transfer.

活检经胞质内单精子注射(intracytoplasmic sperm injection，ICSI)授精的胚胎发育至6～8细胞期的1～2个单卵裂球，采用巢式PCR扩增单卵裂球的FGFR3基因的高发突变位点G380R区域，用限制性内切酶BfmⅠ消化巢式PCR的内扩增产物，再经10%聚丙烯酰胺凝胶电泳检测有无遗传患者的该种突变，从而筛选出正常胚胎移植。

Resembled to control oocytes, MAPK was inactivated after cytoplasts activation. However, after activation the cytoplasts showed fragmentation while pronuclei were formed in the control oocytes. This indicated that nuclei play important role in maintainence of oocytes normal morphology.

成熟胞质的活化与卵母细胞相似，在活化后MAPK被迅速灭活，只是胞质在活化处理后均发生碎裂，而卵母细胞活化后形成原核，这表明核在细胞形态维持中起重要作用。

On the other hand, the more important thing is because the urban housing is a kind of heterogeneity products.

另一方面，更重要的是由于城市住房是一种异质性产品。

Climate histogram is the fall that collects place measure calm value, cent serves as cross axle for a few equal interval, the area that the frequency that the value appears according to place is accumulated and becomes will be determined inside each interval, discharge the graph that rise with post, also be called histogram.

气候直方图是将所收集的降水量测定值，分为几个相等的区间作为横轴，并将各区间内所测定值依所出现的次数累积而成的面积，用柱子排起来的图形，也叫做柱状图。