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Isolation of mammalian oocytes and early embryos is now a routine procedure, and there are many publications reporting oocyte and embryonic gene expression. However, the protocols used to isolate the oocytes and embryos are different in a number of respects, and it is not always clear whether protocols were identical for the different experiments. A further complication with respect to the isolation of mammalian oocytes and embryos is that a number of variables can affect both the quality of the oocytes and the ability of the oocytes to be fertilized. Because the development of these variables cannot be readily controlled for, they invariably exist in oocytes and early embryos, and because it is widely believed that the quality of the oocytes and early embryos affects the expression of genes and the development of the early embryos, it is important to establish whether or not these variables affect the gene expression and developmental ability of the early embryos. Because the expression of many genes in early embryos is controlled by the proximity of nuclei to the mitotic spindle, we have used indirect immunofluorescence to analyse the relative levels of proteins that comprise the mitotic spindle in human, mouse and bovine oocytes and early embryos. Oocytes were cultured for 2, 4, 6, 8 and 16 h and the relative levels of the mitotic spindle proteins were analysed. The results demonstrate that there was no obvious change in the level of the phospho-histone H3, MPM2, CDK1, cyclin B, securin, PRP8, ZW10 and ZWILCH proteins up to 4 h of in vitro culture. By contrast, after 6 h of in vitro culture there was a decrease in the relative levels of all the mitotic spindle proteins. The levels of all proteins tested decreased further after 8 h of in vitro culture, indicating that there was a loss of mitotic spindle integrity after this time. After 16 h of culture, the relative levels of all proteins tested increased, and it seems likely that the loss of spindle integrity and the failure to polarize the cytokinetic asters that normally occur after 16 h of in vitro

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To investigate the protective effect of baicalin on the prefrontal cortex synapse. Forty Sprague-Dawley rats were randomly divided into 4 groups: middle cerebral artery occlusion (MCAO) model group, baicalin group, neurogenin3 (NGN3) group, and control group. Each group included 10 rats. The MCAO model was established via suture embolism, and the occurrence of reperfusion arrhythmia was monitored. The hind-limb sensorimotor deficits were detected by using the modified neurological severity score (mNSS) at 3 h, 6 h, 12 h, 24 h, and 48 h after reperfusion. The pathological changes in the brain tissues were observed under optical microscope, while the ultrastructure of the prefrontal cortex was observed under transmission electron microscopy (TEM). The mNSS scores in MCAO group, baicalin group, and NGN3 group at 3 h, 6 h, 12 h, 24 h, and 48 h after reperfusion were 3.6 ± 0.4, 2.0 ± 0.5, 2.0 ± 0.5, 1.6 ± 0.6, 2.0 ± 0.5, 1.8 ± 0.6, 1.6 ± 0.5, 1.6 ± 0.6, 1.6 ± 0.6, 1.6 ± 0.6, 1.6 ± 0.5, 1.6 ± 0.6, and 1.8 ± 0.5, respectively, significantly lower than that in the control group with the scores of 5.8 ± 1.1 (F = 16.9, P = 0.000). The pathological changes in the brain tissues and the ultrastructural changes of the prefrontal cortex in the MCAO model were obviously in the model group at