激素和添加物对蝴蝶兰组培快繁的影响

以蝴蝶兰人工授粉的果荚种胚和组培苗的叶片、茎尖为外植体,进行组织培养快速繁殖研究.结果表明,果荚种胚是最好的外植体材料,1个果荚可培育出上万个原球茎.种胚诱导原球茎最佳激素组合为:6-BA 0.3mg/L+NAA 0.2 mg/L;原球茎增殖最佳激素组合为:6-BA 0.5 mg/L+NAA 0.2 mg/L;2%蔗糖和10%香蕉泥可以显著促进原球茎增殖;原球茎分化最佳激素组合为:6-BA 1.0 mg/L+NAA 0.2 mg/L;诱导生根IBA 0.5mg/L最佳;炼苗后移栽苗成活率达95%以上.优化后的蝴蝶兰组织培养条件,可为蝴蝶兰快速繁殖工厂化生产提供参考.     

Aberrant DNA methylation patterns in cultured mouse embryos

Mouse early embryos undergo genome-wide demethylation and remethylation events during pre-implantation development. Abnormal methylation reprogramming is thought to be associated with development arrest. Using immunofluorescence staining with an antibody against 5-methylcytosine (MeC), we examined the genome methylation patterns of mouse embryos cultured in vitro. The results did not show the difference in staining patterns between development-blocked two-cell embryos that cultured in vitro and the two-cell embryos that were freshly collected from the donor mice. But in vitro-arrested morulae displayed a strong positive staining when compared to the morulae freshly collected from the donor mice. At the blastocyst stage, although most embryos showed the expected methylation patterns, with highly stained inner cell mass (ICM) and weekly stained trophectoderm (TE), a proportion of embryos were dimly stained in both ICM and TE. These results indicated that the methylation profile of the embryos could be changed by culturing in vitro when the embryos were in the transition from morulae to blastocyst.     

利用玉米胚芽渣制取植酸

利用玉米胚芽渣为原料，采用改良氨菲丁法制取植酸，结果表明，１７ｋｇ玉米胚芽渣可获得５０％植酸水溶液０．３２ｋｇ，其质量符合国内外相应标准，可使玉米胚芽的经济效益提高４０％以上。     

Effect of the calmodulin inhibitor R24571 (calmidazolium) on rat embryos cultured in vitro

Summary The possible effects of inhibition of the calcium-binding protein, calmodulin, on mammalian morphogenesis have been investigated by culturing rat embryos in vitro from 9 1/2 to 11 1/2 days of development in the presence of R24571 (calmidazolium), a specific inhibitor of calmoldulin. Embryos cultured in 10^–2 mM R24571 for 48 h show inhibited development and exhibit a range of morphogenetic abnormalities including assymetry and neural tube defects. Embryos exposed to R24571 for the first 24 h of a 48 h culture are more severely affected than embryos exposed to R24571 for the last 24 h.Acknowledgments. We wish to thank Mr C. Veltkamp and Mr B. lewis for their expert help with the scanning electron microscopy and photography.     

外源视黄酸对小鼠早期胚胎形态及Otx2基因表达的影响

以全反式视黄酸（ａｌｌｔｒａｎｓ－ｒｅｔｉｎｏｉｃａｃｉｄ，ＲＡ）体外处理ＥＢ（ｅａｒｌｙａｌｌａｎｔｏｉｃｂｕｄ）期和ＬＢ（Ｌａｔｅａｌｌａｎｔｏｉｃａｃｉｄ）期的小鼠胚胎，然后用洋地黄毒苷（ｄｉｇｏｘｉｇｅｎｉｎ）标记的Ｏｔｘ２反意义ＲＮＡ探针对整体胚胎进行原位杂交．０．５×１０－６ｍｏｌ～２×１０－６ｍｏｌ的ＲＡ处理ＥＢ和ＬＢ期胚胎１２ｈ后对其形态学和Ｏｔｘ２表达都没有影响；以４×１０－６ｍｏｌ的ＲＡ处理，抑制或减少了胚胎的前肠形成，经处理的胚胎的头褶没有对照的那样明显向腹面突出，其神经沟也不整齐；以４×１０－６ｍｏｌ的ＲＡ处理ＥＢ期的胚胎，其ＯｔＸ２表达范围剧烈地向前退缩或只有很微弱的表达；但以同样浓度处理ＬＢ期胚胎时，与对照胚胎相比，ＯｔＸ２的表达除了在极少数胚胎中变得弱一些外，在绝大多数胚胎中其表达模式没有变化．     

小鼠胚胎移植技术优化及对巨细胞病毒净化研究

目的 建立小鼠巨细胞病毒净化方法,以获得无MCMV感染的小鼠。方法 利用胚胎移植技术,通过使用不同激素、不同发情周期注射激素、受体鼠品系、不同的移植方法、不同时期胚胎移植,以及移植胚胎数量等对比试验,优化了净化条件。利用优化的胚胎移植净化方法,对供体鼠进行了净化。结果 SIGMA生产的激素,在10 IU剂量下超排得到的可用胚胎数量约为17枚/只;在小鼠发情间期超排得到的可用胚胎最多,超排的可用胚胎数约为23枚/只;选取C57雄性小鼠与ICR雌性小鼠交配的子一代作为受体鼠,产仔率达44.5%;输卵管移植较子宫移植效果好,产仔率为40.64%;移植2细胞胚胎的妊娠率为80%,明显优于单细胞和8细胞;受体鼠移植24枚胚胎时,产仔率达到了43.75%。利用优化的小鼠巨细胞病毒胚胎移植净化方法,净化得到了无MCMV感染的小鼠。结论 建立了小鼠巨细胞病毒净化方法,为获得无MCMV感染的小鼠种群提供了可靠的保障。     

Combinational electroporation and transplantation approach to studying gene functions in avian embryos

Gene transfection is an indispensable approach for studying gene function since it provides important information on gain- and/or loss-of-function. Chick embryos are also extensively employed for studying bio- logical function since they are easily accessible and can be maintained alive after manipulation. The combination of both techniques presents a powerful approach to under- standing how genes regulate embryo development. Fur- thermore, combining these approaches with tissue transplant techniques make even more attractive for elu- cidate gene function. Electroporation, employing parallelly fashioned electrodes, has been widely used in chick embryos. However, experimenters have been frustrated by unsuccessfully transfection in some embryonic tissue of interest because the electrodes were improperly positioned.We presently demonstrated the different patterns of orga- nizing and positioning the electrodes, in combination with tissue transplantation, to efficiently and specifically trans- fect the chick embryonic head, trunk neural tube, heart tube, somites and neural crest cells with the GFP reporter gene.     

伽马辐照对斑马鱼胚胎谷胱甘肽还原酶活性及基因表达的影响

谷胱甘肽还原酶(GR)是一种重要的抗氧化酶,其在生物体的抗氧化系统中起着关键作用.为了研究伽马辐照对斑马鱼胚胎GR活性及基因表达的影响,本实验用不同的辐照剂量(0.01、0.05、0.1、0.5和1 Gy)分别对三个发育时期(原肠胚期、体节期和咽囊期)的斑马鱼胚胎进行了辐照处理.分析了各组GR的活性,并采用荧光实时定量PCR技术(qRT-PCR)检测了GR基因的表达水平.结果显示,伽马辐照以剂量依赖的方式调整了斑马鱼胚胎GR的活性及GR基因的表达.随着辐照剂量的增加,GR活性及其mRNA的相对表达量呈现先升高后回落的趋势.这表明,GR是一种可以对伽马辐照作出早期预警的生物标志物.斑马鱼胚胎可以通过上调GR基因的表达,来增加GR的合成,从而提高对伽马射线的防御能力.但是,这种反应和调整是有限度的.咽囊期胚胎比其它两个时期的胚胎对伽马射线的防御能力更强.     

Effects of free sterols on the fluidity of polar lipids in rice dry embryo

Free sterols from rice dry embryo reduce the onset phase transition temperature and widen the transition range of total polar lipids and phosphatidylglycerol, which increases the membrane fluidity below their phase transition temperature and decreases the fluidity of membranes above their phase transition temperature. And 50% of free sterols result in a complete disappearance of phase transition. The fluidity of other main components of polar lipids is decreased by free sterols above 0℃. The results confirm that free sterols can modulate membrane lipid fluidity.