核透明质

Changes of lung ultrastructures: The results of electron microscope showed that in group 1, there were swelling mitochondria was arranged in disorder, less matrix, and hyalomere appeared, thin double-deck membrane in 50% samples: Interalveolar septum stroma was with edema in 60% samples; Polymorphonuclear leukocyte gathered in vessel or emigrated out of vessel in 50% samples; In group 2, double-deck membranes of mitochondria were integral, densely matrix showed micro-granule shape in 90% samples; Pinocytosis in epithelial cells of type Ⅰ lung strengthened, being destroyed, and pinocytosis in endothelial cells strengthened in 10% samples; lnteralveolar septum stroma was with edema, no polymorphonuclear leukocyte gathered in vessel or emigrated out of vessel or corpuscule was empty in type Ⅱ alveolar epithelium in 20% samples.

肺组织超微结构改变观察：纯氧机械通气组50%标本线粒体肿胀、排列紊乱，基质变浅，出现透明区，双层膜变薄，60%标本肺泡隔间质水肿，50%标本多形核白细胞血管内聚集或游出血管外现象；34%氧浓度机械通气组90%线粒体双层膜较完整，基质致密呈细颗粒状；10%标本可观察到Ⅰ型肺上皮细胞有吞饮增强、破坏及内皮细胞吞饮增强；20%标本肺泡隔间质水肿；未见到多形核白细胞血管内聚集或游出血管外现象及Ⅱ型肺泡上皮板层小体排空。

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。

Observed at the ultrastructural level by electron microscope, hyalinocyte had no organelles and contained abundant euchromatin.

颗粒细胞比透明细胞大且多，染色深，但核质比小，在细胞质中有颗粒，有些颗粒细胞具双核。

Following the puncture of zona pellucida with two holes by injection pipette that contained donor nuclei or cells, the injection pipette was pulled back to the perivitelline space while the negative pressure was increased in the holding pipette until the polar body and karyoplasm were wiped off completely. Then a reconstructed embryo was completed by the direct injection of the donor nucleus or cell without pulling out the injection pipette.

首先以预先吸有细胞核或细胞的注射针在固定于持卵针上的卵母细胞透明带上穿刺两个孔，然后一边缓慢地将注射针回拔至卵周隙中，一边逐渐增加持卵针中的负压，直至极体与目标核质被完整吸入持卵针中而完成去核，最后在不拔出注射针的情况下直接注射细胞核或完整细胞进而完成重构胚的构建。

The porine is the polyembryony animal, many obtains the ovocyte from the porine ovary although but the maturity quite to be low; Because the ROSI technology is does not have to grow the mature sole round sperm cell to pour into directly completely in the ovicell nature, jumped over the spermatozoon in to pass through the physiology and the biochemistry, if the ovicell nature mature or the activation degree were insufficient, added the round immature sperm cell maturity quite inferior reason, very possibly Causes the ROSI micro fertilization defeat.

猪是多胎动物，从猪卵巢上获得的卵母细胞数量虽然较多，但成熟度比较低；由于ROSI技术是将没有完全发育成熟的单一圆形精细胞直接注入卵胞质内，跳越了精子在穿过透明带和卵质膜等过程中所发生的生理和生化反应；如果卵胞质成熟或活化程度不够，加之圆形未成熟精细胞成熟度比较差等原因，很可能造成ROSI受精的卵母细胞内部激活因子蓄积减少和生成不足，引起精核解聚困难、精圆核不能形成、第二极体不能排出、卵母细胞孤雌发育率增加等，使ROSI显微受精失败。

RESULT: 1.Ouabain act on lens sodium-pump,under the LM,lens anterior capsular membrane discontinuous,epithelium cells clustered,occluding zonule seperated,lens fiber layers fractured.Under the EM,cells totally hollowed,mitochondria swelling,myelin figure appeared.RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1、α2 and α3-isoform are all decreased.2.Digoxin act on lens sodium-pump,under the LM,lens cell oedema,linkage distructed,extensive exfoliation.Under the EM,plasma appeared little half-transparant hollow region,mitochondria swelling and ridge disappeared. RT-PCR examine,α1、α2 and α3-isoform are all decreased.3.Amphotericin B act on lens sodium-pump,under the LM,lens epithelium cells linked tightly,arranged in-line,lens fiber layers arranged tightly and regularily.Under the EM,abbundant cellular organes,exuberant cells function indicated. RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1 and α3-isoform are increased significantly,demonstrated isoform-specific action.4D-thyroxine act on lens sodium-pump,under the LM,lens plasmalemma integrated,cells arranged tightly and regularily.Under the EM,nucleus fission appeared,desmosome half-desmosome and tensile microfilaments linked the cells. RT-PCR examine,α2 and α3-isoform are increased, also demonstrated isoform-specific action.5.Vitamin E act on lens sodium-pump,under the LM,lens anterior capsular membrane continuous and smooth,epithelium cells tightly linked,lens fiber layers appearede hollow region occasionally.Under the EM,lateral membrane high density belt appeared,abundant nucleolus. RT-PCR examine,onlyα1-isoform are increased, demonstrated significantly isoform-specific action.6.DMSO act on lens sodium-pump,under the LM,lens anterior capsular membrane slightly thicker,cells linkage partly distructed.Under the EM,plasmalemma denaturation,mitochondria swelling.RT-PCR examine,α1、α2 and α3-isoform are all altered slightly and haven't significant meanning.

结果：1、哇巴因作用于晶状体钠泵后，光镜下晶状体前囊膜断裂、上皮细胞聚积、闭合连接分离、纤维板层裂隙，电镜下全层细胞空泡化、线粒体肿胀出现髓样结构，RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达均减弱。2、地高辛作用于晶状体钠泵后，光镜下晶状体细胞水肿、细胞连接破坏、广泛剥离，电镜下胞质见少许半透明空化区、线粒体肿胀嵴消失，RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达均减弱。3、两性霉素B作用于晶状体钠泵后，光镜下晶状体上皮细胞紧密连接、线状排列、纤维板层紧密规整，电镜下细胞器丰富、细胞功能旺盛，RT-PCR法检测α1及α3表达显著增强、具有一定的重整异构作用特异性。4、D甲状腺素作用于晶状体钠泵后，光镜下晶状体质膜完整、细胞排列紧密规整，电镜下胞核见分裂像、细胞间有桥粒、半桥粒及张力微丝，RT-PCR法检测α2及α3表达均增强、亦有一定的重整异构作用特异性。5、维生素E作用于晶状体钠泵后，光镜下晶状体前囊膜连续光滑、上皮细胞紧密连接、纤维板层偶见空化，电镜下囊侧膜内有高电子密度带、细胞核仁丰富，RT-PCR法检测仅有α1的表达显著增强、具有极强的重整异构作用特异性。6、二甲基亚砜作用于晶状体钠泵后，光镜下晶状体前囊膜轻度增厚、细胞连接部分破坏，电镜下质膜变性、线粒体肿胀，RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达无显著改变。

Under the transmission election microscope, we had observed that the state of oocytes arid the cumulus cells around them was tightness, and there were plenty of long and thin microvilli playing the role of connection in the enwrapping. The shape of cumulus cells nucleus was ovate, and the longer and shorter diameters of the cumulus cells were 4.250±0.042μm and 2.750±0.07 μm respectively. The zona pellucida was well proportioned and tightly linking with oocyte. There were great deals of multi-crista mitochondria, lipid droplet and vacuolus. The mitochondria longer and shorter diameters were 0.600±0.106μm and 0.490±0.117μm. Granular nucleoplasma distributed very well in the cell nucleus.

电镜下，卵母细胞外的卵丘细胞紧紧包裹卵母细胞，且与卵母细胞之间有大量细长的微绒毛相联系；卵丘细胞核呈卵圆形，其长径为4250±0.042μm，短径为2.750±0.071μm；透明带均匀，与卵母细胞结合紧密；在皮质区集中分布大量的多峰型线粗体、脂滴和空泡，线拉体长径为0.600±0.106μm，短径为0.490±0.117μm；细胞核中颗粒状的核质分布非常均匀。

I didn't watch TV last night, because it .

昨晚我没有看电视，因为电视机坏了。

Since this year, in a lot of villages of Beijing, TV of elevator liquid crystal was removed.

今年以来，在北京的很多小区里，电梯液晶电视被撤了下来。