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GFP gene eukaryotic express plasmids were transfected into ES cells. The cells were observed under the inverted fluorescent microscope. The clones which expressed the powerful green fluorescence were chosen to be named ES-GFP cells and cultured continuously. The examination of ES cell totipotent including Alkine phosphatase staining, embryonic body formation in vitro and teratomas formation in nude mice was implemented.

1。采用脂质体方法将GFP质粒转染ES-D3细胞株，经筛选后倒置荧光显微镜观察细胞形态及荧光状态，挑取荧光最强的克隆，命名为ES-GFP细胞，进一步扩增培养，观察细胞生长及荧光表达情况，行细胞分化全能性鉴定包括碱性磷酸酶（alkine Phosphtase，AKP）染色、体外胚胎体（embryonic body，EB）形成实验及裸鼠体内成瘤实验。2。

In view of continuous function problems, it is first proved that the-ES algorithm based on uniform distribution converges in probability in a weaker condition by means of the central limit theorem. Then it is proved that the-ES algorithm which adopts the general continuous random variables as its variable operators converges in probability.

针对连续函数优化问题，利用中心极限定理，在较弱的条件下，首先证明了基于均匀分布的-ES算法依概率收效，然后给出了采用一般连续性随机变量作为变异算子的-ES算法依概率收效的证明。

In view of continuous function problems,it is first proved that the-ES algorithm based on uniform distribution converges in probability in a weaker condition by means of the central limit theorem.Then it is proved that the-ES algorithm which adopts the general continuous random variables as its variable operators converges in probability.

针对连续函数优化问题，利用中心极限定理，在较弱的条件下，首先证明了基于均匀分布的-ES算法依概率收敛，然后给出了采用一般连续性随机变量作为变异算子的-ES算法依概率收敛的证明。

These culture conditions not only maintain the undifferentiated state and normal diploid karyotypes of BALB/c mouse ES cells effectively, but also maintain a series of their characteristics of murine stem cells. two different kinds of digestive methods and Two kinds of digestive juice with different concentrations were designed to dissociate proliferous inner cell mass and ES cell clonies deriver from dissociative ICM.

这一培养条件不但有效地维持了ES细胞的未分化状态和正常二倍体核型，而且维持了其作为多能性胚胎干细胞的一系列特征；实验设计了两种离散方法和两种浓度的消化液，用来离散增殖的ICM和ICM离散后出现的ES集落。

Type A band was detected in nearly all the organs and tissues,when coat color chimerism was over 85%, indicated the ES cellhas a fairly high potential to differentiate into cells of endoderm, mesoderm and ectoderm,In addition, only type A band was observed in the muscles of 6 mice which coat color chimerism is over 85%,the results indicated that these muscles were differentiated only from ES cells.

当毛色嵌合率大于85%时，几乎所有的器官组织都检测到A型条带，显示了ES细胞在发育形成内、中、外胚层的细胞方面具有很高的分化潜能。另外，在毛色嵌合率大于85%的其中的6只嵌合鼠的肌肉中，只观察到A型的条带，表明这些肌肉只单独来源于ES细胞。

The results indicated that:The multi\|locus DNA fingerprint with a new synthesized probe\|JL\|02,has enough polymerism and good stability,and should be very useful to monitor the chimerism in different tissues of ES cell chimeric mouse and to check whether an ES cell line has the capacity to enter the germ line,especially when involving strains that can not be discerned with coat color or biochemical markers.

结果表明：采用新型的人工合成的寡核苷酸多聚体探针(JL-02探针)的多位点DNA指纹图谱，具有足够的多态性和很好的稳定性。适用于鉴定ES细胞在嵌合体小鼠各种组织器官中的嵌合情况，也适用于鉴定ES细胞是否具有种系嵌合能力，尤其适用于在嵌合体研究中，最适宜的品种组合具有相同的毛色或相同的酶型。

Chapter 4 uses empirical analysis to test the convexity and subadditivity of VaR, ES and TCE, and test the validity of VaR and ES by back test in China stock market.

第四章——市场风险度量的实证比较分析采用实证分析方法，以我国股票市场的历史数据为样本数据，对VaR、ES和TCE进行凸性和次可加性检验；通过返回检验对VaR和ES两种风险度量在我国股票市场的有效性进行实证检验。

We evaluated 350 tumors: 136 ES, including 64 conventional ES, 64 proximal-type ES, and 8 with hybrid features of conventional and proximal-type ES; 54 metastatic carcinomas (22 from lung, 6 breast, 6 stomach, 5 colorectum, 5 kidney, 5 prostate, 5 pancreas); 12 metastatic testicular embryonal carcinomas; 20 metastatic melanomas; 20 epithelioid mesotheliomas; 20 epithelioid angiosarcomas; 10 epithelioid hemangioendotheliomas; 24 epithelioid malignant peripheral nerve sheath tumors; 22 myoepithelial carcinomas of soft tissue; 7 anaplastic large cell lymphomas; 5 histiocytic sarcomas; and 10 control MRT of infancy (4 brain, 3 liver, 2 soft tissue, 1 kidney).

我们总共研究了350例肿瘤：136例ES，其中包括64例寻常型ES、64例近端型ES和8例混有前两型特征的混合型ES；54例转移癌（22例来自肺、6例来自乳腺、6例来自胃、5例来自结直肠、5例来着肾、5例来着前列腺、5例来着胰腺）；12例转移性睾丸胚胎性癌；20例转移性黑色素瘤；20例上皮样间皮瘤；20例上皮样血管肉瘤；10例上皮样血管内皮瘤；24例上皮样恶性外周神经鞘膜瘤；22例软组织的肌上皮癌；7例间变性大细胞淋巴瘤；5例组织细胞肉瘤；10例作为对照的婴幼儿MRT（4例位于脑、3例位于肝、2例位于软组织、1例位于肾）。

After 6 to 7 days of culture, Eleven ES cell lines were selected and explanted. At last we obtained 4,3,4 and 3 ES cell lines separately. One of the B6C3F_1×B6D2F_1 ES cell lines was selected to future study its competent for germline transmission.We founded the ES cell line has normal karyotype on the chromosome analyse.In teratoma experiment by injected ESCs into C57 mices inguen, we gained teratoma contained cells come from all three embryonic germ layers.

选择一株具有高度杂合背景的B6C3F_1×B6D2F_1 ES细胞系进行核型分析、畸胎瘤实验后与ICR品系2n胚胎构建嵌合体并得到具有生殖系传递能力的嵌合体小鼠，证明此细胞系具有生殖系嵌合能力。

MATERIALS AND METHODS: Arg to Ser mutation was introduced into the 249 position of the p53 gene by knock-in method. These ES cells with this mutation were selected according to the homologues-recombination with PCR and Southern blot. The positive ES cells without a selection marker were injected into blastocysts recovered from Hprt(superscript -/-) mice, which were derived from Hprt-deficient ES cells. The injected blastocysts then were implanted into pseudopregnant females.

材料与方法：利用基因打靶技术在小鼠胚胎干细胞p53基因249编码子中引入点突变，使编码子249由精氨酸变成丝氨酸，然后将含突变的ES细胞显微注射到Hprt小鼠囊胚中，将注射过的囊胚植入假孕的雌性小鼠子宫，到第14d取小鼠胚胎纤维母细胞，用含HAT的培养液筛选出从ES细胞分化而成的鼠EF细胞，经测序证实细胞含有由249Arg到Ser的突变。

By using competitive adsorption of activated carbom to remove unfavorable influence of carbon in carbonaceous gold concentrate on leaching process, the leaching rate of gold concentrate can be improved to 88% as compared with 10%by direct cyanidation.

本文论述了利用活性炭竞争吸附作用，消除含炭金精矿中炭在浸出过程中的影响，使金精矿的浸出率可由直接氰化的10％提高到88％。

The only thing you're ever gonna be king of is King of the Stupids!

你只配做傻瓜的国王！