中国地鼠卵巢细胞培养上清中重组骨形态发生蛋白2的三步纯化法(英文)

骨形态发生蛋白2(BMP-2)是转化生长因子β(TGF-β)超家族的成员,在骨组织工程领域被广泛应用.如今,BMP-2的商业生产所受关注程度与日俱增.这里建立了一种方便且低成本的方法,将人重组骨形态发生蛋白2(rhBMP-2)从中国地鼠卵巢细胞系(CHO)培养上清中纯化出来.该方法由三步组成:肝素亲和层析,Q-Sepharose离子交换层析和SephacrylS-200凝胶过滤,得率约29%.从1L培养上清中能得到约0.26mg的纯度在95%以上的rhBMP-2成熟二聚体,并通过异位成骨实验组织学分析证实了其生物活性.因此,该方法在CHO培养上清中纯化rhBMP-2方面具有潜在的应用价值.     

克隆世界的自我救赎

<正>十年前,黄禹锡正值研究事业的顶峰,却不料生命伦理行为中的造假和欺诈使得他的职业脱离原有的轨道。这个科学的弃儿能否自我救赎?韩国Sooam生物技术研究基金会位于首尔西南郊古罗区的一个林木茂密的山坡上。这个冬日寂静又寒冷,灰白的外表掩饰了山里各种活动的嘈杂,还有其中斯巴达式的生活。     

Production of human lysozyme-transgenic cloned porcine embryos by somatic nuclear transfer

Due to their physiology and organ size, pigs have significant potential as human disease models and as organ transplantation donors. Genetic modification of pigs could provide benefits for both agriculture and human medicine. In this study, five fetal pig fibroblast cell lines from two species (Wuzhishan and Landrace pigs) were transfected using double-marked human lysozyme (HLY) plasmids (pBC1-HLY-GFP-NEO) by a liposome-mediated method. The ratio of green fluorescent protein (GFP)-expressing cells was >95% in sw7, sw8, slw3 and slw6 cell lines, but only 49.3% in slw9 cells. Cells from the four highly transgenic lines were used as nuclear donors to construct embryos, which were then cultured after fusion and activation by electric stimulation. The rate of cleavage was 76.7%, 48 h after activation. After 7 days, 18.5% of cleaved eggs had developed to the blastocyst stage and 93.3% of blastocysts were GFP-positive. These results indicate that transgenic fetal pig fibroblast cell lines could be obtained by a liposome-mediated method, though the transfection efficiency varied between cell lines. Reconstructed embryos derived from transgenic cells could successfully develop into blastocysts, most of which were GFP-positive.     

葡萄耐盐细胞系的超策结构研究

运用透射电子显策镜技术研究葡萄耐盐细胞系和对照系的细胞超微结构。结果表明两类细胞系均由排列紧密的分生细胞团组成。但是，与对照系相比，耐盐细胞有几个显的超微结构变化：液泡较大；质体内积累淀粉粒，形成杯形变形质体；脂滴、高尔基体和平行于胞壁堆积的内质网明显增多，出现包绕着高密度核糖体的环形内质网，细胞壁内突生长，表面凹凸不平，胞间连丝丰富。这些结构改变可能与葡萄细胞的耐盐能力密切相关。     

Iron metabolism in the mononuclear phagocyte system

The maintenance of body iron homeostasis requires the coordination of multiple regulatory mechanisms of iron metabolism. The mononuclear phagocyte system （MPS, composed of monocytes, macrophages, and their precursor cells） is crucial in the maintenance of iron homeostasis. Recycling of iron is carried out by specialized macrophages via engulfment of aged erythrocytes. The iron stores of macrophages depend on the levels of recovered and exported iron. However, the molecular mechanisms underlying iron homeostasis in macrophages are poorly understood. Recent studies characterizing the function and regulation of natural resistance-associated mac- rophage protein 1 （Nrampl）, divalent metal transporter 1 （DMT1）, HLA-linked hemechromatosis gene （HFE）, ferroportin 1 （FPN1）, and hepcidin are rapidly expanding our knowledge on the molecular level of MPS iron handling. These studies are deepening our under- standing about the molecular mechanism of iron homeostasis and iron-related diseases.     

葡萄抗羟脯氨酸变异细胞系和抗盐变异细胞系的生化特性比较研究

对抗ＨＹＰ和抗盐的葡萄变异细胞系进行了比较分析，结果表明抗ＨＹＰ变异细胞的脯氨酸含量是抗盐变异系的２．８６倍。在盐胁迫下，抗ＨＹＰ变异系吸收的Ｋ＾＋和Ｎａ＾＋含量高于抗盐变异系，但二者的Ｋ＾＋／Ｎａ＾＋比值相近。实验表明在筛选葡萄抗逆变异系过程中，以ＨＹＰ作为选择压力更为适合。     

扬子鳄肝脏、心脏和肌肉细胞系构建

扬子鳄属国家一级保护的珍稀濒危动物. 细胞系的构建和保存是濒危野生动物离体保护工程的主要内容之一. 本研究构建了扬子鳄肝脏、心脏和肌肉细胞系. 结果表明, 从3 种不同来源组织块中原代细胞的生长速度表现出明显差异, 分别为11~12, 13~14 和17~18 d; 而传代细胞的生长速度在3 种组织来源的细胞之间没有明显差异, 均在6~7 d, 细胞倍增时间为36 h. 扬子鳄的染色体核型为2n = 32, 无明显的性染色体. 该研究所建立的3 种不同组织来源的细胞系将为扬子鳄的相关保护及研究工作提供可靠的材料平台, 也将为其他两栖爬行动物的细胞系构建提供有益参考.     

不同恶化程度胃癌细胞系的细胞间隙连接通讯功能及Cx43表达的差异

以人正常胃黏膜上皮细胞系GES-1和不同恶化程度的人胃癌细胞系(低恶化AGS细胞系、中恶化SGC-7901细胞系、高恶化BGC-823细胞系)为研究对象,采用激光共聚焦显微镜的荧光漂白恢复技术及划痕标记荧光染料示踪技术研究不同恶化程度的胃癌细胞系细胞间隙连接通讯(Gap junctional intercellular communication,GJIC)功能的差异;采用荧光定量PCR技术和间接免疫荧光技术检测间隙连接蛋白(Connexin43,Cx43)基因在mRNA和蛋白水平的表达差异,探求其与胃癌恶性程度的相关性.结果表明,在正常胃细胞系和低、中及高恶化胃腺癌细胞系中,细胞间的GJIC功能随着恶性程度的升高而减弱(AGS、SGC-7901)或消失(BGC-823),且Cx43蛋白及mRNA的表达量随着恶性程度的升高而下调(AGS、SGC-7901)或缺失(BGC-823).这提示胃癌恶性程度与胃癌细胞的GJIC功能显著相关(P<0.05),GJIC在胃癌发生中起重要作用.