Inhibitory effect of a modified adenovirus type 5 E1A gene on the NF-kB activity in porcine aortic endothelial cells induced by TNF-α

During the DXR process of xenotransplantation, the endothelial cells (EC) are activated, and the expression of NF-kB is immediately up-regulated[1]. NF-kB consisting of p65/p50 heterodimers or homodimers are retained in the cytoplasm by association with IkB (inhibitor of NF-kB) proteins. After stimulation, IkB is degraded, re-leasing NF-kB from these trimeric complexes, and allow-ing NF-kB translocation to the nucleus to play its biology function[2,5]. The promoter and enhancer region…     

胞内表达猪生长激素基因的重组酵母菌构建与鉴定

将猪生长激素(pGH)cDNA定向插入毕赤酵母胞内表达载体pPIC 3.5k质粒，重组构建了pPIC 3.5kpGH/GS115酵母表达系统, 其表达产物的SDSPAGE和Western blot结果表明, pGH蛋白在胞内获得正确表达，表达量达378 mg/L.     

猪β干扰素基因17位丝氨酸突变体重组酵母表达载体的构建

根据猪β干扰素基因序列设计一对引物,并按酵母密码子的偏好性,在引物中对稀有密码子进行同义突变,即第3位的TAT基因突变为TAC,第7位的CGA基因突变为AGA,第164位的CGG基因突变为CGT。同时,利用聚合酶链式反应基因定点突变技术,把17位的半胱氨酸密码子TGT突变为丝氨酸密码子TCT,构建poIFNβSer17...     

猪源纤维蛋白粘合剂对大鼠皮肤创伤的愈合及羟脯氨酸表达的影响

【】 目的 为促进皮肤创伤愈合提供实验依据,最终为临床新应用提供依据,探究猪源纤维蛋白粘合剂(FS)其对大鼠皮肤创伤的愈合及其对创面皮肤羟脯氨酸表达的影响,。方法 72只体重为160-200 g SD雄性大鼠随机分为空白对照组、白凡士林组和FS组,于背部制作直径为1.5 cm的全层圆形皮肤切除创面,每组各24只,空白对照组不做任何处理,白凡士林组和FS组则分别用医用白凡士林和FS涂抹伤口。术后3 d、7 d、14 d和21 d测量各组大鼠的皮肤伤口面积,计算各组皮肤创面愈合率,取术后7d和14d伤口皮肤标本进行HE病理组织学观察;并于术后7d、14d和21d取组织进行酶联免疫法测定愈合组织中羟脯氨酸蛋白含量。结果 在术后3 d、7 d、14 d和21 d时,FS组的伤口愈合率高于空白对照组,且其差异具有统计学意义(p<0.05)。HE染色结果显示在第7d时FS组胶原沉积高于空白对照组,血管新生数亦高于空白组、白凡士林组,差异均具有统计学意义 (p<0.05);在14 d时,与空白对照组相比,FS组胶原沉积较高且差异具有统计学意义(p<0.05),而血管新生数各组之间无统计学意义(p>0.05)。对各组创面组织中HYP含量进行测定,结果发现,在第7 d、14 d和21 d时,FS组大鼠皮肤创伤组织中HYP含量均明显高于空白对照组 (p<0.05)。结论 猪源纤维蛋白粘合剂能有效促进大鼠皮肤创伤的愈合,缩短愈合时间,其具有潜在的应用价值,为临床皮肤创伤治愈提供了新思路。     

Adhesion of subsets of human blood mononuclear cells to porcine endothelial cells

Cellular immune response is a major barrier to xenotransplantation, and cell adhesion is the first step in intercellular recognition. Flow-cytometric adhesion assay has been used to investigate the differential adhesions of monocyte (Mo), natural killer cell (NK) and T lymphocyte (T) present within human peripheral blood mononuclear cells (PBMC) to porcine aortic endothelial cells (PAEC), and to demonstrate the effect of human interferon-γ (hIFN-γ) or/and tumor necrosis factor-α (hTNF-α) pretreatment of PAEC on their adhesiveness for different PBMC subsets. The preferential sequence for PBMC subset binding to resting PAEC is Mo, NK and T cells, among which T cells show the slightest adherence; hTNF-α can act across the species, and augment Mo, NK and T cell adhesion ratios by 40%, 110% and 3 times, respectively. These results confirm at the cell level that host Mo and NK cells are major participants in the cellular xenograft rejection, thereby, providing a prerequisite for further studying the human Mo/NK-PAEC interactive mechanisms.     

猪胃蛋白酶原A在毕赤酵母中的高效表达及其分离纯化

尝试利用毕赤酵母(pichia pastoris)表达体系大量生产制备猪胃蛋白酶,用化学的方法合成猪胃蛋白酶原A基因,并构建表达载体pGAPZα-pepsinogen;利用化学转化技术,将该基因整合到巴斯德毕赤酵母x-33菌株的染色体上.经过发酵,三角摇瓶培养条件下获得的猪胃蛋白酶原A总酶活达5 250U/L.发酵液上清经过DEAE离子交换色谱和Sephacryl S-200分子筛色谱分离,活化酶原后测定其比活达到52 800 U/g.