TGF-β1对不同肺癌细胞生物学行为的影响

用重组人转化生长因子β1（TGF－β1）处理不同的肺癌细胞95C、95D，研究TGF－β1对不同肺癌细胞迁移、粘附等生物学行为的影响，从而揭示肿瘤细胞的恶性行为机制。按Dean Sheppard方法测定细胞迁移能力；用划痕法和琼脂滴法测定细胞迁移能力；同时还测定了各肺癌细胞的铺展。结果表明：95C的迁移能力、细胞铺展率和增殖率最高，95D次之；用TGF－β1处理后，95C、95D与Fn的粘附率和铺展率较对照组都有显著增加，TGF－β1处理24、48h后可以明显增加两株肺癌细胞的迁移率。TGF－β1处理对两株肺癌细胞的增殖和生长无显著影响。提示不同肺癌细胞迁移和粘附能力存在差异，TGD－β1对肺癌细胞生长学行为存在显著影响。TGF－β1对所介导的信号转导通路起着重要的调控作用，从而影响肺癌细胞的若干生物学行为。     

单细胞克隆法快速筛选重组昆虫杆状病毒

首先采用显微操作系统挑取包含有重组病毒的单细胞，成功地获得了重组杆状病毒的单细胞克隆，并得到了重组病毒的纯培养，对有多角体的和无多角体的重组病毒细胞的挑取表明，这一方法对重组杆状病毒的筛选具有普遍适用性，我们发展的这一方法比常规的空斑技术简便、便速，只需两周左右即完成重组病毒的筛选。     

敏捷制造:企业实行科学、系统化经营的有效模式

针对企业在全球市场竞争中面临的挑战,介绍了敏捷制造模式及企业重组的管理思想．指出采用敏捷制造模式是企业实行科学、系统化经营,增强自身竞争力,快速适应严酷而多变的全球市场的有效方法.     

具有AAV载体包装功能的重组HSV的产生

腺病毒伴随病毒（ＡＡＶ）载体有望成为理想的基因治疗载体,常规的共转染法难以大规模制备重组ＡＡＶ（ｒＡＡＶ）,本研究生产了一种具有ｒＡＡＶ包装功能的重组单纯疱疹病毒（ｒＨＳＶ）,为ｒＡＡＶ的大规模制备提供了新的思路。采用一套含有１型单纯疱疹病毒（ＨＳＶ－１）全基因组的粘粒系统（Ｄａｖｉｓｏｎ ＡＪ ｅｔ ａｌ,１９９３）,该系统含有５个粘粒,依次称为ｃｏｓ４８,ｃｏｓ２８,ｃｏｓ６,ｃｏｓ１４和ｃｓ     

重组人脑钠素在大肠杆菌中的表达、纯化与鉴定

人脑钠素(hBNP,humanbrainnatriureticpeptide)是心室分泌的由32个氨基酸组成的多肽激素.临床研究表明,hBNP是治疗充血型心衰竭的一种安全有效的多肽药物.化学合成脑钠素全基因,以pET32a为表达载体,构建了硫氧还蛋白-脑钠素(thioredoxin＿hBNP)融合蛋白表达质粒,并转化至大肠杆菌BL21(DE3),经IPTG诱导,融合蛋白的表达量占全菌蛋白量的25%以上.融合蛋白经肠激酶裂解、Zn＿Sepharose亲和层析和C4反相高效液相层析,从每升培养液中可获取4mgrhBNP,纯度达到95%.经质谱测定,rhBNP样品的分子量为3464Da.体外活性测定结果表明,rhBNP样品对兔胸主动脉条具有显著的血管舒张效应,其ED50为(2.24±0 58)×10-6mg/mL.     

Immunogenicity of recombinant fowl-pox virus co-expressing structural protein precursor P1-2A and proteinase 3C of FMDV

Two recombinant plasmids, pUTA2P1 and pUTAL3CP1, were constructed by inserting structural protein precursor P1-2A and proteinase 3C of foot-and-mouth disease virus (FMDV) into fowl-pox virus (FPV) recombinant vectors pUTA-2 and pUTA-16-LacZ respectively, and two recombinant FPVs (vUTA2P1 and vUTAL3CP1) screened by the RT-PCR, IFA assay and Western blotting assay were obtained successfully. Mice injected respectively with rFPVs were induced high level specific anti-FMDV antibodies, increasing of T subtypes, and higher cytotoxicities of splenocytes than those of control groups. These results indicated that a new method was used to construct a potential candidate vaccine of FMDV.     

GFP标记的GDNF重组腺病毒载体的构建和体内外表达

通过构建绿色荧光蛋白(GFP)和胶质细胞源性神经营养因子(GDNF)共表达的腺病毒载体,讨论其在治疗脊髓和脑有关神经系统疾病的潜在应用价值,了解腺病毒载体在大鼠体内的分布及在体外分泌外源基因产物能力,并对其生物安全性作了初步考察,为GDNF重组腺病毒载体在临床和基础研究中的应用作了一定准备．     

国际互联网技术与旅游业企业过程重组

信息技术、互联网、通讯技术的发展引起了整个旅游业的一场革命,正在深刻地改变着旅游业的经营、管理和运作模式。网络技术的出现则恰好给旅游业的加速发展、企业过程重组带来了新的机遇。文章集中分析了旅游业的信息特征、信息技术、互联网技术引起的旅游业的经营环境变化,提出了旅游服务供应链管理的概念。在此基础上,结合英特网技术在旅游业的功能,提出了旅游业的企业过程重组,建立虚拟旅游公司模式。     

Recombinant PsbF from Synechococcus sp. PCC 7002 forms β:β homodimeric cytochrome b559

All organisms with oxygenic photosynthesis contain two photosystems:photosystem Ⅰ(PSⅠ)and photo-systemⅡ(PSⅡ),The minimal photosystem Ⅱ particles which are photochemically active contain three subunits:D1,D2 and cytochrome b559 (Cyt b559),The function of Cyt b559 remains unclear,We have successfully overxpressed the psbF gene,encoding the β subunit of Cyt b559,from a marine cyanobacterium Synechoccous sp.PCC 7002 as a fusion gene and obtained a redox-active from of Cyt b559,When the N-terminal GST protein of the fusion gene product was removed with thrombin ,the PsbF protein was still redox-active,suggesting that the recombinant PsbF can form dimer in Escherichia coli.The absorption spectra of either the oxidized from or the reduced form of both GST fusion protein and the purified PsbF dimer and the difference Spectra between the two forms are the same as that of the Cyt b559 isolated from the higher plants .Redox titration analysis of recombinant PsbF showed that the mid-point redox potential of the recombinant Cyt b559 was approximately 50 mV, which is close to the low potential of Cyt b559 ,The results are helpful to the understanding of locatlization and function of Cyt b559 on thylakoid membranes.Ⅰ     

Expression of recombinant human lysozyme in the milk of transgenic mice

Human lysozyme is a 130-aa (amino acid) alkaline polypeptide, and has both anti-bacterial and anti-viral properties which make it an important component of human natural immunity system. As a first step toward the ultimate goal of improving the anti-bacterial properties of bovine and ovine milk, a transgenic mouse that contains the genomic DNA sequence of the human lysozme gene has been generated for the first time. From 83 mice generated by microinjection, a total of 6 positive transgenic mice were identified by PCR and Southern blot. F1 mice positive for transgene in lines were also detected by PCR. This shows that transgene could be transmitted from founder transgenic mice to their offspring. Recombinant human lysozyme (rHlys) was found in the whey of 3 female positive transgenic mice by Western blot. The highest concentration of rHlys for transgenic mice was 0.2mg/mL. The antibacterial activity of the whey for transgenic mice was highly enhanced up to 0.4 times as much as that of human, while that of non-transgenic mouse was very low. Although the lysozyme activity of transgenic mice is still lower than that of human, the rHlys exhibits the same specific activity as that of human lysozyme. It provides a strong basis for further studies into the possible application of rHlys express in mammary gland.