Gene therapy for pathological scar with hepatocyte growth factor mediated by recombinant adenovirus vector

A complementary DNA (cDNA) encoding human hepatocyte growth factor was introduced into a replication-defective type 5 adenovirus (lacking E1, E3 domains) vector by homologous recombination of intracellular plasmid DNA, thus a recombinant vector containing HGF (Ad-HGF) was obtained. Ad-HGF and Ad-GFP (adenovirus vector carrying green fluorescence protein gene) were expanded in 293 cells and purified by cesium chloride gradient centrifugation for large-scale preparation, then were infected to the primarily cultured scar fibroblast of rabbit ear to observe the transfer efficiency and expression level of HGF in vitro. To evaluate the effect of Ad-HGF on established scar Ad-HGF solution was injected into excessively formed scar, which bears some clinical and histologic similarities to human hypertrophic scars. The results showed that: (i) the transfer efficiency was 36.8% ±14.1% on day 3 in primarily cultured scar fibroblasts treated with Ad-GFP and lasted more than 20 d; (ii) high-level expression of HGF protein was detected by means of ELISA in supernatant of scar fibroblasts treated with Ad-HGF, the amount of expression was 76 ng/4.0 x 10⁵ cells on day 3; (iii) on day 32 after a single intradermal injection of Ad-HGF at different doses (8.6 x 10⁹ pfu, 8.6 x 10⁸ pfu, 8.6 x 10⁷ pfu, 8.6 x 10⁶ pfu) per scar, most of the scars in the former two dose groups were dramatically flattened, some were even similar to that of the normal skin. The value of Hl (hypertrophie index) showed that there was a therapeutic effect of Ad-HGF on scars at the dose of 10⁹ pfu and 10⁸ pfu. Whereas no therapeutic effects were seen at lower dose (10⁷ pfu and 10⁶ pfu of Ad-HGF) groups. In addition, clusters of hair were observed to different extent on healed wound treated with Ad-HGF. Histopathologic examination revealed that in most healed wounds of Ad-HGF treated group, the dermal layer was thinner, the amount of fibrous tissue was much fewer, and hair follicles growth and sebaceous glands were observed. In Sirius red-stained sections the amount of type I collagen in the Ad-HGF-treated scars was diminished markedly, compared to that in Ad-GFP group, in which a huge amount of type I collagen was still observed; (iv) immune response against HGF was absent. Antibody against HGF was not detectable by ELISA in serum from rabbit treated with Ad-HGF; (v) no local or systemic side-effects and toxicity associated with the gene transfer were found. These results demonstrated the potential use of treating pathologic scar by Ad-HGF, an alterative strategy of gene therapy for scar in clinical practice.     

奶粉中蜡样芽胞杆菌污染状况及生化学和毒力试验

蜡样芽胞杆菌(Bacillus cereus,以下简称B.cereus)广泛分布于土壤、尘埃和水等自然界环境中,为需氧性芽胞杆菌,过去一直认为是一种腐败细菌。其致病性未得到重视。近年来,国内外由B.cereus引起的食物中毒较为多     

中国绵叶蚜属（同翅目：斑蚜科）研究及新种记述

本文了同翅目斑蚜科绵叶蚜属－新种肖朴绵叶蚜Ｓｈｉｖａｐｈｉｓ ｓｉｍｉｌｉｃｅｌｔｉ Ｚｈａｎｇ,Ｗ．Ｙ ａｎｄ Ｇ．ＸＺｈａｎｇ,ｓｐ．ｎｏｖ．和斯氏绵叶蚜Ｓｈｉｖａｐｈｉｓ ｓｚｅｌｅｇｉｅｗｃｚｉ Ｑｕｅｄｎａｕ,１９７９－中国新记录文中系统研究了该属的材料,给出了有翅孤雌蚜的分种检索表。     

肠源性氧化三甲胺在慢性肾病中的作用

氧化三甲胺是由人体正常消化产生并进入血液中的一种物质,一直以来被认为是正常的血液循环物质。但是近年来研究发现,氧化三甲胺具有生物活性,对多种慢性疾病都会产生不利影响。氧化三甲胺的产生主要依靠肠道菌群的代谢。在慢性肾脏疾病中,很多原因可导致肠道菌群发生改变,并且可以代谢生成氧化三甲胺的菌群种类明显升高,这就使慢性肾脏病患者血液中的氧化三甲胺含量上升。由于氧化三甲胺主要通过肾脏排泄,肾脏功能障碍可造成其在体内的积累,从而加剧慢性肾脏病患者的肾脏损伤。     

同义突变SND1慢病毒质粒的构建及回复表达

[目的]构建抵抗shRNA的同义突变SND1慢病毒质粒,并在SND1敲低的卵巢癌细胞系中回复表达SND1。[方法]依据密码子的简并性,针对shRNA识别的序列(TCTCGTCTCAAACTCTATTTG)设计同义突变序列(TAGGTATAACTTTAACCTGCT)并通过重叠延伸PCR的方法将同义突变序列引入SND1的编码序列中,得到recombination SND1(rSND1)目的片段,与pLVX-IRES-Hyg载体连接。测序验证pLVX-FLAG-rSND1质粒序列。pLVX-FLAG-rSND1质粒与包装质粒共转染293T细胞,获得携带FLAG-rSND1的慢病毒毒粒。用慢病毒感染SND1敲低的卵巢癌SKOV3-sh SND1-2细胞株,经潮霉素B筛选后,用Western Blot检测FLAG和SND1的表达。[结果]抵抗shRNA的真核pLVX-FLAG-rSND1重组慢病毒质粒构建成功,可检测到FLAG及SND1的表达。[结论]成功构建了同义突变SND1的pLVX-FLAG-rSND1质粒并在卵巢癌细胞中回复表达,为继续研究SND1蛋白在卵巢癌中的功能奠定了基础。     

新疆伊犁地区蒙古族苯硫脲尝味能力的测定

用 Harris 和 Kalmus 改良阈值法测定伊犁地区1,082名蒙古族的苯硫脲(PTC)尝味阈值。以4号液为味盲界线,在1,082人(男507;女575)中味盲者132人(男71;女61),占12.2%。男女之间或成年与儿童之间的味盲率均无显著差异。尝味者950人,占87.8%,全部950人尝味者的平均尝味阈值为9.38±1.65,其中成年女性的平均尝味阈值比成年男性的低0.52,男女少年儿童的平均尝味阈值比男女成人的低0.42,均有显著差异。Ⅰ—Ⅱ度甲状腺肿患者及吸烟者的 PTC 尝味阈值与上述平均值无显著差异,但绿色色盲者的 PTC 尝味阈值则较高。     

罗布麻（药）茶辐照灭菌研究

＾６０Ｃｏ－γ射线处理罗布麻茶,既可有效杀灭茶叶中的细菌又能保持其风味和药效。研究表明;６ＫＧｙ可作为灭菌工艺的最佳参考剂量。依此剂量批量处理的罗布麻茶,不仅灭菌彻底,而且主要成份无明显的变化。     

混合单克隆抗体ELISA直接法和夹心法检测包虫病人循环抗原

本文对混合单克隆抗体ＥＬＩＳＡ直接法和ＥＬＩＳＡ夹心法检测包虫病人循环抗原的效果进行了比较研究。结果表明,ＥＬＩＳＡ夹心法对细粒棘球蚴囊液纯化抗原的最低检出量（１．２ｎｇ／ｍｌ）较直接法（１０ｎｇ／ｍｌ）低,但检测包虫病人循环抗原时,ＥＬＩＳＡ直接法的阳性检出率为５６．３９％（２７／４８）,ＥＬＩＳＡ夹心法为６０．４％（２９／４８）,二者无显著性差异（Ｐ＞０．０５）。两法同时检测１４份猪囊尾蚴病人血清和４４份正常人血清,均未出现假阳性反应。     

甲基纤维素辐射降解及工艺研究

介绍了^60Co γ－射线对甲基纤维素的辐射降解作用,初步研究了辐射剂量、剂量率对甲基纤维素粘的影响。评价了甲基纤维素辐照后的贮存稳定性,并对辐射降解工艺进行了探讨。     

γ射线辐照灭菌剂量的模糊优化

γ射线辐照灭菌处理的关键性技术是辐照剂量的优化和确定。通过对照射后的SPF试验动物饲料的理化、微生物和感官指标的分析,提出γ辐照灭菌处理的模糊综合质量评价方法,进而实现对辐照剂量的模糊优化。