髓源性防御素的研究进展

防御素是一组含有6个半胱氨酸的阳离子抗微生物多肽。在中性粒细胞内,防御素含量丰富,具有广谱、高效的抗微生物活性,是机体天然免疫机制的重要组成部分。近年来的临床观察表明,测定血浆及各种体液中髓源性防御素的改变有助于某些炎症性疾患的诊断。此外,提纯或重组防御素作为“超级抗生素”则已成为药学界的研究热点。本文着重对髓源性防御素的生化特性、生物学活性及其与疾病的关系作一概述。     

A novel and highly efficient production system for recombinant adeno-associated virus vector

Recombinant adeno-associated virus(rAAV) has proven to be a promising gene delivery vector for human gene therapy. However, its application has been limited by difficulty in obtaining enough quantities of high-titer vector stocks. In this paper, a novel and highly efficient production system for rAAV is described. A recombinant herpes simplex virus type 1(rHSV-1) designated HSV1-rc/△UL2, which expressed adeno-associated virus type2(AAV-2) Rep and Cap proteins, was constructed previously. The data confirmed that its functions were to support rAAV replication and packaging, and the generated rAAV was infectious. Meanwhile, an rAAV proviral cell line designated BHK/SG2, which carried the green fluorescent protein(GFP) gene expression cassette, was established by transfecting BHK-21 cells with rAAV vector plasmid pSNAV-2-GFP. Infecting BHK/SG2 with HSV1-rc/△UL2 at an MOI of 0.1 resulted in the optimal yields of rAAV, reaching 250 transducing unit(TU) or 4.28×104 particles per cell. Therefore, compared with the conventional transfection method, the yield of rAAV using this "one proviral cell line, one helper virus" strategy was increased by two orders of magnitude. Large-scale production of rAAV can be easily achieved using this strategy and might meet the demands for clinical trials of rAAV-mediated gene therapy.     

鹿衔草化学成分的研究：羟基肾叶鹿蹄草甙的结构鉴定

从鹿衔草（Ｐｙｒｏｌａ ｃａｌｌｉａｎｔｈａ Ｈ．Ａｎｄｒｅｓ）分离得到两个化合物。经光谱分析,确定其中１个化合物为新化合物,命名为羟基肾叶鹿蹄草甙（１, ｈｙｄｒｏｘｙｌｒｅｎｉｆｏｌｉｎ）,另一化合物为儿茶素（３）。     

Variation in alpha-L-fucosidase properties among 28 inbred mouse strains: Six strains have high enzyme activity and heat-stabile enzyme with a variant pH-activity curve; twenty-two strains have low activity and heat-labile enzyme

Alpha-L-fucosidase in tissues of 28 inbred mouse strains varied with respect to three properties: high or low heat stability, a pH-activity curve with high or low relative activity at pH 2.8, and high or low activity. Alpha-L-fucosidase from six strains (A/J, BDP/J, LP/J, P/J, SEA/GNJ, and 129/J) had high heat stability, high pH 2.8 relative activity, and high activity, whereas the other 22 strains all had low heat stability, low pH 2.8 relative activity, and low activity. The heat-stability difference was seen in all organs tested (brain, liver, kidney, spleen, heart, skeletal muscle, lung, and testis) for two heat-stabile strains (P/J and 129/J) and four heat-labile strains (C57 BL/6J, C3H/HeJ, DBA/2J, and BALB/cJ) studied in detail. The findings suggested that two structural variants of alpha-L-fucosidase, probably genetically determined, exist in these 28 inbred mouse strains, although the presence of linkage disequilibrium between alleles of tightly linked structural and regulatory genes could not be excluded.This work was supported by grants from the National Institutes of Health (NS-15281 and NS-11766), the Muscular Dystrophy Association (H. Houston Merritt Clinical Center for Muscular Dystrophy and Related Diseases), the March of Dimes Birth Defects Foundation, and a generous gift from the Alexander Rapaport Foundation.     

抗H9亚型流感病毒HA抗血清对小鼠的被动免疫预防和治疗研究

H9亚型流感病毒是引起大流感的潜在威胁.抗血清被动免疫是一种有效的应对流感的方法.采用H9N2禽流感病毒血凝素(hemagglutinin,HA)DNA重组质粒免疫BALB/c小鼠制备抗血清.在致死量同源病毒感染前或后通过尾静脉给小鼠注射不同剂量抗血清,观察小鼠14d内的体重丢失率和死亡率.结果显示,1280血凝抑制单位(hemagglutination Inhibition unit,HIU)抗血清可给小鼠提供至少长达11d的100%预防和1d的80%治疗保护.HA DNA疫苗免疫制备的抗血清可以有效地抵抗同源H9病毒的致死攻击,在H9病毒感染的预防和治疗中发挥作用.     

社鼠组织器官同工酶的研究

用聚丙烯酰胺凝胶等电聚焦电泳方法,分析了社鼠的肝、肾、心肌、骨骼肌、肺、脾和脑等多种组织器官的ＬＤＨ、ＡＤＨ、ＥＳＴ、和ＳＯＤ４种同工酶,对各组织器官的酶带数目和分布,以及酶活性进行了比较研究。结果表明,社鼠的ＬＤＨ同工酶、ＡＤＨ同工酶和ＥＳＴ同工酶具有比较明显的组织特异性,而ＳＯＤ同工酶的组织特异性较低。肺和脾除ＥＳＴ同工酶活性较高外,脑除ＬＤＨ同工酶活性较高外,其它３种同工酶的活性均较低;而肝和肾中４种同工酶的活性普遍很高。心肌和骨骼肌因氧张力不同而使ＬＤＨ同工酶酶谱存在明显差异,但其它３种同工酶酶谱却非常相似。同工酶的组织特异性与各组织器官所执行的生理功能是相一致的     

树载脂蛋白CIcDNA序列及其组织表达

分离提取树（ｔｒｅｅｓｈｒｅｗ,ＴＳ）肝组织ｍＲＮＡ,以此为模板,反转录构建了ＴＳ肝组织ｃＤＮＡ文库。利用制备的兔抗ＴＳ载脂蛋白ＣＩ多抗血清为探针筛选该文库,获得两个阳性克隆。测序及序列分析表明：两个克隆均为ＴＳａｐｏＣＩｃＤＮＡ基因。大的克隆由３８０个核苷酸构成,包括２１ｂｐ、９５ｂｐ组成的５′和３′非编码区,２６４ｂｐ组成的一个完整开放阅读框架,编码８８个氨基酸的ａｐｏＣＩ前体,含２６个氨基酸构成的信号肽和６２肽的成熟蛋白。其成熟肽长度与大鼠、小鼠和狗的相应结构一致,但较人和狒狒ａｐｏＣＩ的成熟肽多５个氨基酸。并对其功能域进行了初步预测。ＲＮＡ印迹显示ＴＳａｐｏＣＩｍＲＮＡ不仅主要在肝组织表达;而且不同于人和其他哺乳类动物,亦可少量在小肠表达。上述结果为进一步研究ＴＳａｐｏＣＩ基因组结构、功能和体外表达奠定了基础。     

内蒙古清水河县姑姑庵汉代墓地人骨研究

本文对内蒙古清水河县姑姑庵墓地出土的13例汉代的颅骨(男性6例, 女性7例)进行了体质人类学的研究, 该组颅骨在种族特征上可归入现代亚洲蒙古人种中的东亚人种范围。在若干古代和现代对比组中,姑姑庵汉代居民的体质特征与将军沟组、近代华北组最为接近,很可能是来自中原地区的移民或是当地早期中原移民的后裔。