Kinetics of retroviral production from the amphotropic ΨCRIP murine producer cell line

Rapidly expanding development and practice of gene therapy requires the availability of large quantities of high titer retroviral supernatants. One way to achieve high retroviral titers is through improved understanding of the kinetics of retroviral production and decay, and the subsequent development of improved cell culture methods. In the present study we investigated the effects of different operational modes on the retroviral production of the NIH 3T3 fibroblast derived amphotropic murine retroviral producing cell line pMFG/CRIP. Semi-continuous culture (exchange of 50% of medium volume daily) was found to promote cell growth and enhance retroviral production. The rapid medium exchange resulted in significantly larger amounts of high titer supernatants and an extended production phase as compared to the batch control cultures. The specific viral productivity of the pMFG/CRIP cells was in the range of 10 to 40 infectious viruses produced per thousand producer cells per day. The CV-1 African Green Monkey kidney cell line was used as the infection target. Lowering the serum level form 20% to 10% improved retroviral production slightly. However, at lower serum levels (1%, 5% and 10% (v/v)) growth of the producer cell line, and thus retroviral production, was directly proportional to the serum level. The half-life of the virus at 37°C was found to be 5.5 hours. Promoting the growth of producer cell lines can improve retroviral vectors titers and viral production. High cell density systems that allow for rapid cell growth and waste product removal are likely to be used to generate high-titer retroviral supernatants.     

包囊游仆虫休眠包囊的超微结构研究

包囊游仆虫休眠包囊中,各类纤毛器的纤毛基体上方的大部分纤毛杆退化,或仅保留毛基体,有时部分额腹棘毛的毛基体也瓦解消失。残留纤毛的纤毛杆周围微管和中央微管仍具有“9 2”结构特征,也有少数纤毛杆出现2套“9 2”微管共处于一层纤毛膜内的现象。毛基体中周围三联体微管的中央形成微管形结构聚合体,基体附属结构仅存在基体间连接及纤毛器托架的残余物;非纤毛区皮层表膜下未见微管层。纤毛区皮层含纤毛器腔周围微管层(相当于表膜下微管层)、纤毛器深部及附近的微管束和分散的微管群。并且,纤毛区皮层囊泡内含有呈不同形态的纤毛杆结构;大核核孔明显变大,核孔数目减少,核孔内膜附着染色质。     

牛病毒性腹泻病毒的成熟和释放

试验中用电镜观察了牛病毒性腹泻病毒ＯｒｅｇｏｎＣ２４Ｖ株在感染新生牛睾丸细胞中的形态发生。成熟的病毒颗粒是直径约为５０ｎｍ的球形颗粒,内含直径约为３０ｎｍ的核心。病毒在宿主细胞的胞质内复制,通过糙面内质网膜出芽成熟。病毒可以通过外排或在细胞死亡后含有病毒颗粒的空泡崩溃而释放到胞外。     

中国茶对变形链球菌的抑菌作用

对３６只ｓｐｒａｑｕｅ－Ｄａｗｌｅｙ鼠分４组给予诱龋饲料（Ｄｉｅｔ２０００,ｋｅｙｓ）,并连续５天在口腔内接种变链菌６７１５菌株,分别给予我国主要产茶区的三种茶浸液：龙井绿茶（Ｂ组）,四川涪陵红茶（Ｃ组）,乌龙茶（Ｄ组）,先用０．５ｇ茶和５０ｍｌ双蒸馏水加热煮沸过滤后浸泡１０分钟以及对照组（Ａ组）用双蒸馏水为饮料喂养实验鼠。实验６０天后,杀鼠观察计算鼠磨牙面的菌落形成单位（ＣＦＵ）和菌斑指数（ｐｌａｑｕｅｓｃｏｒｅ）均显著下降。实验结果,实验组和对照组菌斑指数下降值顺序如下：Ｃ组＞Ｄ组＞Ｂ组＞Ａ组,并与实验组含氟量有相关性。很多研究表明变链菌是较为肯定的致龋菌、茶中氟化物对变链菌生长、粘附都有抑制作用。本研究选用茶来源广泛、价廉、无毒安全等优点,为开拓新的防龋方法进行探索。     

Normal and expanded Huntington's disease gene alleles produce distinguishable proteins due to translation across the CAG repeat.

BACKGROUND: An expanded CAG trinucleotide repeat is the genetic trigger of neuronal degeneration in Huntington's disease (HD), but its mode of action has yet to be discovered. The sequence of the HD gene places the CAG repeat near the 5' end in a region where it may be translated as a variable polyglutamine segment in the protein product, huntingtin. MATERIALS AND METHODS: Antisera directed at amino acid stretches predicted by the DNA sequence upstream and downstream of the CAG repeat were used in Western blot and immunohistochemical analyses to examine huntingtin expression from the normal and the HD allele in lymphoblastoid cells and postmortem brain tissue. RESULTS: CAG repeat segments of both normal and expanded HD alleles are indeed translated, as part of a discrete approximately 350-kD protein that is found primarily in the cytosol. The difference in the length of the N-terminal polyglutamine segment is sufficient to distinguish normal and HD huntingtin in a Western blot assay. CONCLUSIONS: The HD mutation does not eliminate expression of the HD gene but instead produces an altered protein with an expanded polyglutamine stretch near the N terminus. Thus, HD pathogenesis is probably triggered by an effect at the level of huntingtin protein.     

c-fos expression in the amygdala:In vivo antisense modulation and role in anxiety

Summary 1. The amygdaloid complex is a key structure in mechanisms of fear and anxiety. Expression of the immediate-early gene c-fos has been reported in the central nucleus of the amygdala following various stressors, but the functional role of this phenomenon has remained unknown.2. c-fos expression was observed in the central nucleus when rats were subjected to a pharmacologically validated animal model of anxiety, the Vogel conflict test, but not after mere exposure to the test apparatus. Bilateral amygdala injection of a 15-mer phosphorothioate c-fos antisense oligodeoxynucleotide prior to testing blocked conflict-induced c-fos expression and had behavioral effects similar to those of established antianxiety drugs.3. Separate experiments determined that antisense treatment did not affect conflict behavior by acting on shock thresholds or drinking motivation.4. These findings provide evidence that neuronal activation and c-fos induction in the amygdala may be of importance for mechanisms of fear and anxiety.     

Multiple genotypes of mitochondrial DNA within a horse population from a small region in Yunnan province of China

mtDNA genotypes of six domestic horses (three adult short horses whose heights are under 1 m and three common domestic horses) from a small region of 15 km² in Malipo county of Yunnan province of China were investigated by the technique of restriction fragment length polymorphism (RFLP) with 16 restriction endonucleases which recognize 6-bp sequences. An average of 56 fragments for an individual was obtained. Unlike other domestic animals, this population of horses exhibits high mtDNA genetic diversity. Each of the six horses has a specific mtDNA genotype showing a pattern of multiple maternal origins, as suggested by fossil and literature records. We think the population of horses is an amazing seed-resource pool of horses and hence deserves to be paid more attention from the view of conservation genetics. However, it is also remarkable that we did not find any typical mtDNA genetic markers which would discriminate between short horses and common domestic horses.     

湖北南襄盆地枣阳凹陷第三纪轮藻化石及其地层意义

本文研究了南襄盆地枣阳凹陷第三系玉皇顶组到凤凰镇组轮藻化石。共划分出４个组合,其时代分别属早始新世、中始新世、晚始新世和渐新世。     

菜用香椿良种选育性状的模糊综合评判

采用模糊数学方法对菜用香棒１６个不同种源的无性系从形态、颜色、香气、抗病性四个方面进行了综合评判,筛选出了接近选育目标的５个菜用优良无性系。     

酶电泳资料和系统与进化植物学研究综述

酶电泳资料和系统与进化植物学研究综述葛颂（中国科学院植物研究所系统与进化植物学开放研究实验室北京１０００９３）关键词同工酶,电泳,植物系统学,进化ＥＬＥＣＴＲＯＰＨＯＲＥＴＩＣＤＡＴＡＡＮＤＳＴＵＤＩＥＳＯＦＰＬＡＮＴＳＹＳＴＥＭＡＴＩＣＳＡＮＤＥＶ...