艾滋病病毒蛋白酶高效表达载体及体外活性检测系统的构建

艾滋病是本世纪80年代初发现的一种烈性传染病,5年病死率为100％,致病因子为人免疫缺陷病毒,该病毒的蛋白酶在病毒复制和成熟中具有决定性的意义。由于目前国内外尚未获得艾滋病病毒蛋白酶的高效表达的重组子及活性检测系统,限制了它的研究与应用。本文利用PCR技术修饰了艾滋病病毒蛋白酶的基因,使其具有便于克隆及表达用的限制酶切位点及转录终止码,井在其C末端设置了一个可用于检验该酶活性的特殊序列。DNA序列分析揭示上述突变策略成功,将修饰后的艾滋病病毒蛋白酶基因克隆入大肠杆菌表达系统,并获得高效表达(>30％),Western-Bolt鉴定结果表明所表达的蛋白为艾滋病病毒所特有,并具有较好的生物活性。     

棉铃虫单粒包埋型核型多角体病毒(HaSNPV)复制的研究——Ⅰ.细胞病理学和病毒复制的一些特性

本文报道了棉铃虫单粒包埋型核型多角体病毒(Heliothis armigera single nucleocapsid nuclear polyhedrosis virus, HaSNPV)在棉铃虫及棉铃虫蛹卵巢细胞系SFE-HA-8212中的复制。HaSNPV的复制和其他的核型多角体病毒大体相符,复制过程也可分为形成出芽型病毒与形成包埋型病毒这两个时相。研究了影响病毒在细胞中复制的诸因素,包括病毒感染复数、细胞接种密度和细胞生长阶段等。在适宜的条件下平均每细胞可生产出芽型病毒14PFU,多角体24个。生成的病毒具有感染力。这些表明SFE-HA-8212细胞可供HaSNPV有效复制。同时,作为细胞群体该细胞系对HaSNPV感染的反应并非均一,其中有89.65±21.4％的感染细胞释放病毒,但仅有37.85±6.7％的细胞形成多角体。表明HaSNPV的感染并不一定导致形成多角体,在大部分感染细胞中病毒复制进行到产生病毒粒子就停止了。初步讨论了这种不均一性的原因。     

大鼠胰腺γ-氨基丁酸免疫组织化学定位

本文用ABC—GDN免疫组织化学方法,研究了γ-氨基丁酸(Gamma—Aminobutyric Acid,GABA)在大鼠胰腺的定位和分布,并用相邻切片法,观察它与胰岛素的共存关系。结果发现GABA免疫反应阳性细胞主要分布于胰腺内分泌部(胰岛)。在外分泌部亦有少许分布。大部分胰岛细胞呈GABA免疫反应阳性,集中位于胰岛的中央部。相邻连续切片免疫染色证实GABA与胰岛素共存于胰岛B细胞中。外分泌部胰腺GABA免疫反应阳性细胞,呈零散分布于腺泡和导管上皮间。本文为进一步探讨GABA在胰腺的生理作用提供了形态学依据。     

EFFECTS OF VASOACTIVE INTESTINAL PEPTIDE ON ACTIVITIES OF SUCCINIC DEHYDROGENASE AND ALKALINE PHOSPHATASE IN RAT HEPATOMA CELLS

Using cytochemical method,microspectrophotometry and image analysis,effects of va-soactive intestinal peptide(VIP)on activities of succinic dehydrogenase(SDH)and alkalinephosphatase(ALP)in rat hepatoma cells were studied in vitro.The results showed that thehepatoma cell expressed potent positive reactions of SDH and ALP,the positive positionswere located at the cell membranes and/or cytoplasm.Having been treated with VIP,ALPdecreased obviously in activity(P<0. 01,compared with hepatoma cells untreated by VIP).The sites of ALP activty were chiefly located at the cell membranes,particularly at the cell-cell contacts.Cultured rat hepatoma cells had intensive SDH activity in their cytoplasm.Compared with untreated eclls,there was no marked difference in the intensity of SDH activ-ity in VIP-treated hepatoma cells(P>0.05).     

慈竹无性系种群能值特点及其影响能值计测因素的研究

 本文引用Harper(1977)的构件结构理论,从构件结构单位、无性系分株和无性系三个层次,对四川南充市郊慈竹无性系种群的能值特点及其影响能值的计测因素进行了定量研究。研究结果表明：慈竹无性系种群中,各构件单位的去灰分热值(AFCV)分别为：根15349.42J／g、根茎16372.92J／g、秆17106.06J／g、枝18111.99J／g和叶19451.90J／g;慈竹无性系分株的AFCV(J／g)随龄级增大而递增;慈竹无性系水平上的AFCV为：Ⅰ龄占16.47%、Ⅱ龄占25.76%、Ⅲ龄占36.32%、Ⅳ龄为13.08%及Ⅴ龄为8.37%。用恒容燃烧法测定热值时,其能值变化与氧分压密切相关。用经验公式计算的能值较作图法高;用AFCV表示能值较总干重热值(GCV)准确。     

中蜂处女王性成熟日龄及其交尾习性的探讨

本研究对210只人工培育的出房后1,2,3,4,6,8．10日龄的中蜂处女王受精囊液的pH值进行测定．结果表明,1日龄处女王受精囊液的pH值平均为7．7,2日龄为8．5,从3日龄开始达到最高值8.8．并保持稳定状态,由此判断出房后3日龄的中蜂处女王性已成熟．通过对72只中蜂处女王交尾飞行的观察,结果表明,蜂王交尾日龄多为6-8日．最早为4日龄,最迟为16日龄;处女王认巢飞行持续时间平均为7．4分钟,交尾飞行平均为21．5分钟：处女王连续交尾飞行1-3次,有9只处女王在一天里进行2次交尾飞行．     

Diverse flowering responses subjecting to ambient high temperature in soybean under short-day conditions

Flowering time is one of important agronomic traits determining the crop yield and affected by high temperature. When facing high ambient temperature, plants often initiate early flowering as an adaptive strategy to escape the stress and ensure successful reproduction. However, here we find opposing ways in the short-day crop soybean to respond to different levels of high temperatures, in which flowering accelerates when temperature changes from 25 to 30 °C, but delays when temperature reaches 35 °C under short day. phyA-E1, possibly photoperiodic pathway, is crucial for 35 °C-mediated late flowering, however, does not contribute to promoting flowering at 30 °C. 30 °C-induced up-regulation of FT2a and FT5a leads to early flowering, independent of E1. Therefore, distinct responsive mechanisms are adopted by soybean when facing different levels of high temperatures for successful flowering and reproduction.     

Improvement of chemostat performance via nonlinear oscillations

A novel operation strategy employing self-generated oscillation to imrpove the performances of bioreactors is proposed and applied to a model system consisting of two continuous stirredtank bioreators (CSTBs) connected in series. It is demonstrated via computation that the performance of the system (in terms of timeaveraged cell concentration) can be greatly enhanced by adopting the proposed strategy. The process concept presented and the results obtained in this paper are expected to have significant implications beyond the bioprocessing industry.     

Inhibitory role of bone marrow mesenchymal stem cells-derived exosome in non-small-cell lung cancer: microRNA-30b-5p,EZH2 and PI3K/AKT pathway

Exosomal microRNA (miRNA) exerts potential roles in non-small-cell lung cancer (NSCLC). The current study elucidated the role of miR-30b-5p shuttled by bone marrow mesenchymal stem cells (BMSCs)-derived exosomes in treating NSCLC. Bioinformatics analysis was performed with NSCLC-related miRNA microarray GSE169587 and mRNA data GSE74706 obtained for collection of the differentially expressed miRNAs and mRNAs. The relationship between miR-30b-5p and EZH2 was predicted and confirmed. Exosomes were isolated from BMSCs and identified. BMSCs-derived exosomes overexpressing miR-30b-5p were used to establish subcutaneous tumorigenesis models to study the effects of miR-30b-5p, EZH2 and PI3K/AKT signalling pathway on tumour growth. A total of 86 BMSC-exo-miRNAs were differentially expressed in NSCLC. Bioinfomatics analysis found that BMSC-exo-miR-30b-5p could regulate NSCLC progression by targeting EZH2, which was verified by in vitro cell experiments. Besides, the target genes of miR-30b-5p were enriched in PI3K/AKT signalling pathway. Animal experiments validated that BMSC-exo-miR-30b-5p promoted NSCLC cell apoptosis and prevented tumorigenesis in nude mice via EZH2/PI3K/AKT axis. Collectively, the inhibitory role of BMSC-derived exosomes-loaded miR-30b-5p in NSCLC was achieved through blocking the EZH2/PI3K/AKT axis.