水痘-带状疱疹病毒糖蛋白gE在昆虫细胞中的表达鉴定及其晶体培养

目的利用杆状病毒-昆虫表达系统建立纯化水痘带状疱疹病毒(VZV)包膜糖蛋白gE的方法,筛选gE蛋白的晶体培养条件,以期用于结构解析。方法将VZV糖蛋白gE基因序列克隆至杆状表达载体pAcgp67B载体中,利用High FiveTM昆虫细胞表达gE蛋白并进行TALON亲和层析纯化;利用WAVE生物波浪反应器建立含硒代甲硫氨酸的gE蛋白方法,以便在晶体结构解析中利用单波长或多波长异常散射进行相位解析;通过分子排阻色谱、分析超离,差示扫描量热法和酶联免疫吸附试验等分析gE蛋白的理化性质;利用结晶试剂盒对gE498和gE354蛋白的结晶条件进行初筛。结果获得的gE498和gE354蛋白纯度约为90%、产量为8～10 mg/L。理化分析显示两种gE蛋白在溶液中主要以均一稳定的单体形式存在,并呈现出良好的反应原性。在gE354晶体初筛中获得3个结晶条件:CrystalH3、PEGH12和IndexB10。结论建立了VZV糖蛋白gE表达和纯化方法,制备的蛋白纯度高,且具有反应原性,并筛选出CrystalH3、PEGH12和IndexB10这3个结晶条件,为gE糖蛋白的结构与功能研究及新型疫苗开发奠定了基础。国家自然科学基金项目(No.81871648);;\n新药创制专项项目(No.2018ZX09711003-005

Application of ARM and Linux in Deep Sea ROV Control System

利用ARM处理器功耗低、速度快等特点,采用耐压电子技术,设计了深海ROV下位机控制系统,有效地解决了随ROV下潜深度增大电子舱体积增大的问题。详细介绍了控制系统软、硬件设计方法。系统采用耐压电子技术设计,抗压能力强、性能可靠、速度快,在深海ROV有良好的应用前景

Nanofiltration Separation of Monosaccharide Solution Under High Concentrations

以葡萄糖溶液为主要原料,用商业卷式纳滤膜研究高浓度单糖溶液纳滤分离过程中料液浓度、操作压力、操作温度3个主要因素对膜分离的影响.在较低浓度下,渗透流率受操作压力影响较大,表观截留率受操作压力影响较小;而在较高浓度下,结果刚好相反.随着温度的升高,压力增大可以加快渗透流率的增加速率,降低表观截留率的下降速率;浓度增大将降低渗透流率的增加速率,将加快表观截留率的降低速率.本研究可为工业生产中综合选择工艺条件以提高处理效率并减少原料损失提供指导.In this paper,the influence of three operation factors namely feed concentration,operating pressure and temperature on the performance of nanofiltration membrane separation at high solution concentrations is evaluated using glucose aqueous as solution with a commercial spiral-wound nanofiltration membrane.At lower solution concentrations,operating pressure has more impact on permeate volume flux,while less on the rejection.On the other hand,the results are opposite at higher solution concentrations.As the operating pressure increased,the raising rate of permeate volume flux increases with the operating temperature,while the falling rate of rejection reduces with the operating pressure.In the cases of high solution concentrations,the increasing rate of permeate volume flux changing with temperature decreases with operating pressure,and the declining rate of rejection increases with operating pressure.The investigation results in this work provide guidance for optimizing the design and operation conditions for industrial nanofiltration processes thus to enhance the efficiency and reduce the raw material wastage.国家重点基础研究发展计划(973)项目(2010CB732201);科技部国际科技合作项目(2009DFA60930

程海鱼类区系的来源及其物种的分化

程海共有土著鱼类15种,它位于青藏高原横断山区的东缘,云南高原的西北,鱼类区系却独具一格,表现出江河平原鱼类区系的特点,显示程海鱼类区系来自长江的事实。程海自与金沙江隔离之后,湖内存在空白生态灶(niche)是产生物种分化的重要因素,尤以鲤鱼Cyprinus carpio L.的形态,表现出一系列的适应性变异,这些变异发生在近300年内,与处于海拔和纬度较高的甘子河裸鲤Gymnocypris przewalskii ganzihoensis Zhu et Wu形成的时间相比,相对而言是比较快的,然而却慢

Apoptosis of Human Gastric Cancer Cell Induced by Photochemical Riboflavin

背景与目的:核黄素在光照下具有抑癌活性,并且被认为与核黄素光化学反应产生的活性氧自由基有关。本研究探讨核黄素在光照处理下,体外诱导人胃癌MGC80-3细胞凋亡效应。方法:以台盼蓝拒染法计数细胞存活率,Giemsa染色观察形态改变,DNA琼脂糖电泳分析DNA片段化,测定断裂及未断裂DNA含量定量凋亡效率,流式细胞术及Westernblot分别检测细胞周期及凋亡相关基因表达变化。结果:细胞存活率依浓度及处理时间而下降;光照核黄素处理24h,细胞呈典型的凋亡形态;以10、20、30和40μmol/L的核黄素光照处理MGC80-3细胞,DNA电泳显示20μmol/L以上处理组均出现典型的凋亡DNA梯形带;且相应凋亡效率分别为35.4%、54.1%、70.6%和86.8%,与核黄素的浓度呈正相关;经10及20μmol/L核黄素光照处理后细胞周期主要阻滞于G2/M期;Westernblot结果显示凋亡相关基因p53、c-myc及Bax表达上调,而Bcl-2表达下调。结论:核黄素光化学反应抑制胃癌MGC80-3细胞生长的作用是以诱导细胞凋亡为主要途径,并与诱导细胞阻滞于G2/M期有关。BACKGROUND &OBJECTIVE: Photochemical riboflavin, the production of reactive oxygen species (ROS) , has been reported having cytotoxity on some cancer cells. The aim of this study was to investigate the effect of photochemical riboflavin on inducing apoptosis in human gastric cancer cell line MGC80 3. METHODS: Trypan blue exclusion method, Giemsa staining, DNA electrophoresis, DNA quantification, Western blot analysis, and flow cytometry were conducted to determine the effect of photochemical riboflavin on cell survival, morphology, DNA fragmentation, gene expression, and cell cycle arresting. RESULTS: The cell viability dropped down according to the riboflavin concentration and treating time. Exposure of the cells in 20 μmol/L riboflavin for 24 hours resulted in typical apoptotic morphology and G2/M arresting. As MGC80 3 cells were separately treated with 10, 20, 30, and 40 μmol/L photochemical riboflavin, DNA electrophoresis showed that the ladder bands, a typical feature of apoptotic cell, appeared in the groups treated by over 20 μmol/L photochemical riboflavin. The efficiency rates of DNA fragmentation were 35.4%, 54.1%, 70.6%, and 86.8%, respectively. Western blot analysis showed that the expression of apoptosis related proteins p53, C myc, and Bax were up regulated whereas the expression of Bcl 2 was down regulated. CONCLUSION: These results indicate that photochemical riboflavin has high efficiency of inducing cell apoptosis on MGC 80 3 cells in vitro and there is a correlation between apoptosis and G2/M arresting.福建省自然科学基金项目(C97108