腺病毒介导的人抑瘤素M基因对A375人黑色素瘤细胞的抑制作用

目的:构建抑瘤素M(OSM)重组腺病毒载体,研究其对人黑色素瘤细胞A375的抑制作用。方法:以PEGZ-OSM重组质粒为模板,通过PCR技术扩增出OSM片段,采用腺病毒载体的基因重组和体外包装技术获得表达与人OSM氨基酸序列相同的重组腺病毒子Ad-OSM,感染A375细胞,用荧光显微镜、RT-PCR、Western blot法检测OSM在A375细胞中的转录和表达;荧光显微镜观察A375细胞的形态学改变;MTT法和流式细胞术(FCM)检测Ad-OSM对A375细胞的生长抑制和细胞周期的抑制效应;半定量RT-PCR法检测OSM基因表达对A375细胞中的Bax、Bcl-2基因表达的影响。结果:基因测序和PCR分析结果显示,成功构建了Ad-OSM腺病毒表达载体;RT-PCR和Western blot法检测到OSM基因在A375细胞中的转录和表达;OSM基因的表达对A375细胞增殖有明显抑制作用,并可诱导细胞凋亡,OSM基因可通过上调细胞中Bax和下调Bcl-2基因表达诱导细胞凋亡。结论:成功构建了Ad-OSM腺病毒表达载体,感染OSM基因可明显抑制A375人黑色素瘤细胞的生长,诱导其凋亡,该现象可能是通过改变Bax、Bcl-2基因表达水平来发挥抗肿瘤作用。     

响应面法优化獐牙菜苦苷生物转化条件

采用响应面法对獐牙菜苦苷的生物转化条件进行优化,首先应用Plackett-Burman方法筛选出对转化具有显著性影响的因素,再用中心响应面法的Response Optimizer工具对最优转化条件进行了分析和预测,最后对预测结果进行了验证。结果表明,獐牙菜新碱的最优化转化条件(mg/mL)为MgSO4.7H2O 5.14、MnSO4.4H2O 3.42、葡萄糖9.95、蛋白胨9.24、pH5.80。在此条件下獐牙菜新碱的最大转化产率为17.64%;红百金花内酯的最优转化条件(mg/mL)为MgSO4.7H2O 4.09、MnSO4.4H2O6.07、葡萄糖7.37、酵母膏7.37,pH5.73。在此条件下红百金花内酯的最大转化产率为8.81%。     

保留特殊种质材料的核心库构建方法

本研究提出了能保留特殊种质材料的多次聚类构建种质资源核心库方法,对种质材料的表现型数据采用合适的遗传模型及混合线性模型统计分析方法无偏预测基因型值,用基因型值构建核心库,计算材料间的马氏距离,用不加权类平均法进行聚类,根据聚类图选取材料构建核心库时,优先保留特殊遗传材料,用方差F测验、均值t测验、极差比和变异系数评价核心库代表原有种质资源群体遗传多样性的程度,以168个棉花基因型的5个纤维性状构建核心库。     

利用流式细胞仪分选拟南芥根尖发育早期非根毛细胞

建立了应用流式细胞仪分选植物特定类型细胞的方法。以拟南芥（Arabidopsis thaliana）Wer：：GFP转基因株系为材料,用激光共聚焦显微镜鉴定GFP的表达位置,采用酶解法制备拟南芥根尖原生质体,应用流式细胞仪荧光激活细胞分选技术（FACS）分选收集GFP阳性细胞,并提取细胞的RNA。结果表明,Wer：：GFP转基因株系仅在根表皮发育早期的非根毛细胞中表达GFP;利用酶解法制备的根尖原生质体数目较多;从FACS分选收集的细胞中提取的RNA质量较好,可用于研究特定类型细胞的基因表达谱。应用流式细胞仪分选拟南芥非根毛细胞的方法为研究植物特定类型细胞的基因表达谱及基因功能奠定了技术基础。     

稳定转染Dicer基因对HELF细胞功能的影响

目的观察正常人胚肺成纤维细胞（HELF）在稳定转染Dicer基因后增殖能力及迁移能力的变化。方法应用脂质体介导方法转染Dicer基因表达载体于HELF细胞,G418筛选,PCR检测整合情况,RT-PCR检测表达情况,Western-blot检测蛋白表达水平,确定稳定转染细胞株,MTT方法检测细胞增值能力,Transwell方法检测细胞迁移能力。结果Dicer基因稳定转染HELF细胞系建立,用RT-PCR和Western-blot方法检测到Dicer基因mRNA和蛋白表达水平都明显增强。与转染空载体的HELF细胞相比,转染Dicer基因的HELF细胞48、72、96h的MTT光吸收值明显升高（P〈0.05）,并且穿过人工重构基底膜的细胞数明显增多。结论Dicer基因稳定转染HELF细胞后,HELF细胞增殖能力和迁移能力都明显增强。     

Glutamine synthetase and glutamate synthase activities during growth and sporulation in Bacillus subtilis.

Glutamine synthetase in Bacillus subtilis 168 was repressed to a greater extent by L-glutamine or L-arginine than by ammonia when each was used as sole nitrogen source. It was derepressed when either L-glutamate or nitrate was used as nitrogen source. Glutamate synthase was repressed by L-glutamate or L-arginine and, to a lesser extent, by L-glutamine but was derepressed during growth with ammonia or nitrate. Glutamine synthetase activity was unaltered during the onset of sporulation. Glutamate synthase activity, however, underwent a small and apparently transient increase in bacteria induced to sporulate by nitrogen limitation.     

A cell pattern generation model based on an extended artificial regulatory network

Cell pattern generation has a fundamental role in both artificial and natural development. This paper presents results from a model in which a genetic algorithm (GA) was used to evolve an artificial regulatory network (ARN) to produce predefined 2D cell patterns through the selective activation and inhibition of genes. The ARN used in this work is an extension of a model previously used to create simple geometrical patterns. The GA worked by evolving the gene regulatory network that was used to control cell reproduction, which took place in a testbed based on cellular automata (CA). After the final chromosomes were produced, a single cell in the middle of the CA lattice was allowed to replicate controlled by the ARN found by the GA, until the desired cell pattern was formed. The model was applied to the problem of generating a French flag pattern.     

Kinetics and mathematical modeling of homoacetic fermentation of lactate by Clostridium formicoaceticum

Fermentation kinetics of Clostridium formicoaceticum grown on lactate at pH 7.0 and 35 degrees C was studied. Acetate was the only fermentation product and its production was growth associated. The growth of this bacterium was insensitive to the lactate concentrations studied, but was inhibited by acetic acid. A Monod-type expression with product inhibition similar to the noncompetitive inhibition of enzyme kinetics was used to model the batch fermentation. An integrated equation was developed and used to help estimating the kinetic parameters in the model. This mathematical model can be used to simulate the homoacetic fermentation of lactate by C. formicoaceticum at pH 7.0 and 35 degrees C.     

Following the biochemical and morphological changes of Bacillus atrophaeus cells during the sporulation process using Bioaerosol Mass Spectrometry

Bioaerosol Mass Spectrometry (BAMS), a real-time single cell analytical technique, was used to follow the biochemical and morphological changes within a group of Bacillus atrophaeus cells by measuring individual cells during the process of sporulation. A mutant of B. atrophaeus that lacks the ability to produce dipicolinic acid (DPA) was also analyzed. Single cell aerodynamic sizing was used to follow gross morphological changes, and chemical analysis of single cells by mass spectrometry was used to follow some biochemical changes of B. atrophaeus cells during endospore formation.     

The social behaviour of the Pin-tailed Whydah Vidua macroura in northern Ghana

This paper describes the pattern of territory usage by male Pin-tailed Whydahs Vidua macroura , and the display postures used by territorial males, intruding males and visiting females are described. Each territory holder used one or two favoured display sites at which he was visited by, and displayed towards females in groups of one to eight. The frequency of display flights and song output reflected the diurnal pattern of visits by females. Each female was courted by up to six males, but the number of copulations observed was too few to indicate the type of mating system used. The operative sex ratio was estimated at 1:6 (females to territory holders), and the results of a removal experiment suggested that a proportion of males was prevented from breeding.     

Survey of parasite control programs used in captive wild ruminants

The purpose of this study was to document, through a mailed questionnaire survey, the methods of parasite control currently used in captive wild ruminants. There was a 69% overall response to the survey. The majority of respondents indicated that they used parasite surveillance and identification techniques similar to those used for domestic ruminants. Of the four major anthelmintic drug classes, the benzimidazoles and ivermectin were used most commonly. Many of the parasite control programs had high treatment frequencies which were similar to dosing frequencies reported to result in anthelmintic resistance in domestic animals. Lack of effectiveness by benzimidazoles was perceived to be a problem by many respondents.     

Peroxidation of mullet and rat liver lipids in vitro: effects of pyridine nucleotides, iron, incubation buffer, and xenobiotics

1. Lipid peroxidation was observed in homogenates of mullet and rat livers and was affected by the specific conditions used for the in vitro assays. 2. Lipid peroxidation was altered by the buffer used in the assays and was dependent on the amount of Fe added to the assay. 3. Lipid peroxidation was stimulated by both NADH and NADPH and by the xenobiotics, CCL4 and DEM. 4. Lipid peroxidation was observed in microsomes of mullet livers and was inhibited by addition of cytosol.     

不同碳氮源对糙皮侧耳木质纤维素酶活性的影响

以糙皮侧耳Pleurotus ostreatus为材料,研究简单碳氮源及木质素纯品诱导条件对其木质纤维素酶活性的影响。结果表明,不同的碳源培养基和氮源培养基对糙皮侧耳漆酶活性、羧甲基纤维素酶活性和木聚糖酶活性均具有极显著的影响（P<0.001）,且对糙皮侧耳菌丝生物量也有极显著的影响（P<0.001）。以蔗糖作主要碳源诱导物时,有利于提高糙皮侧耳漆酶活性;以果糖作主要碳源诱导物时,有利于提高糙皮侧耳羧甲基纤维素酶活性和菌丝生物量的积累;以葡萄糖作主要碳源诱导物时,有利于提高糙皮侧耳木聚糖酶活性。以酵母浸粉作主要氮源诱导物时,有利于提高糙皮侧耳漆酶活性和菌丝生物量的积累;以硝酸钾作为主要氮源诱导物时,有利于提高糙皮侧耳羧甲基纤维素酶活性;以硫酸铵作为主要氮源诱导物时,有利于提高糙皮侧耳木聚糖酶活性。碱性木素的存在,有利于提高糙皮侧耳漆酶活性,但不利于菌丝生物量的积累。与此同时,碱性木素的存在对糙皮侧耳羧甲基纤维素酶和木聚糖酶活性并没有促进作用。     

Application of chemical ionization mass spectrometry and the twin-ion technique to better define a mechanism in acetylhydrazine toxicity.

Chemical ionization mass spectrometry was used to show that N-acetylcysteine is formed in microsomal incubations containing NADPH, oxygen, cysteine and acetylhydrazine. The twin-ion technique was used to compare the ratio of hydrogen and deuterium in the N-acetylcysteine product with the initial ratio in a mixture of acetylhydrazine-trideuteroacetylhydrazine used as substrate. The results showed that the entire acetyl group was trapped by cysteine, thereby eliminating ketene as the reactive acylating agent formed during the oxidation of acetylhydrazine by liver microsomes.     

Diffuse reflectance infrared Fourier transform spectroscopic characterization of a silica-immobilized N-hydroxysuccinimide active ester crosslinking agent and its precursors

Diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy was used to characterize the product of each step in the preparation of a silica-immobilized N-hydroxysuccinimide (NHS) active ester. The preparation of this NHS active ester linkage was based on a literature procedure for the immobilization of proteins. The DRIFT method was used to guide modification of this literature procedure. The DRIFT method also was used to indicate an impurity entrapped in the 60-A diameter pores of the silica support during the formation of the immobilized active ester. Degradation of the immobilized NHS active ester, stored under either argon or dioxane, can be followed by the DRIFT method. Myoglobin and glycine were allowed to react with the active ester, and the result for this silica support was evaluated by the DRIFT method. Elemental analysis was used to provide information on the loading of the silica-immobilized moieties that were presented for DRIFT analysis.     

Oxygen supply to immobilized cells: 2. Studies on a coimmobilized algae—bacteria preparation with in situ oxygen generation

A coimmobilized mixed culture of algae, Chlorella pyrenoidosa, and bacteria, Gluconobacter oxydans, has been studied. The conversion of glycerol to dihydroxyacetone(1,3-dihydroxy-2-propanone), catalysed by the bacteria, was used to indicate the oxygen supply in the immobilized preparation. The oxygen produced by the algae in the coimmobilized preparation was used by the bacteria more effectively than when the cells were immobilized separately and mixed within the reactor. A preparation consisting of only bacteria and no algae was much less effective. The coimmobilized preparation was used in the continuous production of dihydroxyacetone for six days without any significant loss of activity.     

Genetic Programming as Alternative for Predicting Development Effort of Individual Software Projects

Statistical and genetic programming techniques have been used to predict the software development effort of large software projects. In this paper, a genetic programming model was used for predicting the effort required in individually developed projects. Accuracy obtained from a genetic programming model was compared against one generated from the application of a statistical regression model. A sample of 219 projects developed by 71 practitioners was used for generating the two models, whereas another sample of 130 projects developed by 38 practitioners was used for validating them. The models used two kinds of lines of code as well as programming language experience as independent variables. Accuracy results from the model obtained with genetic programming suggest that it could be used to predict the software development effort of individual projects when these projects have been developed in a disciplined manner within a development-controlled environment.     

冬瓜蛋白酶的分离纯化及其部分性质研究(英文)

冬瓜的果肉中发现了丰富的蛋白水解酶.用硫酸铵将冬瓜果肉汁分级盐析,得到粗酶液.再经DEAE-Sepharose FF离子交换层析和Superdex-75柱层析等步骤得到一种电泳纯的冬瓜蛋白酶.SDS-PAGE测得其分子量为64 kDa.以酪蛋白做底物时,该酶的最适反应温度为70℃,最适作用pH为6.5,在pH 4.5～10.5,40～70℃范围内较稳定.PMSF强烈抑制该蛋白酶的活性.另外,Hg~(2+)对该酶有强烈的抑制作用,Mn~(2+)离子对其有保护作用,Zn~(2+)、Ca~(2+)和Cu~(2+)等离子对其活性没有影响.     

An improved method for anti-glycolipid antibody and glycolipid determination by an enzyme-linked immunosorbent assay using polystyrene beads

Sensitive determination of anti-glycolipid antibody titer and glycolipid content by an enzyme-linked immunosorbent assay (ELISA) using polystyrene beads was achieved. Glycolipid-coated polystyrene beads were used as the immobilized antigen. As antigen glycolipids, gangliotetraosylceramide (GA1), gangliotriosylceramide (GA2) and neolactotetraosylceramide (paragloboside) were used. Concentrations of 1-500 ng glycolipid in liposomes/ml or 0.1-100 micrograms glycolipid/ml could be used for the glycolipid determination. Glycolipid determination by the competitive inhibition method was not influenced by the presence of other glycolipids. A great advantage of this method is that the glycolipid-coated beads can be used repeatedly by washing the used beads with 3M NaSCN solution. The method was applied to the detection of auto-antibody against GA1 in ascitic fluid from cancer patients.