植物个体远程辐射信号早期传递过程的研究体系创建

自2005年以来,活体中的远程辐射旁效应逐渐成为辐射生物学的研究热点,然而其早期信号传递过程的相关研究却鲜见报道,主要原因是由于早期信号传递过程研究所需的旁区与辐射区的自由分离与组合在动物模型上无法实现。本研究是基于植物个体可以切割和嫁接的特性,借鉴离体细胞培养基转移方法研究旁效应早期过程的思想,在植物个体上实现旁区与辐照区的"分离"与"组合",构建了一种研究个体远程辐射信号早期传递过程的植物实验体系。具体是以模式植物拟南芥菜转基因系(AtRAD54promoter∶∶GUS)为材料,同源重组修复相关基因AtRAD54表达水平为生物学检测终点,人为地将辐照区的组织(或器官)与旁区部分"分离"或"嫁接",通过测定旁区组织(或器官)的AtRAD54基因表达水平变化,研究其辐射信号传递的早期过程。该研究体系的创建为活体旁效应早期过程的研究提供了一种可行的研究方法。     

基于大肠杆菌的重离子辐射靶点序列定位和LacI突变分析的实验体系构建及验证

重离子辐射具有独特的物理和生物学特性,在诱变育种等领域有着广泛的应用,但其诱变的机制并不完全清楚。不同于传统的X和γ射线,重离子辐射具有较高的线传能密度(Linear energy transfer,LET),主要诱导团簇状的DNA损伤,其演化为遗传变异的过程更为复杂,突变类型也更难预测。目前的实验技术很难在序列水平对重离子击中DNA的靶点进行定位,这致使重离子辐射诱变机制的研究相对滞后。针对这一问题,根据重离子辐射诱导的团簇损伤核心区域富含DNA双链断裂(Double-strand break,DSB)以及同源重组机制对DSB特异性响应的特性,首先构建了四环素抗性基因(TetA)同源重组元件用于确定DNA团簇损伤的序列定位,并在重组原件侧翼连接反向突变筛选基因LacI用于团簇损伤—突变的检测,最后把该质粒转化到大肠杆菌E.coll。在此基础上,比较分析γ射线与碳重离子(80 MeV/u)辐照后同源重组和报告基因突变的情况,验证了该体系用于重离子辐射靶点序列定位及突变检测的可行性,为进一步研究重离子辐射诱变的相关机制奠定了方法学基础。     

模型化研究两细胞间基因、蛋白耦合振荡中的噪声效应

本文基于Hill动力学与Michaelis-Menten方程,建立理论模型研究两细胞间基因、蛋白耦合振荡中的噪声效应.研究发现,在Notch信号通路中,两细胞间基因、蛋白耦合振荡呈现了周期振荡特性,表明了细胞间信号传导的同步振荡特性.“内在”噪声和“外在”噪声对两细胞间基因、蛋白耦合振荡有着不同的作用.内噪声有利于细胞间Notch信号通路中各基因、蛋白表达再次提升.外噪声诱导通路中基因、蛋白的表达水平降低,周期振荡变得阻尼.内、外噪声共同作用不仅可使得基因表达适当并呈现出持续振荡模式,而且还可使得细胞间基因转录合成相应的蛋白过程呈现出持续振荡模式.从而表明了基因表达的内、外噪声共同作用有利于控制细胞间基因激活、蛋白合成保持周期节律性.本文理论结果揭示了内外噪声对细胞间Notch信号通路动力学的一种调控机制,确定了内外噪声各自的调控效应,澄清了内外噪声共同作用调控体系持续周期振荡的物理机制,理论结果符合实验,可为设计阻止Notch体系基因、蛋白变异导致的多种疾病和癌症的通路治疗方案提供理论依据.     

基因芯片技术及其在放射治疗中的应用

基因芯片技术是建立在杂交序列基本理论上的分子生物学技术,它以一种全面、综合和系统的思维方式研究生命现象。基因芯片技术可以完整地研究整个细胞或器官全部基因变化,可以通过基因分析发现对电离辐射的基因反应差异,从而建立一种新的分子放射生物学方法。综述了基因芯片技术及应用领域,重点介绍了基因芯片技术在辐射治疗癌症中的应用。概述了重离子治疗肿瘤优于其它射线的原因。展望了利用基因芯片技术的优势探索肿瘤经重离子辐照前、中、后期的生物学效应。     

基因芯片技术及其在放射治疗中的应用

基因芯片技术是建立在杂交序列基本理论上的分子生物学技术, 它以一种全面、 综合和系统的思维方式研究生命现象。 基因芯片技术可以完整地研究整个细胞或器官全部基因变化, 可以通过基因分析发现对电离辐射的基因反应差异, 从而建立一种新的分子放射生物学方法。 综述了基因芯片技术及应用领域, 重点介绍了基因芯片技术在辐射治疗癌症中的应用。 概述了重离子治疗肿瘤优于其它射线的原因。 展望了利用基因芯片技术的优势探索肿瘤经重离子辐照前、 中、 后期的生物学效应。     

线性共轭齐聚物共轭单元依赖的光学特性

研究三种线性共轭齐聚物(F-P、F-P-F和P-F-P-F-P)的光学特性,其中吩噻嗪单元(F)和芴单元(P)交替排列. 随着共轭体系的增强,它们的吸收和发射带都逐渐红移,双光子吸收特性明显的改进. 利用连续速率分布模型分析荧光动力学的结果显示,它们的弛豫速率在逐渐增加,速率分布宽度也在变窄. 量子化学计算给出了它们的分子结构和跃迁机制,显示π共轭系统的增强促使了双光子特性的改进.     

上海光源在生命科学中的应用

<正>生命科学包含涉及生命和生物有机体科学研究的许多分支,研究对象包含微生物、植物和动物,包括人类。经过几个世纪的发展,生命科学已经从单纯的描述逐渐发展成了一门理论、计算、实验等多领域交叉学科。自DNA双螺旋结构模型的提出以来,生命科学迈进了崭新的层次——分子生物学的层次,迄今为止,生命科学已经发展成了一门包含众多分支学科的科学体系。生命科学和生物技术,包括基因工程、合成生物学、基因组学和蛋白质     

离子束介导技术在生物体遗传改良上的发展趋势

在利用离子束介导技术对生物体进行遗传改良的研究领域内仍然存在着7大研究难题值得注意。 明确提出了离子束介导技术进一步发展的技术思路, 即立足于离子束介导技术这一物理学技术平台, 注重研究两个关键性问题(即离子束介导异源遗传物质进入受体基因组的机理和受体发生异源基因重组的机制)。 在研究中完成3个有效转变(从形态学鉴定为主有效地转变为对其遗传学规律进行研究, 从对介导当代群体的变异效应的研究为主有效地转变为研究突变性状在多个世代内所表现的后效性, 从对单一性状的研究为主有效地转变为研究突变性状的综合表现), 寻找4个方面的实验证据(形态学、 生理生化、 细胞和分子生物学的证据), 研究5大生物学特性(生殖特性、 发育特性、 光合特性、 抗逆性和品质等特性)。 The 7 research puzzles in the genetic improvement of biological bodies made by ion beam mediated technique, are worth noticed. The technical ideas, including one mediated technique in physics, 2 significant subjects, 3 effective changes, the mediated evidences of 4 aspects and 5 biological characteristics, were particularly put forward according to the existing states in the field. The 2 significant subjects consist of the mechanics of the allogenetic materials entering into the acceptor and they being to be recombined. The 3 effective changes include from studying morphology to genetic laws, from researching M1 generation to the next generations, from determining the single character to the synthetic traits. The mediated evidences of 4 aspects come from morphology, physiology and biochemistry, molecule biology. The 5 biological characteristics are mainly reproduction, development, photosynthesis, bad condition resistant and quality.     

模型化研究药物小分子阻滞细胞周期

基于扩散动力学与细胞信号传导动力学,研究药物小分子对细胞周期的阻滞特性.理论模型考虑药物小分子穿越细胞膜输运的动力学特性,以及进入细胞内的药物分子对细胞周期的阻滞效应.研究发现,穿越细胞膜内层进入细胞内的药物分子,将会在很大程度上决定药物分子对相关的靶向基因、蛋白的阻滞功效.细胞膜对药物分子的输运特性,是影响药物分子阻滞细胞周期的关键因素.另外,药物分子的降解程度,将会改变药物分子与靶向基因、蛋白作用时间,进而改变对相关细胞生长增殖的抑制效应.通过对模型中各参数的敏感度分析,我们确认了药物分子穿越细胞膜、进入细胞内过程的多种因素对细胞周期的抑制效应.本文理论结果符合模拟、实验观测,进一步深刻揭示了药物小分子阻滞细胞周期的物理机制,可为设计确切的疗法药物提供必要的参考和新方案.     

岩白菜属植物研究的新进展

从植物分布、栽培技术、生物学特性、化学成分以及药理作用5个方面综述了岩白菜研究取得的新进展,同时分析了当前研究中存在的问题,并对岩白菜今后的研究和发展方向进行了展望.     

激光技术在林木和园艺植物育种及基因工程中的应用

激光具有极高的功率密度，因而可被应用于种子催芽和诱变育种，可作为林木和园艺植物育种和基因改良的新技术手段。文章从四个方面进行了探讨：（1）激光辐照的生物效应机理；（2）激光在林木和园艺植物种子催芽中的应用；（3）激光在林木和园艺植物诱变育种中的应用；（4）激光在林木和园艺植物基因工程中的应用。     

Adamantane Derivatives Functionalized Gold Nanoparticles for Colorimetric Detection of MiRNA

MiRNAs are riveting RNA molecules due to their close relevance to the regulation of gene expression and certain physiological or pathological processes. Rapid and sensitive methods for miRNA assay are essential for biological researches and clinical diagnosis of many diseases. In this work, gold nanoparticles (AuNPs) have been modified with adamantane derivatives and then Dwith NA probes for colorimetric detection of miRNA. Target miRNA induces a change in DNA conformation and initiates strand displacement amplification, eventually leading to massive aggregations of AuNPs. By comparing the UV–vis absorption spectra, miRNA concentration can be determined. This developed method can detect miRNA as low as 3.7 × 10^?15m with remarkable specificity. Moreover, it is successfully used to inspect the expression of miRNA in biological samples. Therefore, adamantane derivatives functionalized AuNPs are demonstrated to offer a novel platform for biosensing and the miRNA sensing strategy may find a broad spectrum of practical applications.     

Two-color, double-slit in vivo flow cytometer

The in vivo flow cytometer enables the real-time detection and quantification of fluorescent cells circulating within a live animal without the need for incisions or extraction of blood. It has been used in demonstrating flow velocity disparities in biological flows, and in the investigation of the circulation kinetics of various types of cells. However, a shortcoming of this in vivo flow cytometer is that it provides only one excitation slit at one wavelength, resulting in several performance limitations. Therefore, a second in vivo flow cytometer that provides two different laser wavelengths, 473 and 633 nm, and one or two excitation slits has been designed and built. Thus far, the two-color system has been used to acquire circulation kinetics data of two different cell populations each labeled with a different marker, one cell population labeled with two different markers, and one cell population expressing the green-fluorescent protein gene. In addition, accurate arterial red blood cell velocities within a mouse have been determined using the cytometer.     

DNA微阵列制备的新进展

随着人类基因组测序的完成和蛋白组学工程的开展，具有高产出测序的特性的DNA微阵列技术的发展日新月异，其应用已经深入到了生命科学研究的很多方面．与此同时，几种新的DNA微阵列制备技术迅速发展起来，文章介绍了滚轮放大技术、聚酰胺胺表面法、三甲氧基对胺苯基硅烷／重氮化法、化学纳米印迹法等四种制备方法的新进展。     

Simultaneous Expression of GUS and Actin Genes by Using the Multiplex RT-PCR and Multiplex Gold Nanoparticle Probes

Gene expression analysis is considered to be extremely important in many different biological researches. DNA-based diagnostic test, which contributes to DNA identification, has higher specificity, cost, and speed than some biochemical and molecular methods. In this study, we try to use the novel nano technology approach with Multiplex RT-PCR and Gold nano particular probes (GNPs-probes) in order to get gene expression in Curcumas melons. We used Agrobacterium tumefactions for gene transfer and GUS reporter gene as a reporter. After cDNA synthesis, Multiplex PCR and Multiplex RT-PCR techniques were used. Finally, probes were designed for RNA of GUS and Actin genes, and then the analysis of the gene expression using the probes attached to GNPs was carried out and the color changes in the GNPs were applied. In the following, probes hybridization was checked with DNA between 400 to 700 nm wavelengths and the highest rate was observed in the 550 to 650 nm. The results show that the simultaneous use of GNP-attached detectors and Multiplex RT-PCRcan reduce time and costmore considerably than somelaboratory methods for gene expiration investigation. Additionally, it can be seen thatthere is an increase in sensitivity and specificity of our investigation. Based on our findings, this can bea novel study doneusingMultiplex RT-PCRand unmodified AuNPs for gene transfer and expression detection to plants. We can claim that this assay has a remarkable advantage including rapid, cost-effectiveness, specificity and accuracy to detect transfer and expression genes in plants. Also,we can use this technique from other gene expressionsin many different biology samples.     

Measuring technique for thermal ionisation mass spectrometry of human tracer kinetic study with stable cerium isotopes

Thermal ionisation mass spectrometry (TIMS) method has been developed for the simultaneous detection of different cerium isotopes in biological samples (i.e., blood and urine) at very low concentrations. The work has been done in the frame of a biokinetic study, where different stable cerium isotopes have been administered orally and intravenously as tracers to the human body. In order to develop an appropriate detection method for the tracers in the biological samples, an optimum sample preparation technique has been set and adapted to the specific requirements of the analysis technique used, i.e., TIMS. For sample evaporation and ionisation, the double tantalum filament technique showed the best results. The ions produced were simultaneously collected on a secondary electron multiplier so that the isotopic ratios of the cerium isotopes in the biological samples could be measured. The technique has been optimised for the determination of cerium down to 1?ng loaded on the evaporation filament corresponding to cerium concentrations of down to 1?ng?ml(-1) in the blood or urine samples. It has been shown that the technique is reliable in application and enables studies on cerium metabolism and biokinetics in humans without employing radioactive tracers.     

Study on the Interaction of the CpG Alternating DNA with CdTe Quantum Dots

A novel sensitive method for detection of DNA methylation was developed with thioglycollic acid (TGA)-capped CdTe quantum dots (QDs) as fluorescence probes. Recognition of methylated DNA sites would be useful strategy due to the important roles of methylation in disease occurrence and developmental processes. DNA methylation occurs most often at cytosine-guanine sites (CpG dinucleotides) of gene promoters. The QDs significantly interacted with hybridized unmethylated and methylated DNA. The interaction of CpG rich methylated and unmethylated DNA hybrid with quantum dots as an optical probe has been investigated by fluorescence spectroscopy and electrophoresis assay. The fluorescence intensity of QDs was highly dependent to unmethylated and methylated DNA. Specific site of CpG islands of Adenomatous polyposis coli (APC), a well-studied tumor suppressor gene, was used as the detection target. Under optimum conditions, upon the addition of unmethylated dsDNA, the fluorescence intensity increased in linear range from 1.0?×?10^??10 to 1.0?×?10^??6M with detection limit of 6.2?×?10^??11 M and on the other hand, the intensity of QDs showed no changes with addition of methylated dsDNA. We also demonstrated that the unmethylated and methylated DNA and QDs complexes showed different mobility in electrophoresis assay. This easy and reliable method could distinguish between methylated and unmethylated DNA sequences.     

高效液相色谱及联用技术在砷形态分析中的应用

砷与人类的生活息息相关,是环境中存在的一种极为重要的非金属元素。由于不同形态的砷其毒性各不同,因此无论对环境样品、食品、药物还是对人体的体液进行砷的形态分析都是至关重要的。高效液相色谱因具有较高的分离能力而被广泛地应用于砷形态的分离研究中。随着对分析灵敏度与准确度要求的不断提高,作为砷形态分析的检测器不断发展,不同的联用技术也日益完善。其中,电感耦合等离子体质谱法以其极高的灵敏度、多元素同时检测能力、极宽的动态范围以及同位素比检测能力而被成功地作为高效液相色谱的检测器,广泛地应用于砷的形态分析中。     

OCT技术在发育生物学中的应用

基于光学低相干反射测量而发展起来的光学相干层析技术（ Optica1 Coherence Tomography,OCT）是一种新型的成像技术，它可以对强散射介质如生物组织实施非侵入性的快速活体成像。与传统的组织切片相比,OTC可以大大减少对发育形态成像所需的时间、复杂程度及成本。非接触和非侵入式的成像方式有助于研究个体发育中由基因变异所引起的形态和功能变化，因而在发育生物学的研究当中有着广阔的应用前景。概述了OCT技术在发育生物学当中诸方面的应用情况。     

Measuring technique for thermal ionisation mass spectrometry of human tracer kinetic study with stable cerium isotopes

Thermal ionisation mass spectrometry (TIMS) method has been developed for the simultaneous detection of different cerium isotopes in biological samples (i.e., blood and urine) at very low concentrations. The work has been done in the frame of a biokinetic study, where different stable cerium isotopes have been administered orally and intravenously as tracers to the human body.

In order to develop an appropriate detection method for the tracers in the biological samples, an optimum sample preparation technique has been set and adapted to the specific requirements of the analysis technique used, i.e., TIMS. For sample evaporation and ionisation, the double tantalum filament technique showed the best results. The ions produced were simultaneously collected on a secondary electron multiplier so that the isotopic ratios of the cerium isotopes in the biological samples could be measured.

The technique has been optimised for the determination of cerium down to 1 ng loaded on the evaporation filament corresponding to cerium concentrations of down to 1 ng ml^?1 in the blood or urine samples. It has been shown that the technique is reliable in application and enables studies on cerium metabolism and biokinetics in humans without employing radioactive tracers.