THE CRYSTALLIZATION BEHAVIOUR OF BLOCK COPOLYMER/HOMOPOLYMER BLENDS I. INFLUENCE OF NONSPHERICAL MICELLE ON CRYSTALLIZATION BEHAVIOUR IN DILUTE SOLID SOLUTIONS

The effect of micelle on crystallization behaviour of dilute poly(methyl methacrylate-b-tetrahydrofuran) diblock copolymer/tetrahydrofuran homopolymer, dilute poly (ethylene-b-styrene-b-ethylene) triblock copolymer/ethylene homopolymer solutions has been studied. The results show that with the structural teansitions from spherical to nonspherical micelle in the blends, great changes in the nucleation and spherulite morphologies take place.

Kinetic and Size Control of Polystyrene and Polyacrylic Octadecyl Ester Lattices Via Polymerization in O/W Microemulsions

The kinetic studies of the acrylic octadecyl ester and styrene polymerization in microemulsion systems, (1) cetyl pyridine bromide (CPDB)/t-butanol/styrene/water; (2) CPDB/t-butanol/toluene + acrylic octadecyl ester (1:1, w/v)/ water; (3) cetyl pyridine bromide/styrene/formamide, were made by using dynamic laser light scattering techniques (DLS). The mechanisms of nucleation of latex particles were discussed. The most possible nucleation location of the styrene and acrylic octadecyl ester microlatex particles in aqueous microemulsion system is in aqueous phase via homogeneous nucleation. Meanwhile, parts of microlatex particles are possibly produced via swollen micelles (microemulsions) and monomer droplets nucleation. On the other hand, the most possible nucleation location of the styrene microlatex particles in nonaqueous microemulsion system is inside monomer droplets. The relationship between the amount of monomer and the size of microlatex was also investigated. It has been found that the size of microlatex particles could be controlled by changing the amount of monomer. (C) 2002 Elsevier Science B.V. All rights reserved.

配液结晶法制备溶菌酶蛋白质晶体的生长机理研究

采用配液结晶法制取了溶菌酶蛋白质晶体,使用动态光散射测量了溶液中聚集体的颗粒度几率分布;使用Zeiss显微镜测定了溶菌酶(110)晶面的生长速度.实验表明:随着蛋白质和NaCl浓度的增加,溶液中聚集体的颗粒尺寸也相应增加.随着反应时间的增加,溶菌酶分子在溶液中的聚集反应,逐渐达到平衡;在蛋白质和NaCl浓度较高时,溶菌酶晶体的(110)面生长较快,而在蛋白质和NaCl浓度较低时,该晶面生长较慢.基于二维成核生长机理,从晶体生长动力学理论方程出发,计算了二维成核的形成能a=4.01×10-8J?cm-2.

Detection of constitutive heterodimerization of the integrin Mac-1 subunits by fluorescence resonance energy transfer in living cells

Macrophage differentiation antigen associated with complement three receptor function (Mac-1) belongs to beta(2) subfamily of integrins that mediate important cell-cell and cell-extracellular matrix interactions. Biochemical studies have indicated that Mac-1 is a constitutive heterodimer in vitro. Here, we detected the heterodimerization of Mac-1 subunits in living cells by means of two fluorescence resonance energy transfer (FRET) techniques (fluorescence microscopy and fluorescence spectroscopy) and our results demonstrated that there is constitutive heterodimerization of the Mac-1 subunits and this constitutive heterodimerization of the Mac-1 subunits is cell-type independent. Through FRET imaging, we found that heterodimers of Mac-1 mainly localized in plasma membrane, perinuclear, and Golgi area in living cells. Furthermore, through analysis of the estimated physical distances between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) fused to Mac-1 subunits, we suggested that the conformation of Mac-1 subunits is not affected by the fusion of CFP or YFP and inferred that Mac-1 subunits take different conformation when expressed in Chinese hamster ovary (CHO) and human embryonic kidney (HEK) 293T cells, respectively. (c) 2006 Elsevier Inc. All rights reserved.

Detection of constitutive homomeric associations of the integrins Mac-1 subunits by fluorescence resonance energy transfer in living cells

Integrins alpha(M)beta(2) plays important role on leukocytes, such as adhesion, migration, phagocytosis, and apoptosis. It was hypothesized that homomeric associations of integrin subunits provide a driving force for integrins activation, and simultaneously inducing the formation of integrins clusters. However, experimental reports on homomeric associations between integrin subunits are still controversial. Here, we proved the homomeric associations of the isolated Mac-1 subunits in living cells using three-channel fluorescence resonance energy transfer (FRET) microscopy and FRET spectra methods. We found that the extent of homomeric associations between beta(2) subunits is higher than alpha(M) subunits. Furthermore, FRET imaging indicated that the extent of homomeric associations of the Mac-1 subunits is higher along the plasma membrane than in the cytoplasm. Finally, we suggested that homomeric associations of the transmernbrane domains or/and cytoplasmic domains may provide the driving force for the formation of constitutive homomeric associations between alpha(M) or beta(2) subunits. (c) 2006 Elsevier Inc. All rights reserved.

脂筏极化参与活性氧介导的白杄花粉管顶端生长

质膜上存在一种富含甾醇物质的液相有序膜脂微区，被称作脂筏 (lipid rafts或lipid microdomains)。这种小的膜微区可以通过在质膜上的侧向移动，聚集形成较大的片状结构，而与微区相关联的蛋白可以通过脂筏的这种聚合作用而凝聚分布于特定的亚细胞结构上。脂筏区域在真菌和动物质膜上具极性分布，并参与细胞的极性形态建成和运动。最近，通过生物化学研究证实，脂筏也存在于植物细胞，然而迄今为止，脂筏与植物细胞极性生长相关联的直接功能证据尚未见报道。 NADPH氧化酶 (NOX，在植物中又称为 Rboh) 产生的活性氧 (Reactive oxygen species, ROS) 可能是调控植物细胞（包括花粉管、根毛和墨角藻合子等）极性生长的通用信号机制。花粉管作为研究细胞极性控制的一种理想模式系统，已被许多信号转导调控研究所采用。在本研究中，我们使用一种能螯合甾醇类物质的多烯类抗生素filipin破坏脂筏结构，以探讨脂筏极化对ROS介导的白杄花粉管极性生长的作用。 我们首次在白杄 (Picea meyeri) 花粉管上应用一种全新的苯乙烯基染料di-4-ANEPPDHQ，成功地在活体细胞上观察到脂筏在花粉管生长顶端的极性分布模式。通过脂筏和甾醇在质膜上的相似定位清楚表明：在花粉管极性生长过程中，存在富含甾醇类物质的质膜微区在花粉管生长顶端的极化现象。 氮蓝四唑（NBT）的还原和二氯二氢荧光素（H2DCF）的氧化均显示，在活跃生长的花粉管顶端区域存在一个以顶端为基底的陡峭ROS梯度，从而进一步验证了ROS在细胞极性生长过程中的信号作用。此外，我们还发现在生长花粉管的亚顶端位置有另一类性质的活性氧组分存在，该ROS组分与线粒体的能量代谢相关。研究结果首次揭示，在快速生长的花粉管中同时存在两类性质不同的ROS组分。 ROS是一种寿命很短而且容易扩散的分子，NADPH氧化酶产生的ROS信号在细胞伸长位点的准确定位是调控极性生长的必要条件。免疫共定位实验显示，NOX成簇极化分布于花粉管的生长顶端。使用filipin进行甾醇的螯合会破坏膜的异质性，干扰NOX簇在生长顶端的定位，减少了顶端的ROS形成，消弱了胞质Ca2+ 浓度梯度，进而抑制了花粉管的顶端生长。 在纯化质膜的基础上，我们使用Triton去垢剂处理结合Optiprep密度梯度离心，分离纯化了抗去垢剂抽提的质膜微区 (Detergent-resistant microdomains, DRMs)。通过免疫印迹分析证实，NADPH氧化酶部分地存在于DRMs中。非变性胶活性实验证明，该酶需要脂筏定位来保持酶活性。因此我们认为，在正常的细胞极性生长中，脂筏招募并运载NADPH氧化酶到花粉管的生长顶端，并为NOX及其活性亚基的有效互作提供了适宜的微环境，由此保证了NOX蛋白产生ROS的较高酶活性，进而维持花粉管的极性顶端生长。 总之，甾醇螯合对白杄花粉管生长影响的研究，为脂筏极化在花粉管极性生长中的作用提供了证据。基于以上生物化学和细胞生物学的结果，我们针对花粉管中富含甾醇的脂筏微区和NOX功能之间的联系，提出了一种假说模式：(1) 植物细胞质膜上的脂筏为信号分子ROS在特定位点的聚集提供了物理载体；(2) 脂筏的完整性和甾醇依赖性对NOX的定位和活性是必要的，并为花粉管细胞极性产生和维持所必需。上述研究结果表明，脂筏在花粉管顶端的极化，以及作为关键生长因子的NOX在质膜脂筏中的定位，对花粉管的高度极性生长具有重要作用。

Contributions of phosphatase and microbial activity to internal phosphorus loading and their relation to lake eutrophication

Phosphatase may accelerate the process of lake eutrophication through improving phosphorus bioavailability. This mechanism was studied in three Chinese eutrophic shallow lakes (Lake Taihu, Lake Longyang,and Lake Lianhua). Phosphatase activity was related to the concentration of soluble reactive phosphorus (SRP) and chlorophyll a. Stability of dissolved phosphatase in reverse micelles may be attributed to molecular size, conformation and active residues of the enzyme. At the site with Microcystis bloomed in Lake Taihu, dissolved phosphatase activity was higher and more stable in micelles, SRP concentrations were lower in interstitial water, the contents of different forms of phosphorus and the amounts of aerobic bacteria were lower while respiration efficiency was higher in sediments. Phosphobacteria, both inorganic and organic and other microorganisms were abundant in surface water but rare in sediments. Therefore, internal phosphorus may substantially flux into water column by enzymatic hydrolysis and anaerobic release, together with mobility of bacteria, thereby initiating the bloom. In short, biological mechanism may act in concert with physical and chemical factors to drive the internal phosphorus release and accelerate lake eutrophication.

Vertical variation in dissolved alkaline phosphatase activity in a shallow eutrophic lake determined in reverse micelles

We evaluated the feasibility of microencapsulating dissolved alkaline phosphatase of a water body into reverse micelle systems prepared by hexadecyltrimethylammonium bromide as a surfactant in cyclohexane and 1-butanol as co-surfactant. The dissolved alkaline phosphatase activity within the micelle was described, including its kinetic parameters and the effects of pH and temperature on catalytic activity in surface, overlying and interstitial water of Lake Donghu. We found the similarities on the behavior of dissolved alkaline phosphatase of surface and interstitial water in reverse micelles, which was distinctly different from its behavior in the overlying water. This difference likely reflected the different origins of the dissolved alkaline phosphatase in the vertical profile of the lake. This system provides a novel tool with which to study the diversity and ecological significance of extracellular enzymes in aquatic environments.

Water solubility enhancement of phthalates by cetyltrimethylammonium bromide and beta-cyclodextrin

Water solubility enhancements of six phthalates (five aliphatic phthalates and one phenyl phthalate) by cetyltrimethylammonium bromide (CTAB) and beta-cyclodextrin (beta-CD) were studied at 25 degreesC. The solubilities of these plithalates are remarkably enhanced by CTAB solutions above the critical micelle concentration (cmc). Only marginal enhancement of phthalate solubility was observed in solutions containing CTAB below its cmc and beta-CD at low concentrations (less than 5 mM). The solubility enhancements of the plithalates are proportional to the added amount of CTAB and beta-CD. Partition coefficients of the plithalates between monomeric CTAB surfactant and water (K-MN) and between CTAB micelle and water K-MC) were estimated from the experimental data. The mechanisms of solubility enhancements by CTAB and beta-CD were discussed. A log-linear equation was proposed and evaluated for the solubilization by CTAB below cmc, while the previously proposed linear partitioning model was questioned. The structures of the complexes formed between plithalates and beta-CD were proposed, and the formation constants were estimated. The values of log K-MC, log K-MN, and log Kbeta-CD of the plithalates were found to correlate linearly with the log K-OW of plithalates, with the exception of the solid phenyl phthalate.

INFLUENCE OF LINEAR ALKYLBENZENE SULFONATE (LAS) AS ORGANIC COSOLVENT ON LEACHING BEHAVIOR OF PCDD/FS FROM FLY-ASH AND SOIL

The leaching of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) was measured in soil and standard fly ash column eluted with pure water and linear alkylbenzene sulfonate (LAS)- water. The data obtained were used to evaluate the leachability of PCDD/Fs from waste dump like incineration residual slag and fly ash deposition. The leaching rate was shown to be increased significantly by using LAS water. The leachate contents of PCDD/Fs were above their known water solubility. Concentration of PCDD/Fs in the leachates as well as the relative leaching (calculated on the fly ash content) increased with increasing chlorinating degree and decreasing water solubility. LAS above the critical micelle concentration (CMC) probably enhances PCDD/Fs solubility.

一个支持可信主体特权最小化的多级安全模型

对已有多级安全模型的可信主体支持进行回顾和分析,提出了DLS(离散标记序列)多级安全模型.该模型将可信主体的生命周期分解为一系列非可信状态,对每一个状态赋予一个敏感标记.可信主体的当前敏感标记等于当前非可信状态的敏感标记,非可信状态的切换由预定义的可信请求事件触发.从而可信主体的当前敏感标记可以根据其应用逻辑而动态调整.同时给出了模型保持系统安全性的安全状态和规则.与Bell模型等可信主体敏感标记范围模型相比,该模型在多级安全的策略范围内实现了可信主体的特权最小化.

水凝胶的制备及壳聚糖的改性和应用

水凝胶用作创口敷料时有很多的优点。壳聚糖是天然的阳离子多糖,已经在很多领域得到了应用。我们制备了几种具有抑菌能力的水凝胶敷料，同时利用可生物降解聚酯和多聚肽对壳聚糖进行了修饰，得到了一些新的结果。 本论文的工作主要包括以下两部分： 一.含有抗生素或纳米银粒子的水凝胶的制备和表征。 1．载药聚乙烯醇/聚乙烯基吡咯烷酮/壳聚糖（PVA-PVP-Chitosan）复合水凝胶的制备和抑菌性能评价。采用循环冷冻－熔融与辐射交联相结合的方法制备出了PVA-PVP-chitosan复合水凝胶，测定了凝胶的溶胀率、凝胶含量、力学强度和结晶性能，并考察了所用壳聚糖分子量，冷冻－熔融循环处理次数和辐射剂量对凝胶性能的影响。然后通过溶胀法将抗生素（环丙沙星）和壳寡糖载入水凝胶中，考察了它们的体外释放行为。同时利用琼脂糖平板法检验了载药凝胶的抑菌能力，结果表明它们可以有效抑制革兰氏阳性菌和革兰氏阴性菌的繁殖和生长。 2．含纳米银-PVA-PVP水凝胶的制备和抑菌性能评价。通过循环冷冻－熔融法制备了含银的PVA-PVP复合水凝胶。以含银的PVA-PVP复合膜为模型，利用DSC、红外光谱、紫外－可见光谱和X-射线衍射考察了掺入的纳米银与PVA-PVP网络的相互作用。利用扫描电镜观察了PVA-PVP凝胶的网络结构和银纳米粒子在网络中的分布。利用琼脂糖平板法和LB肉汤法对含银PVA-PVP凝胶的抑菌性能进行了评价，结果证明这种水凝胶可以有效抑制大肠杆菌和金黄色葡萄球菌的生长和繁殖。 二. 壳聚糖的改性和应用。 1．发展了一种制备聚ε－己内酯（PCL）接枝壳聚糖共聚物的新方法，并且由合成的接枝共聚物在水中自组装得到纳米粒子。利用DLS、AFM和SEM对纳米粒子进行了表征，结果显示制备的纳米粒子为圆球形或椭圆形。利用茚三酮显色法准确测定了活性胺基的含量，发现纳米粒子表面活性胺基的数量可以通过改变PCL接枝度进行调节。 2．由6-O-三苯甲基壳聚糖引发赖氨酸NCA的开环聚合得到了聚赖氨酸（PLL）接枝壳聚糖阳离子共聚物。利用红外光谱、核磁共振和GPC对接枝共聚物的分子结构和分子量进行了表征。通过动态光散射测量发现接枝共聚物可以和小牛胸腺DNA形成粒径在100~340 nm之间的复合物。凝胶阻滞电泳证明接枝共聚物对质粒DNA的结合和保护能力要优于壳聚糖。细胞转染实验证明接枝共聚物对293T和Hela细胞的转染效率要远远高于壳聚糖原料和PLL。同时，接枝共聚物的转染效率对PLL的聚合度有明显的依赖性，增加PLL聚合度有助于提高接枝共聚物的转染效率。这种新型的阳离子接枝共聚物有望用作高效基因载体。