Enhancement of transfection efficiency for HeLa cells via incorporating arsinine moiety into chitosan

Arginine-rich peptides have attracted considerable attention due to their distinct internalization mechanism. It was reported that arginine and guanidino moieties were able to translocate through cell membranes and played a critical role in the process of membrane permeation. In this work, arginine was conjugated to the backbone of chitosan to form a novel chitosan derivative, arginine modified chitosan （Arg-CS）. Arg-CS/DNA complexes were prepared according to the method of coacervation process. The physicochemical properties of Arg-CS and Arg-CS/DNA complexes were characterized and the transfection activity and efficiency mediated by Arg-CS/DNA complexes were investigated taking HeLa cells as target cells. Arg-CS was characterized by FTIR and ^13C NMR. Arg-CS/DNA polyelectrolyte complexes were investigated by agarose gel retardation, dynamic light scattering （DLS） and atomic force microscopy （AFM）. The results revealed that the Arg-CS/DNA complexes started to form at N/P ratio of 2：1, and the size of particles varied from 100 to 180 nm. The cytotoxicity of Arg-CS and their complexes with plasmid DNA were determined by MTT assay for HeLa cells, and the results suggested that Arg-CS/DNA complexes were slightly less toxic than Arg-CS. Moreover, the derivative alone and their complexes showed significantly lower toxicity than PEI and PEI/DNA complexes, respectively. Taking HeLa cells as target cells and using pGL3-control as reporter gene, the luciferase expression mediated by Arg-CS was greatly enhanced to about 100 folds compared with the luciferase expression mediated by chitosan at different pH media. These results suggest that Arg-CS is a promising candidate as a safe and efficient vector for gene delivery and transfection.     

Self-assembly of insulated molecular wires of a watersoluble cationic PPV and anionic dendrons

Insulated molecular wires of poly（phenylenevinylene） （PPV） were prepared by wrapping the conjugated backbones with dendrons through a noncovalent approach. It was found that electrostatic interaction between the quaternary ammonium groups of PPV-1 and the carboxylate moieties in dendrons induced the packing of dendrons along PPV-1 conjugated backbones. Absorption and emission spectroscopic examinations in solution and solid film indicated that the PPV-1 backbones adopted a more planar and isolated conformation in the complexes. Furthermore, interchain interactions in the complexes could be greatly reduced, improving the quantum yield of PPV-1.     

Enhancement of transfection efficiency for HeLa cells via incorporating arginine moiety into chitosan

Arginine-rich peptides have attracted considerable attention due to their distinct internalization mechanism. It was reported that arginine and guanidino moieties were able to translocate through cell membranes and played a critical role in the process of membrane permeation. In this work, arginine was conjugated to the backbone of chitosan to form a novel chitosan derivative, arginine modified chitosan (Arg-CS). Arg-CS/DNA complexes were prepared according to the method of coacervation process. The physicochemical properties of Arg-CS and Arg-CS/DNA complexes were characterized and the transfection activity and efficiency mediated by Arg-CS/DNA complexes were investigated taking HeLa cells as target cells. Arg-CS was characterized by FTIR and 13C NMR. Arg-CS/DNA polye- lectrolyte complexes were investigated by agarose gel retardation, dynamic light scattering (DLS) and atomic force microscopy (AFM). The results revealed that the Arg-CS/DNA complexes started to form at N/P ratio of 2:1, and the size of particles varied from 100 to 180 nm. The cytotoxicity of Arg-CS and their complexes with plasmid DNA were determined by MTT assay for HeLa cells, and the results suggested that Arg-CS/DNA complexes were slightly less toxic than Arg-CS. Moreover, the derivative alone and their complexes showed significantly lower toxicity than PEI and PEI/DNA complexes, respectively. Taking HeLa cells as target cells and using pGL3-control as reporter gene, the luciferase expression mediated by Arg-CS was greatly enhanced to about 100 folds compared with the luciferase expression mediated by chitosan at different pH media. These results suggest that Arg-CS is a promising candi- date as a safe and efficient vector for gene delivery and transfection.     

Synthesis,structure and catalytic behavior of yttrium complexes bearing a diaminobis（phenolate） ligand

Yttrium complexes stabilized by a diaminobis（phenolate） ligand were synthesized and their catalytic behavior was explored. Reaction of YCI3 with 1 equiv of LNa2 [L= Me2NCH2CH2N{CH2-（2-O-C6H2-^1Bu2-3,5）}2] gave the yttrium chloride LYCI（THF） （1） in 92% yield. Complex I can be used as starting material to prepare the yttrium amido derivative. Complex 1 reacted with 1 equiv of LiNPh2 in THF to afford the expected yttrium amido complex LYNPh2 （2） in high yield. Both of complexes 1 and 2 have been well detected by elemental analysis, NMR spectra and single-crystal X-ray analysis. It was found that complex 2 can efficiently initiate the ring-opening polymerization of L-lactide and ε-caprolactone, and a controlled manner is observed in the former case.     

Evaluation of virus removal in MBR using coliphages T4

A membrane bioreactor (MBR) with gravity drain was tested for domestic wastewater for 65 days. Resultss howed that the effluent quality was excellent, and met with the reuse water standard of China (GB/T 18920-2002). Virus removal in the membrane separation process was investigated by employing coliphages T4 as a tracer. Two microfiltration membrane modules, with pore sizes of 0.22 and 0.1 μm, were used to investigate their effects on virus rejection at the transmembrane pressure of 8.5 kPa. It was found that 0.1 μm membrane had complete rejection of virus, and 0.22 μm membrane had significant rejection of virus. In the longterm operation of this MBR, no significant difference was observed between both pore sizes because the virus concentrations of the effluent in both cases were in the same order. Effluent virus concentration at steady state of MBR running was less than 2 PFU/mL. The removal ratios of coliphage T4 in MF processes were more than 10^5.5. The membrane surface deposits played an important role in the rejection of virus. The formation of cake clay on the membrane surface was the main cause of high rejection of colipbage T4 with MF of 0.22 μm.     

Cyclic voltammetry,convolutive voltammetry,chronopotentiometry and disital simulation studies of [Pt（C≡C tol）2（dppm）2lr（CO）2]^＋PF^-6 complex

The electrochemical behaviour of the heterobimetallic complex [Pt（C≡C tol）2（dppm）2lr（CO）2]^＋PF^-6 was studied via cyclic voltammetry, convolutive voltammetry and chronopotentiometry at glassy carbon electrode in dichloromethane solution. The electrochemical parameters calculated from experimental data were tested and confirmed by matching the experimental cyclic voltammograms with the simulated data. It was found that convolutive voltammetry provided higher sensitivity, better resolution and more accurate method for determination of the electrochemical parameters than ordinary cyclic voltammetry.     

Influence of Dispersion of Nano-Zn0 Particles in Polymer Matrices on Properties of Relevant Nano Composite Fibers

The surface-passivated and non-surface-passivated zinc oxide nano-particles （marked as s-nanoZnO and ns-nanoZnO respectively） were evenly dispersed in polymer solutions with the aid of ultrasonic vibration to prepare nanocomposite film by free casting and to prepare nanocomposite fibers by wet spinning and to prepare nancomposites coating by surface smearing. The dispersion of s-nanoZnO and nsnanoZnO in PAN matrix were observed by transmittance electron microscopy, the mechanical properties of the relevant compesite samples were studied by INSRTON tensile strength tester. It was found that s-nanoZnO behaves a well-dispersed morphology in PAN films and fibers when its concentration was 2 wt% but ns-nanoZnO nano particles agglomerate into larger congeries in PAN films. It means that the surface-passivated process oft zinc oxide nano. particles was effective to disperse. The relative intensity and elonsation at break of s-nanoZnO-PAN composite fibers show maximum values with the increase of nano particle content in compesites （from 0 wt% to 2 wt% of s- nanoZnO）. The elasticity of the composite fibers increases whereas their modulus declines. Balanced the changes of the properties mentioned above, 2 wt% s-nanoZnO in PAN matrix is a proper content for the composite fibers spun by wet spinning. The result of surface smearing test means that the reactim between s-nanoZnO and polymer can be indicated by the color of nanocomposite surface coat on fibers.     

A strategy to obtain a high-density and high-strength zirconia ceramic via ceramic injection molding by the modification of oleic acid

Despite its unique high efficiency and good environmental compatibility, the water-soluble binder system still encounters problems achieving a desired sintered part via ceramic injection molding because of the poor compatibility and the powder-binder segregation between ceramic powders and binders. The objective of this study was to obtain a sintered part with excellent properties by introducing a small quantity of oleic acid to the surface of zirconia powders before the mixing process. As opposed to many previous investigations that focused only on the rheological behavior and modification mechanism, the sintering behavior and densification process were systematically investigated in this study. With the modified powders, debound parts with a more homogeneous and smaller pore size distribution were fabricated. Also, a higher density and greater flexural strength were achieved in the sintered parts fabricated using the modified powders.     

Effect of Extractant and Cold-drawing on the Structure and Performance of FIDPE Hollow Fiber Membranes Fabricated via Thermally Induced Phase Separation Method

Microporous polyolefin hollow fiber membranes were prepared from high density polyethylene （HDPE）-paraffin solution via thermally induced phase separation （TIPS） method. Effects of extraction and cold-drawing condition on membrane structure and performance were investigated.Five volatile solvents were used as extractant. Dimension of hollow fiber and gas permeation rate of membrane were measured. Mierostructure of membrane was examined by Scanning Electronic Microscope （SEM）. The results show that the membrane treated by pentane possesses a higher porosity, nitrogen permeability and lower shrinkage than those of membranes extracted by other three extractants. It is also found that the membrane stretched 133% shows the highest porosity and gas permeability in this study.     

Properties and Working Principle of Airstop Membranes

In this paper, the surface and cross-section, pore size and distribution, contact angle and hydrostatic pressure of the membrane were tested. Then the membrane＇s working principle was analyzed and the stopping height of membranes with different pore sizes was calculated. It was found that the surface tension and pore size of the membrane were key factors to determine its stopping height.     

甲壳三糖-g-壳聚糖/累托石纳米复合材料作为基因载体的研究

本文合成了甲壳三糖-g-壳聚糖（TCS），并与累托石插层复合制备纳米复合材料，采用XRD和TEM对其进行了表征。粒径分析结果表明，两种分子量的TCS与pDNA复合物的粒径大小都在75nm左右，而累托石加入后，其粒径都不同程度的增加，最大达到191nm。在PBS中的聚集动力学及琼脂糖凝胶电泳实验发现，高分子量TCS /pDNA复合物较低分子量复合物稳定，而累托石的加入降低了其稳定性。体外转染实验初步表明，甲壳三糖-g-壳聚糖/累托石-DNA复合物能介入肝癌细胞中并表达荧光。该纳米复合材料可作为潜在的非病毒基因载体。     

基于壳聚糖纳米颗粒的基因枪法转化洋葱细胞研究

以交联法制备壳聚糖纳米颗粒,用透射电子显微镜和Zeta电位仪对壳聚糖纳米颗粒进行表征,发现颗粒呈球形,粒径约为50rim,微球表面光滑、球形团整、颗粒比较均匀、分散性好,电位约为11．1mv;琼脂糖凝胶电泳结果显示,壳聚糖纳米颗粒能有效地结合质粒DNA,并能保护所结合的DNA防止DNaseⅠ的酶切;将含GFP基因的壳聚糖纳米颗粒复合物使用基因枪转化洋葱表皮细胞,在倒置荧光显微镜下观察,发现细胞表达了绿色荧光蛋白,表达效率为8％．     

Identification of the haemoglobin scavenger receptor

Intravascular haemolysis is a physiological phenomenon as well as a severe pathological complication when accelerated in various autoimmune, infectious (such as malaria) and inherited (such as sickle cell disease) disorders. Haemoglobin released into plasma is captured by the acute phase protein haptoglobin, which is depleted from plasma during elevated haemolysis. Here we report the identification of the acute phase-regulated and signal-inducing macrophage protein, CD163, as a receptor that scavenges haemoglobin by mediating endocytosis of haptoglobin-haemoglobin complexes. CD163 binds only haptoglobin and haemoglobin in complex, which indicates the exposure of a receptor-binding neoepitope. The receptor-ligand interaction is Ca2+-dependent and of high affinity. Complexes of haemoglobin and multimeric haptoglobin (the 2-2 phenotype) exhibit higher functional affinity for CD 163 than do complexes of haemoglobin and dimeric haptoglobin (the 1-1 phenotype). Specific CD163-mediated endocytosis of haptoglobin-haemoglobin complexes is measurable in cells transfected with CD163 complementary DNA and in CD163-expressing myelo-monocytic lymphoma cells.     

高铁酸钾对水中锑的去除机理研究

采用实验室制备的高铁酸钾对水溶液中的三价锑进行去除性能研究. 高铁酸钾对水中三价锑的去除包含2步:一是高铁酸钾对三价锑的氧化,二是原位生成的高铁酸钾分解产物对锑的吸附. XRD和FTIR检测表明高铁酸钾的分解产物符合无定型的2线水铁矿,比表面积大,孔隙率高. 吸附实验结果表明:低pH值更有利于锑的吸附;而离子强度对锑的吸附,在pH 3.5 ~5.5与pH 5.5~7.0这2个区间有着截然相反的影响;吸附动力学过程拟合以准二级动力学拟合最好(R2=0.999 1),说明以化学吸附为主;吸附等温线拟合以Freundlich模型最优(R2=0.980 4),而使用Langmuir-Freundlich模型拟合时最大吸附量的理论值可达到129.93 mg/g. 通过吸附前后样品的FTIR表征表明在吸附锑的过程中形成了内层络合物和外层络合物.     

Surface modification of nanogold particles in DNA detection with quartz crystal microbalance

Nanogold particles in different sizes, from 5 to 60 nm, were utilized to modify the surface of the quartz crystal microbalance (QCM). It was found that the gold-particle size of the modified QCM affects both the amount of the immobilization of the probe on the surface of QCM and the hybridization rate of the target DNA. 20 nm was determined to be the optimal size for the surface modification and it can maximally increase the sensitivity of the DNA detection.     

Study of the interaction of DNA and histones by spin-stretching and droplet evaporation

Molecular combing is a powerful and simple method for aligning DNA molecules onto a surface. Using this technique combined with fluorescence microscopy, DNA-histone complexes are stretched on a hydrophobic polymethyl methacrylate (PMMA) surface and observed directly. We have developed a new method to stretch single DNA-histone complexes, termed spin-stretching. The results show that the histones markedly enhance DNA binding to the PMMA surface. DNA winds around the histones and therefore decreases in length. The number of histones that bind to each DNA molecule is found to correlate with the histone concentration. The combed DNA-histone complexes are found to depend on two factors: the binding force on the surface and the centrifugal force at its local position. Na+ ions should compete with histones for binding to DNA; however, the observed competitive binding effect of Na+ ions at low concentrations was negligible.     

氨基化纳米银的制备及抗菌性研究

利用原位修饰法合成了表面氨基化纳米银,优化了纳米银的制备条件.通过紫外-可见光谱、傅里叶变换-红外光谱、X射线衍射、Zeta电势及透射电子显微镜等对其进行了分析和表征.对纳米银的抗菌性能进行了研究.结果表明:氨基化纳米银带有正电荷,能通过静电吸引作用结合表面带负电荷的细菌,使抗菌活性显著提高;氨基化的纳米银可有效抑制革兰氏阴性细菌和阳性细菌的生长;该材料可被应用于医学器件和细菌控制领域.     

Ectopic beta-chain of ATP synthase is an apolipoprotein A-I receptor in hepatic HDL endocytosis

The effect of high-density lipoprotein (HDL) in protecting against atherosclerosis is usually attributed to its role in 'reverse cholesterol transport'. In this process, HDL particles mediate the efflux and the transport of cholesterol from peripheral cells to the liver for further metabolism and bile excretion. Thus, cell-surface receptors for HDL on hepatocytes are chief partners in the regulation of cholesterol homeostasis. A high-affinity HDL receptor for apolipoprotein A-I (apoA-I) was previously identified on the surface of hepatocytes. Here we show that this receptor is identical to the beta-chain of ATP synthase, a principal protein complex of the mitochondrial inner membrane. Different experimental approaches confirm this ectopic localization of components of the ATP synthase complex and the presence of ATP hydrolase activity at the hepatocyte cell surface. Receptor stimulation by apoA-I triggers the endocytosis of holo-HDL particles (protein plus lipid) by a mechanism that depends strictly on the generation of ADP. We confirm this effect on endocytosis in perfused rat liver ex vivo by using a specific inhibitor of ATP synthase. Thus, membrane-bound ATP synthase has a previously unsuspected role in modulating the concentrations of extracellular ADP and is regulated by a principal plasma apolipoprotein.