12C6+束和Mitomycin C 诱导的DNA损伤效应

本文旨在分别研究重离子束及MMC在诱导细胞的DNA损伤效应中一些具体的分子机制，为治疗的进行以及相关辅助药物的开发提供理论依据。本文探索的重点有两个，第一个是重离子束辐射诱导的DNA损伤效应及p53在其中的激活，第二个是MMC诱导的DNA损伤效应及p53和BRCA1、H2AX等分子在其中的角色。 1. 12C6+离子束诱导HeLa细胞DNA损伤效应为了研究HeLa细胞经过12C6+ 束辐照之后的DNA损伤效应，及这个过程中p53激活的分子机制。我们运用中性单细胞电泳技术，检测了HeLa细胞经过4Gy 12C6+ 束辐照0h、3h、6h和12h之后DNA的损伤情况，以及0.5Gy、1Gy、2Gy和4Gy 12C6+ 束辐照0h后的DNA损伤情况。同时运用细胞生长实时监测仪监测了HeLa细胞在经过0Gy、0.5Gy和1Gy 12C6+ 束辐照之后的生长变化，并运用AO/EB双染检测了辐照24小时后的凋亡情况。另外，利用8mmol/L的caffeine（抑制ATM和ATR）和20μmol/L的wortmannin（抑制ATM和DNA-PK）处理HeLa细胞后再进行1Gy 12C6+ 束辐照，通过western blot检测p53的表达。结果显示，12C6+ 束辐照可造成HeLa细胞的DNA损伤，损伤随剂量升高而升高但随时间降低；并诱导HeLa细胞发生凋亡；而且辐照后p53表达升高，但经过caffeine或者wortmannin预先处理的细胞p53均没有显著升高。我们的结论是：12C6+ 束辐照可造成HeLa细胞的DNA损伤并诱导损伤修复及凋亡等效应，损伤效应相关的分子p53被激活，并且激活依赖于ATM。 2. MMC诱导的DNA损伤效应在这一部分研究中，首先，我们利用与上面相同的研究方法，探讨了p53在MMC诱导的DNA损伤效应中的激活情况，结果显示，MMC诱导的DNA损伤效应并不依赖于p53。另外，我们还探讨了， BRCA1在FANCD2的γ-H2AX依赖性转移中的作用。MMC可造成DNA的ICL（interstrand cross-link）损伤，ICL可通过FA(Fanconi Anemia)通路进行修复。FANCD2是FA通路的核心分子，在DNA产生ICL时被各种分子修饰然后转移到损伤部分，这个过程的涉及到ATR、γ-H2AX及BRCA1等，本文试图探讨BRCA1在其中的作用方式。研究中，我们监测了不同处理（包括对照、caffeine（可抑制ATR）、MMC及MMC ＋caffeine）的HCC1937(BRCA1缺陷型)和MCF7（BRCA1野生型）细胞的生长；并用Western blot检测MMC处理之后HCC1937细胞γ-H2AX的表达情况。结果表明，MMC和caffeine均可以抑制HCC1937的生长，但caffeine和MMC+caffeine的抑制效果是一样的；MMC和caffeine均可以抑制MCF7的生长，且MMC+caffeine处理比仅进行caffeine处理的抑制作用强；MMC处理之后，HCC1937的γ-H2AX表达显著升高。我们的结论是，在FANCD2的γ-H2AX依赖性转移中，H2AX的磷酸化并不依赖于BRCA1，不过，BRCA1和ATR应该参与一个相同的分子事件，可能是FANCD2的磷酸化。这个有待进一步的实验验证

Preparation and performance of cellulose acetate/polyethyleneimine blend microfiltration membranes and their applications

A modified microfiltration membrane has been prepared by blending a matrix polymer with a functional polymer. Cellulose acetate (CA) was blended with polyethyleneimine (PEI), which was then crosslinked by polyisocyanate, in a mixture of solvents. In the membrane, PEI can supply coupling sites for ligands in affinity separation or be used as ligands for metal chelating, removal of endotoxin or ion exchange. The effects of the time of phase inversion induced by water vapor, blended amount of PEI and amount of crosslinking agent on membrane performance were investigated. The prepared blend membranes have specific surface area of 12.04-24.11 m(2)/g and pure water flux (PWF) of 10-50 ml/cm(2) min with porosity of 63-75%. The membranes, made of 0.15 50 wt.% PEI/CA ratio and 0.5 crosslinking agent/PEI ratio, were applied to adsorbing Cu2+ and bovine serum albumin (BSA) individually. The maximum adsorption capacity of Cu2+ ion on the blend membrane is 7.42 mg/g dry membrane. The maximum adsorption capacities of BSA on the membranes with and without chelating Cu2+ ion are 86.6 and 43.8 mg/g dry membrane, respectively. (C) 2004 Elsevier B.V. All rights reserved.

Preparation and Characterization of Interpenetration Polymer Network Films Based on Poly(vinyl alcohol) and Poly(acrylic acid) for Drug Delivery

A series of full interpenetrating polymer network (full-IPN) films of poly(acrylic acid) (PAA)/poly (vinyl alcohol) (PVA) were prepared by radical solution polymerization and sequential IPN technology. Attenuated total reflectance-Fourier transform infrared spectroscopy, swelling properties, mechanical properties, morphology, and glass transition temperature of the films were investigated. FTIR spectra analysis showed that new interaction hydrogen bonds between PVA and PAA were formed. Swelling property of the films in distilled water and different pH buffer solution was studied. Swelling ratio increased with increasing PAA content of IPN films in all media, and swelling ratio decreased with increasing PVA crosslink degree. Tensile strength and elongation at break related not only to the constitution of IPNs but also to the swelling ratio of IPNs.

Nucleic Acid-Induced Aggregation and Pyrene Excimer Formation

Nucleic acid was found to induce the aggregation of the positively charged pyrene probe (compound 1); as a result, strong pyrene excimer emission was observed. The intensity of the excimer emission was dependent on the concentration of the pyrene probe and the oligonucleotide length, sequence, and concentration. These results suggest a new strategy for label-free nucleic acid-based biosensing applications.

Crosslinking of poly[(3-hydroxybutyrate)-co-(3-hydroxyvalerate)] using dicumyl peroxide as initiator

In order to modify poly [(3-hydroxybutyrate)-co-(3-hydroxyvalerate)] (PHBV), the crosslinking of this copolymer was carried out at 160degreesC using dicumyl peroxide (DCP) as the initiator. The torque of the PHBV melt showed an abrupt upturn when DCP was added. Appropriate values for the gel fraction and crosslink density were obtained when the DCP content was up to 1 wt% of the PHBV. According to the NMR spectroscopic data, the location of the free radical reaction was determined to be at the tertiary carbons in the PHBV chains. The melting point, crystallization temperature and crystallinity of PHBV decreased significantly with increasing DCP content. The effect of crosslinking on the melt viscosity of PHBV was confirmed as being positive. Moreover, the mechanical properties of PHBV were improved by curing with DCP. When 1 wt% DCP was used, the ultimate elongation of PHBV increased from 4 to 11 %. A preliminary biodegradation study confirmed the total biodegradability of crosslinked PHBV.

Grafting of maleic anhydride onto linear polyethylene: A Monte Carlo study

Monte Carlo simulation was used to study the graft of maleic anhydride (MAH) onto linear polyethylene (PE-g-MAH) initiated by dicumyl peroxide (DCP). Simulation results revealed that major MAH monomers attached onto PE chains as branched graft at higher MAH content. However, at extremely low MAH content, the fraction of bridged graft was very close to that of branched graft. This conclusion was somewhat different from the conventional viewpoint, namely, the fraction of bridged graft was always much lower than that of branched graft under any condition. Moreover, the results indicated that the grafting degree increased almost linearly to MAH and DCP concentrations. On the other hand, it was found that the amount of grafted MAH dropped sharply with increasing the length of grafted MAH, indicating that MAH monomers were mainly attached onto the PE chain as single MAH groups or very short oligomers. With respect to the crosslink of PE, the results showed that the fraction of PE-(MAH)(n)-PE crosslink structure increased continuously, and hence the fraction of PE-PE crosslink decreased with increasing MAH concentration.

Surface modification of single-walled carbon nanotubes with polyethylene via in situ Ziegler-Natta polymerization

Single-walled carbon nanotubes (SWNTs) were modified with polyethylene (PE) prepared by in situ Ziegler-Natta polymerization. Because of the catalyst pre-treated on the surface of the SWNTs, the ethylene was expected to polymerize there. Scanning electron microscopy images and solubility measurements showed that the surface of the SWNTs was covered with a PE layer, and a crosslink may have formed between the SWNTs and PE. When the SWNTs covered with a PE layer were mixed with commercialized PE by melt blending, the resulting composite had better mechanical properties than the composite from the SWNTs without a PE layer. The yield strength, the tensile strength and modulus, the strain at break, and the fracture energy of the modified-SWNT/PE composites were improved by 25, 15.2, 25.4, 21, and 38% in comparison with those of the raw-SWNT/PE composites.

Quantitative FTIR study of PHBV/bisphenol A blends

FTIR spectroscopy was used to verify the presence of intermolecular hydrogen bond (inter-H-bond) between poly-(3-hydroxybutyrate co-3-hydroxyvalerate) (PHBV) and bisphenol A (BPA). By monitoring the spectral changes during PHBV crystallization and blends dissociation, the absorptivity ratio of C=O bonds in crystalline and amorphous regions in PHBV and that of H-bonded and free C=O in PHBV/BPA blends were experimentally determined as 1.40 and 1.68, respectively. Using curve-fitting program, the C=O absorptions in spectra of blends were ascribed to three types of bonds: amorphous, crystalline and H-bonded C=O. The crystallinity of PHBV and the fraction of H-bonded C=O were calculated. These results indicated that the H-bond clearly suppressed the PHBV crystallization. Furthermore, the fraction of BPA molecules that simultaneously formed two hydrogen bonds (H-bonds) with C=O was estimated. It revealed that there existed a H-bond network in PHBV/BPA blends. This network was compared with the covalent network by estimating the number of atoms between every two adjacent crosslink points in chain. Up to the high density of H-bond discussed in this paper, there was always a certain part in PHBV that crystallized due to the dynamic character of hydrogen bonds; however, the hydrogen bonds significantly reduced the crystallization rate of PHBV.

The effect of chain flexibility and chain mobility on radiation crosslinking of polymers

Radiation crosslinking of polymers mainly depends on the structure of polymer chain. The flexibility and mobility of chain directly influence the possibility of the reactive radicals recombination. Flexible chain is easier to crosslink than rigid-chain polymer. The latter must be crosslinked at high temperature, as most polymers can only crosslink above their melting point. Structural effect also influences the mechanism of radiation crosslinking of polymers. We find from the results in literature and in our laboratory that, the flexibility chain polymer mainly crosslinked with H type, but the rigid chain polymer mainly crosslinked with Y type. (C) 2001 Published by Elsevier Science Ltd.

Effect of gamma-irradiation on brittle-tough transition of PBT/EPDM blends

Irradiation can be applied to crosslink the dispersed elastomer phase to increase the modulus and decrease the voiding ability of the elastomer, which results in increasing critical brittle-tough transition elastomer content at constant temperature. The experimental results show that at 25 degrees C the critical elastomer content of EPDM shifts to higher composition (shift of about 4 wt%) for PBT/EPDM blends after 100 kGy gamma-irradiation. (C) 1997 Elsevier Science Ltd.

The effect of formulated molecular weight on temperature resistance and mechanical properties in polyimide based composites

This experimental study examines the role of formulated molecular weight between crosslink sites on the temperature resistance and mechanical properties of composites based on a polyimide containing a diphenyl thioether unit (PTI). The composites are fabricated by in situ polymerization of monomer reactants (PMR) using three monomeric ingredients: bis(3,4-dicarboxyphenyl) sulfide dianhydride (TDPA); 4,4'-methylene dianiline (MDA); and the monomethyl ester of norbornene anhydride (NE). By changing monomeric molar ratio, three formulations are prepared, in which formulated molecular weight between crosslink sites varies from 1487 to 3446 g mol(-1). Unidirectional composite laminates from each formulation and T300 carbon fibres are compression moulded and cut into a series of test specimens. By measuring the glass transition temperature (T-g), Mode I interlaminar fracture toughness (G(IC)) and other mechanical properties at room and elevated temperatures, the influences of formulated molecular weight on the temperature resistance and mechanical properties of PTI-based composites are investigated.

MORPHOLOGY AND DYNAMIC MECHANICAL-PROPERTIES OF AB CROSS-LINKED POLYMERS BASED ON POLYURETHANES

Polyoxypropylene glycol (PPG) (or castor oil) and toluene diisocyanate (TDI) were mixed, and the prepolymer polyurethane (PU) (I) was formed. Vinyl-terminated polyurethane (II) was prepared from (I), and hydroxyethyl acrylate, AB crosslinked polymers (ABCPs) were synthesized from (II) and vinyl monomers such as styrene, methyl methacrylate, vinyl acetate, etc. The dynamic mechanical properties and morphology of ABCPs were measured. The ABCPs based on PPG have double glass transition temperatures (T(g)) on the sigma-vs. temperature curves. They display a two-phase morphology with plastic components forming the continous phase and PU-rich domains forming the separated phase on the electron micrographs. Irregular shapes and a highly polydisperse distribution of PU-rich domain sizes were observed. The crosslink density of ABCPs has a notable effect on the morphology and properties. The average diameter of the PU-rich domains depends on the molecular weight of prepolymer PPG. The highly crosslinked structures will produce large numbers of very small domains. ABCPs based on castor oil show a single T(g) relaxation on the dynamic mechanical spectra. The compatibility between the two components is much better in ABCPs based on castor oil than in those based on PPG, because there is a high crosslink density in the former. Comparison of the dynamic mechanical spectra of ABCP and interpenetrating networks (IPN) based on castor oil with similar crosslink density and composition imply that the two components in ABCP are compatible whereas microphase separation occurs in IPN. An improvement in the compatibility is achieved by the crosslinking between the two networks.

Comparative studies on the efficiency of energy transfer of two R-phycoerythrin-C-phycocyanin conjugates constructed from sodium periodate and glutaraldehyde respectively

The C-phycocyanin and the R-phycoerythrin were purified from the blue-green alga Spirulina platensis and red alga Polysiphonia urceolata respectively. Both sodium periodate and glutaraldehyde are effective coupling agents being capable of constructing the R-phycoerythrin-C-phycocyanin conjugate, which was also called phycobiliproteins energy transfer model. The two artificial conjugates constructed with different methods were purified by Sephadex G-200 chromatography respectively. Spectra analysis indicated that energy transfer occurred in the two conjugates. The conjugate with sodium periodate had the higher efficiency of energy transfer than that with glutaraldehyde conjugate.