[Association of interleukin-1B gene polymorphisms with coronary heart disease].

OBJECTIVE: To determine whether polymorphisms in interleukin-1B gene promoter -31 and exon 5 +3953 loci are associated with coronary heart diseases (CHD) in Chengdu Han population. METHODS: Two SNPs of IL-1B gene (+3953C/T and -31T/C) in 100 patients with CHD (CHD group) and 144 healthy controls in Chengdu were analysed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Two genotypes at IL-1B + 3953 locus (CC and CT) and 3 genotypes at IL-1B -31 locus (CC, TC and TT) were identified. The -31T alleles carriers were associated with a significantly increased risk of CHD as compared with the non-carriers (OR = 2.12, 95% CI: 1.45-3.09, P < 0.01). Genotypes and allele frequencies at IL-1B + 3953 locus in CHD cases did not differ from the controls. CONCLUSION: IL-1B -31T/C polymorphism may contribute to the risk of developing CHD in Chengdu Han population.

Theoretical study of binding interactions and vibrational Raman spectra of water in hydrogen-bonded anionic complexes: (H2O)n- (n = 2 and 3), H2O...X- (X = F, Cl, Br, and I), and H2O...M- (M = Cu, Ag, and Au).

Binding interactions and Raman spectra of water in hydrogen-bonded anionic complexes have been studied by using the hybrid density functional theory method (B3LYP) and ab initio (MP2) method. In order to explore the influence of hydrogen bond interactions and the anionic effect on the Raman intensities of water, model complexes, such as the negatively charged water clusters ((H2O)n-, n = 2 and 3), the water...halide anions (H2O...X-, X = F, Cl, Br, and I), and the water-metal atom anionic complexes (H2O...M-, M = Cu, Ag, and Au), have been employed in the present calculations. These model complexes contained different types of hydrogen bonds, such as O-H...X-, O-H...M-, O-H...O, and O-H...e-. In particular, the last one is a dipole-bound electron involved in the anionic water clusters. Our results showed that there exists a large enhancement in the off-resonance Raman intensities of both the H-O-H bending mode and the hydrogen-bonded O-H stretching mode, and the enhancement factor is more significant for the former than for the latter. The reasons for these spectral properties can be attributed to the strong polarization effect of the proton acceptors (X-, M-, O, and e-) in these hydrogen-bonded complexes. We proposed that the strong Raman signal of the H-O-H bending mode may be used as a fingerprint to address the local microstructures of water molecules in the chemical and biological systems.

[Preparation and identification of monoclonal antibody against human GST-Rta150 fusion protein from Epstein-Barr virus].

OBJECTIVE: To prepare a monoclonal antibody (mAb) against GST-Rta150 fusion protein from EB virus and identify its characteristics. METHODS: BALB/c mouse was rendered immune by GST-Rta150 fusion protein; after that, the spleen cells of immunized mouse were taken to fuse with SP2/0 myeloma cells by a routine method. Then screening for positive hybridoma cells and subclone. Following that, the hybridoma cells were injected into the peritoneal cavity of BALB/c mouse. The ascites was collected and purified. The prepared mAb was identified by double immunodiffusion method for its subclass; in addition, its titer and specificity were identified by ELISA and Western-blot, respectively. RESULTS: One hybridoma cell line 1A9 which stably secreted mAb against GST-Rta150 fusion protein was obtained. Its Ig subclass belonged to IgG1, the titer degree of the purified mAb being 10(-6). Western-blot analysis showed the mAb could combine with GST-Rta150 fusion protein specially. The titer degree of this hybridoma cell line remained unchanged after frozen in liquid nitrogen (LN) for three months. CONCLUSION: The mAb obtained by this method has powerful specificity and high stability. This work could serve as a foundation on which to study the Rta protein from EB virus and the diagnosis and treatment of EB virus correlated diseases.

[Expression and purification of GST-Rta fusion protein from EB virus and preparation of the polyclonal antibody against GST-Rta].

OBJECTIVE: To obtain the recombinant fusion proteins GST-Rta185 and GST-Rta150 from EB virus and prepare two Rta protein specific polyclonal antibodies, respectively. METHODS: Plasmids pGEX-R1501, pGEX-R1851 and pGEX-5X-3 were separately transformed into Escherichia coli BL21 (DE3). Expressions of the recombinant proteins R150-GST, R185-GST and free GST were induced by 0.1 mmol/L IPTG in LB medium. The expressed proteins were purified from lysates with Glutathione Sepharose 4B. Purified proteins were mixed with Freund's adjuvant and then were used to immunize rabbits. RESULTS: High levels of expression of target proteins were detected in the lysates and the purified proteins were obtained by affinity chromatography with Glutathione Sepharose 4B. Western blot and ELISA analysis suggested that the polyclonal antibodies against GST-R185 and GST-R150 were specific. CONCLUSION: The antiserums have good specificity. They are important for the research on Rta fusion proteins from EB virus and for the diagnosis or treatment of EB virus associated diseases.