Glycation of high-density lipoprotein triggers oxidative stress and promotes the proliferation and migration of vascular smooth muscle cells

Background In type 2 diabetes mellitus （T2DM）, high-density lipoprotein （HDL） impairs its anti-atherogenic properties and even develops to a pro-inflammatory and pro-atherogenic phenotype because of abnormal compositions and modifications. In this study, we ex- amined the effects and the related mechanisms of glycation of HDL on the proliferation and migration of vascular smooth muscle cells （VSMCs）. Methods ＆ Results Glycated HDL （G-HDL） was modified with D-glucose （25 mmol/L） in vitro. Diabetic HDL （D-HDL） was isolated from T2DM patients. Rat VSMCs were isolated from the thoracic aortas. Human VSMCs were obtained from ScienCell Research Laboratories. Alpha-actin was detected through immunofiuorescence. VSMC proliferation was assayed by Cell Count. VSMC migration was determined by transwell chamber and scratch-wound assay. Intracellular reactive oxygen species （ROS） was detected based on ROS-medi- ated 2＇,7＇-dichlorofluorescein （DCFH-DA） fluorescence. Compared to native HDL （N-HDL）, G-HDL remarkably promoted VSMC prolif- eration and migration in the dose and time-dependent manners. In addition, G-HDL enhanced ROS generation in VSMCs. However, the ROS scavenger, N-acetylcysteine, efficiently decreased ROS production and subsequently inhibited the proliferation of VSMCs induced by G-HDL. Similarly, D-HDL from T2DM patients also promoted ROS release and VSMC proliferation and migration. Conclusions HDL either glycated in vitro or isolated from T2DM patients triggered VSMC proliferation, migration, and oxidative stress. These results might partly interpret the higher morbidity of cardiovascular disease in T2DM patients.

光学相干层析成像技术确定组织光学特性

光学相干层析成像(OCT)是近年来发展较快的一种新型成像技术,能对生物组织内部的微观结构进行高分辨率的横断面层析成像,具有快速、非侵入及高分辨率等特点,在体生物组织微观结构分析和疾病诊断等方面具有重要的应用价值。综述了采用光学相干层析成像技术确定生物组织光学特性的研究。

A Comparison between Neutron Activation Analysis and X-Ray Fluorescence Methods in Analyzing the Granite Samples

This study focused on the performance of where elements analysing techniques were used to detect the elements in granite stones. These techniques are NAA （neutron activation analysis） and XRF （X-ray fluorescence）. They were applied to detect the elements in samples which had been chosen from different areas of Pulua Penang in Malaysia collected by geophysics group which helped to describe and identify the elements found in the granite stone that were used in the study procedures to control the analytical results. The integration of both methods has enabled the researcher to determine 40 elements in the samples. The numbers of elements detected by XRF analysis method are 12 elements （Ar, K, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu and Zn）; while, the elements detected by NAA method have three folds of elements with XRF analysis method were 35 elements （Na, AI, Si, K, Ca, Sc,Ti, Mn, Fe, Co, Ga, Ce, As, Br, Rb, Zr, Sb, I, Cs, Ba, La, Nd, Sm, Eu,Tb, Dy, Yb, Lu, Hf, Ta, W, Au, Pa and Np）. Seven common elements were detected in both techniques： K, Sc, Ti, V, Mn, Fe and Co. Si has a higher concentration in NAA technique which is 331.8 ppm. Sc has a lower concentration in XRF technique which is 0.25 ppm. Nd has a lower concentration in NAA technique which is 3.09 - 10-5 ppm. Finally, it is found that the NAA is better to detect the elements than XRF.

Sensitive detection of DNA methyltransferase activity based on supercharged fluorescent protein and template-free DNA polymerization

DNA methylation, catalyzed by DNA methyltransferases(MTases), is a key component of genetic regulation, and DNA MTases have been regarded as potential targets in anticancer therapy. Herein, based on our previously developed DNA-mediated supercharged green fluorescent protein(Sc GFP)/graphene oxide(GO) interaction, coupled with methylation-initiated template-free DNA polymerization, we propose a novel fluorescence assay strategy for sensitive detection of DNA MTase activity. A hairpin DNA with a methylation-sensitive site and an amino-modified 3′-terminal(DNA-1) was designed and worked as a starting molecule. In the presence of DNA MTase, methylation-sensitive restriction endonuclease, and terminal deoxynucleotidyl transferase(Td T), DNA-1 can be sequentially methylated, cleaved, and further elongated. The resulting long DNA fragments quickly bind with Sc GFP and form the Sc GFP/DNA nanocomplex. Such nanocomplex can effectively protect Sc GFP from being adsorbed and quenched by GO. Without the methylation-initiated DNA polymerization, the fluorescence of Sc GFP will be quenched by GO. Thus, the DNA MTase activity, which is proportional to the amount of DNA polymerization products, can be measured by reading the fluorescence of Sc GFP/GO. The method was successfully used to detect the activity of DNA adenine methylation(Dam) MTase with a wide linear range(0.1–100 U/m L) and a low detection limit of 0.1 U/m L. In addition, the method showed high selectivity and the potential to be applied in a complex sample. Furthermore, this study was successfully extended to evaluate the inhibition effect of 5-fluorouracil on Dam MTase activity and detect Td T activity.

Homogeneous label-free fluorescent assay of small molecule-protein interactions using protein binding-inhibited transcription nanomachine

Quantitative analysis of interactions between small molecules and proteins is a central challenge in chemical genetics, molecular diagnostics and drug developments. Here, we developed a RNA transcription nanomachine by assembling T7 RNA polymerase on a small molecule-labeled DNA heteroduplex. The nanomachine, of which the RNA transcription activity can be quantitatively inhibited by protein binding, showed a great potential for small molecule-protein interaction assay. This finding enabled us to develop a novel homogeneous label-free strategy for assays of interactions between small molecules and their protein receptors. Three small molecule compounds and their protein receptors have been used to demonstrate the developed strategy. The results revealed that the protein-small molecule interaction assay strategy shows dynamic responses in the concentration range from 0.5 to 64 nM with a detection limit of 0.2 nM. Due to its label-free, homogeneous, and fluorescence-based detection format, besides its desirable sensitivity this technique could be greatly robust, cost-efficient and readily automated, implying that the developed small molecule-protein interaction assay strategy might create a new methodology for developing intrinsically robust, sensitive and selective platforms for homogeneous protein detection.

Detecting the intrinsic Baldwin Effect of the optical FeII complex in Luminous Seyfert 1 Galaxy Fairall 9

We analyze the optical spectra of Fairall 9,a luminous Seyfert 1 galaxy.They are obtained from the AGN Watch program that has monitored this object for 9 months.The flux and variation of the optical FeII complex is measured using multi-component profile modeling with FeⅡ templates.A convincing intrinsic Baldwin Effect(BEff) of optical FeⅡ emission blends in Fairall 9 is discovered for the first time.The variation of FeⅡ/Hβ(RFe) in Fairall 9 is obtained.We also detect a marginal anti-correla tion between RFe and the continuum flux in its spectra.This anti-correlation and the relatively broad profile of Hβ in this galaxy support the dichotomy reported by Wang et al.(2005).