Statin Treatment Improves Plasma Lipid Levels but not HDL Subclass Distribution in Patients Undergoing Percutaneous Coronary Intervention

Despite the established efficacy of statin therapy, the risk of cardiovascular events remains high in many patients. We examined high-density lipoprotein (HDL) subclass distribution profiles among statin-treated coronary heart disease (CHD) patients undergoing percutaneous coronary intervention (PCI). Plasma HDL subclasses were measured in 85 patients with established CHD and quantified by two-dimensional gel electrophoresis and immunoblotting. In CHD patients with statin treatment, the mean value of total cholesterol (TC) reached the desirable level and the triacylglycerol level (TAG) was borderline high. Moreover, low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), apolipoproteinA-I, and apolipoproteinB-100 levels in these patients resembled those in normolipidemic healthy subjects. The HDL subclass did not show a normal distribution and was characterized by the lower large-sized HDL_2b contents and higher contents of small-sized preβ₁-HDL in CHD patients, compared to those in normolipidemic control subjects. Multiple stepwise regression analysis revealed that the severity of coronary stenosis, determined by the Gensini Score, was significantly and independently predicted by HDL_2b and HDL_3b. Statin therapy was effective in modifying plasma lipids levels, but not adequate as a monotherapy to normalize the HDL subclass distribution phenotype of patients with CHD undergoing PCI. The HDL subclass distribution may aid in risk stratification, especially in patients with CHD and therapeutic LDL-C and HDL-C levels.     

金属纳米颗粒在表面等离激元共振生物传感器上的应用

文章研究不同形状和长径比的金属纳米颗粒应用在表面等离激元共振（SPR）生物传感器中对传感信号的影响,自制金属纳米颗粒并进行物理表征,以金纳米颗粒与抗兔IgG进行生物偶联,利用自制角度检测型SPR生物传感器对兔IgG抗体进行检测,结果表明,金属颗粒的形状和长径比对SPR传感器的共振角都有影响,金纳米棒能够明显提高SPR生物传感器的检测灵敏度。     

Pre-screening for antigen detectability in cells: a TEM-based solid phase digital immunogold detection method utilizing ultra low volumes of reagents

Rapid and sensitive pre‐screening for the presence of antigens in cell samples and confirmation of reactivity of antibodies, before proceeding with electron microscopy, is highly desirable. Most of the methods developed for this purpose are generally not very efficient and suitable for dealing with very small volumes of sample and reagents. In this work we present a simple, sensitive and rapid solid phase transmission electron microscope (TEM) based method for the detection of picogram (pg) levels of soluble antigens using as little as 10 µL of reagents. Protein was adsorbed onto grids coated with polystyrene films to form the solid phase. The presence of antigen was detected using immunogold labelling. Gold particles adhering to the film were visualized and counted in a TEM providing a digital signal. This method was 100‐fold more sensitive than dot blot in detection of rabbit IgG. We have demonstrated the utility of this technique by screening for Vitreoscilla haemoglobin (VHb) antigen in cell lysates and confirming the results directly with immunogold labelling transmission electron microscopy of cell sections.     

Gender and age differences in the distribution of the HDL subclasses among the Chinese population

Background and aims: to analyze the gender and age differences in the distribution of the high‐density lipoprotein (HDL) subclasses among the Chinese population, and to clarify the mechanism of these changes. Methods and results: the apoA‐I contents of the plasma HDL subclasses were determined by 2‐DE coupled with immunodetection in 324 men (including 186 normolipidemic subjects) and 186 women (including 114 normolipidemic subjects). The contents of preβ₁‐HDL and HDL₃ (HDL_3c, HDL_3b, HDL_3a) were significantly lower, whereas the contents of HDL_2a and HDL_2b were higher for women than for men in the <50 years age group. Moreover, the contents of preβ₁‐HDL and HDL₃ were higher for female subjects; the HDL_2a and HDL_2b contents were lower for both female and male subjects in the 50–59, 60–69, and ≥70 years age groups versus the subjects of the same gender in the <50 years age group. When compared to the normolipidemic premenopausal women, preβ₁‐HDL, HDL_3b, and HDL_3a increased while HDL_2b decreased significantly in normolipidemic men and postmenopausal women. Conclusions: the contents of the large‐sized HDL particles HDL_2b were higher, but the contents of the small‐sized HDL particles (preβ₁‐HDL, HDL_3b, HDL_3a) were lower for women versus men in the <50 years age group. Meanwhile, the gender difference in distribution of the HDL subclass narrowed obviously with advancing age. Moreover, the characteristics of the HDL subclass distribution profile for the normolipidemic postmenopausal women resembled those for the normolipidemic men.     

Characterization of a 60‐kDa Thermally Stable Antigenic Protein as a Marker for the Immunodetection of Bovine Plasma‐Derived Food Ingredients

A sandwich enzyme‐linked immunosorbent assay (sELISA) based on 2 monoclonal antibodies (Bb3D6 and Bb6G12) that recognize a 60‐kDa antigenic protein in bovine blood was previously developed for detecting bovine blood in animal feed for the prevention of mad cow disease. This study sought to establish the identity of this 60‐kDa antigenic protein and consequently determine the suitability of the sELISA for detecting bovine plasma‐derived food ingredients (BPFIs), which are widely used in dietary products without explicit labeling. Results from western blot confirmed the 60‐kDa protein to be present in the plasma fraction of bovine blood. Further proteomic analyses involving 2‐dimensional gel electrophoresis (2‐D GE) and amino acid sequencing revealed the 60‐kDa protein to be bovine serum albumin (BSA). The sELISA proved capable of detecting BPFIs in all the commercial dietary supplements tested, including those that were formulated with hydrolyzed BPFIs. The assay could also detect 0.01% and 0.5% of different BPFIs in spiked raw and cooked ground beef, respectively. This assay based on the detection of BSA therefore has the potential to become a valuable analytical tool to protect consumers who avoid consuming BPFIs for religious, health, or ethical reasons.     

Multipurpose vectors designed for the fast generation of N- or C-terminal epitope-tagged proteins

In this paper are described a set of new high-copy-number yeast vectors, which are specially designed for the conditional expression of epitope-tagged proteins in vivo. One of the major advantages of these plasmids is that they allow polymerase chain reaction-amplified open reading frames to be automatically fused in frame with the epitope-coding sequence, avoiding longer procedures such as site-directed mutagenesis. This heterologous construction can be realized either at the 5′-end of the coding sequence, in the pYeF1 vector, or at its 3′-end, in pYeF2, generating N- or C-terminal tagged proteins, respectively. Moreover, to increase the usefulness of the method, derivatives of the two basic URA3-borne pYeF1 and pYeF2 were constructed, carrying either the HIS3 or TRP1 gene as a marker of selection. These vectors could be of use for the purpose of functional analysis of the newly discovered genes resulting from the systematic sequencing of the yeast genome. Here, we present results showing the functional expression and the efficient immunoprecipitation of the epitope-tagged Rna15 protein, which is involved in Saccharomyces cerevisiae mRNA stability.     

应用荧光微球技术进行免疫检测研究

以聚苯乙烯羧基荧光微球为固相载体,将人免疫球蛋白G(IgG)和猪血红蛋白(Hb)分别与微球偶联,应用Luminex-100TM荧光微球检测系统检测抗体IgG和抗原Hb与荧光微球的偶联效率.结果表明,采用100μg·mL-1抗体或抗原浓度可以获得较高的偶联效率,且检测快速灵敏.应用荧光微球进行抗原抗体免疫检测可行.     

表面等离子共振生物传感器技术及在生命研究中的应用

表面等离子体共振（Surface Plasmon Resonance,SPR）是一种入射光照射引起金属表面产生等离子体共振而导致反射光衰减的现象,反射光衰减程度与入射角和金属表面物质质量相关。根据这一原理研制的表面等离子体传感器,在检测、分析生物分子问的相互作用等方面有广泛的应用前景。本文对表面等离子共振生物传感器的工作原理、技术参数、检测方法进行了详细介绍,同时介绍了表面等离子共振技术在免疫检测、药物代谢及其蛋白质动力学等生命研究中的应用。