Protein array for assist diagnosis of acute myocardial infarction

A nanogold probe immunoassay for cardiac troponin I (cTnI) combining the concepts of the one-step dual monoclonal antibody “sandwich” principle, the low density protein array, and silver enhancement on the gold particle is described. Two main substrates, namely the capture antibody (IgG1) coated supporting nitrocellulose membrane and the colloidal gold-labeled detection antibody (cAu–IgG2), were prepared before the detection. The detection procedure involved two steps, i.e. immunoreaction and silver amplification. The assay needs only small amounts of serum samples of patients. The detection results could be easily imaged with a simple flatbed scanner or even the naked eye. The whole detection procedure of the assay could be fulfilled within 40 min (much faster than the routine enzyme-linked immunosorbent assay (ELISA) that takes usually at least 3 h for a turnaround test). The detection limit of cTnI was found to be 1 ng/ml. The detecting results of cTnI in serum samples were similar to those detected by ELISA.     

Myocardial infarction patients show altered lipoprotein properties and functions when compared with stable angina pectoris patients

Several parameters and risk factors were compared between Korean male myocardial infarction (MI) patients (n = 10) and angina pectoris (AP) patients (n = 17) to search unique biomarkers for myocardial infarction (MI) in lipoprotein level. Individual serum and lipoprotein fractions (VLDL, LDL, HDL₂, HDL₃) were isolated and analyzed by lipid and protein determination and enzyme assay. The MI group was found to have a 25 and 30% higher serum cholesterol and triacylglycerol (TG) level than the AP group, respectively, however, their body mass index (BMI), LDL-cholesterol (C), HDL-C, and glucose levels fell within the normal range. MI patients were found to have an approximately two-fold higher level of serum IL-6 and an 18% lower serum apoA-I level than that of the AP group. LDL and HDL₂ fraction of the MI group were more enriched with TG than those of AP group. The increased TG was correlated well with the increased level of apoC-III in the same fraction. Cholesteryl ester transfer protein (CETP) activity and protein level were greatly increased in MI patients in the LDL and HDL₃ fractions. MI patients showed more severely oxidized LDL fraction than patients in the AP group, as well as the weakest antioxidant ability of serum. Conclusively, MI patients were found to have unique serum and lipoprotein characteristics including increased IL-6 and TG in serum, with CETP and apoC-III in the LDL and HDL fractions, as well as severely impaired antioxidant ability of HDL.     

通脑活络针刺法治疗脑梗死患者的BI与NIHSS指数的协方差分析

利用协方差分析将那些认为很难控制的因素,即患者入院时的BI得分或NIHSS得分作为协变量x,在排除协变量x对观测变量y,即治疗90天后患者的BI、NIHSS得分影响的条件下,分析治疗方法对治疗90天后患者的BI、NIHSS得分的影响,从而更加准确地对治疗方法进行评价.     

Development of a myoglobin impedimetric immunosensor based on mixed self-assembled monolayer onto gold

Immunosensors rely on antibody-antigen binding with a range of possible detection methodologies. In this study, electrochemical impedance spectroscopy was used to monitor the sensor surface assembly and recognition of the analyte (myoglobin). Myoglobin is rapidly released into the circulatory system after an acute myocardial infarction and rapidly rising levels make it the first biochemical marker of myocardial damage. The immunosensor fabrication steps comprised the steps of (a) formation of mixed self-assembled monolayers (mSAM) on gold electrodes using a mixture of biotinyl-phospholipid and mer captohexadecanoic acid; (b) neutravidin functionalisation and (c) attachment of biotinyl anti-myoglobin antibodies. A range of analyte concentration (10⁻¹²–10⁻⁶ M) was successfully detected in phosphate buffered saline and in serum concentration ranging from 10% (v/v) to 100% (v/v) serum. Quartz crystal microbalance and atomic force microscopy studies were carried out to study each step of fabrication to elucidate binding characteristics and surface topography. Correspondence: Morsaline Billah, Institute of Membrane and Systems Biology, Garstang Building, University of Leeds, Woodhouse Lane, Leeds LS2 9JT, United Kingdom     

基于χ~2检验法的通脑活络针刺法对脑梗死患者BI指数的影响

用r×s列连表的Pearson x~2检验法,讨论通脑活络针刺法、溶栓剂法与常规法在各救治时间窗对脑梗死患者BI评分的影响,并得到结论,通脑活络针刺法在各救治时间窗的"治愈率"及较好的康复程度方面显示了显著疗效.     

Mesenchymal stem cells reciprocally regulate the M1/M2 balance in mouse bone marrow-derived macrophages

Mesenchymal stem cells (MSCs) have been widely studied for their applications in stem cell-based regeneration. During myocardial infarction (MI), infiltrated macrophages have pivotal roles in inflammation, angiogenesis and cardiac remodeling. We hypothesized that MSCs may modulate the immunologic environment to accelerate regeneration. This study was designed to assess the functional relationship between the macrophage phenotype and MSCs. MSCs isolated from bone marrow and bone marrow-derived macrophages (BMDMs) underwent differentiation induced by macrophage colony-stimulating factor. To determine the macrophage phenotype, classical M1 markers and alternative M2 markers were analyzed with or without co-culturing with MSCs in a transwell system. For animal studies, MI was induced by the ligation of the rat coronary artery. MSCs were injected within the infarct myocardium, and we analyzed the phenotype of the infiltrated macrophages by immunostaining. In the MSC-injected myocardium, the macrophages adjacent to the MSCs showed strong expression of arginase-1 (Arg1), an M2 marker. In BMDMs co-cultured with MSCs, the M1 markers such as interleukin-6 (IL-6), IL-1β, monocyte chemoattractant protein-1 and inducible nitric oxide synthase (iNOS) were significantly reduced. In contrast, the M2 markers such as IL-10, IL-4, CD206 and Arg1 were markedly increased by co-culturing with MSCs. Specifically, the ratio of iNOS to Arg1 in BMDMs was notably downregulated by co-culturing with MSCs. These results suggest that the preferential shift of the macrophage phenotype from M1 to M2 may be related to the immune-modulating characteristics of MSCs that contribute to cardiac repair.     

替罗非班在老年STEMI患者PCI治疗中的作用

目的 观察小剂量替罗非班在老年急性ST段抬高型心肌梗死(STEMI)急诊经皮冠状动脉介入（PCI）术中的治疗效果, 并探讨其临床安全性。方法 选取急性STEMI并行急诊PCI治疗的老年患者150 例,使用随机数字表法分为替罗非班组（80 例）和对照组（70例）,两组患者术前均予以阿司匹林300mg 及氯吡格雷300mg顿服,术后长期服用阿司匹林100mg、氯吡格雷75mg,1 次/d,替罗非班组在常规治疗基础上给予替罗非班负荷量（5μg/kg）在3min内静脉推注后以0.075μg/（kg·min）静脉滴注维持24h。观察两组手术前后梗死相关动脉TIMI血流情况、术后心电图相关导联抬高的ST 段回落幅度、住院期间出血及继发血小板减少症等事件发生率、术后3个月左心室射血分数（LVEF）及主要不良心血管事件（MACE）发生情况。结果 PCI术后,替罗非班组TIMI 血流2～3 级者75例（93.75%）,ST 段回落幅度为（69.96±15.53）%,均显著高于对照组的58 例（82.86%）和（64.18±14.02）%,差异均有统计学意义（均P＜0.05）;住院期间替罗非班组出血事件发生6例（7.5%）,低于对照组（P＜0.05）,而血小板减少情况未见明显增多（P＞0.05）。3个月随访结果显示,替罗非班组LVEF改善情况显著优于对照组（P＜0.01）,而MACE 发生率则降低（P＜0.05）。结论 小剂量替罗非班能有效提高老年急性STEMI患者PCI术后冠状动脉血流灌注,且不增加出血及血小板减少的风险,同时可改善心功能、降低MACE的发生。     

In vivo relaxation of N-acetyl-aspartate, creatine plus phosphocreatine, and choline containing compounds during the course of brain infarction: a proton MRS study.

Localized water suppressed proton spectroscopy has opened up a new field of pathophysiological studies of severe brain ischemia. The signals obtained with the pulse sequences used so far are both T₁ and T₂ weighted. In order to evaluate the extent to which changes in metabolite signals during the course of infarction can be explained by changes in T₁ and T₂ relaxation times, eight patients with acute stroke were studied. STEAM sequences with varying echo delay times and repetition times were used to measure T₁ and T₂ of N-acetyl-aspartate (NAA), creatine plus phosphocreatine (Cr+PCr) and choline containing compounds (CHO) in a 27-ml voxel located in the affected area of the brain. Ten healthy volunteers served as controls. We found no difference in T₁ or T₂ of the metabolites between the patients and the normal controls. The T₂ of CHO was longer than that of NAA and Cr+PCr. Our results indicate that spectra obtained in brain infarcts and normal tissue with the same acquisition parameters are directly comparable with respect to relative signal intensities as well as signals scaled with internal and external standards.     

Surface plasmon resonance biosensor with high anti-fouling ability for the detection of cardiac marker troponin T

Designing a surface recognition layer with high anti-fouling ability, high affinity, and high specificity is an important issue to produce high sensitivity biosensing transducers. In this study, a self-assembled monolayer (SAM) consisting of a homogeneous mixture of oligo(ethylene glycol) (OEG)-terminated alkanethiolate and mercaptohexadecanoic acid (MHDA) on Au was employed for immobilizing troponin T antibody and applied in detecting cardiac troponin T by using surface plasmon resonance (SPR). The mixed SAM showed no phase segregation and exhibited human serum albumin resistance, particularly with an antibody-immobilized surface. X-ray photoemission spectra revealed that the chemical composition ratio of OEG to the mixed SAM was 69% and the OEG packing density was 82%. The specific binding of troponin T on the designed surface indicated a good linear correlation (R = 0.991, P < 0.0009) at concentrations lower than 50 μg mL⁻¹ with the limit of detection of 100 ng mL⁻¹ using a SPR measuring instrument. It is concluded that the mixed SAM functions as designed since it has high detection capability, high accuracy and reproducibility, as well as shows strong potential to be applied in rapid clinical diagnosis for label-free detection within 2 min.     

心肌梗死患者红细胞的光镊拉曼光谱

利用激光光镊拉曼光谱系统,测定了健康者红细胞和心肌梗死患者的红细胞拉曼光谱,研究其拉曼光谱之间存在的差异性,探讨其红细胞内容物的改变情况及其变化机理,为疾病的诊断提供一种新的、快速简便的光谱分析手段,为临床诊断提供有力的实验依据.