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Jiajia Dong Leiliang He Fei Yu Songcheng Yu Lie Liu Yongmei Tian Yilin Wang Jia Wang Lingbo Qu Yongjun Wu Runping Han 《Luminescence》2019,34(3):368-374
The occurrence of many diseases is closely related to the high expression of DNA methyltransferase 1 (DNMT1). However, most studies are focused on the detection of DNMT1 activity, a few are concerned with the detection of DNMT1 content. In this study, we developed a simple and highly sensitive chemiluminescence (CL) assay for the detection of DNMT1 content. In this method, anti‐DNMT1 monoclonal antibody was coated on a polystyrene microplate to capture DNMT1. Then anti‐DNMT1 polyclonal antibody and goat anti‐rabbit immunoglobulin G with horseradish peroxidase (IgG‐HRP) were respectively added to combine with captured DNMT1 to form a sandwich structure. Finally, the HRP could catalyze CL substrate and achieve CL signal response. Based on this novel sensitive strategy, the recovery percents were in the ranges from 71.5% to 91.0%. The precision of intra‐assays and inter‐assays were 5.45%–11.29% and 7.03%–11.25%, respectively. The method was successfully applied for the determination of DNMT1 in human serum. The detection results of serum samples showed that the proposed assay had a high correlation with enzyme‐linked immunosorbent assay (ELISA) kit. Compared with the ELISA kit (limit of detection = 0.1 ng/mL), the method has a lower limit of detection of 0.042 ng/mL. Therefore, our method has the potential for the detection of DNMT1 content in clinical diagnosis. 相似文献
93.
Lescano AR Blazes DL Montano SM Moran Z Naquira C Ramirez E Lie R Martin GJ Lescano AG Zunt JR 《PloS one》2008,3(9):e3274
With the rapidly increasing number of health care professionals seeking international research experience, comes an urgent need for enhanced capacity of host country institutional review boards (IRB) to review research proposals and ensure research activities are both ethical and relevant to the host country customs and needs. A successful combination of distance learning, interactive courses and expert course instructors has been applied in Peru since 2004 through collaborations between the U.S. Naval Medical Research Center Detachment, the University of Washington and the Department of Clinical Bioethics of the National Institutes of Health to provide training in ethical conduct of research to IRB members and researchers from Peru and other Latin American countries. All training activities were conducted under the auspices of the Peruvian National Institute of Health (INS), Ministry of Health. To date, 927 people from 12 different Latin American countries have participated in several of these training activities. In this article we describe our training model. 相似文献
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东方鲎C因子的分段克隆及表达 总被引:1,自引:0,他引:1
东方鲎C因子 (factorCfromTachypleustridentatus)是鲎血细胞中的一种对内毒素敏感的丝氨酸蛋白酶原 ,它与内毒素特异结合的特性 ,使之具有很大的应用价值。根据报道的C因子序列 ,设计了两对引物 ,以中国福建沿海的东方鲎 (Tachypleustridentatus)为材料抽提其血细胞总RNA ,首次用RT PCR的方法分两段扩增了鲎血中编码C因子蛋白的基因全序列。序列分析表明 ,得到的FC基因与文献报道的来自日本的东方鲎的FC有很高的同源性。将分段克隆得到的两段FC片段经相应酶切后 ,与含T7启动子的质粒 pET 2 8a( )在一个体系中作连接反应 ,构建表达质粒pET2 8a FC ,转化大肠杆菌BL2 1(DE3) ,筛选表达菌株。表达菌株经 1mmol/LIPTG诱导表达 2 .5h后 ,收集菌体并超声破菌。经SDS PAGE分析表明 ,在 110kD左右处有明显的表达条带 ,大小与FC的计算分子量相吻合。但表达产物以包涵体形式存在。经洗涤与变复性后 ,重组FC在体外表现出明显的抑菌活性。同时蛋白质印迹实验也提示了在大肠杆菌中FC的单基因表达物可能发生部分自剪切反应 ,而形成了额外的免疫印迹条带 相似文献
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The effect of pelleting seeds of lucerne with lime was studied in an acid sandy soil. In pot experiments, the fraction of seedlings with crown nodules, i.e. nodules on the upper 10 mm of the taproot, increased from 26% to 71%. In rhizotrons, the application of CaCO3 resulted in an even stronger response.An agar-contact method was used to study pH changes in the rhizosphere during a period up to 12 days. Application of 1.0 µmol of CaCO3, in drops of 12 µL volume, resulted in an initial soil pH of 6.1 and yielded 75% crown nodulation. In the absence of CaCO3, roots induced a pH increase from 5.1 (day 0) to 5.7 (day 12). However, this did not increase nodulation (5%). Obviously, this type of alkalinization does not overcome the acid-sensitive step of the nodulation process. 相似文献
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A lime-pellet around seeds of lucerne significantly increased crown nodulation in an acid soil. To investigate whether neutralization or calcium were of importance when lime was supplied, experiments with plants were done either in pots or in rhizotrons. Crown nodulation was used to quantify the effect of these two parameters.For the neutralization of the soil, KOH (in pots) or K2CO3 (in rhizotrons) was added. The crown nodulation of pot-grown plants increased from 31% to 53%. In rhizotrons, the number of crown-nodulated seedlings increased from 9% to 53%. If calcium was supplied additionally (as CaCl2 or CaSO4), 63% crown nodulation was found in pots, and 68% in rhizotrons. These numbers are close to the crown nodulation with lime (CaCO3) alone: 70% in pots and 71% in rhizotrons. In the soil studied, the beneficial effect of lime is largely due to neutralization (80%), and only a minor part (20%) is due to the input of calcium.Using rhizotrons, the dynamics of the pH in the rhizosphere of lime-treated and untreated seedlings was followed during a period of 12 days. It was found that, even in the absence of lime, the pH along the taproot increased from 5.1 to 5.7. However, this did not result in the formation of root nodules. Nodulation was obtained only by adding neutralizing chemicals, which increased the pH during the initial 3 days, the acid sensitive period of the process. 相似文献
100.
Shift from facilitative to neutral interactions by the cushion plant Silene acaulis along a primary succession gradient
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