表皮生长因子受体基因突变检测方法改进及初步临床验证

目的:改进现有的检测表皮生长因子受体(EGFR)基因突变的荧光PCR法并开发出新的试剂盒,将其与直接测序法和ARMS法进行对比,验证该试剂盒用于临床诊断的敏感性、特异性和准确性。方法:收集2013年6月至2015年8月手术确诊的141例非小细胞肺癌(NSCLC)的石蜡包埋组织标本。采用盲法分别使用直接测序法、ARMS法和新试剂盒检测EGFR突变,比较新试剂盒与其他两种检测方法的差异,结果不一致时采用三种方法分别重复检验一次。结果:三种方法检测成功率均为100%,新试剂盒与直接测序法测得结果完全一致的比率达75.9%(107/141),在直接测序法测得的96例突变阳性中,92例在新试剂盒检测中得到验证(95.8%)。而直接测序法显示突变阴性的45例中,新试剂盒检测发现了23例突变阳性,两种检测方法的结果存在统计学差异(x2=40.745,P0.05)。与直接测序法进行比较,新试剂盒检测EGFR突变的敏感性、特异性分别为95.8%、48.9%,阳性预测值、阴性预测值分别为80.0%、84.6%,检测准确度为80.9%。以ARMS检测法为金标准,新试剂盒测得结果完全一致的比率达84.4%(119/141),两者的一致性比较好(K=0.749,P0.05),敏感性、特异性分别为94.1%、86.4%。结论:改进后EGFR基因突变检测的试剂盒在技术上较好地控制了检测结果的假阳性和假阴性,该检测方法较直接测序法具有更好的敏感性和准确性,与现有的ARMS法一致性较高。     

Vegetation mapping with the aid of low-altitude aerial photography

Abstract. A technique for fine-scale vegetation mapping with the aid of low-altitude aerial photography was developed. The procedure is as follows: 1. The site is divided into a lattice pattern - in case the site is too large to fit into a single photograph with satisfactory resolution. The coordinates of every lattice point are surveyed to be used as control points for geometric correction. A photograph of each block of the lattice is taken using a remote-controlled camera system lifted by a captive helium balloon. 2. The vegetation is classified on the basis of a phytosociological survey. 3. The shapes and locations of vegetation patches appearing in the photographs are entered into a computer, using a digitizer. A geometric correction is carried out through coordinate transformation referring to the coordinates of the control points and subsequently a draft vegetation map is produced. Finally, discrepancies are corrected and the map is coloured to produce the final version of the vegetation map. This technique was applied to vegetation mapping at a bar, 500 m wide and 2 km long, in the river Yoshino in Shikoku, Japan. A fine-scale vegetation map was obtained and used to analyse the influence of plants on geomorphic processes and community-specific hydrogeomorphic conditions on the bar.     

Stimulation of tissue plasminogen activator production from epithelial cell lines

The aim of this study was to investigate the possibility of enhancing the yield of tissue plasminogen activator (tPA) from two epithelial cell lines of normal (non-malignant) derivation grown in tissue culture. The three agents used in this investigation were chosen because of their proven enhancing effect on analogous cells or products. The anabolic hormone stanozolol was found to have no significant stimulatory effect on these cell lines. A phorbol acetate (12-O-tetradecanoylphorbol 13-acetate) caused a twofold enhancement in tPA yield but the most significant results were obtained with 5-azacytidine. This agent increased the yield by up to fourfold in small stationary cultures and threefold in large-scale microcarrier cultures. A combination of azacytidine and phorbol acetate did not have an additive effect on total yield but did alter the kinetics of tPA expression with time. Indications were that the maximum yield with these types of potentiating agents was achieved as it could not be increased by using a combination of two different agents.     

Physical and chemical variability in sugarcane fields infested with the red-striped soft scale insect,Pulvinaria Tenuivalvata (Newstead)

The red-striped soft scale insect Pulvinaria tenuivalvata (Newstead) (Hemiptera: Coccidae) started to infest sugarcane plants (Saccharum officinarum L.) in different districts in Egypt during the last decade. The percentage of infestation was recorded in El-Wakf area, Qena Governorate (Naghhamadi mill zone) Upper Egypt in some fields. There are three levels of infestation, low, intermediate and high. From these fields, samples were selected for physical and chemical studies. The results obtained show that the stalks of infested plants decreased in weight, the sugar content (glucose and sucrose) drastically reduced and as the percentage of infestation increased the percentage of glucose and sucrose content significantly decreased. The primary and secondary humidity and the cellulose content also increased in the healthy plants compared to the infested ones. All the physical character of the infested plants was significantly affected in comparison with the healthy ones.     

The Everyday Life Context of Increasing Energy Demands: Time Use Survey Data in a Decomposition Analysis

Industrial ecologists have modeled with precision the material foundations of industrial systems, but given less attention to the demand for products and the drivers of structural changes in these systems. This article suggests that time use data complement data on monetary expenditure and can be used to elucidate the everyday life context in which the changes in the economy take place. It builds upon the claim that goods are not direct sources of utility, but enter specific household activities as inputs. A second argument for the proposed approach is that it can be used to introduce and foster human agency in analyses of production systems. The article uses Finnish time use survey data, consumption expenditure data, and data on the sectoral energy intensities of financial output in the Finnish economy. First, a measure of the energy intensity of activities is derived by relating consumer time use and the required direct and indirect energy requirements. Second, the results include a decomposition of changes in the energy requirements of private consumption in Finland during the 1990s. It is shown that although the same activities on average require increasing energy inputs per unit of time, Finns have simultaneously changed the structure of their everyday life toward less energy-intensive activities.     

Screening costramid libraries for chromosomal genes: an alternative interspecific hybridization method

Abstract Broad host-range RK2-based cosmid vectors ('costramids') are increasingly used in molecular genetic studies of Gram-negative soil bacteria such as Rhizobium spp. we describe a simple modification of existing methods, whereby a genomic library constructed in a stringently replicated vector can be screened for genes which are undetectable by colony hybridization due to background cross-hybridization. This method allows the use of 'heterologous' probes (interspecies hybridization) to isolate several presumptive genes of interest from a gene bank of Rhizobium sp. NGR234 made in the costramid pRK7813. These are a gene with homology to the citrate synthase gene ( gltA ) or Escherichia coli , the gene encoding δ-aminol evulinic acid synthase ( hemA ), and a gene or genes regulating dicarboxylate transport.