Novel experimental strategy for high resolution AFM imaging of membrane-associated bacterial toxins

Bacterial pore-forming toxins （PFTs） are essential virulence factors of many human pathogens. Knowl- edge of their structure within the membrane is critical for an understanding of their function in pathogenesis and for the development of useful therapy. Atomic force microscopy （AFM） has often been employed to structurally interrogate many membrane proteins, including PFTs, owing to its ability to produce sub-nanometer resolution images of samples under aqueous solution. However, an absolute prerequisite for AFM studies is that the samples are single-layered and closely-packed, which is frequently challenging with PFTs. Here, using the prototypical member of the cholesterol-dependent cytolysin family of PFTs, perfringolysin O （PFO）, as a test sample, we have developed a simple, highly robust method that routinely produces clean, closely-packed samples across the entire specimen surface. In this approach, we first use a small Teflon well to prepare the supported lipid bilayer, remove the sample from the well, and then directly apply the proteins to the bilayer. For reasons that are not clear, bilayer preparation in the Teflon well is essential. We anticipate that this simple method will prove widely useful for the preparation of similar samples, and thereby enable AFM imaging of the greatest range of bacterial PFTs to the highest possible resolution.     

Re-evaluation of the widely applied force-frequency relation for frequency-modulation AFM under solution

Frequency-modulation atomic force microscopy （FM-AFM） is a highly versatile tool for surface science. Besides imaging surfaces, FM-AFM is capable of measuring interactions between the AFM probe and the surface with high sensitivity, which can provide chemical information at sub-nanometer resolution. This is achieved by deconvoluting the frequency shift, which is directly measured in experiments, into the force between the probe and sample. At present, the widely used method to perform this deconvolution has been shown to be accurate under high quality （high-Q） factor vacuum conditions. However, under low quality （low-Q） factor conditions, such as in solution, it is not clear if this method is valid. A previous study apparently verified this relation for experiments in solution by comparing the force calculated by this equation with that obtained in separate experiments using the surface force apparatus （SFA）. Here we show that, in solution, a more direct comparison of the force calculated by this relation with that directly measured by the cantilever deflection in AFM reveals significant differences, both qualitative and quantitative. However, we also find that there are complications that hinder this comparison. Namely, while contact with the surface is clear in the direct measurements （including the SFA data）, it is less certain in the FM-AFM case. Hence, it is not clear if the two methods are measuring the same tip-sample distance regimes. Thus, our results suggest that a more thorough verification of this relation is required, as application of this formulation for experiments in solution may not be valid.     

INFLUENCE OF SURFACE MORPHOLOGY ON THE MICROHARDNESS OF ELECTROLEESS NI-B DEPOSIT REVEALED BY AFM OBSERVATION

A crystalline and non-crystalline two-phase Ni-B coating was prepared by electroless deposition with potassium borohydride as reducing agent. When the deposit was heated from 10 C to 500 C, it took place three kinds of phase transformation and formed corresponding nickel borides. An Atomic Force Microscopy (AFM) was used to directly view the surface morphologies of the samples in various treated states. The AFM images indicate that nano-dimensional protrusions grow on the surface of the deposit grain during some heat treatment processes,and they are distinguished greatly from the small bulges existing in the as-plated deposit. Further studies show that the microhardness related to the surface topography of thecoating to some extent. Especially when a two-step heat treatment process was performed, the nano-dimensional protrusions on the grain surface grew upt to 40～100nm in diameter and 10～20 nm in height, and the deposit microhardness reached up to HV1500～ 1600 in the meantime.     

Improvement of Two-Dimensional Gel Electrophoresis for Study of Corm Formation Related Proteins in vitro from Taro (Colocasia esculenta)

It is an important step in understanding a bio-logical activity to profile proteins and peptides incells and tissues[1]. Two-dimensional polyacry-lamide gel electrophoresis (2D-PAGE) is very use-ful method for separating complex protein mix-tures[2]. Because of high resolution, improved re-producibility of 2D-PAGE with immobilized pHgradients, it has become one of the standard proce-dures for proteome research. Proteomic analyses are now emerged in manyareas of plant biology, such as genet…     

TRANSCRIPTION ACTIVATION BY HBV PRE SI PROTEIN

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Nc-Si Thin Film Deposited at Low Temperature and Nc-Si Heterojunction Solar Cel

IntroductionMuchworkhasbeendonetostudythestruc-tureandpropertiesofthehydrogenatednano-crystallinesilicon(nc-Si∶H)thinfilmsinthepastfewyears[1～3].Ithasbeenreportedthatthesefilmshavemanygoodopticalandelectricalproper-ties[4～6].Severaldepositiontechniqueshavebeenestablishedtopreparenc-Si∶Hthinfilms,includ-ingplasmaenhancedchemicalvapordeposition(PECVD),hotwirechemicalvapordeposition(HWCVD)[7,8]andreactivemagnetronsputter-ing[9]withthesubstratetemperaturevaryingfrom150to250°C.Comparedwi…     

NIR Hyperspectral Imaging Measurement of Sugar Content in Peach Using PLS Regression

Near infrared (NIR) hyperspectral imaging measurement of sugar content in peach was introduced. NIR spectral images (650～1 000 nm, resolution: 2 nm) of peach samples were captured with developed hyperspectral imaging setup. Partial least square (PLS) regression prediction model was developed to estimate the sugar content in peach; step-wise backward method was utilized to determine optimal wavelength subsets. Experimental results show that the calibration model with optimal wavelength subsets has a correlation coefficient of prediction of 0.97 and a standard error of prediction of 0.19, the prediction accuracy is higher than the calibration model applied over the whole wavelength, which proves that variable selection plays an important role in improving the prediction accuracy of PLS regression model.     

层次极限学习机用于高光谱图像预测绝缘子污秽度

高光谱图像具有图谱合一、光谱范围广及分辨率高等优势，能精细化地反映物质微观特性. 为此，引入高光谱成像技术以非接触式预测绝缘子污秽度. 考虑到极限学习机具有学习效率高和泛化能力强等优点，提出基于正则化约束极限学习机的绝缘子污秽度预测（extreme learning machine-insulator pollution degree prediction，ELM-IPDP）模型. 此外，为进一步提升预测性能，引入层次极限学习机从复杂的高光谱图像中学习出有效、抽象、判决性特征表示，继而建立基于层次极限学习机的绝缘子污秽度预测（hierarchical ELM-IPDP，HELM-IPDP）模型. 在不同的训练集与测试集比例和不同隐含层神经元个数的情况下分别进行实验，从实验结果可知:ELM-IPDP模型和HELM-IPDP模型的预测性能基本上随着隐含层神经元个数和训练样本的增加而不断提高；当训练集与测试集比例为9∶1时，ELM-IPDP模型的均方根误差和相关系数分别为0.040 3和0.944 7，而HELM-IPDP模型的均方根误差和相关系数分别提升到0.022 3和0.972 0.      

肝细胞肝癌患者的血清蛋白质组学研究

目的分析肝细胞肝癌患者的血清蛋白质谱,寻找诊断肝癌和检测治疗的特异性标志物。方法收集了50例肝癌患者血清,在去除血清中6种高丰度蛋白后进行双向凝胶电泳分离及银染,将凝胶扫描图像与正常人血清比较分析,然后经质谱鉴定20个差异表达蛋白质点。结果共鉴定了11种差异表达蛋白质,其中7种蛋白质在肝癌患者血清中表达水平上调,其余4种蛋白质表达水平下调。有7个点被同时鉴定为α1-抗胰蛋白酶,均在肝癌患者血清中表达上调,提示α1-抗胰蛋白酶在肝癌中降解加快。Western blotting检测进一步证实多数肝癌血清中的α1-抗胰蛋白酶小片段增多。结论血清是诊断肝癌和检测治疗非常重要的临床标本,本实验中鉴定的蛋白质虽不能作为临床诊断标志物,但对分析肝癌血清蛋白谱的改变有一定的参考价值。     

酞菁铜薄膜制备及其光学性能

采用真空蒸发的方法在ITO玻璃上制备了CuPc薄膜,并用分光光度计（U-3310）测试了四种不同厚度的CuPc薄膜的透射/吸收/反射率随波长变化情况,重点分析了其中的吸收规律.结果显示波段在340~370 nm和570~720 nm光吸收率基本上在90%左右,在480 nm吸收率最低,大部分光都已透射.同时用扫描电子显微镜对薄膜进行了成分分析,对照能谱图可以看出样品中含有碳、氧、铜等元素,与酞菁铜薄膜元素组成成分相符.     

近代企业经营战略的历史演变

本文研究了近代企业生产战略、营销战略、多元化战略、虚拟经营战略的演变历程。虚拟经营战略一方面关注市场迅速变化的需求 ,另一方面又关注企业自身核心能力建设 ,它是未来经营战略的主流     

Screening of hepatocyte proteins binding to NS5ABP37 protein by yeast-two hybrid system

Objective To investigate the biological function of NS5ABP37 and to look for proteins interacting with NS5ABP37 protein in hepatocytes. Methods We constructed bait plasmid expressing NS5ABP37 protein of hepatitis C virus (HCV) by cloning the gene of NS5ABP37 protein into pGBKT7, then the recombinant plasmid DNA was transformed into yeast AH109 (α type). The transformed yeast AH109 was mated with yeast Y187 (α type) containing liver cDNA library plasmid in 2×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-α-gal for selection and screening. After extracting and sequencing of plasmids from positive (blue) colonies, we made a sequence analysis by bioinformatics. Results We screened twenty-five proteins binding to NS5ABP37, including Homo sapiens cyclin Ⅰ (CCNI) gene, Homo sapiens matrix metallopeptidase 25 (MMP25) and Homo sapiens talin 1. Conclusion The yeast-two hybrid system is an effective method for identifying hepatocyte proteins interacting with NS5ABP37 of HCV. And the biological function of NS5ABP37 may be associated with glycometabolism, lipid metabolism and apoptosis.     

V(D)J重组起始阶段的研究进展与展望

重组激活基因(recombination activating gene)1和2编码的RAG1和RAG2蛋白通过对V(D)J重组起始阶段的调节作用,使得抗原受体基因重排严格地按组织、细胞发育阶段进行。本文将就RAG蛋白结合和催化断裂抗原受体基因机制作简要介绍。这些新发现明确了体内V(D)J重组发生的部位以及其出现错误时可能发生的地方,提示V(D)J重组机制不仅对淋巴细胞正常发育起关键性作用,而且对基因组不稳定性和淋巴系统恶性肿瘤的发生也具有重要意义。     

Proteomics Related to Resistance to the Cucumber Powdery Mildew Disease in F2Generation of Cucumber

Cucumber powdery mildew disease caused bySphaerotheca fuliginea is a serious disease of culti-vated cucumber in most cucumber-growing regionsof the world. Understanding resistant mechanismof cucumber against powdery mildew disease is in-creasingly crucial for breeding resistant cultivars toeffectively overcome the disease. In the case, agreat deal of researches on single proteins or en-zymes encoded by the resistant genes have beenperformed to uncover defense mechanism. Howev-er, so far there …     

STUDY ON ION-POLYMER INTERACTION AND MORPHOLOGIC STRUCTURE OF POLYURETHANE/SALT SYSTEMS

IntroductionRecently,many chemical and physical scien-tists have focused their investigations on solidpolymer electrolytes(SPE) because of their poten-tial applications in high technology fields,includ-ing high energy- density batteries,electrochromicdevices,chemical sensors,smart windows and soon.It is well known that,many factors,such aschemical structure,salt concentration,tempera-ture,morphologic structure,etc.,would have ef-fect on ionic conductive properties of SPE[1,2 ] .Inorder to ob…     

重塑黄土的湿化变形规律及细观结构演化特性

为了研究填土在三轴浸水过程中的湿化变形规律及其细观结构演化特性,用改进的非饱和土CT-三轴仪对延安新区的重塑Q₂黄土做了3组共17个偏压固结浸水试验. 在三轴浸水过程中对试样的两个断面进行了多次CT扫描,得到了固结和湿化过程中土样的宏观变形以及土样内部细观结构演化的CT图像和相应的CT数据,基于CT数定义了土的结构性参数和结构演化变量. 研究结果表明:干密度、净围压、基质吸力和偏应力均对试样的湿化变形特性有显著影响;提高干密度可有效减小湿化变形量和降低湿剪破坏的风险（干密度1.52、1.69 g/cm3的试样浸水过程中体应变分别为–0.58%～4.66%、–0.58%～2.43%,干密度1.79 g/cm3的试样体应变为0.019%）;湿化过程中试样越来越密实,试样的CT数均增大;浸水初期,试样原有结构发生破坏,CT数变化较剧烈,均能达到总变化量的60%;同时干密度、净围压、基质吸力、偏应力及含水率对土的结构性参数和结构演化变量有显著影响. 研究成果对填土工程的设计具有重要参考价值,为建立非饱和重塑黄土的结构损伤演化方程与结构性模型提供了科学依据.      

Combined Effects of Cadmium and Phenanthrene on DNA Sequence Diversity of Soil Microbial Community

Pollution of soil environment has become a se-rious problem in many countries, including China.Heavy metal (HM) and polycyclic aromatic hydro-carbons (PAHs) are two of the most abundant andpotentially harmful pollutants found in most pol-luted soil[1, 2]. Besides their natural occurrence,heavy metals may enter the soil environmentthrough anthropogenic activities, such as mining,smelting, sewage sludge disposal, application ofpesticides and inorganic fertilizers, and atmospher-ic deposition. …     

REGULATED PHOSPHORYLATION OF THE GATA－2 DNA BINDINGPROTEIN IN ENDOTHELIAL CELLS

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Adsorption models and structural characterization for activated carbon fibers

The nitrogen adsorption isotherms at 77.69 K were measured for two samples of activated carbon fibers and their microstructures were investigated. Among established isotherm equations, the Dubinin-Radushkevich equation showed the best agreement with the experimental data, while the Langmuir equation showed a large deviation when employed at low relative pressures. The MP method, t-method and αs-method were used to analyze the pore size distribution. The calculated average pore widths and BET (Brunauer-Emmett-Teller) surface areas for the sample A-13 were 0.86 nm and 1 286.60 m2/g, while for the sample A-16, they were 0.82 nm and 1 490.64 m2/g. The sample with larger pore width was more suitable to be used as additive in chemical heat pumps, while the other one could be used as adsorbent in adsorption refrigeration systems.     

Application of Small Sample Analysis in Life Estimation of Aeroengine Components

The samples of fatigue life tests for aeroengine components are usually less than 5, so the evaluation of these samples belongs to small sample analysis. The Weibull distribution is known to describe the life data accurately, and the Weibayes method (developed from Bayesian method) expands on the experiential data in the small sample analysis of fatigue life in aeroengine. Based on the Weibull analysis, a program was developed to improve the efficiency of the reliability analysis for aeroengine compgnents. This program has complete cycle fatigue life was evaluated by this program. From the results, the following conclusions were drawn: (a) that this program could be used for the engineering applications, and (b) while a lack of former test data lowered the validity of evaluation results, the Weibayes method ensured the results of small sample analysis did not deviate from the truth.