A simple method for DNA extraction from sporophyte in the brown alga Laminaria japonica

A simple method was developed for extracting DNA from brown algae Laminaria japonica, which possess large amounts of acidic polysaccharides. Firstly, the sporophyte were washed by eliminating polysaccaride buffer to remove the polysaccharides and then ground in liquid nitrogen. Secondly, the powders were treated with lysing buffer. Thirdly, KAc was used to eliminate the remaining acidic polysaccharides. The extracted DNA was purified using a chloroform-isoamyl alcohol ( 24: 1 v/v), and precipitated in cold isopropanol. The yield was from 18.7 to 37.5 mu g g(-1) (wet weight) and the purity of total DNA was determined spectrophotometrically as the ratio of A(260)/A(280), which was about 1.7 - 1.9. The extracted DNA was of high quality and suitable for molecular analyses, such as PCR, restriction enzyme digestion. This method is a reproducible, simple, and rapid technique for routine DNA extraction from sporophyte in Laminaria japonica. Furthermore, the low cost of this method makes it attractive for large-scale studies.

The experimental research of R-phycoerythrin subunits on cancer treatment: A new photosensitizer in PDT

The mouse tumor cell 5180 and human liver carcinoma cell SMC 7721 cells were first treated with R-PE and its subunits (alpha, beta, gamma subunits), then irradiated with Argon laser (496 nm, 28.8 J/cm(2)). Survival rate was measured by MTT method. In order to compare the phototoxicity in normal cells, the mouse marrow cells were treated with photofrin II and beta-subunit, irradiated with 45 J/cm(2) of light; survival rate was also measured by MTT method. The result showed that R-PE subunits had better PDT effect on s180 cells than R-PE and lower phototoxicity in marrow cells than photofrin II Flow cytometric analysis showed that PDT results in a growth inhibition and a G(0)-G(1) cell cycle arrest in SMC 7721 cells. The tumor cells inhibited by PDT in vivo were morphologically observed by TEM, the tumor cell death was daze to the occlusion of tumor blood vessels and inducement of cell programmed death in nuclei. Therefore, with the advantage in special fluorescence activity, loth molecular weight, good light absorbent character and weak phototoxicity, R-PE subunit is art attractive option for improving the selectivity of PDT.

靶向斑马鱼VEGF的shRNA表达载体的构建及其对斑马鱼胚胎血管发育的影响

血管内皮生长因子(vascular endothelial growth factor, VEGF)是一种多功能的细胞因子，其主要作用是促进血管内皮细胞增殖和增加血管通透性，是肿瘤及正常组织血管生成的中心调控因素，以VEGF为靶点的肿瘤血管靶向性治疗成为近几年肿瘤治疗的新途径。RNAi是近年来新发展的一项反向遗传学技术，是一种研究基因功能的有力工具。斑马鱼作为一种重要的模式生物，被广泛用于胚胎的分子发育机制、疾病模型的构建以及药物筛选等研究中。然而在斑马鱼中运用RNAi技术进行基因功能研究是一个相对较新的领域，研究资料较少，并且目前进行的斑马鱼RNAi实验中，siRNA大都是通过化学方法或体外转录合成的。体外合成的siRNA在进入体内后会被降解而无法达到持久阻抑基因表达的目的。因此本研究旨在探讨VEGF特异性siRNA表达载体对斑马鱼VEGF基因的沉默作用，通过分析表型及相关细胞因子的变化，阐明VEGF对斑马鱼胚胎血管生成的影响及作用机制。 研究通过计算机辅助设计软件，针对斑马鱼VEGF mRNA不同位点设计合成了4段含siRNA特异序列的DNA单链，经退火，克隆入pSilencer 4.1-CMV neo载体CMV启动子下游，构建了重组质粒pS1-VEGF、pS2-VEGF、pS3-VEGF及pS4-VEGF。 通过显微注射的方法将载体导入1-2细胞期斑马鱼体内，于胚胎发育的48 h采用RT-PCR的方法检测VEGF基因的表达量，研究不同干扰序列对VEGF基因表达的干涉作用。结果显示，针对不同位点的表达载体对VEGF基因表达的抑制效率有显著差异。它们对VEGF mRNA的抑制率分别为80.5%，42.8%，12.5%，40.7%。通过筛选我们得到了一条具有高效抑制作用的载体pS1-VEGF，该载体的相应序列靶向斑马鱼两个主要异构体VEGF165和VEGF121的共有外显子序列。 形态学检测结果显示，注射了pS1-VEGF的胚胎出现了心包膜水肿、血流速度减慢、循环红细胞堆积等症状。定量碱性磷酸酶染色显示，注射pS1-VEGF能够抑制斑马鱼胚胎新生血管的形成，当注射剂量为0.4 ng时，血管生成的抑制率为31.8%。NBT/BCIP血管染色显示，注射该载体后72 h，50%的斑马鱼肠下静脉、节间血管以及其它血管的发育受到不同程度的抑制。随着注射剂量的加大，血管发育受抑制的情况也随之加重，当注射剂量为1 ng时，只有心脏、头部及卵黄有血液循环。对干扰效果的特异性进行了研究，结果表明pS1-VEGF对斑马鱼内源基因胸苷酸合成酶（thymidylate synthase, TS）基因的表达没有明显的抑制作用。针对TS基因的shRNA表达载体及与斑马鱼没有同源性的对照载体对VEGF基因表达也没有明显的抑制作用。浓度梯度实验表明在0-1.2 ng的范围内干扰效果具有剂量依赖性。 以胚胎整体原位杂交的方法检测质粒对VEGF基因受体NRP1基因表达的影响，发现VEGF特异性shRNA表达载体能够引起NRP1基因表达的降低，说明斑马鱼中VEGF所介导的血管生成作用至少在部分上是依赖于NRP通路所调节的。 本研究工作为进一步研究斑马鱼基因功能、VEGF调控网络提供了一个快速、有效的手段，为阐明斑马鱼的血管生成机制提供了新的资料，为采用RNAi技术，以VEGF为靶点，以斑马鱼为模型对肿瘤进行基因治疗研究奠定了基础。

基于改进势场法的可变形机器人路径规划研究

采用传统的势场法对机器人进行路径规划时,如果目标位置处在障碍物产生的斥力影响范围内,由于斥力大于引力机器人很难到达目标,为此本文提出了一种改进的势场法。该算法通过机器人向着合势能最小的方向运动完成路径规划。并对目标产生的引力做了进一步的改进,解决了目标不可达问题。同时改进的方法不会增加计算量,保证了算法的实时性。仿真实验证明了机器人能够避开障碍顺利到达目标,验证了改进算法的有效性。

连续型联想细胞神经网络局部指数稳定性研究

由于细胞神经网络的潜在应用前景 ,它现已成为神经网络研究的新热点。首先给出连续型联想细胞神经网络的数学模型 ,得到了连续型细胞神经网络平衡点局部指数稳定的充要条件及平衡点指数吸引域的估计 ,研究表明对平衡点的指数吸引域的估计 ,只要计算平衡点处的导算子矩阵的对数范数即可。该研究对连续型联想细胞神经网络的设计和应用均有重要的作用。

机器人机械手的离散时间学习控制

本文为机器人机械手提出了一种基于离散时间的重复学习控制法，这种学习控制法利用机器人动力学模型的部分知识，从它的特性和实用观点看，这种控制法比现有的其它学习控制法更有吸引力．本文还给出了学习控制法的收敛性证明和计算机仿真结果。

Fourier有限差分法深度偏移与正演模拟

With the development of seismic exploration, the target becomes more and more complex, which leads to a higher demand for the accuracy and efficiency in 3D exploration. Fourier finite-difference (FFD) method is one of the most valuable methods in complex structure exploration, which keeps the ability of finite-differenc method in dealing with laterally varing media and inherits the predominance of the phase-screen method in stablility and efficiency. In this thesis, the accuracy of the FFD operator is highly improved by using simulated annealing algorithm. This method takes the extrapolation step and band width into account, which is more suitable to various band width and discrete scale than the commonely-used optimized method based on velocity contrast alone. In this thesis, the FFD method is extended to viscoacoustic modeling. Based on one-way wave equation, the presented method is implemented in frequency domain; thus, it is more efficient than two-way methods, and is more convenient than time domain methods in handling attenuation and dispersion effects. The proposed method can handle large velocity contrast and has a high efficiency, which is helpful to further research on earth absorption and seismic resolution. Starting from the frequency dispersion of the acoustic VTI wave equation, this thesis extends the FFD migration method to the acoustic VTI media. Compared with the convetional FFD method, the presented method has a similar computational efficiency, and keeps the abilities of dealing with large velocity contrasts and steep dips. The numerical experiments based on the SEG salt model show that the presented method is a practical migration method for complex acoustical VTI media, because it can handle both large velocity contrasts and large anisotropy variations, and its accuracy is relatively high even in strong anisotropic media. In 3D case, the two-way splitting technique of FFD operator causes artificial azimuthal anisotropy. These artifacts become apparent with increasing dip angles and velocity contrasts, which prevent the application of the FFD method in 3D complex media. The current methods proposed to reduce the azimuthal anisotropy significantly increase the computational cost. In this thesis, the alternating-direction-implicit plus interpolation scheme is incorporated into the 3D FFD method to reduce the azimuthal anisotropy. By subtly utilizing the Fourier based scheme of the FFD method, the improved fast algorithm takes approximately no extra computation time. The resulting operator keeps both the accuracy and the efficiency of the FFD method, which is helpful to the inhancements of both the accuracy and the efficiency for prestack depth migration. The general comparison is presented between the FFD operator and the generalized-screen operator, which is valuable to choose the suitable method in practice. The percentage relative error curves and migration impulse responses show that the generalized-screen operator is much sensiutive to the velocity contrasts than the FFD operator. The FFD operator can handle various velocity contrasts, while the generalized-screen operator can only handle some range of the velocity contrasts. Both in large and weak velocity contrasts, the higher order term of the generalized-screen operator has little effect on improving accuracy. The FFD operator is more suitable to large velocity contrasts, while the generalized-screen operator is more suitable to middle velocity contrasts. Both the one-way implicit finite-difference migration and the two-way explicit finite-differenc modeling have been implemented, and then they are compared with the corresponding FFD methods respectively. This work gives a reference to the choosen of proper method. The FFD migration is illustrated to be more attractive in accuracy, efficiency and frequency dispertion than the widely-used implicit finite-difference migration. The FFD modeling can handle relatively coarse grids than the commonly-used explicit finite-differenc modeling, thus it is much faster in 3D modeling, especially for large-scale complex media.

指数矩阵近似计算在地震波场延拓中的应用

Seismic wave field numerical modeling and seismic migration imaging based on wave equation have become useful and absolutely necessarily tools for imaging of complex geological objects. An important task for numerical modeling is to deal with the matrix exponential approximation in wave field extrapolation. For small value size matrix exponential, we can approximate the square root operator in exponential using different splitting algorithms. Splitting algorithms are usually used on the order or the dimension of one-way wave equation to reduce the complexity of the question. In this paper, we achieve approximate equation of 2-D Helmholtz operator inversion using multi-way splitting operation. Analysis on Gauss integral and coefficient of optimized partial fraction show that dispersion may accumulate by splitting algorithms for steep dipping imaging. High-order symplectic Pade approximation may deal with this problem, However, approximation of square root operator in exponential using splitting algorithm cannot solve dispersion problem during one-way wave field migration imaging. We try to implement exact approximation through eigenfunction expansion in matrix. Fast Fourier Transformation (FFT) method is selected because of its lowest computation. An 8-order Laplace matrix splitting is performed to achieve a assemblage of small matrixes using FFT method. Along with the introduction of Lie group and symplectic method into seismic wave-field extrapolation, accurate approximation of matrix exponential based on Lie group and symplectic method becomes the hot research field. To solve matrix exponential approximation problem, the Second-kind Coordinates (SKC) method and Generalized Polar Decompositions (GPD) method of Lie group are of choice. SKC method utilizes generalized Strang-splitting algorithm. While GPD method utilizes polar-type splitting and symmetric polar-type splitting algorithm. Comparing to Pade approximation, these two methods are less in computation, but they can both assure the Lie group structure. We think SKC and GPD methods are prospective and attractive in research and practice.

大城市内部商业环境满意度评价与消费区位决策研究-以北京市西城区和海淀区为例

Since 1990s, commercial conditions in China including commercial environment, retail types, scale of retail enterprises, spatial structure of retail and shopping decision making factors have changed. In order to keep up with these changes, commercial geography should set up new perspectives, theories and methods to analyze its internal mechanism and changing rules, and thus provide reasonable and practical scientific basis to commercial planning, location decision of retail enterprises and commercial environment construction. Taking Xicheng and Haidian District of Beijing as research case, which is a sector region from city center to rural region, this paper selects 12 commercial centers as most important study objects of this sector. This paper mainly makes use of the methods of Modeling, Pearson Bivaiiate Correlations Analysis, Factor Analysis and Logit model. Based on 1300 questionnaires and fieldwork, this paper focuses on modeling of Consumer Satisfaction of Commercial Environment (CSCE), evaluation of commercial environment and driving factors of consumers' shopping location decision. Firstly, this paper discusses the development of commercial geography and commercial environment evaluation, the new characteristics and trends of commercial development in Beijing and physical commercial environment of Xicheng and Haidian District of Beijing from chapter 1 to chapter 4. Secondly, this paper summarizes characteristics of residents' shopping behavior in chapter 5. Thirdly, this paper sets up an evaluative model of CSCE, and analyzes consumer satisfaction indexes of commercial environment and their spatial features in chapter 6. Fourthly, this paper infers how residents' attributes and shopping behaviors affect their preferences of shopping location and what are residents' shopping location decisions and their influencing factors in chapter 7. Fifthly, this paper constructs a significant index model and a pyramidal framework of CSCE, and further analyzes the diversity and competitive advantage of commercial environment in chapter 8. Finally, some conclusions are drawn as follows: 1. Characteristics of residents' shopping behavior mostly embody residents' time distance preference, commodity consumption preference, shopping time distribution and shopping activity characteristics. The important factors that influence shopping location choice of residents are distance, transportation, commodity price, commodity types and commodity quality. However, the important factors, which influence shopping location re-choice of residents, are commodity price, commodity quality, commodity types and transportation. 2. CSCE indexes of 12 commercial centers show us significant spatial characteristics, such as spatial differences of "Center-fringe region", spatial characteristics of axes, spatial diversity of ring roads and so on. 3. Influencing factors including factor endowments, relative establishment factor and location and transportation factor of commercial environment are of importance for CSCE. 4. Logit model 1 indicates that shopping behavior of residents is significantly and positively related to working in high-tech companies, high income and by car and positively related to high school diploma, by bus and subway. 5. Logit model 2 indicates that residents' shopping location decision is significantly and positively related to leisure establishment and relative restaurant and entertainment establishment and negatively related to commercial location, commodity price, service quality, parking site. 6. The significant index model and the pyramidal framework of CSCE indicate competitive advantages are crucial to attractive capability of commercial center, and competitive weakness limits development of commercial centers, in particular the weakness of service quality and parking site now is the chief factors restricting development of commercial centers

Extracting Salient Curves from Images: An Analysis of the Saliency Network

The Saliency Network proposed by Shashua and Ullman is a well-known approach to the problem of extracting salient curves from images while performing gap completion. This paper analyzes the Saliency Network. The Saliency Network is attractive for several reasons. First, the network generally prefers long and smooth curves over short or wiggly ones. While computing saliencies, the network also fills in gaps with smooth completions and tolerates noise. Finally, the network is locally connected, and its size is proportional to the size of the image. Nevertheless, our analysis reveals certain weaknesses with the method. In particular, we show cases in which the most salient element does not lie on the perceptually most salient curve. Furthermore, in some cases the saliency measure changes its preferences when curves are scaled uniformly. Also, we show that for certain fragmented curves the measure prefers large gaps over a few small gaps of the same total size. In addition, we analyze the time complexity required by the method. We show that the number of steps required for convergence in serial implementations is quadratic in the size of the network, and in parallel implementations is linear in the size of the network. We discuss problems due to coarse sampling of the range of possible orientations. We show that with proper sampling the complexity of the network becomes cubic in the size of the network. Finally, we consider the possibility of using the Saliency Network for grouping. We show that the Saliency Network recovers the most salient curve efficiently, but it has problems with identifying any salient curve other than the most salient one.

Large-pore mesoporous SBA-15 silica particles with submicrometer size as stationary phases for high-speed CEC separation

A new mesoporous sphere-like SBA-15 silica was synthesized and evaluated in terms of its suitability as stationary phases for CEC. The unique and attractive properties of the silica particle are its submicrometer particle size of 400 nm and highly ordered cylindrical mesopores with uniform pore size of 12 nm running along the same direction. The bare silica particles with submicrometer size have been successfully employed for the normal-phase electrochromatographic separation of polar compounds with high efficiency (e.g., 210 000 for thiourea), which is matched well with its submicrometer particle size. The Van Deemeter plot showed the hindrance to mass transfer because of the existence of pore structure. The lowest plate height of 2.0 mu m was obtained at the linear velocity of 1.1 mm/s. On the other hand, because of the relatively high linear velocity (e.g., 4.0 mm/s) can be generated, high-speed separation of neutral compounds, anilines, and basic pharmaceuticals in CEC with C-18-modified SBA-15 silica as stationary phases was achieved within 36, 60, and 34 s, respectively.

Mandibular Manipulation for the Treatment of Temporomandibular Disorder

The aim of this study was to conduct a systematic review to identify the randomized clinical studies that had investigated the following research question: Is the mandibular manipulation technique an effective and safe technique for the treatment of the temporomandibular joint disk displacement without reduction? the systematic search was conducted in the electronic databases: PubMed (Medical Publications), LILACS (Latin American and Caribbean Literature in Health Sciences), EMBASE (Excerpta Medica Database), PEDro (Physiotherapy Evidence Database), BBO (Brazilian Library of Odontology), CENTRAL (Library Cochrane), and SciELO (Scientific Electronic Library Online). the abstracts of presentations in physical therapy meetings were manually selected, and the articles of the ones that meet the requirements were investigated. No language restrictions were considered. Only randomized and controlled clinical studies were included. Two studies of medium quality fulfilled all the inclusion criteria. There is no sufficient evidence to support the effectiveness of the mandibular manipulation therapy, and therefore its use remains questionable. Being minimally invasive, this therapy is attractive as an initial approach, especially considering the cost of the alternative approaches. the analysis of the results suggests that additional high-quality randomized clinical trials are necessary on the topic, and they should focus on methods for data randomization and allocation, on clearly defined outcomes, on a priori calculated sample size, and on an adequate follow-up strategy.

Time-dependent Hyperstar algorithm for robust vehicle navigation in time-dependent stochastic road networks.

The vehicle navigation problem studied in Bell (2009) is revisited and a time-dependent reverse Hyperstar algorithm is presented. This minimises the expected time of arrival at the destination, and all intermediate nodes, where expectation is based on a pessimistic (or risk-averse) view of unknown link delays. This may also be regarded as a hyperpath version of the Chabini and Lan (2002) algorithm, which itself is a time-dependent A* algorithm. Links are assigned undelayed travel times and maximum delays, both of which are potentially functions of the time of arrival at the respective link. The driver seeks probabilities for link use that minimise his/her maximum exposure to delay on the approach to each node, leading to the determination of the pessimistic expected time of arrival. Since the context considered is vehicle navigation where the driver is not making repeated trips, the probability of link use may be interpreted as a measure of link attractiveness, so a link with a zero probability of use is unattractive while a link with a probability of use equal to one will have no attractive alternatives. A solution algorithm is presented and proven to solve the problem provided the node potentials are feasible and a FIFO condition applies for undelayed link travel times. The paper concludes with a numerical example.

Exploitation of Ebola Virus VP35 Protein to identify new drugs counteracting its type I IFN antagonism

Ebolaviruses (EBOVs) are among the most virulent and deadly pathogens ever known, causing fulminant haemorrhagic fevers in humans and non-human primates. The 2014 outbreak of Ebola virus disease (EVD) in West Africa has claimed more lives than all previous EVD outbreaks combined. The EBOV high mortality rates have been related to the virus-induced impairment of the host innate immunity reaction due to two virus-coded proteins, VP24 and VP35. EBOV VP35 is a multifunctional protein, it is essential for viral replication as a component of the viral RNA polymerase and it also participates in nucleocapsid assembly. Early during EBOV infection, alpha-beta interferon (IFN-α/β) production would be triggered upon recognition of viral dsRNA products by cytoplasmic retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs). However, this recognition is efficiently prevented by the double-stranded RNA (dsRNA) binding activity of the EBOV VP35 protein, which hides RLRs binding sites on the dsRNA phosphate backbone as well the 5’-triphosphate (5’-ppp) dsRNA ends to RIG-I recognition. In addition to dsRNA binding and sequestration, EBOV VP35 inhibits IFN-α/β production preventing the activation of the IFN regulatory factor 3 (IRF-3) by direct interaction with cellular proteins. Previous studies demonstrated that single amino acid changes in the VP35 dsRNA binding domain reduce EBOV virulence, indicating that VP35 is an attractive target for antiviral drugs development. Within this context, here we report the establishment of a novel method to characterize the EBOV VP35 inhibitory function of the dsRNA-dependent RIG-I-mediated IFN-β signaling pathway in a BLS2 cell culture setting. In such system, a plasmid containing the promoter region of IFN-β gene linked with a luciferase reporter gene was transfected, together with a EBOV VP35 mammalian expression plasmid, into the IFN-sensitive A549 cell line, and the IFN-induction was stimulated through dsRNA transfection. Through alanine scanning mutational studies with biochemical, cellular and computational methods we highlighted the importance of some VP35 residues involved in dsRNA end-capping binding, such as R312, K282 and R322, that may serve as target for the development of small-molecule inhibitors against EBOV. Furthermore, we identified a synthetic compound that increased IFN-induction only under antiviral response stimulation and subverted VP35 inhibition, proving to be very attractive for the development of an antiviral drug. In conclusion, our results provide the establishment of a new assay as a straightforward tool for the screening of antiviral compounds that target i) dsRNA-VP35 or cellular protein-VP35 interaction and ii) dsRNA-dependent RIG-I-mediated IFN signaling pathway, in order to potentiate the IFN response against VP35 inhibition, setting the bases for further drug development.

Novelty and Habituation: the Driving Forces in Early Stage Learning for Developmental Robotics

Meng, Q., & Lee, M. (2005). Novelty and Habituation: the Driving Forces in Early Stage Learning for Developmental Robotics. Wermter, S., Palm, G., & Elshaw, M. (Eds.), In: Biomimetic Neural Learning for Intelligent Robots: Intelligent Systems, Cognitive Robotics, and Neuroscience. (pp. 315-332). (Lecture Notes in Computer Science). Springer Berlin Heidelberg.