Identification, expression, function and localization of a DUF985 domain-containing hypothetical gene from amphioxus Branchiostoma belcheri

The progress in genome sequencing has led to an increasing submission of uncharacterized hypothetical genes with the domain of unknown function, DUF985, in GenBank, and none of these genes is related to a known protein. We therefore underwent an experimental study to identify the function of a DUF985 domain-containing hypothetical gene BbDUF985 (GenBank Accession No. AY273818) isolated from amphioxus Branchiostoma belcheri (B. belcheri). BbDUF985 was successfully expressed in both prokaryotic and eukaryotic systems, and its recombinant proteins expressed in both systems definitely exhibited an activity of phosphoglucose isomerase (PGI). Both tissue-section in situ hybridization and immunohistochemistry demonstrated that BbDUF985 was expressed in a tissue-specific manner, with most abundant levels in the hepatic caecum and ovary. In CHO cells transfected with the expression plasmid pEGFP-N1/BbDUF985, the fusion protein was targeted in the cytoplasm of CHO cells, suggesting that BbDUF985 is a cytosolic protein. In contrast, Western blotting indicated that BbDUF985 was also present in amphioxus humoral fluids, suggesting that it exists as a secreted protein as well. Our study provided a framework for further understanding the biochemical properties and physiological function of DUF985-containing hypothetical proteins in other species. (c) 2008 Elsevier Inc. All rights reserved.

中国明对虾免疫系统中抗氧化相关基因的克隆与表达分析

对虾病害在世界范围内的广泛传播，给水产养殖和沿海农村经济造成了重大损失。深入开展对虾免疫机制研究并在此基础上寻找对虾疾病防治的有效方法已成为当务之急。研究表明，当对虾等甲壳动物受到外界病原刺激时，其体内的吞噬细胞在吞噬活动中会激活磷酸己糖支路的代谢，引起呼吸爆发，产生多种活性氧分子。另外，受到病原侵染的对虾还会产生其他多种免疫反应，这些免疫反应将消耗大量的能量（ATP），产能的呼吸链会加速运转，由此也会引发大量活性氧的产生。这些活性氧分子可以杀灭入侵的病原微生物，但同时由于活性氧分子反应的非特异性，它们也会对宿主的细胞、组织和器官造成严重伤害，进而导致对虾生理机能的损伤和免疫系统的破坏。所以，消除对虾体内因过度免疫反应产生的过量氧自由基将能够增强其抵御病原侵染的能力，提高免疫力。本论文从中国明对虾体内克隆了线粒体型超氧化物歧化酶（mMnSOD）、胞质型超氧化物歧化酶（cMnSOD）、过氧化氢酶（Catalase）和过氧化物还原酶（Peroxiredoxin）等四种与免疫系统相关的抗氧化酶基因，分析了它们的分子结构特征，组织分布及应答不同病原刺激的表达变化模式，并对其中的mMnSOD基因和Peroxiredoxin基因进行了体外重组表达、分离纯化和酶活性分析。 采用RACE技术从中国明对虾血细胞中克隆了两个超氧化物歧化酶（SOD）基因，通过序列比对分析发现，其中一个为mMnSOD基因，另一个为cMnSOD基因。mMnSOD基因的cDNA全长为1185个碱基，其中开放阅读框为660个碱基，编码220个氨基酸，其中推测的信号肽为20个氨基酸。多序列比对结果显示中国明对虾mMnSOD基因的推导氨基酸序列与罗氏沼虾、蓝蟹的推导氨基酸序列同源性分别为88%和82%。Northern blot结果表明，该基因在对虾的肝胰脏、血细胞、淋巴器官、肠、卵巢、肌肉和鳃等组织中均有表达。半定量RT-PCR结果显示，对虾感染病毒3 h时，该基因在血细胞和肝胰脏中的转录水平显著升高。此外，通过构建原核表达载体，本研究对该基因进行了体外重组表达，并对纯化的重组蛋白进行了质谱鉴定和酶活分析。cMnSOD基因的cDNA全长为1284个碱基，其中开放阅读框为861个碱基，编码287个氨基酸。多序列比对结果显示中国明对虾cMnSOD基因的推导氨基酸序列与斑节对虾和凡纳滨对虾的同源性高达98%和94%。组织半定量结果显示，cMnSOD基因在对虾被检测的各个组织中均有表达。 另外，半定量RT-PCR结果表明，对虾感染病毒23h时，该基因在肝胰脏中的转录上升到正常水平的3.5倍；而感染后59 h时，该基因在血细胞中的转录上升到正常水平的2.5倍。 利用根据其他生物过氧化氢酶保守氨基酸序列设计的简并引物，结合RACE技术，从中国明对虾肝胰脏中克隆到了过氧化氢酶基因的部分片段，片段长1725个碱基。多序列比对结果发现目前所得中国明对虾Catalase基因部分片段的推导氨基酸序列与罗氏沼虾和皱纹盘鲍Catalase氨基酸序列的同源性分别达到95%和73%。通过实时荧光定量PCR技术对中国明对虾Catalase基因在各个组织中的分布情况及病毒感染后该基因在血细胞和肝胰脏中的转录变化进行了研究。结果发现，该基因在肝胰脏、鳃、肠和血细胞中表达水平较高，在卵巢、淋巴器官和肌肉中的表达水平相对较弱；感染病毒23 h和37 h时，对虾血细胞和肝胰脏中该基因mRNA的表达量分别出现显著性上升。 依据中国明对虾头胸部cDNA文库提供的部分片段信息，结合SMART-RACE技术，从中国明对虾肝胰脏中克隆到了过氧化物还原酶基因（Peroxiredoxin）， 该基因的cDNA全长为942个碱基，其中开放阅读框为594个碱基，编码198个氨基酸。中国明对虾Peroxiredoxin基因的推断氨基酸序列与伊蚊、文昌鱼和果蝇等Peroxiredoxin基因的推断氨基酸序列同源性分别为77%、76%和73%。其蛋白理论分子量为22041.17 Da，pI为5.17。Northern blot结果表明，Peroxiredoxin基因在对虾的肝胰脏、血细胞、淋巴器官、肠、卵巢、肌肉和鳃等组织中均有表达。实时荧光定量PCR结果显示，弧菌感染后，该基因在对虾血细胞和肝胰脏中的转录水平都有明显变化并且表达模式不同。另外，对该基因进行了体外重组表达，并对纯化的重组蛋白进行了质谱鉴定和酶活性分析。酶活性分析表明，复性后的重组蛋白能在DTT存在的条件下还原H2O2。

Embryology of Swertia cincta (Gentianaceae) and Its Systematic Value

This paper reports the mega-, micro-sporogenesis and female-, male-gametogenesis of Swertia cincta for the first time, with the aim of discussing the systematic position of section Platynema and section Ophelia of Swertia. Anthers are tetrasporangiate. The development of anther walls conforms to the dicotyledonous type. The tapetum cells have dual origin and are similar to the glandular type. There are two middle layers. The endothecium and epidermis persist. Cytokinesis in the microsporrocyte meiosis is simultaneous type and the microscpore teads are tetrahedral. Pollen grains are 3-celled. The ovary is bicarpellum and unilocular. The placentation is of suparietal placentation with 12 series of ovules. The ovules. The ovule is unitegmic, tenuinucellar and ana-campylotropous, The embryo sac orignates from the single-archesporial cell. The one chalazal megaspore in lienar tetrad become the functional megasore. The development of embryo sac is of the polygonum type. Before fertilization, two polar nuclei fuse into one secondary nucleus. Three antipodal cells persisted and divided into 5-8 cells. A comparison between two sections indicates that section Plathnema is better treated as distinct section and is more advanced than section Ophelia according to the evolutionary trends of embryological characters.

Embryology of Crawfurdia delavayi (Gentianaceae) and its systematic value

The embryological characters of Crawfurdia delavayi Frabnch. are described and the systematic relationships of Crawfurdia discussed. Anthers are tetrasporangiate. The development of anther walls conforms to the Dicotyledonous type. The tapetum is of single origin. The development of the tapetum with uninucleate cells is of the glandular type. The tapetal cells on the connective side show radial elongation or periclinal division and intrude into the anther locule. The epidermis of anther walls persists and its cells become pillar and fibrous, and the endothecium degenerates. The ovary is bicarpellary and unilocular. The placentation is typically parietal with 8 rows of anatropous ovules. The development of embryo sac is of the polygonum type. Before fertilization, two polar nuclei fuse into a secondary nucleus. Three antipodal cells persist. Flowers are protandrous. Fertilization is porogamous. The development of the endosperm is of the nuclear type. The embryogeny corresponds to the solanad type physalis II variation. The embryological data indicate that it is better to separate Crawfurdia from Gentiana as an independent genus.

基于SP95标准的工厂模型设计与建模方法研究

为解决当前数字化工厂模型所存在的实时性不强和集成性不足等问题,提出了使用管控系统间集成通用标准--SP95标准进行数字化工厂建模。在应用SP95标准对工厂模型进行定义和描述的基础上,结合国内离散企业中的实际情况,提出了一种扩展的工厂模型。该模型在SP95标准的基础上扩展了料架和变量两个元素,并介绍了其实现方式和作用。通过SP95标准提供的功能,扩展了基于可扩展标记语言技术的制造标记语言的Schema结构,为业务系统集成提供统一的数据共享格式。最后给出了一个构建工厂模型的实例,验证了所提出的模型。

Pg震相走时的各向异性影响及各向异性结构成像

As the first arrival of seismic phase in deep seismic sounding, Pg is the important data for studying the attributes of the sedimentary layers and the shape of crystalline basement because of its high intensity and reliable detection. Conventionally, the sedimentary cover is expressed as isotropic, linear increasing model in the interpretation of Pg event. Actually, the sedimentary medium should be anisotropic as preferred cracks or fractures and thin layers are common features in the upper crust, so the interpretation of Pg event needs to be taken account of seismic velocity anisotropy. Traveltime calculation is the base of data processing and interpretation. Here, we only study the type of elliptical anisotropy for the poor quality and insufficiency of DSS data. In this thesis, we first investigate the meaning of elliptical anisotropy in the study of crustal structure and attribute, then derive Pg event’s traveltime-offset relationship by assuming a linear increasing velocity model with elliptical anisotropy and present the invert scheme from Pg traveltime-offset dataset to seismic velocity and its anisotropy of shallow crustal structure. We compare the Pg traveltime calculated by our analytic formula with numerical calculating method to test the accuracy. To get the lateral variation of elliptical anisotropy along the profiling, a tomography inversion method with the derived formula is presented, where the profile is divided into rectangles. Anisotropic imaging of crustal structure and attribute is efficient method for crust study. The imaging result can help us interprete the seismic data and discover the attribute of the rock to analyze the interaction between layers. Traveltime calculation is the base of image. Base on the ray tracing equations, the paper present a realization of three dimension of layer model with arbitrary anisotropic type and an example of Pg traveltime calculation in arbitrary anisotropic type is presented. The traveltime calculation method is complex and it only adapts to nonlinear inversion. Perturbation method of travel-time calculation in anisotropy is the linearization approach. It establishes the direct relation between seismic parameters and travetime and it is fit for inversion in anisotropic structural imaging. The thesis presents a P-wave imaging method of layer media for TTI. Southeastern China is an important part of the tectonic framework concerning the continental margin of eastern China and is commonly assumed to comprise the Yangtze block and the Cathaysia block, the two major tectonic units in the region. It’s a typical geological and geophysical zone. In this part, we fit the traveltime of Pg phase by the raytracing numerical method. But the method is not suitable here because the inefficiency of numerical method and the method itself. By the analytic method, we fit the Pg and Sg and get the lateral variation of elliptical anisotropy and then discuss its implication. The northeastern margin of Qinghai-Tibetan plateau is typical because it is the joint area of Eurasian plate and Indian plate and many strong earthquakes have occurred there in recent years．We use the Pg data to get elliptical anisotropic variation and discuss the possible meaning.

HHT在电磁信号处理中的应用

In modem signal Processing，non－linear，non－Gaussian and non－stable signals are usually the analyzed and Processed objects，especially non－stable signals. The convention always to analyze and Process non－stable signals are： short time Fourier transform，Wigner－Ville distribution，wavelet Transform and so on. But the above three algorithms are all based on Fourier Transform，so they all have the shortcoming of Fourier Analysis and cannot get rid of the localization of it. Hilbert－Huang Transform is a new non－stable signal processing technology，proposed by N. E. Huang in 1998. It is composed of Empirical Mode Decomposition (referred to as EMD) and Hilbert Spectral Analysis (referred to as HSA). After EMD Processing，any non－stable signal will be decomposed to a series of data sequences with different scales. Each sequence is called an Intrinsic Mode Function (referred to as IMF). And then the energy distribution plots of the original non－stable signal can be found by summing all the Hilbert spectrums of each IMF. In essence，this algorithm makes the non－stable signals become stable and decomposes the fluctuations and tendencies of different scales by degrees and at last describes the frequency components with instantaneous frequency and energy instead of the total frequency and energy in Fourier Spectral Analysis. In this case，the shortcoming of using many fake harmonic waves to describe non－linear and non－stable signals in Fourier Transform can be avoided. This Paper researches in the following parts： Firstly，This paper introduce the history and development of HHT，subsequently the characters and main issues of HHT. This paper briefly introduced the basic realization principles and algorithms of Hilbert－Huang transformation and confirms its validity by simulations. Secondly, This paper discuss on some shortcoming of HHT. By using FFT interpolation, we solve the problem of IMF instability and instantaneous frequency undulate which are caused by the insufficiency of sampling rate. As to the bound effect caused by the limitation of envelop algorithm of HHT, we use the wave characteristic matching method, and have good result. Thirdly, This paper do some deeply research on the application of HHT in electromagnetism signals processing. Based on the analysis of actual data examples, we discussed its application in electromagnetism signals processing and noise suppression. Using empirical mode decomposition method and multi-scale filter characteristics can effectively analyze the noise distribution of electromagnetism signal and suppress interference processing and information interpretability. It has been founded that selecting electromagnetism signal sessions using Hilbert time-frequency energy spectrum is helpful to improve signal quality and enhance the quality of data.

攻击性、情绪调节评价对青少年攻击行为的作用

The aggressive behavior and the violent crime of the Adolenscents have aroused social widespread attention. The Social Implicit Cognition Method could overcome the insufficiency of explicit research methods such as Self-Report Scale, and carry on the thorough discussion to the intrinsic mechanism of the Adolenscent’s aggressive behavior. In this research, Implicit Association Test and Explicit Questionnaire are synthetically applied to inspect the relations among the aggression, Evaluation of Emotion Regulation and the aggressive behavior of the normal middle-school students and the Adolenscent violent criminals under the explicit and implicit conditions, and have carried the supplement to the internal condition of the General aggression model (GAM). The result indicates, 1. The concept of implicit aggression exists independently and separates from explicit aggression. However the relations between implicit aggression and the groups are remarkable, i.e. both the implicit and explicit aggressions of the Adolenscent violent criminals are remarkably higher than those of the normal middle-school students, but not influenced by sex factors. 2. Implicit Evaluation of Emotion Regulation is suitable to Chinese Adolenscents. The correlation of groups and sex factors exist to Implicit Evaluation of Emotion Regulation, i.e. in the group of criminals, the female inclines to the mood control compared with the male in the aspect of Implicit Evaluation of Emotion Regulation; however, in the group of normal persons, the difference between male and female is not remarkable in the aspect of Implicit Evaluation of Emotion Regulation. 3. Under the explicit conditions, it is discovered that the remarkable explicit aggression can predict violent criminals. But it is adjusted by Explicit Evaluation of Emotion Regulation, i.e. the predicting function is not remarkable when the Explicit Evaluation inclines to expression. Meanwhile, when checking the functions and the effects of the implicit variables after the explicit conditions are under control, it is discovered that the predicting function of implicit aggression is still remarkable, and the predicting function of Implicit Evaluation of Emotion Regulation is remarkable at the edge and also adjusted by sex factors. These findings help us not only understanding thoroughly the intrinsic mechanism of the Adolenscent’s aggressive behavior, discovering the moderating role of Evaluation of Emotion Regulation on aggression, but also providing an academic foundation on the prevention and intervention of the Adolenscent’s aggressive behavior.