Molecular cloning, characterization and expression analysis of a putative C-type lectin (Fclectin) gene in Chinese shrimp Fenneropenaeus chinensis

Lectin is regarded as a potential molecule involved in immune recognition and phagocytosis through opsonization in crustacean. Knowledge on lectin at molecular level would help us to understand its regulation mechanism in crustacean immune system. A novel C-type lectin gene (Fclectin) was cloned from hemocytes of Chinese shrimp Fenneropenaeus chinensis by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA consists of 1482 bp with an 861 bp open reading frame, encoding 287 amino acids. The deduced amino acid sequence contains a putative signal peptide of 19 amino acids. It also contains two carbohydrate recognition domains/C-type lectin-like domains (CRD1 and CRD2), which share 78% identity with each other. CRD1 and CRD2 showed 34% and 30% identity with that of mannose-binding lectin from Japanese lamprey (Lethenteron japonicum), respectively. Both CRD1 and CRD2 of Fclectin have I I amino acids residues, which are relatively invariant in animals' C-type lectin CRDs. Five residues at Ca2+ binding site I are conserved in Fclectin. The potential Ca2+/carbohydrate-binding (site 2) motif QPD, E, NP (Gln-Pro-Asp, Glu, Asn-Pro) presented in the two CRDs of Fclectin may support its ability to bind galactose-type sugars. It could be deduced that Fclectin is a member of C-type lectin superfamily. Transcripts of Fclectin were found only in hemocytes by Northern blotting and RNA in situ hybridization. The variation of mRNA transcription level in hemocytes during artificial infection with bacteria and white spot syndrome virus (WSSV) was quantitated by capillary electrophoresis after RT-PCR. An exploration of mRNA expression variation after LPS stimulation was carried out in primarily cultured hemocytes in vitro. Expression profiles of Fclectin gene were greatly modified after bacteria, LPS or WSSV challenge. The above-stated data can provide us clues to understand the probable role of C-type lectin in innate immunity of shrimp and would be helpful to shrimp disease control. (c) 2006 Elsevier Ltd. All rights reserved.

左侧梭状回内文字的字形加工：中文正常阅读者，中文文盲、非中文母语被试的对比研究

Reading is an important human-specific skill obtained through extensive learning experience and is reliance on the ability to rapidly recognize single words. According to the behavioral studies, the most important stage of reading is the representation of “visual word form”, which is independent on surface visual features of the reading materials. The prelexical visual word form representation is characterized by the abstractive and highly effective and precise processing. Neuroimaging and neuropsychological studies have investigated the neural basis underlying the visual word form processing. On the basis of summary of the existing literature, the current thesis aimed to address three fundamental questions involving neural basis of word recognition. First, is there a dedicated neural network that is specialized for word recognition? Second, is the orthographic information represented in the putative word/character selective region (VWFA)? Third, what is the role of reading experience in the genesis of the VWFA, is experience a main driver to shape VWFA instead of evolutionary selectivity? Nineteen Chinese literate volunteers, 5 Chinese illiterates and 4 native English speakers participated in this study, and performed perceptual tasks during fMRI scanning. To address the first question, we compared the differential responses to three categories of visual objects, i.e., faces, line drawings of objects and Chinese characters, and defined the region of interesting (ROI) for the next experiment. To address the second question, Chinese character orthography was manipulated to reveal possible differential responses to real characters, false characters, radical combinations, and stroke combinations in the regions defined by the first experiment. To examine the role of reading experience in genesis of specialization for character, the responses for unfamiliar Chinese characters in Chinese illiterates and native English speakers were compared with that in the Chinese literates, and tracked the change in cortical activation after a short-term reading training in the illiterates. Data were analyzed in two dimensions. Both BOLD signal amplitude and spatial distribution pattern among multi-voxels were used to systematically investigate the responsiveness of the left fusiform gyrus to Chinese characters. Our results provide strong and clear evidence for the existence of functionally specialized regions in the human ventral occipital-temporal cortex. In the skilled readers a region specialized for written words could be consistently found in the lateral part of the left fusiform gyrus, line drawings in the median part and faces in the middle. Our results further show that spatial distribution analysis, a method that was not commonly used in neuroimaging of reading, appears to be a more effective measurement for category specialization for visual objects processing. Although we failed to provide evidence that VWFA processes orthographic information in terms of signal intensitiy, we do show that response pattern of real characters and radical collections in this area is different from that of false characters and random stroke combinations. Our last set of experiments suggests that the selective bias to reading material is clearly experience dependent. The response to unknown characters in both English speakers/readers and Chinese illiterates is fundamentally different from that of the skilled Chinese readers. The response pattern for unknown characters is more similar to that for line drawings rather as a weak version of character in skilled Chinese readers. Short-term training is not sufficient to produce VWFA bias even when tested with learned characters, rather the learned characters generated a overall upward shift of the activation of the left fusiform region. Formation of a dedicated region specialized for visual word/character might depend on long-term extensive reading experience, or there might be a critical period for reading acquisition.

真实和虚拟现实环境下场景识别中的空间更新

Currently，one of the important research areas in Spatial updating is the role of external (for instance visual) and internal (for instance proprioceptive or vestibular) information in spatial updating of scene recognition. Our study uses the paradigm of classic spatial updating research and the experimental design of investigation of Burgess(2004),first, we will explore the concrete influence of locomotion on scene recognition in real world; next, we will use virtual reality technology, which can control many spatial learning parameters and exclude the influence of extra irrelevant variables, to explore the influence of pure locomotion without visual cue on scene recognition, and furthermore, we will explore whether the ability of spatial updating can be transferred to new situations in a short period of time and compare the result pattern in real word with that in virtual reality to test the validity of virtual reality technology in spatial updating of scene recognition research. The main results of this paper can be summarized as follows： 1. In real world, we found two effects: the spatial updating effect and the viewpoint dependent effect, this result indicated that the spatial updating effect based on locomotion does not eliminate the viewpoint dependent effect during the scene recognition process in physical environment. 2. In virtual reality environment, we still found two effects: the spatial updating effect and the viewpoint dependent effect, this result showed us that the spatial updating effect based on locomotion does not eliminate the viewpoint dependent effect during the scene recognition process in virtual reality environment either. 3. The spatial updating effect based on locomotion plays double role in scene recognition: When subjects were tested in different viewpoint, spatial updating based on locomotion promoted scene recognition; while subjected were tested in same viewpoint, spatial updating based on locomotion had a negative influence on scene recognition, these results show us that spatial updating based on locomotion is automated and can not be ignored. 4. The ability of spatial updating can be transferred to new situations in a short period of time , and the experiment in the immersed virtual reality environment got the same result pattern with that in the physical environment, suggesting VR technology is a very effective method to do research on spatial updating of the scene recognition studies. 5. This study about scene recognition provides evidence to double system model of spatial updating in the immersed virtual reality environment.

利用本征脸方法减少冷原子云吸收成像照片中的干涉条纹

本征脸方法是广泛应用于人脸识别的一种图像处理方法，本文将其引入到原子芯片上囚禁的冷原子云吸收成像照片的图像处理中，以减少其中的干涉条纹，增加信噪比。本文首先介绍了吸收成像照片的标准处理方法以及干涉条纹的产生原因，由于参考照片和吸收成像照片中的干涉条纹会发生随机的相对变化，处理后干涉条纹难以消除。和标准的处理方法相比，本征脸方法不是使用1张而是50张参考照片，利用这些照片重构出一张新的参考照片，这张照片比那50张中的任何一张都更近似于吸收成像照片，因此和只使用1张参考照片相比，处理之后的干涉条纹对比度明显降

Trimeresurus stejnegeri snake venom plasminogen activator - Site-directed mutagenesis and molecular modeling

The specific plasminogen activator from Trimeresurus stejnegeri venom (TSV-PA) is a serine proteinase presenting 23% sequence identity with the proteinase domain of tissue type plasminogen activator, and 63% with batroxobin, a fibrinogen clotting enzyme from Bothrops atrox venom that does not activate plasminogen. TSV-PA contains six disulfide bonds and has been successfully overexpressed in Escherichia coli (Zhang, Y., Wisner, A., Xiong, Y. L,, and Bon, C, (1995) J. Biol. Chem. 270, 10246-10255), To identify the functional domains of TSV-PA, we focused on three short peptide fragments of TSV-PA showing important sequence differences with batroxobin and other venom serine proteinases. Molecular modeling shows that these sequences are located in surface loop regions, one of which is next to the catalytic site, When these sequences were replaced in TSV-PA by the equivalent batroxobin residues none generated either fibrinogen-clotting or direct fibrinogenolytic activity, Two of the replacements had little effect in general and are not critical to the specificity of TSV-PA for plasminogen. Nevertheless, the third replacement, produced by the conversion of the sequence DDE 96a-98 to NVI, significantly increased the K-m for some tripeptide chromogenic substrates and resulted in undetectable plasminogen activation, indicating the key role that the sequence plays in substrate recognition by the enzyme.

Intelectin gene from the grass carp Ctenopharyngodon idella: cDNA cloning, tissue expression, and immunohistochemical localization

The cDNA encoding grass carp intelectin was isolated from a head kidney cDNA library, and termed gcIntL. The deduced amino acid sequence of gcIntL consists of 318 amino acids, and about 55% identical and 74% similar to human intelectin, which is a new type of lectin recognizing galactofuranose, and plays a role in the recognition of bacteria-specific components in animal hosts. The gcIntL gene consists of seven exons and six introns, spacing over approximately 3 kb of genomic sequence. Phylogenetic analysis clearly demonstrated that the gcIntL formed a clade with Danio rerio intelectin and 35 kDa serum lectin. By real-time quantitative RT-PCR analysis, gcIntL transcripts were significantly induced in head kidney, trunk kidney, spleen, and intestine from LPS-stimulated fish. RT-PCR and Western blotting analysis demonstrated that the mRNA and protein of gcIntL gene have the same expression pattern, and both were detected in brain, gill, intestine, head kidney, trunk kidney, spleen, and heart. Furthermore, gcIntL protein could be detected in gill, intestine, trunk kidney, head kidney, spleen, heart, and brain including medulla oblongata and optic lobe, as determined by immunohistochemistry. This is the first report of intelectin expression pattern in fish, and of recombinant gcIntL and polyclonal antibody against gcIntL. (c) 2006 Elsevier Ltd. All rights reserved.

Double synaptic weight neuron theory and its application

In this paper, a novel mathematical model of neuron-Double Synaptic Weight Neuron (DSWN)(l) is presented. The DSWN can simulate many kinds of neuron architectures, including Radial-Basis-Function (RBF), Hyper Sausage and Hyper Ellipsoid models, etc. Moreover, this new model has been implemented in the new CASSANN-II neurocomputer that can be used to form various types of neural networks with multiple mathematical models of neurons. The flexibility of the DSWN has also been described in constructing neural networks. Based on the theory of Biomimetic Pattern Recognition (BPR) and high-dimensional space covering, a recognition system of omni directionally oriented rigid objects on the horizontal surface and a face recognition system had been implemented on CASSANN-II neurocomputer. In these two special cases, the result showed DSWN neural network had great potential in pattern recognition.

Spectral response of winter wheat to crop development and various agronomic limitations

For maximizing the effective applications of remote sensing in crop recognition, crop performance assessment and canopy variables estimation at large areas, it is essential to fully understand the spectral response of canopy to crop development and varying growing conditions. In this paper, the spectral properties of winter wheat canopy under different growth stages and different agronomic conditions were investigated at the field level based on reflectance measurements. It was proved that crop growth and development, nitrogen fertilization rates, nutrient deficit (e.g. lacking any kind of nitrogen, phosphorus and kalium fertilizer or lacking all of them), irrigation frequency and plant density had direct influence on canopy reflectance in 400-900 nm which including the visible/near infrared bands, and resulted in great changes of spectral curves. It was suggested that spectral reflectance of crop canopy can well reflect the growth and development of crop and the impacts from various factors, and was feasible to provide vital information for crop monitoring and assessment. ©2010 IEEE.

Context and conformation dictate function of a transcription antitermination switch

In bacteriophage, transcription elongation is regulated by the N protein, which binds a nascent mRNA hairpin ( termed boxB) and enables RNA polymerase to read through distal terminators. We have examined the structure, energetics and in vivo function of a number of N boxB complexes derived from in vitro protein selection. Trp18 fully stacks on the RNA loop in the wild-type structure, and can become partially or completely unstacked when the sequence context is changed three or four residues away, resulting in a recognition interface in which the best binding residues depend on the sequence context. Notably, in vivo antitermination activity correlates with the presence of a stacked aromatic residue at position 18, but not with N boxB binding affinity. Our work demonstrates that RNA polymerase responds to subtle conformational changes in cis-acting regulatory complexes and that approximation of components is not sufficient to generate a fully functional transcription switch.

Analysis of the effect of copper on the virulence of a pathogenic Edwardsiella tarda strain

Aim: To investigate the effect of copper on the virulence of Edwardsiella tarda. Methods and Results: The pathogenic Edw. tarda strain TX5 was cultured under copper-stressed conditions and examined for any potential alteration in capacities that are associated with pathogenicity. The results showed that compared to untreated TX5, Cu-treated TX5 exhibits reduced planktonic and biofilm growth, an impaired ability to adhere to host mucus, modulation of host immune response, and dissemination in host blood and liver. Consistent with these observations, the overall bacterial virulence of Cu-treated TX5 is significantly attenuated. SDS-PAGE analyses of whole cell protein production showed that Cu-treated TX5 differs from the untreated TX5 in its production of at least one protein. Quantitative real time reverse transcriptase PCR analyses showed that copper treatment decreased the expression of virulence-associated genes encoding components of the type III and type VI secretion systems, the Eth haemolysin system, and the LuxS/AI-2 quorum-sensing system. Conclusions: Prolonged exposure to copper has multiple effects on TX5 and results in significant attenuation of bacterial virulence. Significance and Impact of the Study: The results of this study demonstrate that copper treatment has a broad and profound effect on the virulence-associated capacities of TX5, which is exerted at least in part at the transcription level. These findings provide new insights to the antimicrobial mechanism of copper.

A novel C1q-domain-containing protein from Zhikong scallop Chlamys farreri with lipopolysaccharide binding activity

The C1q-domain-containing (C1qDC) proteins are a family of proteins characterized by a globular C1q (gC1q) domain in their C-terminus. They are involved in various processes of vertebrates and supposed to be an important pattern recognition receptor in innate immunity of invertebrates. In this study, a novel member of C1q-domain-containing protein family was identified from Zhikong scallop Chlamys farreri (designated as CfC1qDC) by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of CfC1qDC was of 777 bp, consisting of a T-terminal untranslated region (UTR) of 62 bp and a 3' UTR of 178 bp with a polyadenylation signal sequence AATAAA and a poly (A) tail. The CfC1qDC cDNA encoded a polypeptide of 178 amino acids, including a signal peptide and a C1q-domain of 158 amino acids with the theoretical isoelectric point of 5.19 and the predicted molecular weight of 17.2 kDa. The C1q-domain in CfC1qDC exhibited homology with those in sialic acid binding lectin from mollusks and C1qDC proteins from higher vertebrates. The typical 10 beta-strand jelly-roll folding topology structure of C1q-domain and the residues essential for effective packing of the hydrophobic core were well conserved in CfC1qDC. By fluorescent quantitative real-time PCR, mRNA transcripts of CfC1qDC were mainly detected in kidney, mantle, adductor muscle and gill, and also marginally detectable in hemocytes. In the bacterial challenge experiment, after the scallops were challenged by Listonella anguillarum, there was a significant up-regulation in the relative expression level of CfC1qDC and at 6 h post-injection, the mRNA expression reached the maximum level and was 4.55-fold higher than that of control scallops. Similarly, the expression of CfC1qDC mRNA in mixed primary cultures of hemocytes stimulated by lipopolysaccharides (LPS) was up-regulated and reached the maximum level at 6 h post-stimulation, and then dropped back to the original level gradually. In order to investigate its function, the cDNA fragment encoding the mature peptide of CfC1qDC was recombined and expressed in Escherichia coli BL21 (DE3). The recombinant CfC1qDC protein displayed a significantly strong activity to bind LIDS from E. coli, although no obvious antibacterial or agglutinating activity toward Gram-negative bacteria E. coli JM109, L. anguillarum and Gram-positive bacteria Micrococcus luteus was observed. These results suggested that CfC1qDC was absolutely a novel member of the C1qDC protein family and was involved in the recognition of invading microorganisms probably as a pattern recognition molecule in mollusk. (c) 2008 Elsevier Ltd. All rights reserved.

A lectin (CfLec-2) aggregating Staphylococcus haemolyticus from scallop Chlamys farreri

Lectins are a family of carbohydrate-recognition proteins which play crucial roles in innate immunity. In this study, a new lectin (CfLec-2) gene was cloned from Chlamys farreri by EST and RACE approaches. The full-length cDNA of CfLec-2 was composed of 708 bp, encoding a typical Long form carbohydrate-recognition domain of 130 residues. The deduced amino acid sequence showed high similarity to Brevican in Homo sapiens, C-type lectin-1 and lectin-2 in Anguilla japonica. The cDNA fragment encoding the mature peptide of CfLec-2 was recombined into plasmid pET-32a (+) and expressed in Escherichia coli Rosseta-Gami (DE3). The recombinant CfLec-2 (rCfLec-2) protein exhibited aggregative activity toward Staphylococcus haemolyticus, and the agglutination could be inhibited by D-mannose but not EDTA or D-galactose, indicating that CfLec-2 was a Ca2+ independent lectin. Moreover, rCfLec-2 could suppress the growth of E. coli TOP10F'. These results suggested that CfLec-2 was perhaps involved in the recognition and clearance of bacterial. pathogens in scallop. (C) 2007 Elsevier Ltd. All rights reserved.

基于修正的最小均方误差分类算法的移动机器人运动规划

研究了不确定性环境下移动机器人躲避运动轨迹未知的移动障碍物的一种新方法．通过实时最小均方误差估计算法预测每个障碍物的位置及运动轨迹，并利用模式识别中最小均方误差分类器的修正模型计算出机器人的局部避障路径，再运用船舶导航中使用的操纵盘技术来确定每个导航周期中移动机器人的速．度仿真结果表明了该方法的可行性

图象噪声的模糊聚类平滑方法

模糊C-means算法在聚类分析中已得到了成功的应用,本文提出一种利用模糊C-means算法消除噪声的新方法。一般来说,图象中的噪声点就是其灰度值与其周围象素的灰度值之差超过某个门限值的点。根据这个事实,首先利用模糊C-means算法分类,再利用标准核函数检测出噪声点,然后将噪声点去掉。由于只修改噪声点处的象素灰度值,而对于其它象素的灰度值不予改变,所以本算法能够很好地保护细节和边缘。本方法每次处理3×3个点,而以往的方法只能每次处理一个点,所以本方法能提高运算速度。文中给出了利用本方法对实际图象处理的结果,并与梯度倒数权值法进行了定量的比较。