无指盘臭蛙皮肤抗感染多肽组学研究

在过去的一个多世纪里，两栖类动物皮肤分泌液作为它们的第一道防御屏障引起了研究者们极大的兴趣，同时也开展了相关的许对研究，到目前为止，已从中分离鉴定出了百余种的活性物质。无指盘臭蛙是我国的一种特有两栖动物，初步的活性检测发现，无指盘臭蛙皮肤分泌液具有很强的抗菌，溶血以及蛋白酶抑制剂活性。 在本论文中，我们利用多肽组学与基因组学的方法对无指盘臭蛙皮肤的抗感染多肽组进行了研究。 通过三步分离纯化过程：一步Sephadex G-50分子筛和两步反相高压液相（RP-HPLC）的方法，从无指盘臭蛙皮肤分泌液中分离纯化得到了21条新的抗菌肽，它们分别属于17个不同的抗菌肽家族，其中8个分别属于已知的5个抗菌肽家族，它们是：Brevinin-1E(2个)、Brevinin-2E（1个）、Esculentin-1（1个）、Esculentin-2（1个）和Nigrocin（3个）抗菌肽家族。另外的13个抗菌肽与已发现的抗菌肽表现出较低的相似性，我们将它们归类到12种新的抗菌肽家族，它们是：Odorranain-A (1个)、Odorranain-B(1个)、 Odorranain-C(1个)、 Odorranain-G (1个)、Odorranain-H (2个)、 Odorranain-J (1个)、Odorranain-L (1个)、Odorranain-M (1个)、 Odorranain-N (1个)、 Odorranain-O (1个)、Odorranain-Q(1个)、 Odorranain-T(1个)。 从单个无指盘臭蛙皮肤里面，我们克隆得到了372条抗菌肽序列，它们编码107条新的抗菌肽，这一发现使得目前发现的两栖类抗菌肽的数目几乎增加了1倍。这也是目前发现的抗菌肽最为丰富的物种。这107条抗菌肽分别属于30个不同的抗菌肽家族，其中有24个为新的抗菌肽家族。这些抗菌肽的多样性可能是通过点突变、碱基的插入或删除、结构域的穿梭以及拼接等多种机制形成的。这些抗菌肽多样性的形成可能与无指盘臭蛙生活环境中微生物的组成有关。30个家族抗菌肽前体序列的SPD区域（包括信号肽和前导肽序列，Signal and Propiece Domain, SPD）非常保守，表明它们可能起源于同一个祖先基因。对7个抗菌肽家族的非同义碱基替代率Dn与同义碱基替代率Ds进行检测发现，它们可能经受着不同选择压力的作用。无指盘臭蛙皮肤抗菌肽在二级结构和功能上都表现出丰富的多样性。在一个两栖类个体里面发现如此丰富的抗菌肽甚是让人惊讶。这一发现也使得我们不得不重新认识先天性免疫在两栖类动物防御系统中的重要性，对两栖类生态环境的分子基础以及那种认为一种两栖类只需要20-30种抗菌肽就足以抵御环境中的微生物的看法重新审视。我们的研究还显示：无指盘臭蛙抗菌肽之间还存在着协同效用。 我们对无指盘臭蛙皮肤抗菌肽的去极化作用进行了研究，发现所检测的7个抗菌肽都可使金黄色葡萄球菌发生去极化，但是它们使细菌发生去极化的能力不同。对12种抗菌肽的抗菌机制研究发现，它们通过多种不同的机制发挥作用：有些在细菌内形成片层样的囊泡状结构，有些导致细胞质壁的分离，有些在细菌的膜上形成穿孔，有些则导致了细菌染色质的固缩。 总之，这些研究为设计新型的抗菌肽提供了有用的参考资料。

金藻吞噬微囊藻产生的无毒变异株与产毒原始株的比较

分别从喂食三株原始产毒铜绿微囊藻Microcystis aeruginosa(AC、DS和PCC 7820)的金藻Poterioochromonassp.培养物中获得三株藻,以Nest PCR方法(引物对CC/CG和CH/CI)确定此三株藻均为微囊藻属藻株。HPLC测试结果显示这三株藻均不产生微囊藻毒素。显示Poterioochromonassp.具有将产毒微囊藻转化为无毒微囊藻的能力。比较产毒原始株与无毒变异株的生理特性发现,变异株的类胡萝卜素/叶绿素比值高于原始株;而光反应曲线结果表明,变异株的PSⅡ

微囊藻毒素对几种淡水微藻的生长和光合活性的影响

微囊藻毒素(MC)是富营养化淡水水体中最常见的藻类毒素,而MC对藻类生长效应的影响却鲜见报道。通过模拟培养实验,研究了不同质量浓度的MC-RR对淡水藻类的生长和光合效能的影响。结果显示,100μg·L-1以下的MC-RR对产毒铜绿微囊藻Ds(MicrocystisaeruginosaDs)作用并不明显;相反,100μg·L-1MC-RR对铜绿微囊藻无毒株854(Microcystisaeruginosa854)有显著的杀藻效应,表明MC可能改变浮游植物种群中产毒与非产毒微囊藻的比例。MC-RR对其它藻类的

八角枫挥发油化学成分研究

八角枫是八角枫科八角枫属药用植物 ,分布较广。用气相色谱 /质谱 /计算机联用技术 ( GC/MS/DS)分析了水蒸气蒸馏得到的八角枫挥发油 ,鉴定出 5 9种化学成分 ,其主要成分为 :1 ,8-桉叶素 ( 43.32 5 % )、β-侧柏烯 ( 1 0 .71 3% )、丁香酚甲醚 ( 7.0 88% )、α-松油醇( 7.0 1 7% )、α-蒎烯 ( 5 .830 % )等 5种化合物。被鉴定的 5 9种成分 ,共占挥发油总量的97.0 4 %。

Molecular evolution of connective tissue growth factor in Cyprinidae (Teleostei : Cypriniformes)

Connective tissue growth factor (CTGF) plays an important role in regulation of cell growth, differentiation, apoptosis and individual development in animals. The study of sequences variation and molecular evolution of CTGF gene across various species of the cyprinid could be helpful for understanding of speciation and gene divergence in this kind of fish. In this study, 19 novel sequences of CTGF gene were obtained from the representative species of the family Cyprinidae using PCR amplification, cloning and sequencing. Phylogenetic relationships of Cyprinidae were reconstructed by neighbor-joining (NJ), maximum parsimony (MP), maximum likelihood (ML) and Bayesian method. Oryzias latipes from the family Cyprinodontidae was assigned to be the outgroup taxon. Leuciscini and Barbini were clustered into the monophyletic lineages, respectively, with the high nodal supports. The estimation of the ratio of non-synonymous to synonymous substitution (dN/dS) for the various branches indicated that there stood the different evolution rates between the Leuciscini and the Barbini. With the ratio of dN/dS of the Leuciscini being lower than that of the Barbini, species within the Barbini were demonstrated to be subjected to the relatively less selection pressure and under the relaxable evolution background. A 6 by indel (insertion/deletion) was found at the 5' end of CTGF gene of Cyprinidae, and this 6 by deletion only appeared in the Leuciscini, which is a typical characteristic of the Leuciscini and provides evidence for the monophylogeny of the Leuciscini. For the amino acid sequences of CTGF protein, the most variations and indels were distributed in the signal region and IGFBP region of this protein, implying that these variations were correlated with the regulation of the CTGF gene expression and protein activity. (c) 2007 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.

The innate immune response to grass carp hemorrhagic virus (GCHV) in cultured Carassius auratus blastulae (CAB) cells

Virus infection of mammalian cells activates an innate antiviral immune response characterized by production of interferon (IFN) and the subsequent transcriptional upregulation of IFN-stimulated genes (ISGs) by the JAK-STAT signaling pathway. Here, we report that a fish cell line, crucian carp (Carassius auratus L.) blastulae embryonic (CAB) cells, can produce IFN activity and then form an antiviral state after infection with UV-inactivated grass carp hemorrhagic virus (GCHV), a double-stranded (ds) RNA virus. From UV-inactivated GCHV-infected CAB cells, 15 pivotal genes were cloned and sequenced, and all of them were shown to be involved in IFN antiviral innate immune response. These IFN system genes include the dsRNA signal sensing factor TLR3, IFN, IFN signal transduction factor STAT1, IFN regulatory factor IRF7, putative IFN antiviral effectors Mx1, Mx2, PKR-like, Viperin, IFI56, and other IFN stimulated genes (ISGs) IFI58, ISG15-1, ISG15-2, USP18, Gig1 and Gig2. The identified fish IFN system genes were highly induced by active GCHV, UV-inactivated GCHV, CAB IFN or poly(I).poly(C), and showed similar expression patterns to mammals. The data indicate that an IFN antiviral innate immune response similar to that in mammals exists in the UV-inactivated GCHV-infected CAB cells, and the IFN response contributes to the formation of an antiviral state probably through JAK-STAT signaling pathway. This study provides strong evidence for existence of IFN antiviral innate immune response in fish, and will assist in elucidating the origin and evolution of vertebrate IFN system. (c) 2006 Elsevier Ltd. All rights reserved.

An experimental study about the influence of well thickness on the electroluminescence of InGaN/GaN multiple quantum wells

The influence of well thickness on the electroluminescence (EL) of InGaN/GaN multiple quantum wells (MQWs) grown by metalorganic chemical vapor deposition is investigated. It is found that the peak wavelength of EL increases with the increase of well thickness when the latter is located in the range of 3.0-5.1 nm. The redshift is mainly attributed to the quantum confined Stark effect (QCSE). As a contrast, it is found that the EL intensity of InGaN/GaN MQWs increases with the increase of well thickness in spite of QCSE. The result of X-ray diffraction demonstrates that the interface become smoother with the increase of well thickness and suggests that the reduced interface roughness can be an important factor leading to the increase of EL intensity of InGaN/GaN MQWs. (C) 2009 Elsevier B.V. All rights reserved.

Effect of surface treatment of GaN based light emitting diode wafers on the leakage current of light emitting diode devices

To form low-resistance Ohmic contact to p-type GaN, InGaN/GaN multiple quantum well light emitting diode wafers are treated with boiled aqua regia prior to Ni/Au (5 nm/5 nm) film deposition. The surface morphology of wafers and the current-voltage characteristics of fabricated light emitting diode devices are investigated. It is shown that surface treatment with boiled aqua regia could effectively remove oxide from the surface of the p-GaN layer, and reveal defect-pits whose density is almost the same as the screw dislocation density estimated by x-ray rocking curve measurement. It suggests that the metal atoms of the Ni/Au transparent electrode of light emitting diode devices may diffuse into the p-GaN layer along threading dislocation lines and form additional leakage current channels. Therefore, the surface treatment time with boiled aqua regia should not be too long so as to avoid the increase of threading dislocation-induced leakage current and the degradation of electrical properties of light emitting diodes

Influence of AlN buffer layer thickness on structural properties of GaN epilayer grown on Si (111) substrate with AlGaN interlayer

We present the growth of GaN epilayer on Si (111) substrate with a single AlGaN interlayer sandwiched between the GaN epilayer and AlN buffer layer by using the metalorganic chemical vapour deposition. The influence of the AlN buffer layer thickness on structural properties of the GaN epilayer has been investigated by scanning electron microscopy, atomic force microscopy, optical microscopy and high-resolution x-ray diffraction. It is found that an AlN buffer layer with the appropriate thickness plays an important role in increasing compressive strain and improving crystal quality during the growth of AlGaN interlayer, which can introduce a more compressive strain into the subsequent grown GaN layer, and reduce the crack density and threading dislocation density in GaN film.

Hole concentration test of p-type GaN by analyzing the spectral response of p-n(+) structure GaN ultraviolet photodetector

A new method to test the hole concentration of p-type GaN is proposed, which is carried out by analyzing the spectral response of p-n(+) structure GaN ultraviolet photodetector. It is shown that the spectral response of the photodetector changes considerably with reversed bias. It is found that the difference between photodetector's quantum efficiency at two wavelengths, i.e. 250 and 361 nm, varies remarkably with increasing reversed bias. According to the simulation calculation, the most characteristic change occurs at a reversed voltage under which the p-GaN layer starts to be completely depleted. Based on this effect the carrier concentration of p-GaN can be derived.

Structural and optical properties of Al1-xInxN epilayers on GaN template grown by metalorganic chemical vapor deposition

This paper reports that Al1-xInxN epilayers were grown on GaN template by metalorganic chemical vapor deposition with an In content of 7%-20%. X-ray diffraction results indicate that all these Al1-xInxN epilayers have a relatively low density of threading dislocations. Rutherford backscattering/channeling measurements provide the exact compositional information and show that a gradual variation in composition of the Al1-xInxN epilayer happens along the growth direction. The experimental results of optical reflection clearly show the bandgap energies of Al1-xInxN epilayers. A bowing parameter of 6.5 eV is obtained from the compositional dependence of the energy gap. The cathodoluminescence peak energy of the Al1-xInxN epilayer is much lower than its bandgap, indicating a relatively large Stokes shift in the Al1-xInxN sample.

Carrier capture by threading dislocations in (In,Ga)N/GaN heteroepitaxial layers

Using spatially resolved cathodoluminescence spectroscopy, we investigate the spatial luminescence distribution in a fully strained (In,Ga)N layer, in particular, its correlation with the distribution of threading dislocations (TDs). Regarding the impact of TDs on the luminescence properties, we can clearly distinguish between pure edge-type TDs and TDs with screw component. At the positions of both types of TDs, we establish nonradiative recombination sinks. The radius for carrier capture is at least four times larger for TDs with screw component as for pure edge-type TDs. The large capture radius of the former is due to a spiral-like growth mode resulting in an increase in the In content in the center of the spiral domains in comparison to their periphery.

Fabrication and Characterization of High Power InGaN Blue-Violet Lasers with an Array Structure

InGaN/GaN multi-quantum-well-structure laser diodes with an array structure are successfully fabricated on sapphire substrates. The laser diode consists of four emitter stripes which share common electrodes on one laser chip. An 800-mu m-long cavity is formed by cleaving the substrate along the < 1 (1) over bar 00 >. orientation using laser scriber. The threshold current and voltage of the laser array diode are 2A and 10.5 V, respectively. A light output peak power of 12W under pulsed current injection at room temperature is achieved. We simulate the electric properties of GaN based laser diode in a co-planar structure and the results show that minimizing the difference of distances between the different ridges and the n-electrode and increasing the electrical conductivity of the n-type GaN are two effective ways to improve the uniformity of carrier distribution in emitter stripes. Two pairs of emitters on a chip are arranged to be located near the two n-electrode pads on the left and right sides, and the four stripe emitters can laser together. The laser diode shows two sharp peaks of light output at 408 and 409 nm above the threshold current. The full widths at half maximum for the parallel and perpendicular far field patterns are 8 degrees and 32 degrees, respectively.

Role of Ga vacancies in enhancing the leakage current of GaN Schottky barrier ultraviolet photodetectors

The leakage current of GaN Schottky barrier ultraviolet photodetectors is investigated. It is found that the photodetectors adopting undoped GaN instead of lightly Si-doped GaN as an active layer show a much lower leakage current even when they have a higher dislocation density. It is also found that the density of Ga vacancies in undoped GaN is much lower than in Si-doped GaN. The Ga vacancies may enhance tunneling and reduce effective Schottky barrier height, leading to an increase of leakage current. It suggests that when undoped GaN is used as the active layer, it is necessary to reduce the leakage current of GaN Schottky barrier ultraviolet photodetector.

Effective recombination velocity of textured surfaces

Surface texturization is an effective way to enhance the absorption of light for optoelectronic devices but it also aggravates the surface recombination by enlarging the surface area. In order to evaluate the influence of texture structures on the surface recombination, an effective surface recombination velocity is defined which is assumed to have an equivalent recombination effect on a flat surface. Based on numerical and analytical calculation, the dependences of effective surface recombination on the pattern geometry, the surface recombination velocity, and the diffusion length are analyzed.