Identification of differentially expressed immune-relevant genes in Chinese soft-shelled turtle (Trionyx sinensis) infected with Aeromonas hydrophila

Expressed sequence tag (EST) analysis is an efficient tool for gene discovery and profiling gene expression. Aeromonas hydrophila, a ubiquitous waterborne bacterium, is one of the most frequent pathogens isolated from diseased aquatic organisms. In order to understand the molecular mechanism of anti-bacteria immune response in reptile, we have investigated the differentially expressed genes in Chinese soft-shelled turtle (Trionyx sinensis) experimentally infected with A. hydrophila by suppression subtractive hybridization (SSH). Forty-two genes were identified from more than 200 clones, of which 25 genes are found for the first time in reptiles, and classified into 6 categories: 18 in defense/immunity. 4 in catalysis, 2 in retrotransposon; 2 in cell signal transduction, 5 in cell metabolism, 10 in protein expression, and 1 in cell structure. Of the 42 differentially expressed genes, 6 genes, IL-8, serum amyloid A (SAA), CD9, CD59, activating transcription factor 4 (ATF4) and cathepsin L genes, were further observed to be up-regulated in the infected turtles by virtual Northern hybridization and RT-PCR assays. (C) 2008 Elsevier B.V. All rights reserved.

细鳞鲃类的系统发育与裂腹鱼类起源关系的探讨

裂腹鱼类起源鳃亚科鱼类的观点得到了多数学者的支持。但是，鱼巴亚科鱼类是一个非常庞杂的类群，裂腹鱼类究竟起源于鳃亚科的哪一个小类群或裂腹鱼类与哪个小类群更为接近？这样的小类群又怎样随着急剧隆起的青藏高原峥演化发展成为现今的裂腹鱼类？不同的学者从不同的研究领域出发，得出了较为不一致甚至是相互对立的结论。裂腹鱼类（我国有11属，76种和亚种）均为小型鳞片的鱼类，与鳃亚科中一些具有小型鳞片的属种较为相似，即，妒鲤属（凡rCO～）的3个亚种、似鳃属（Lztciocyprinus）的2个种，金线纪属（Sinocyclocheilus）的26种或亚种，本文将这些具细小鳞片的鳃亚科鱼类统称为细鳞鳃鱼类（small-scaledbarbids）。为了探讨细鳞鱼巴类和裂腹鱼类的系统发育关界本文应用分支系统学的方法和原理，选择小裂腹鱼S．（R）parvus、中甸叶须鱼P.c.chunglienensis、松藩裸鲤。Gp.potanini、单纹似鳡L.langsoni、花妒鲤P.pingiregani、抚仙金线鱼巴s．师分别作为裂腹鱼类和细鳞鱼巴类中各分类：阶元的代，作为内a为了确考险状的进化极向并进一步探讨细鳞鱼巴各分类阶元与其他纪亚科和鲤亚科鱼类的系统关系，又选择了鳃亚科中的保山四须纪B．baoshanensis、云南四作为内群，并以螂鱼C.auratus、祀麓鲤C．(C)chilia作为外类群，用来对裂腹鱼类和细鳞鳃类进行性状的极化和系统发育分析。通过对12种24尾标本的外部形态和骨骼特征的全面观察，选择了63个性状进行描述和比较，并构建了供系统发育分析的特征矩阵。应用PAUP程序对特征矩阵进行运算，得出了包括外类群在内最简约的系统发育分支图。主要研究结论如下：（l）鲤亚科与包括裂腹鱼类在内的鳃亚科鱼类之间在很多特征方面存在着比较明显的差异，除了臀鳍最末一根不分枝鳍条坚硬、具锯齿；背鳍分枝多于1。根；第一椎体横突退化等特征外，还有镖的前后比例、前鳄骨的特征、尾舌骨的形状，基枕骨的骨质盘特征等诸多特征存在着显著差异。支持了陈湘舞等（1984）将鲤亚科和纪亚科分别作为独立亚科的观点。卿通过选取不同的外类群，在各种不同的条件下对特征数据矩阵进行运算和分析，除极少数（1个）结果外（当以祀麓鲤为唯一外群时，由34来看，它们都是第三纪末期以来由原始的鳃亚科鱼类演化而来的，适应过共同的寒冷环境（青藏高原的急剧隆升导致的寒冷气候和河川急流和青藏高原的大幅度隆起，引起了地貌环境的急剧改变，云贵高原一带也发生了差异性的升降运动。同时，全球性气温下降，常年性冰盖由北向南大幅度推进的寒冷气候环境）（曹文宣等，1981；王大忠等，2000）。裂腹鱼类和似鳃、妒鲤的起源时间可能稍晚一些。可以设想，第三纪末期的鳃亚科鱼类物种分化不是很多，相互之间的亲缘关系较近，分布于'青藏高原的原始鳃亚科鱼类和分布于云贵高原的原始鳃亚科鱼类分布经历了各自独特的地质、气候等环境条件，演化成为当今的裂腹鱼类和金线纪鱼类。至于似媳和妒鲤，除了经历与鳃亚科鱼类和金线鳃鱼类共同的寒冷水环境外，它们还向着肉食性的方向进行演化。（3）裂腹鱼类的3个代表种都聚在一起，其内部关系和与其他种的关系都较为稳定，为鳃亚科鱼类中较为特化的一支。与陈湘舞等（1984）、曹文宣等（1981）的观点较为一致，即，裂腹鱼类起源于纪亚科鱼类。晗（4）似媳和妒鲤之间的亲缘关系最近，它们很可能起源于最近的共同祖先。尽管二者作为一个单系的支持率并不是很高（大于50％，小于60％），但它们在个别特征（咽喉齿的排列方式、咽骨的形态特征）上表现其他鲤科鱼类所不具有的独特特征（附图1的。（5：）裂腹鱼类与妒鲤＋似鳃的系统关系最近，这意味着裂腹鱼类和妒鲤＋似鳃有最近的共同祖先或它们之间的关系较它们与金线鳃鱼类的关系更a（e）应用特征分析和系统发育分析所得的结论，综合对鳃系鱼类染色体特征的分析，认为裂腹鱼类和细鳞鳃鱼类在染色体分组组成、哪值等方面的相似特征，极有可能是在进化过程中经历了类似的寒冷水环境。它们之间的核型特征相似，只是说明了它们之间较近的系统发育关系，不支持妒鲤和金线靶是鳃亚科和裂腹鱼亚科之间的中间演化类型的观点。染色体的进化和外部形态特征的进化在纪系鱼类中存在着不平行的现象。（7）通过国内外鱼类学者对鲤科鱼类和纪亚科鱼类各分类单元定义的比较，结合本文的研究结果，认为国外学者的一些分类系统仍然存在一些尚待解决的问题，故建议目前暂不宜将之引入到我国的鱼类分类系统中。

美国国际空间站的经历与探索及对我国的启示

建设空间站是人类探索外层空间的重要步骤。美、俄、欧、日、加等国联合运营的国际空间站将于2010年建成,并延续到2020年。中国的载人空间站将于2020年升空。本文回顾了空间站的主要历史,概述了美国在国际空间站上10年(1998—2008)的工作,分析了空间站可能对人类做出的贡献,介绍了美国新政府对载人航天任务的新选择,最后借鉴国外发展空间站的经验教训,提出了我国研制空间站应注意的问题。

有机质谱裂解规律及智能解析方法研究

本学位论文分为四个部分，第一部分报道了用串联质谱快速分析合成药物中的微量杂质成分以及分析中药材中的化学成分。第二部分报道了通过质谱和串联质谱发现并合成新型的PdPincer 催化剂，同时对其活性进行测试。第三部分为串联质谱自动解析软件的设计及应用。第四部分概述了应用在质谱中的各种碎裂方式。 第一部分首先总结了5-溴粉防己碱及其类似物的裂解规律，并以此为根据推测出2 个微量杂质的结构。随后针对无患子（Sapindus mukurossi Gatren.）中的皂苷成分，由ESI-QTOF 得到各个皂苷成份的高分辨质量数据进而得到其分子式，然后利用ESI-IT 电喷雾串联质谱对无患子总皂苷中各皂苷成分的结构进行进一步的鉴定。进而以同样的方式，先通过ESI-QTOF 得到黄山药（Dioscoreapanthaica）总皂苷中各个组分化合物的分子式，然后对已有的薯蓣皂苷标准品做串联质谱分析，以得到该类化合物的裂解规律并给出解析该类化合物的流程图。在此利用计算化学的方法讨论了离子的丰度与裂解活化能之间的关系。然后应用APCI-MS/MS 方法探讨了四对同分异构体和几个已知的化合物，并最后用液质联用对其进行确认，同时还给出了4 个未知化合物的可能结构。 第二部分报道通过质谱和串联质谱发现并合成新型的PdPincer 催化剂，同时对其活性进行测试。钯催化的交联反应是有机合成中C-C 键形成的最有效的方法，且硫脲是一类对空气和水都稳定的化合物，因此我们设计并合成了一系列的硫脲钯催化剂并得到了很好的催化活性。我们在对其中一类环状双硫脲化合物进行质谱实验的时候，在正离子模式下发现了反常的[M.H]+，通过串联质谱进一步确定了它是一种新型的PdPincer 结构。我们将其合成出来并通过X-ray 衍射实验确定了它的结构。同时测定其催化活性并与未形成pincer 的类似物进行比较发现该类化合物具有较宽的底物适用性。 第三部分为串联质谱自动解析软件的设计及应用。通过前面两部分的启示，独立设计开发了AuMass（1.0）。其算法是：先通过查找特殊的碎片离子，中性丢失或碎片离子质量差来确定某类化合物的骨架结构，然后利用该类化合物的自动解析流程来对其周边取代基进行确认。通过它快速地对白芍中的化学成分进行解析，并对未知的化合物进行了推测。为了增加它的解析能力，我又对其它类型的化合物裂解规律进行总结，并给出了自动解析流程。实践证明该软件具有相当好的应用价值。 第四部分综述了应用在质谱上的各类母离子的碎裂技术。这里包括了碰撞诱导裂解（CID）、光诱导碎裂（LID）、电子捕获裂解/电子转移裂解（ECD/ETD）、红外多光子解离（IRMPD）、黑体辐射解离（BIRD）和PQD 裂解技术。 This dissertation consists of four chapters. The first chapter reports the rapidanalysis of trace impurities from synthetical medicine and analysis of the chemicalconstitutents from Chinese herb medicines. The second chapter elaborates the studieson the discorvery and synthesis of new type of Pd Pincer catalyst by using MS andtandem MS together with the testing of its catalyst activity. The third chapter dwellson the designation and development of automatic tandem mass spectrometry analysissoftware. The last chapter presents a review on the dissociation technique of massspectrometry. The first chapter reports the rapid analysis of trace impurities from synthesismedicine and analysis of the chemical constitutents from Chinese herb medicines. The fission mechanism of 5-bromotetrandrine was obtained by analysis of the dissociationpathways of major product, by using which the possible structure of the two traceimpurties was assumed. There are lots of saponins in Sapindus mukurossi. Except forthe good spumescence and decontamination，it possesses the bioactivity of antigenand antitch. First of all, the high resolution mass information was obtained by ESI-QTOF. Hence the possible molecular formulars were acquired too. Then weconducted the further detection of the structures of its saponins by using ESI-ITtechnology. In the same manner, first the molecular formulars of every constituentfrom Dioscorea panthaica in total saponins were obtained by ESI-QTOF, and thenacquired the fission mechanism of this type of compounds by tandem massexperiment on a series of known and available saponins. In the same time, theanalysis flowchart was concluded. Here the relationship between the ion intensity andthe corresponding dissiociation activation energy was studied by computer chemistry.Then the four pairs of isomers were differentiated by APCI-MS/MS, as well as thecharacterization of known and unknown compounds. The assumption was confirmed by HPLC-MS/MS. Among them the possible structures of four unknown saponinswas presented. The second part was discovery and synthesis of a new type of Pd pincer catalystby MS and tandem MS. The coupling reaction catalyzed by Pd is the most effectivemethod in C-C formation in organic synthesis. Apart from that, thiourea is type ofcompounds that are stable to atmosphere and moisture. Hence we designed a series ofPd thiourea catalysts. Some of them show the excellent catalyst activity. The abnormalparent ion [M.H]+ was founded in positive ESI mode when we conduct some massspectrometry experiments on the bicyclical thiourea Pd complex. The structure wasproposed by mass and tandem mass spectrometry. Because it was a new type of pincer,we want to test its catalyst activity. So the Pd pincer was synthesized and the detailstructure was obtained by x-ray experiment. It shows the more fitness in catalysis ofSuzuki reaction by comparison with the analogue. The third chapter dwells on the design and development of automatic tandemmass spectrometry analysis software. Inspired by the former two chapters, theAuMass (version 1.0) was developed. Its algorithm is: first check the diagnostic ion,diagnostic neutral loss or diagnostic ions mass intervals in database to find out whatthe analyst’s skeleton belongs to, then identify the peripheral functional group by thecorresponding analysis flowchart. The chemical constituents of Paeonia lactiflorawere identified rapidly by using AuMass. To increase the analysis ability, the othertypes of compounds from Chinese herbs was concluded. Actually, the software isproven to have the much valuable application. The last chapter presented the review on the some kinds of fission technique ofmass spectrometry. It involves the collision induced dissociation (CID), laser induceddissociation (LID), electron capture dissociation/electron transfer dissociation(ECD/ETD), infrared multiple photons dissiociation, black body irraditiondissociation and PQD fission technique from Finnigan.

吲哚生物碱Desbromoarborescidine A 的全合成及烯胺C-亚磺酰化反应研究

从新几内亚核桃木的树皮中分离得到的吲哚类喹诺里西定生物碱10-Desbromoarborescidine A，因发现其具有阻滞钙离子通道的活性而倍受关注。10-Desbromoarborescidine A由A、B、C、D四个环组成，只有一个手性中心，是吲哚生物碱中结构较简单的一种，常作为此类生物碱全合成方法的模型化合物。但迄今为止，能高效而简便的实现手性10-Desbromoarborescidine A不对称全合成方法线路不多，大多数以不对称诱导的方式建立其手性中心，手性催化的方式仅有一例金属催化。从逆合成分析可知，Desbromoarborescidine A的全合成可以通过亚胺不对称催化还原进行关键的手性中心构建，而本课题组在之前的研究中通过手性有机小分子催化剂的发展，已将三氯硅烷氢转移还原亚胺发展成了一类简便实用、高效、高对映选择性并具有优良底物适应范围的不对称催化反应，我们希望以这一反应作为关键手段，发展一条Desbromoarborescidine A及其类似物不对称合成新路线。 根据我们设计的新路线，首先成功合成了其关键中间体，然后我们进行了关键的不对称催化尝试。用本实验室已有的高性能有机小分子催化剂虽得到了较好的对应选择性，但是产率很低。同时，为了验证整条线路的可行性，我们也用消旋的中间体进行拉通线路的尝试。但不幸的是，在脱除保护基时遇到了很大困难。尝试换不同的保护基，或改变脱保护基的顺序，都未能成功合成目标产物。究其原因可能是由于吲哚的特殊性造成的，吲哚类亚胺与常规的芳香亚胺有较大的差异，其NH基团无论保护还是不保护，对与其2位相联接的C=N双键均有很大的影响，导致其不对称催化还原难以进行。另外，由于所设计的还原产物含有处在吲哚苄位的胺基，稳定性较差，造成保护基脱除困难。 烯胺C-亚磺酰化反应是本课题组最近发现的一个新反应，之前未见文献报道。本研究对该反应进行了反应条件优化和底物扩展，发现带Cbz，Ac，COt-Bu，CO2Et，Bz等保护基的一系列环状和非环状烯胺在亚磺酸钠、DMAc和MeSiCl3的共同作用下能高效高产率生成β-胺基烯基亚砜类新化合物，为合成多官能团化的烯基亚砜新化合物提供了一条简便实用的途径。 The main constituent of Dracontomelum mangiferum B1, indoloquinolizidine alkaloid 10-Desbromoarborescidine A, has drawn great attention due to its calcium channel blocking activity. Its molecular structure is relatively simple compared with the other alkaloids of the same type, which has only one chiral center, albeit with four cycles A, B, C, and D. This compound is often used as a model target for exploring different strategies for the total synthesis of indole alkaloids. Nevertheless, so far there still lack practical and highly efficient methods for the asymmetric total synthesis of 10-Desbromoarborescidine A. Most of the current available methods rely on stoichiometric asymmetric synthesis for the construction of the chiral center. There is only one example reporting utilization of asymmetric catalysis, but with transition metal complex as the catalyst. Our retrosynthetic analysis shows that catalytic asymmetric reduction of imine could be used as the key step for the construction of the chiral center of Desbromoarborescidine A. Since in the previous studies our group has developed the asymmetric reduction of imines by trichlorosilane into a practical and highly efficient and enantioselective method using newly designed chiral organocatalysts, we hope to apply this method to develop a novel synthetic route for the total synthesis of Desbromoarborescidine A and its analogues in this study. According to the newly designed synthetic route, we first accomplished the synthesis of the key intermediates which was then examined for the critical asymmetric catalysis. The asymmetric reduction using the highly efficient organocatalysts, developed in our lab afforded high ee but poor yield. We tried different reaction conditions to improve the yield, but failed to get any good results. Simultaneously, to vertify the feasibility of the synthetic route we designed, we also tired to go through the route toward the racemic synthesis of Desbromoarborescidine A. But unfortunately, protection and deprotection proved to be big hurdles. All the different protection groups and different sequences of protection and deprotection we tried failed to get us through the designed synthetic sequence and furnish the final product. Most likely, the indole part is the culprit behind the failures.The NH group of the indole, no matter protected or not, may impact the catalytic asymmetric reduction of C-N double bond connected with 2-C. Additionally, the reduction product we designed contains an amino group in the β-position of the indole, which may cause problems due to its instability. C-sulfenylation of enamines is a novel reaction discovered recently by our group, which has not been seen before in the literature. In this study, optimization of the reaction conditions and exploration of the substrate scope were further undertaken for this reaction, which reveal that a series of enamines with N-Cbz, Ac, COt-Bu, CO2Et protection groups could all undergo smooth C-sulfinylations with the comined use of sodium benzene sulphinate, DAMc and MeSiCl3, efficiently furnishing the β-amino vinylsulfoxide products in high yield, affording a practical and highly efficient methods for synthesis of functional vinylsulfoxides.

青藏高原栽培青稞遗传多样性研究

青稞，是我国藏区居民对裸大麦的称谓，它不仅是藏民的主要食粮、燃料和牲畜饲料，而且也是啤酒、医药和保健品生产的原料；青稞不仅为藏区人民的健康和经济发展做出了很大的贡献，而且对人类健康和社会经济的可持续发展都有重要的意义。青藏高原是我国及世界上青稞分布和种植面积最大的地区，资源极其丰富。虽然从经典遗传直到分子标记对我国大麦遗传多样性都有研究，但研究手段、数量仍然不够深入，对我国大麦资源遗传多样性研究的信息非常有限，不能很好地满足大麦遗传研究和育种应用的需要，尤其是对西藏栽培大麦的遗传多样性的研究还只是刚刚开始，关于栽培青稞多态性的研究报道很少。本研究采用SSR标记和蛋白质电泳两类技术，从SSR标记位点、单体醇溶蛋白、B组醇溶蛋白和淀粉粒结合蛋白（SGP）等四个方面对我国青藏高原栽培青稞的遗传多样性进行了综合评价。 SSR标记具有基因组分布广泛、数量丰富、多态性高、容易检测、共显性、结果稳定可靠、实验重现性好、操作简单、经济、易于高通量分析等许多优点，被认为是用于遗传多样性、品种鉴定、物种的系统发育、亲缘关系及起源等研究的非常有效的分子标记。本研究采用SSR标记分析了64份青藏高原栽培青稞的遗传多样性，同时评估SSR标记在我国大麦育种和品种鉴定中的应用潜力。选择了30个已知作图位点SSR标记，其中25个标记与重要性状的控制位点连锁紧密。选择的30个SSR标记，5个未得到很好的扩增产物，3个无多态性。22个多态性SSR标记位点中，每位点检测出等位基因2~15个，共检测出等位基因132个，平均每位点6.0 个。各多态位点检测出基因型为2~11种，位点HVM33的基因型最多。各多态位点的多态信息指数为0.16~0.91, 平均为0.65。根据PIC值选择了13个SSR标记用于我国青藏高原栽培青稞基因型鉴定，这些标记的PIC值为0.6以上。结合PIC值和基因型差异，选择了8个多态信息含量高的SSR标记，构建了高效指纹图谱，此图谱能把64份材料完全区分。 贮藏蛋白电泳分析是研究相关编码蛋白基因多态性的非常有效的方法。大麦单体蛋白与小麦醇溶蛋白相对应，具有丰富的多态性，可用于大麦遗传多样性、品种鉴定和群体进化等研究。本研究通过A-PAGE电泳技术研究了84份青藏高原栽培青稞的单体醇溶蛋白多态性。大麦单体醇溶蛋白图谱与小麦醇溶蛋白电泳图谱类似，所分离的蛋白清晰地分为ω－，γ－，β－和α－四个部分。青藏高原栽培青稞单体醇溶蛋白具有丰富的多态性，84份青稞材料中存在43条不同的蛋白带，75种组合带谱；其中67种为单一材料所独有，另8种则分别包含了2-3份材料。每份材料中拥有醇溶蛋白带为6－16条，含有6-10条单体醇溶蛋白带材料较多。西藏和四川材料群体单体醇溶蛋白多态性不同，具有区域特异性。西藏材料中发现了40条不同蛋白带，3条特异带，46 种蛋白组合；四川材料中出现了40种不同蛋白带，26种条带组合, 3条特异带。基于单体蛋白多态性的聚类与材料的来源有一定的相关性。A-PAGE单体蛋白具有丰富的多态性，可作为遗传研究和品种鉴定的标记。 大麦醇溶蛋白（hordein）是大麦籽粒的主要贮藏蛋白，与大麦的营养品质和加工品质密切相关，而且具有丰富的多态性，广泛用于品种鉴定、种质筛选、遗传多样性和亲缘关系研究。B组醇溶蛋白是主要的醇溶蛋白组份，约占总醇溶蛋白的80%，而且具有丰富的多态性。本研究采用SDS-PAGE分析了72份青藏高原栽培青稞B组醇溶蛋白的遗传多样性。青藏高原栽培青稞B组醇溶蛋白具有丰富的多态性，72份青稞材料中存在15种蛋白带，30种组合带谱，其中15种为单一材料所独有，另15种则分别包含了2-10份材料。每份材料中B组醇溶蛋白条带数为4-8条，含5、6条的材料较常见。不同来源的群体材料间B组醇溶蛋白组成存在差异，西藏青稞含有26种蛋白组合带谱，其中有19种特异带谱；四川群体中共发现11种蛋白组合带型，其中有4种特有带谱。两群体中都存在稀有条带。聚类分析将材料分成三组，材料聚类与材料来源地没有明显的相关性。 淀粉粒蛋白（Starch granule proteins, SGPs）是一类与淀粉粒结合的微量蛋白，一些淀粉粒蛋白具有淀粉生化合成中主要的酶蛋白功能，其变异会影响淀粉含量和特性，从而影响淀粉的应用。关于我国大麦淀粉粒组成研究还未见报道。本实验首次开创了我国大麦淀粉粒结合蛋白的研究工作。采用SDS-PAGE电泳技术研究了青藏高原栽培青稞的SGP组成，并分析了不同SGP组合间淀粉含量的差异，初步探索了所分离的SGP蛋白与淀粉合成的关系。66份青稞材料中分离了10种主要的SGP，其表观分子量为40-100KD，低于60KD的SGP带有7条，共有16种组合带谱；各SGP蛋白和组合带谱出现的频率存在差异，青藏高原青稞的SGP组成存在多态性。西藏青稞和四川青稞的SGP组成有很大差异，SGP组成具有地域差异性，西藏青稞含有12种蛋白组合带谱，其中有9种特异带谱；四川群体中共发现7种蛋白组合带型，其中有4种特有带谱；两群体中仅有3种共同的蛋白组合带谱。SGP蛋白特性将66份青稞分为三组, 即Ⅰ、Ⅱ、Ⅲ，材料聚类与材料来源具有一定的相关性。不同组合带谱材料间淀粉含量差异显著性检验结果显示，不同带谱间材料的总淀粉含量、直链淀粉含量和支链淀粉含量有差异，带谱2（SGP1+3+7+9+10）和8（SGP1+2+4+6+8）的总淀粉含量及支链淀粉含量显著大于组合带谱3（SGP1+3+7+10）的总淀粉含量。组合带谱7（SGP1+2+6+8）的直链淀粉含量显著低于带谱11（SGP1+5+8）的直链淀粉。带谱SGP2、3、4、5、6、7、8、9、10可能参与淀粉合成，SGP9可能与高支链淀粉的合成相关。 SSR标记位点、单体醇溶蛋白、B组醇溶蛋白、淀粉结合蛋白等四个方面的研究结果表明青藏高原SSR标记多态性、单体醇溶蛋白多态性、B组醇溶蛋白多态性和SGP多态性都非常丰富，与青藏高原是栽培青稞的多样性分布中心的观点一致。 青藏高原栽培青稞的SSR标记、单体醇溶蛋白、B组醇溶蛋白和SGP多态性表现出很大差异。SSR标记覆盖了整个基因组，多态性非常高。单体蛋白、B组醇溶蛋白、SGP蛋白是育种中非常关注的性状，他们只是代表基因组中的某一区域或位点，多态性相对较低。但单体蛋白多态性很高，84份材料中检测出43条不同蛋白带，75种不同的组合带谱。SSR标记技术和单体蛋白技术都是遗传多样性研究的有力工具，但单体蛋白技术不仅多态性高，而且经济、操作简便，是种质鉴定的理想方法。 对不同标记的多态性材料数据进行聚类，聚类图能为我们提供各材料间的遗传相似信息，为材料选择提供参考。但材料聚类与材料来源的地理区域的相关性表现不一致。SSR聚类和B组醇溶蛋白聚类与材料的来源地无相关性，而单体醇溶蛋白和SGP聚类与材料来源地有一定相关性，即西藏群体和四川群体分别有集中类群，这可能是人为选择的附加效应。 不同来源的群体材料的遗传多样性不同，具有区域特异稀有基因，加强不同地区间资源的交换和配合使用，有利于增加群体遗传多样性和新品种培育。 青藏高原栽培青稞的麦芽浸提性状、淀粉性状、病虫及裸粒等重要农艺性状控制位点存在丰富的变异，遗传基础宽广，可能蕴藏着多种不同的等位基因，是研究重要性状遗传特性、基因资源挖掘和遗传育种的宝贵资源库。 Hulless barley, due to its favorable attributes such as high feed value, good human nutrition，rich dietary fiber and ease processing, attracts people,s attention . Hulless barley plays a very important role in Tibetan life, used as essential food crop, main animal feed and important fuel. In addition to tsampa (roasted barley flour), a main food for Tibetan, hulless barley is also made into cake, soup, porridge, recent naked barley liquor and cornmeal. Qinghai-Tibet Plateau is one of a few areas which plant naked barley widely in the world and also has a long growing history. Genetic diversity of the cultivated hulless barley in this region , however, has not been documented. The study of genetic diversity existing within this population is of particular interest in germplasm identification, preservation, and new cultivar development. This study analyzed the genetic diversity of the cultivated naked barley from Qinghai-Tibet plateau through the study of SSR marker loci and monomeric prolamins, B-horden and starch granule proteins. SSRs are present abundantly in genomes of higher organisms and have become a popular marker system in plant studies. SSRs offer a number of advantages, such as the high level of polymorphisms, locus specificity, co-dominance, reproducibility, ease of use through PCRand random distribution throughout the genome. In barley, several hundred SSRs have been developed and genetically mapped and can therefore be selected from specific genomic regions. The genetic diversity of 64 cultivated naked barley from Tibet and Sichuan was studied with 30 SSRs of known map location.Among the selected SSR markers, PCR products of 5 SSR markers were not obtained and 3 SSR marker loci were monomeric. A total of 132 alleles were identified at 22 polyomeric SSR loci. The number of alleles per locus ranged from 2 to 15, with an average of 6.0. The polymorphism information content values for the SSRs ranged from 0.08 to 0.94, with an average of 0.65. 13 SSR markers with the PIC value >0.6 have been selected for discrimination of Qinghai-Tibet naked barley genotypews. A finger Print map was developed through 7 SSR markers with the high PIC value. It could be used as an efficient tool for gene discovery and identification of gernplasm. Hordeins, the main storage proteins of the barley seed, are composed of momomeric and polymeric prolamins and divided into -A, B, C and D groups in order of decreasing electrophoretic mobility. Hordeins show high inter-genotypic variation and have been extensively used as markers for cultivar identification and analyzing the genetic diversity. This study analyzed the genetic diversity of B-hordein in 72 naked barley from Qinqhai-Tibet Plateau. Extensive diversity was observed. A total of 15 different bands and 30 distinct patterns were found. Jaccard's coefficient of similarity was calculated, and the accessions were divided into three　main groups by cluster analysis using UPGMA. Differentiation among the populations from different collecting regions based on the polymorphism of B-hordein was investigated. Monomeric prolamins show high inter-genotypic variation and have been used as molecular markers for cultivar identification, analyzing the genetic diversity in collections and investigating the evolution processes and structure of populations However, the cultivated hulless accessions from Qinghai-Tibet Pateau in China have never been examined with respect to monomeric prolamins. This study analyzed the genetic diversity of monomeric prolamins (protein fraction corresponding to wheat gliadins) using the Acid -PAGE technique in eighty-four cultivated hulless barley from Qinqhai-Tibet Plateau in China. Extensive diversity was observed. A total of 43 different bands were found, of which 21 different bands were in the region of ω group, 8 in the region of γ, 8 in the region of β, and 6 in the region of α group. Among the 86 accessions, 75 distinct patterns were identified. The number of bands ranged from 6 to 16, depending on the variety. Jaccard’s coefficient of similarity was calculated, and the lines were grouped by cluster analysis using UPGMA. A dendrogram was obtained from the analysis of the groups and five main clusters were identified. No relationship between the distribution in the dendrogram and growth habits and origins of the cultivars could be detected. Starch is the major constituent of the cereal endosperm, comprising approximately 65% of the dry weight of the mature wheat grain. The starch formed in all organs of plants is packaged into starch granules, which vary widely between species and cultivars in size and shape. Wheat endosperm starch granules contain about corresponding to the main biosynthase of starch. This report firstly dealed with intraspecific variation of the major SGPs in cultivated naked barley from Qinghai-Tibet plateau. A total of 10 major SGPs were observed in the range of 40KD-100KD and 16 types of patterns were found. Based on the variation of SGPs, accessions studied were classified into 3 groups. A geographical cline of electrophoregram was observed. In addition, significance test of the difference of starch content among groups and types of patterns were done, and the results indicated those SGPs could be related to the content of starch. Diagram obtained through cluster analysis exhibited a structuration of diversity and genetic relationship among cultivated hulless accessions. In breeding program, parents with genetically distant relationship for hybridization will increase genetic diversity of progenies. In conclusion, cultivated naked barley from Qinghai-Tibet Plateau in China presents a high variability with respect to monomeric prolamins，SSR markers , B- hordeins and SGPs. The result of this study supports Qinghai-Tibet Plateau is the center of cultivated hulless barley and the cultivated naked barley is considered to be a gene pool with large diversity and could be applied to breeding for cereal.

基于语义的Web服务匹配及其框架模型研究

随着Internet上异构应用系统的大量增加和SOA技术的空前发展，Web服务技术变得越来越重要，已经成为了学术界和工业界关注的热点。在Web服务技术中，服务发现为Web服务消费者调用Web服务提供者提供的服务提供了桥梁，起到非常重要的承接作用，成为了Web服务技术中的重点。目前的Web服务发现机制主要有两种，第一种是传统的Web服务发现方式，主要基于UDDI（Universal Description， Discovery and Integration）的纯粹关键字查找；第二种是基于Web服务的语义信息，进行Web服务间的语义匹配。 第一种方法的基础UDDI是国际标准，而且应用也最为广泛，但UDDI中对于Web服务的描述是基于语法的，而且缺乏Web服务所特有的I/O属性和服务质量属性等信息。第二种方法基于Web服务的语义信息，包括Web服务所特有的I/O属性，但因为缺乏灵活有效的Web服务匹配方法和与之对应的Web服务匹配框架，限制了其应用。 基于此，本文在对当前语义Web服务匹配技术分析和研究的基础上，对当前的语义Web服务匹配方法进行了改进，同时提出了基于过滤器（filter）的语义Web服务匹配框架模型。本文的主要工作有： 1）对目前的语义Web服务匹配技术进行了较为全面深入的探讨和综述。 2）对当前的语义Web服务匹配的各个阶段进行了详细的分析，对其中的匹配方法进行了改进。提出了基于向量空间模型(VSM)和TF-IDF(Term Frequency–Inverse Document Frequency)思想的本体权重的计算方法，本体层次关系图中边的权重的计算方法和本体之间相似度的计算方法。 3）提出了基于web服务黑盒属性的语义。 4）语义Web服务匹配框架方面，本文提出了基于filter的语义Web服务匹配框架，并将其延伸到非语义Web服务系统中。

Tuning the Energy Level of Organic Sensitizers for High-Performance Dye-Sensitized Solar Cells

Cost-effective organic sensitizers will play a pivotal role in the future large-scale production and application of dye-sensitized solar cells. Here we report two new organic D-pi-A dyes featuring electron-rich 3,4-ethylenedioxythiophene- and 2,2'-bis(3,4-ethylenedioxythiophene)-conjugated linkers, showing a remarkable red-shifting of photocurrent action spectra compared with their thiophene and bithiophene counterparts. On the basis of the 3-f{5'-[N,N-bis(9,9-dimethylfluorene-2-yl)phenyl]-2,2'-bis(3,4-ethylenedioxythiophene)-5-yl}2-cyanoacrylic acid dye, we have set a new efficiency record of 7.6% for solvent-free dye-sensitized solar cells based on metal-free organic sensitizers. Importantly, the cell exhibits an excellent stability, keeping over 92% of its initial efficiency after 1000 h accelerated tests under full sunlight soaking at 60 degrees C. This achievement will considerably encourage further design and exploration of metal-free organic dyes for higher performance dye-sensitized solar cells.

Development of expressed sequence tags from the bay scallop, Argopecten irradians irradians

The bay scallop, Argopecten irradians irradians, introduced from North America, has become one of the most important aquaculture species in China. Inan effort to identify scallop genes involved in host defense, a high-quality cDNA library was constructed from whole body tissues of the bay scallop. A total of 5828 successful sequencing reactions yielded 4995 expressed sequence tags (ESTs) longer than 100 bp. Cluster and assembly analyses of the ESTs identified 637 contigs (consisting of 2853 sequences) and 2142 singletons, totaling 2779 unique sequences. Basic Local Alignment Search Tool (BLAST) analysis showed that the majority (73%) of the unique sequences had no significant homology (E-value >= 0.005) to sequences in GenBank. Among the 748 sequences with significant GenBank matches, 160 (21.4%) were for genes related to metabolism, 131 (17.5%) for cell/organism defense, 124 (16.6%) for gene/protein expression, 83 (11.1%) for cell structure/motility, 70 (9.4%) for cell signaling/communication, 17 (2.3%) for cell division, and 163 (21.8%) matched to genes of unknown functions. The list of host-defense genes included many genes with known and important roles in innate defense such as lectins, defensins, proteases, protease inhibitors, heat shock proteins, antioxidants, and Toll-like receptors. The study provides a significant number of ESTs for gene discovery and candidate genes for studying host defense in scallops and other molluscs.

Winter counter-wind currents off the southeastern China coast: A review

This review covers the discovery and studies of the year-round northeastward currents off the southeastern China coast, paying special attention to its upwind characteristic in winter, mainly focusing on work by Chinese oceanographers. This current system is a prominent and unique phenomenon in the shelf circulation of the world ocean. The general features of the current system are summarized. The evidence for the existence and the variation of the three parts of the currents-the South China Sea Warm Current, the Taiwan Strait Warm Current and the Taiwan Warm Current-are separately elucidated. The formation mechanisms of the current as a whole are explained using dynamic analysis and numerical simulation results. Some suggestions for further studies are also made.

转炉钢渣对海洋生态环境影响及碳的生物利用探索研究

利用“铁肥”方法增加海洋初级生产力、吸收大气中的CO2、降低温室效应是目前国际上的一个研究热点。为此，本文利用炼钢企业产生的废料转炉钢渣，系统研究了其对海洋微藻生长的影响， 同时研究了转炉钢渣对海水水质及海洋生物的影响，探讨了其作为“铁肥”材料的可行性。 本文首先研究了转炉钢渣对海水水质的影响，包括金属、营养盐、pH、COD及浊度等理化参数。研究发现，在天然海水中加入转炉钢渣后营养盐均有不同程度的溶出；实验条件下，在天然海水和人工海水中加入钢渣，金属大部份被吸附，个别金属溶出，但基本都达到了一、二类海水标准，而pH明显增高，大多超过了海水水质三、四级（6.88.8）的范围，COD低于一类海水水质标准。实验结果显示，钢渣粒度越小、浓度越大，水质浊度越大，但钢渣的比重一般较大，在水体中可以较快的沉降，所以在实际应用过程中不会对水体的透明度造成不良影响。 研究了转炉钢渣对典型海洋底栖生物（紫贻贝）、海洋游泳生物（日本对虾和中国对虾幼虾）的影响。在实验浓度范围内，钢渣对紫贻贝的急性毒性作用基本可以忽略，但紫贻贝的生长率和摄食率随钢渣浓度的增加有所下降，血细胞个数有所下降，但形态没有显著变化，对紫贻贝的免疫功能没有太大影响。钢渣对日本对虾幼虾的半致死浓度为1.62g/L，对中国对虾幼虾的半致死浓度要高于日本对虾，说明在实验条件下中国对虾幼虾的适应能力高于日本对虾幼虾。 另外，重点考察了转炉钢渣对海洋浮游植物－赤潮异弯藻和中肋骨条藻生长的影响，发现加入钢渣以后，相对于对照组微藻生物量显著增加。当钢渣浓度为124248mg/L时赤潮异弯藻的生物量达到最大，当钢渣浓度达到61124mg/L时，中肋骨条藻的生物量最大；钢渣对两种微藻生长的影响规律是相同的，但是影响程度因不同藻种而有所不同，其中对中肋骨条藻的促进作用更加显著，可能与钢渣中能溶出较大量的硅酸盐有关。在室内实验的基础上，初步估算了通过钢渣对微藻生长的促进作用，近海每平方公里海域对二氧化碳吸收的增加量。

First record of the family Spengeliidae (Hemichordata : Enteropneusta) from Chinese waters, with description of a new species

Six species belonging to two families of Hemichordata have previously been recorded in Chinese waters. This paper records the discovery and description of a new species of the genus Glandiceps found in Jiaozhou Bay, Qingdao, Shandong Province, named Glandiceps qingdaoensis. The new species has a long proboscis with dorsal and ventral grooves, a stomochord with a long vermiform process, a proboscis cavity with a dorsal median, right and left glomeruli, right and left glomeruli very large and encircling the stomochord, a proboscis skeleton in the cavity extends into the median posterior of the collar, a well-developed dorsal ventral muscular septum in the proboscis cavity dividing the cavity completely into two separate parts. The collar cord is without giant nerve roots. The trunk with four distinct regions that can be recognized externally: branchial-genital region, genital region, hepatic region, and intestinal region. The dorsal pharynx is large and the gill pores are small. The tongue bars are encircled by vesicles, and the first gonad commences at the level of the second or third gill slit.

地球介质非线性与各向异性的地震波响应及其成像效应

The practice of geophysical prospecting shows us the complex interior earth. The studies of the complexity play an important role and practical guide for the subsurface structure. At present, the complexity of the earth mainly means lateral and vertical homogeneity, anisotropy and non-linear quality. And the anisotropy and non-linear media studies become the frontier in seismology and exploration seismology. This paper summarizes the development of complexities and presents the forward and inverse in the non-linear and anisotropic media. Firstly, the paper introduces the theory of seismic wave propagation in the non-linear and anisotropic media, the theoretical basis for simulation and inversion research. Secondly, high quality numerical simulation method with little dispersion has been developed to investigate the influence of complexity including anisotropy and non-linear multi-component seismograms. Because most real data in seismology have a single component, we developed two aspects work on anisotropic multi-component imaging. One is prestack reflection migration. The result show that distorted images are obtained if data from anisotropic media are migrated using isotropic extrapolation. Moreover, image quality will be improved greatly after considering anisotropy in subsurface layers. The other one is the we take advantage of multi-component data to inversion of the anisotropic parameters jointly seimic reflection travel time and polarization information. Based on these research works, we get the following results: 1.Combing numerical simulation, systematical studies indicate that anisotropy and non-linear seismograms characters are significant to detect cracked belts in the earth and to understand deformation field and mechanism. 2.Based on anisotropic media models, we developed an efficient prestack migration method for subsurface structure and different observation methods seismic data, which improving the imaging quality with VSP, seismograms and real data. 3.Jointly seismic inversion combining seismic anisotropic reflection traveltimes and polarizations data show that the complete wrong inversion and the following explanation will be resulted by ignoring anisotropy.

东北亚构造-盆地演化及油气资源控制作用

The oil and gas potential of Northeast Asia is enormous, but the degree of exploration is very low in Northeast Asia (the degree is below 3%-10%).The reasons are as follows: First, it is relatively difficult to study the oil and gas bearing basins(OGB), which are of multiple types, in different tectonic settings, with complex geologic frameworks and with long-term geologic evolution. Secondly, because of the non-equilibrium in development of economy and regional market, application of theories and techniques and the research levels in different countries, the conclusions are not conformable, and even contradictory. Thirdly, most of the former researches were limited to one territory or one basin, and lack of systematical and in-depth study on geotectonic evolution, classification of basins, and the evaluation of hydrocarbon resources. In this thesis, integrated study of the regional tectonic feature and basin features of Northeast Asia was done, to understand the basin evolution history and the controlling action on oil and gas. Then, new conclusions are and exploration proposals are as following: 1. Geotectonic evolution in Northeast Asia: The main structural motion system in Paleozoic Era was longitudinal, and in Meso-cenozoic was latitudinal with the Pacific Ocean. The whole evolution history was just the one of pulling-apart, cutting-out, underthrusting and collision of the Central Asia- Mongolia Ocean and the Pacific Ocean. 2. The evolution characteristics of basins in Northeast Asia: mainly developed from longitudinal paste-up, collision and relaxation rifting motion in Paleozoic-Early Mesozoic Era and from underthrust, accretion, and receding of subducted zone of the Pacific Ocean in Late Mesozoic Era-Cenozoic Era. 3. The research in basin classification of Northeast Asia: According to geotectonic system, the basins can be classified into three types: intracratonic, pericratonic and active zone basin. And they can be further classified into 18 different types according to genetic mechanism and dynamic features. 4. The master control factors of oil and gas accumulation in Northeast Asia: high quality cap-rock for craton and pericrationic basin, the effective source rock and high quality cap-rock for Mesozoic rifted basins, intra-arc, fore-arc and back-arc basins. Graded exploration potential of oil and gas for basin in Northeast Asia according to 7 factor, hereby, divided the oil and gas potential of basins into 5 levels. 5. Evaluation of hydrocarbon resources: The difference of resource potential among these basins is huge in Northeast Asia. The evaluation of Mesozoic rifted basin and Pacific Ocean basin showed that the large scale rifted basin and retroarc basin(including backarc marginal sea basin) have great resource potential. 6. The writer believes that the next step should pay more attention to the evaluation of petroleum resource in Far East part of Russia and trace them. On the other hand, according to integrated analysis of oil/gas resource potential and the operation difficulty in this area, suggests that East-Siberia basin, East-Gobi-Tamchag basin, Sakhalin basin, North-Okhotck basin, West-Kamchatka basin could be as cooperation priority basins in future.

松辽盆地长岭断陷火山岩储层识别与勘探目标评价

Changling fault depression is the biggest fault subsidence in south of Songliao Basin. In its Lower Cretaceous Yingcheng and Shahezi formations developed thick source rocks of deep lake facies and developed poly-phase volcanic rock reservoirs as well. In recent years, significant breakthroughs have been obtained in hydrocarbon exploration of volcanic rock reservoir in the different fault depressions in Songliao basin. Lately, I have been involved in hydrocarbon exploration in the Changling rift depression, especially volcanic rock reservoirs and exploration targets research, participating in the deployment of well Yaoshen 1 which gained over 40 × 104m3 natural gas flow. As quick changes of lithology and facies in Changling area in the south of Songliao basin, and the volcanic rock interludes distribution in continental clastic rock and shale in 3D space, so the identification of volcanic rock types and distribution become a difficult problem. Thus, based on the integrated research of the wild outcrop observation, gravity, magnetic and seismic data, geophysical logging, drilling and coring, laboratory test, this paper carried out the reservoir identification, description and prediction of volcanic rocks in Changling fault depression. In this area, this paper analyzed the volcanic rocks litho-facies, the eruption period, and characteristics of cycles. At the same time, tried to know how to use logging, seismic data to separate volcanic rocks from sandstone and shale, distinguish between volcanic reservoir and non-reservoir, distinguish between intermediate-basic and acidic volcanic rocks, and how to identify traps of volcanic rocks and its gas-bearing properties, etc. Also it is summarized forming conditions and distribution of traps, and possible gas-bearing traps were optimized queuing management. Conclusions as follows: There are two faulted basements in Changling fault depression, granite basement in the southeast and upper paleozoic epimetamorphic basement in the northwest. The main volcanic reservoirs developed in Yingcheng period, which was the intermediate-basic and acidic volcanic eruptions, from the south to north by the intermediate-basic to acid conversion. The volcanic vents are gradually young from south to north. According to information of the re-processing 3D seismic data and gravity-magnetic data, the large volcanic vent or conduit was mainly beaded-distributed along the main fault. The volcanic rocks thickness in Yingcheng formation was changed by the deep faults and basement boundary line. Compared with the clastic rocks, volcanic rocks in Changling area are with high resistance and velocity (4900-5800), abnormal Gamma. All kinds of volcanic rocks are with abnormal strong amplitude reflection on the seismic stacked section except tuff. By analyzing the seismic facies characteristics of volcanic rocks, optimizing seismic attributes constrained by logging, using seismic amplitude and waveforms and other attributes divided volcanic rocks of Yingcheng formation into four seismic zones in map. Currently, most volcanic gas reservoirs are fault-anticline and fault-nose structure. But the volcanic dome lithologic gas reservoirs with large quantity and size are the main gas reservoir types to be found.