西藏胡黄连和鸡矢藤的化学成分及黄芪多糖的提取工艺研究

本论文由四部分组成，前三部分为实验论文，第四部分为文献综述。第一、二部分分别报道了中药西藏胡黄连和鸡矢藤的化学成分研究结果。从两种药用植物中共分离和鉴定了32个化学成分，其中3个为新化合物。第三部分为黄芪多糖的提取工艺研究。第四部分概述了近年来植物多糖的研究进展。 第一章为西藏胡黄连化学成分研究。通过正、反相硅胶柱层析等分离方法从药用植物西藏胡黄连（Picrorhiza scrophulariiflora Pennell）的根茎中共分离纯化出7个化合物。运用MS、1H-NMR、13C-NMR、DEPT、HSQC和HMBC等现代谱学方法，结合理化分析对这些化合物的结构进行了分析鉴定。7个化合物中有两个是酚性的葡萄糖苷类成分：西藏胡黄连酚苷D (1)、4-O-β-D-(6-O-vanilloyl glucopyranosyl) vanillic acid (6)；四个苯乙基苷类化合物：plantamajoside (2)、plantainoside D (3)、西藏胡黄连苷A (4) 和西藏胡黄连苷F (5)；一个苯基小分子化合物：香豆酸甲酯 (7)。其中化合物1和5未见文献报道，确定为新化合物；化合物3为首次从该种植物中分到。 第二章为鸡矢藤化学成分研究。从鸡矢藤（Paederia scandense (Lour) Merrill）全草中分离出25个化合物，通过理化常数和波谱数据鉴定了它们的结构。25个化合物中包括一个蒽醌类成分：茜根定-1-甲醚 (1)；两个香豆素：异东莨菪香豆素 (2)和5-羟基-8-甲氧基吡喃香豆素 (3)；两个香豆素-木脂素化合物：臭矢菜素 B (4)和臭矢菜素 D (5)；一个木脂素：异落叶松树脂醇 (6)；两个黄酮：diadzein (7)和蒙花苷 (8)；三个三萜类化合物：齐墩果酸 (9)、乌苏酸 (10)和 3-O-β-D-吡喃葡萄糖基乌苏烷 (11)；三个甾体及其糖苷：b-谷甾醇 (12)、胡萝卜苷 (13)和(24R)-豆甾-4-烯-3-酮 (14)；六个小分子化合物：对羟基苯甲酸 (15)，咖啡酸 (16)，香豆酸 (17)，丁烯二酸 (18)，3,5-二甲氧基-4-羟基苯甲酸(19)，咖啡酸-4-O-β-D-吡喃葡萄糖苷(20)；五个环烯醚萜类化合物：鸡矢藤苷 (21)，鸡矢藤酸 (22)，鸡矢藤酸甲酯 (23)，saprosmoside E (24)和paederoside B (25)。其中化合物25未见文献报道，为新化合物。化合物1～8、11、14、15～20为首次从该化合物中分离得到。同时对鸡矢藤中环烯醚萜类化合物做了高效液相-串联质谱(HPLC-MSn)分析，探讨了这类化合物的质谱裂解规律。 第三章为黄芪多糖的提取工艺研究。首先确定了黄芪多糖含量的测定方法，并进行了方法学验证；其次探讨了黄芪中黄芪多糖的提取工艺，确定以酶法-Sevag法联用来去除黄芪多糖中的蛋白质，可使其提取物中黄芪多糖总含量达到70％以上。 第四章为近年来植物多糖的研究进展。主要包括植物多糖的提取纯化、多糖的定性定量检测方法、多糖的结构分析和多糖的药理活性。 This dissertation consists of four parts. The first and second parts reports the studies on the chemical constituents of medicinal plants of Picrorhiza Scrophulariiflora and Paederia scandens. The third part is about the extract technique of Astragalan Polysaccharide (APS). The last part reviews the progress of the studies on plant polysaccharides. 　 The first chapter is about the chemical constituents of P. Scrophulariiflora which is widely used as an important medicine to treat various immune-related diseases. A new phenyl glycoside, scrophenoside D (1) and a new phenylethyl glycoside, scroside F (5), together with five known compounds, plantamajoside (2), plantainoside D (3), scroside A (4), 4-O-β-D-(6-O-vanilloylglucopyranosyl) vanillic acid (6); and methyl-p-coumarate (7) were isolated from the stems of P. scrophulariiflora. Their structures were elucidated by spectroscopic and chemical methods. The second chapter is about the chemical constituents of medicinal herb of P. scandens. Twenty-five compounds were isolated and purified by normal and reversed phase silica gel column chromatography. By physicochemical properties and spectral analysis, their structures were identified as rubiadin-1-methylether (1), isoscopoletin (2), 5-hydroxyl-8-methoxyl-coumarin (3), cleomiscosin B (4), cleomiscosin D (5), isolariciresinol (6), diadzein (7), linarin (8), oleanolic acid (9), ursolic acid (10), 3-O-β-D-glucopyranosyloxyl-ursane (11), b-sitosterol (12), b-daucosterol (13), (24R)-stigmast-4-ene-3-one (14), p-hydroxyl-benzoic acid (15), caffic acid (16), coumaric acid (17), trans-butenedioic acid (18), 3,5-dimethoxyl-4-hydroxylbenzoic acid (19), caffeic acid 4-O-β-D-glucopyranoside (20), paederoside (21), paederosidic acid (22), paederosidic acid methyl ester (23), saprosmoside E (24), paederoside B (25). Among them, compound 25 is a new compound. Compounds 1～8、11、14、15～20 were isolated from this plant for the first time. Futhermore, we studied the HPLC-MSn analysis and investigation of fragmentation behavior of the sulfur-containing iridoid glucosides. The third chapter is about the extracting process of Astragalan Polysaccharide (APS). The method of the content determination is built. The optimum condition of extraction of polysaccharides from Radix Astragali is defined and the more effective way to remove protein is combined enzyme method with Sevag method, by which the content of polysaccharides extract can be up to 70%. The last part is a review of the research progress of the plant polysaccharides, which includes its extraction, isolation, purification, determination, structure analysis, and pharmacology.

硫手性有机小分子催化剂的设计、合成及其在不对称亚胺氢转移还原中的应用

过去十多年，世界手性药物市场需求迅速增长，手性制药工业的发展壮大，已经引起了各国政府、学术界，特别是企业界的高度重视。手性药物中含有大量的手性胺单元，因此研究高效构建手性胺结构单元的方法具有重要的意义和实用价值，而亚胺的不对称还原是合成手性胺最便捷的方法。 手性有机小分子路易斯碱催化三氯氢硅不对称还原亚胺是最近几年才发展起来的一类新的亚胺不对称还原方法。尽管在对映选择性和底物适用范围等方面已经获得了突破性的进展，但是，高性能的路易斯碱催化剂仅局限于N-甲酰氨基酸酰胺一种类型，而且其底物适用范围和催化活性仍不够理想。因此，发展新型催化剂很有必要。 手性硫氧化物作为手性诱导剂的应用已经有数十年的时间，广泛应用在不对称合成及天然产物的全合成中。理论上，硫氧结构单元也可以作为路易斯碱，对硅烷类试剂进行活化，而且硫氧键还有碳氧键难以比拟的先天优势，硫原子自带手性特征，在反应过程中，手性中心离反应位点更近，因此，从手性硫氧化合物出发，极有可能开发出新的高效手性路易斯碱催化剂。最近，Kobayashi和Khiar在亚胺的不对称烯丙基化反应中用手性亚砜活化烯丙基三氯硅烷，获得了较好的ee值，但反应中手性亚砜的用量都需要化学计量以上，因此还不能算做真正意义上的催化剂，进一步的文献调研也未见真正意义上的硫手性有机小分子催化剂。 本文首次成功将硫手性亚磺酰胺衍生物应用于催化三氯氢硅对亚胺的不对称还原，在经过对亚磺酰胺衍生物的多次结构优化，开发出了合成容易，催化活性和立体选择性都很优良，并且有着前所未有的底物普适性的新型手性路易斯碱催化剂。 我们首先尝试将商品化的20mol%叔丁基亚磺酰胺和对甲基亚磺酰胺直接用作催化剂催化三氯氢硅对亚胺的不对称还原，尽管仅获得中等的收率和很低的对映选择性，但证明我们的设计思路是可行的。在此基础上，我们以叔丁基亚磺酰胺为原料和基本骨架，设计合成了一系列的亚磺酰胺类催化剂，通过对催化剂的结构改造，发现当催化剂中存在较强酸性的酚羟基时，催化效果得到大幅提高。随着对催化剂的进一步结构优化，我们找到了一个结构简单，催化效果还不错的催化剂，经过反应条件优化以后，催化反应的收率最高能达到98%，对映选择性最高达93%，并且这个催化剂的底物适应范围比之前报道的催化剂都要广泛。针对酚羟基在催化剂中的重要作用，我们进行了仔细的机理研究后发现，在催化反应中，催化剂极有可能是通过双分子机理去活化三氯氢硅从而实现不对称催化的，而酚羟基的作用就是通过分子间氢键促进双分子催化剂与三氯氢硅的络合。受此启发，我们设计了一系列具有双齿结构的催化剂，通过对双齿催化剂的结构优化，最终筛选出了一个结构更加简单，但催化效果更好的双齿催化剂。10mol%该催化剂催化亚胺还原最高获得95%的收率和96%的ee值。这一结果也进一步验证了我们先前对催化剂机理的推测。 随后，我们还尝试将这些催化剂用于二级胺和芳香酮的直接还原胺化反应中，虽然能获得不错的收率，但对映选择性却很差，我们对反应条件进行了仔细的摸索，仍然没有获得突破。但这些实验为进一步研究二级胺和酮的不对称直接还原胺化反应奠定了良好的基础。 In the past decade, the rapid growth of the global chiral drug market and the significant development of the chiral pharmaceutical industry have attracted a great deal of attention from government, academia and enterprises. Chiral amine is an important structural motif of chiral drugs. Therefore, development of methods for the construction of this motif is of great importance. Catalytic enantioselective reduction of imines represents one of the most straightforward and efficient methods for the preparation of chiral amines. The chiral Lewis base organocatalysts promoted asymmetric reduction of imines by HSiCl3 has recently achieved significant advancements. Although big breakthroughs have been made in terms of substrate generality and enantioselectivity, the highly effective catalysts are limited to N-formyl amino acid amides, of which the efficiency and substrate scope remain unsatisfactory. Therefore, development of novel organocatalysts for this transformation is in great demand. Chiral sulfoxides have been well established as efficient and versatile stereocontrollers and have been extensively used in asymmetric synthesis and total synthesis of natural products. The S=O structural motif of sulfoxide could also behave as Lewis base activator for cholorsilane reagents, which, moreover, could be even better than caboxamide considering that the sulfur atom is chiral and thus the chirality center is closer to the reaction center. There exist great potentials that highly effective novel Lewis base organocatalysts could be developed starting from S-chiral sulfoxides. Recently, several S-chiral sulfoxides were reported by Kobayashi and Khiar to be used as Lewis base catalyst to activate allyltrichlorosilanes in asymmetric allylations and good enantioselectivities were obtained. However, these S-chiral sulfoxides were all used at a more than stoichiometric amount and were thus not authentically catalytic. A careful literature survey further revealed that there has been so far no S-chiral organocatalyst available. In this study, we, for the first time, successfully used S-chiral sulfinamides as Lewis base organocatalysts for the asymmetric reduction of ketimines by HSiCl3. After several rounds of structural optimization, we developed the first example of highly effective S-chiral organocatalysts, which promoted the asymmetric reduction of ketimines with trichlorosilane in high yield and excellent enantioselectivity with unprecedented substrate spectrum. In our initial practice, we examined 20mol% of the commercially available (R)-tert-butanesulfinamide and (S)-toluenesulfinamide as the catalyst in the hydrosilylation of ketimine. Although the product was only furnished in moderate yield and low ee, these results demonstrated that our strategy of catalyst design is on the right way. Next, starting from chiral tert-butanesulfinamide, we prepared a series of tert-butanesulfinamide derivatives via simple reductive amination and examined their catalytic efficiencies in the reduction of ketimine. We found that the catalyst bearing a phenolic hydroxyl group exhibited good reactivity and enantioselectivity. On the basis of which, we obtained a structurally simple and highly effective novel organocatalyst, affording the product in 98% yield and 93% ee under optimal reaction conditions. After careful exploration on the role of phenolic hydroxyl group in the catalyst, we speculated that two molecules of the catalyst be involved in the course of reaction, of which the assembly around the silicon center is facilitated by the intermolecular hydrogen bonding through the phenolic hydroxyl groups. Thus, we incorporated two units of sulfonamide into one molecular and prepared a new type of bissulfinamides organocatalysts and examined their catalytic efficiencies in the reduction of ketimine. After optimizing the structure of these catalysts, we finally obtained a novel organocatalyst which has even simpler molecular structure but showed better efficacies, 10mol% of which afforded up to 97% yield and 96% ee under optimal reaction conditions. These results further proved our speculation about the catalytic mechanism. We also examined the newly developed S-chiral organocatalysts in direct asymmetric reductive amination of secondary amines with aromatic ketone. The product was furnished in good yield but in low ee. No better results could be obtained despite our intense opimization efforts. Nevertheless, these experiments laid excellent foundations for eventual success.

岷江上游人工针叶、阔叶林土壤可溶性有机物质特征及其对凋落物分解的响应

土壤可溶性有机物质（Dissolved organic matter，DOM）作为土壤有机质的活性组分，在陆地生态系统物质循环中扮演非常重要的角色。土壤DOM的主要成分可溶性有机碳（Dissolved organic carbon，DOC）和氮（Dissolved organic nitrogen，DON）参与C、N循环过程。为深入揭示全球C、N循环过程机制，在未开展DOC和DON的地区进行相关研究是有必要的。森林土壤（包括枯枝落叶层）DOC、DON动态及调控机理的研究是目前国际上森林生态系统C、N循环研究热点之一。本研究立足于暖温带岷江上游茂县地区人工林植被，对土壤DOC和DON的库容量，季节动态及其与其它养分之间的关系进行了系统研究，旨在了解DOC和DON在该区生态系统中的重要作用，并探讨作为DOM主要来源的叶凋落物对DOC和DON的动态影响，研究有助于更加详细地了解该区生态系统C和N循环过程。本论文主要研究结论如下： 1研究了岷江上游地区两大主要土壤类型（棕壤和褐土）不同植物群落下土壤的DOC和DON含量及特征，结果表明：DOC和DON在两种土壤类型中均有库容量存在，DOC在0-10cm和10-20cm土层的含量幅度分别111.96~159.95 mg kg-1和69.02~100.84 mg kg-1。DON在0-10cm和10-20cm土层的含量幅度分别11.88~23.08 mg kg-1和4.70~10.77 mg kg-1。游离氨基酸在0-10cm和10-20cm土层的含量幅度分别0.84~1.66 mg kg-1和0.39~0.73 mg kg-1。DOC、DON与土壤中的一些养分因子表现出了显著的相关关系，共同反映了土壤的状况和质量，在该区开展DOC和DON的系统研究是有必要的。 2 对油松与连香树林地土壤DOC、DON以及其它化学指标的季节动态进行了研究，结果表明：油松与连香树林地土壤DOC和DON的季节动态变化表现了类似的规律，DOC和DON的含量均以秋季最高。DOC和DON的季节动态变化主要受凋落物生物因素的影响，但其微生物活力的生物因素以及降雨、温度等非生物因素也是控制土壤DOC和DON含量的重要因素。土壤DON在土壤中的行为不同于矿质氮，其季节动态不同于NO3--N和NH4+-N的季节动态，在研究N循环过程中，应考虑DON的变化情况。 3 对油松与连香树林地分解层和表层土壤（0-10cm）氨基酸周转动态进行了研究，结果表明：油松林地和连香树林地均以分解层的氨基酸含量高于矿质表层土壤的含量。每个取样时期，油松林地内各层次的氨基酸含量高于连香树林地内相应层次的含量。两林地各层次无机氮含量均超过了氨基酸的含量，并且室内培养30天后无机N的含量仍然高于氨基酸的含量，所以可以认为该区立地条件下无论是在有机分解层还是矿质土层植物吸收利用的氮素仍是以无机N为主。 4 松林下松针凋落物易于累积，这与松针凋落物分解缓慢有关，从而导致松林内养分周转缓慢。通过用不同性质凋落物和灌丛地土壤构建微生态系统，比较油松、辐射松、连香树、灌丛虎榛子凋落物分解对C、N循环过程的影响，结果显示油松和辐射松针叶凋落物比连香树、虎榛子凋落物分解更慢，减缓了养分循环过程。然而将针叶凋落物与阔叶凋落物混合后，油松和辐射松针叶凋落物的分解加快，C、N元素的循环过程也加速。此结果表明在松林内维持具有高质量凋落物的灌丛植被或在松林内栽植一些阔叶树种如连香树对维持和增进松树人工林的土壤肥力有重大的作用。室内培养的结果还显示添加凋落物后土壤DOC和DON的含量显著增加，表明凋落物是土壤DOM的直接来源。然而不同物种凋落物处理下土壤DOC和DON的含量有所不同，并随时间发生改变。混合凋落物处理下土壤DOC和DON的含量均高于松针凋落物单独处理下土壤DOC和DON的含量。DON是一个主要的水溶性N库，随时间的变化趋势与无机N的变化趋势不同，在土壤N循环过程中起到了中间N库的作用。 As a labile fraction of soil organic matter, dissolved organic matter (DOM) plays a very important role in material cycling of terrestrial ecosystem. The turnover of DOM is now being considered as main components in nutrient cycling. DOM mainly includes dissolved organic carbon (DOC), -nitrogen (DON), -phosphorous (DOP) and –sulfur (DOS). Among these constituents, DOC and DON directly participate in C and N cycling. It is essential to study DOC and DON dynamics and their controlling factors in the areas where no related study has ever been carried out. Study about them can provide data supports on understanding the mechanism of the global C and N cycling. DOC and DON dynamics and their controlling factors have been focused on in the research of C and N cycling of forest ecosystems. Based on forest plantations of Maoxian, Minjiang River in warm temperate zone, soil DOC and DON pool size, their seasonal dynamics, and the correlation between DOC, DON and other nutrients were studied in order to understand the importance of DOC and DON in the study area. Soil DOC and DON dynamics induced by leaf litter decomposition were also studied. The study contributed to comprehensively understanding C and N cycling processes and providing baseline data for including DOC and DON into the indices system of evaluating nutrient conditions. The results were as follows: 1 Several different plant communities under brown soil and Cinnamon soil were chosen as sampling plots. The contents and features of soil DOC and DON were evaluated. The results showed that DOC and DON were present under the two soil types. DOC contents in the top soil (0-10 cm) and the subsoil (10-20 cm) respectively varied from 111.96 mg kg-1to 159.95 mg kg-1, and 69.02 mg kg-1 to 100.84 mg kg-1. DON contents in the top soil (0-10 cm) and the subsoil (10-20 cm) respectively varied from 11.88 mg kg-1to 23.08 mg kg-1, and 4.70 mg kg-1 to 10.77 mg kg-1. Free amino acid contents in the top soil (0-10 cm) and the subsoil (10-20 cm) respectively varied from 0.84 mg kg-1to 1.66 mg kg-1, and 0.39 mg kg-1 to 0.73 mg kg-1. Significant correlations were found between DOC, DON and some nutrient indices, which together reflected soil condition and quality. It was hence essential to study DOC and DON in the study area. 2 Seasonal dynamics of DOC, DON, inorganic N, microbial biomass C and N were studied under Pinus tabulaeformis and Cercidiphyllum japonicum plantation. The results indicated that seasonal dynamics of soil DOC and DON under the two plantations performed similar change pattern, with the highest values in autumn. The seasonal dynamics of soil DOC and DON were mainly influenced by the litterfall. However, biotic factors such as soil microbial activities and abiotic factors such as precipitation and temperature also controlled the dynamics of soil DOC and DON. The seasonal dynamic of DON was different from that of NO3--N and NH4+-N, which showed that the behavioral differences between DON and inorganic nitrogen. And hence, it was proposed to include DON into soil N cycling in the study area. 3 Amino acid dynamics in Oa and topsoil (0-10 cm) under P. tabulaeformis and C. japonicum plantation were studied. The results showed that amino acid content in Oa was significantly higher than that in mineral soil. At each sampling time, significantly higher amino acid contents were found in P. tabulaeformis plantation than in C. japonicum plantation. The content of inorganic nitrogen was much higher than the content of amino acid in each sampling layer at each sampling time. After a 30-days laboratory incubation the content of amino acid was still lower than the content of inorganic nitrogen. The results implicated that the form of N absorbed by plants in these study sites were mainly inorganic nitrogen. 4 Usually needle litter is more resistant to decomposition, which leads to needle litter accumulation in pure coniferous stands and slows down the rate of nutrient circulation. By constructing microcosms with local shrubland soil and containing the four single-species (P. tabulaeformis, P. radiata, C. japonicum, Ostryopsis davidiana) litters, the decomposition rates and related C and N dynamics of needle litters and broadleaved litters during the early stage were compared. The results showed that the decomposition rates of pine needles were lower than those of broadleaved litters, which descended C and N cycling processes. However, the presence of C. japonicum or O. davidiana litter into pine needles increased the decomposition rates of pine needles and also dramatically promoted C and N cycling processes. It should be appropriate for plantation managers to consider C. japonicum as an ameliorative species or remain O. davidiana in pine plantations to improve soil conditions and help maintain soil fertility. The laboratory incubation still showed that DOC and DON contents in all litter-amended treatments were significantly higher than no litter-amended treatment, which proved that litter could be a direct source of DOM in soils. Different species litters induced different soil DOC and DON contents, which correspondingly changed over time. DOC and DON contents in mixed litter treatments were higher than those in pine needle litter treatments. As a major soluble N pool, DON developed a different changing pattern over time compared with inorganic N and played a role of interim N pool in soil N cycling.

中国沙棘(Hippophae rhamnoides subsp. sinensis)不同种群对干旱胁迫的响应差异与干旱诱导蛋白的分析

沙棘广泛分布于亚欧大陆的温带地区和亚洲亚热带的高海拔地区。沙棘能适应多种生态环境，能耐受多种逆境(如干旱、低温、高温和盐害等)。在中国，沙棘常常被用作植被恢复中的先锋树种而大量栽培。本文以中国沙棘为试验材料，探索沙棘适应干旱机制，以及沙棘对干旱胁迫的适应机制是否存在种群间的差异，同时试图通过分析干旱胁迫下沙棘叶片蛋白质表达变化探索沙棘适应干旱胁迫的分子机理。 对三个分别来自低海拔湿润地区、低海拔干旱地区和高海拔湿润地区的中国沙棘种群进行干旱胁迫处理。干旱胁迫能提高根冠比，比叶面积，降低平均叶面积和总生物量，提高沙棘的抗氧化性酶活性、脯氨酸含量、脱落酸(ABA)含量、降低光合作用，提高长期用水效率。实验中的这两个低海拔种群比高海拔种群抵抗干旱的能力更强,不同的种群采用了不同的策略来耐受干旱胁迫和过氧化胁迫。 在2004 年度的实验中，干旱胁迫处理下，高海拔湿润种群(道孚种群)严重失水，生长也受到更大的抑制，非气孔因素在抑制光合作用方面占支配地位，抗坏血酸含量下降，ABA和脯氨酸含量增加幅度比九寨沟种群的要高，这可能是因为道孚种群严重失水而引起的；而低海拔湿润种群(九寨沟种群)的体内水分状况几乎不受干旱的影响，生长情况也较道孚种群要好。 在2005 年度的试验中，和高海拔湿润地区种群(道孚)相比较，低海拔干旱地区种群(定西)在叶片相对水含量、根冠比、抗氧化酶活性(过氧化氢酶、抗坏血酸过氧化物酶和谷胱甘肽过氧化物酶)、保护性物质(脯氨酸，脱落酸)含量等方面都要高，光能热耗散能力也更强，而且气体交换参数(气孔扩散阻力和胞间CO2浓度等)对干旱也更不敏感。 分析了干旱胁迫下沙棘叶片蛋白质表达的变化。共发现319 个蛋白质，有4 个蛋白在干旱胁迫下消失(Putative ABCtransporter ATP-binding protein 、Hypothetical proteinXP-515578，热激蛋白Hslu219 和一个没得到鉴定的蛋白)，4 个只在干旱胁迫下出现(没命名的蛋白质产物，对甲基苯-丙酮酸双加氧酶，NTrX 和一个没得到鉴定的蛋白)，46 个蛋白质的表达丰度变化显著，包括32 个干旱负调蛋白，14 个干旱正调蛋白(3 个Rubisco 的大亚基、J-type–co-chaperone Hsc20、putative protein DSM3645-2335、putative acyl-COA 脱氢酶、nesprin-2 和两个没有得到鉴定的蛋白质)。这些蛋白质参与了氮代谢调控、抗氧化行物质的合成、脂肪酸β－氧化、核骨架构造、[Fe-S]基团组装、物质跨膜运输、细胞分裂或作为分子伴侣和蛋白质酶起作用。putative ABC transporter ATP-binging protein、NtrX、nesprin-2 和Hslu 是本试验新发现的高等植物蛋白，我们主要从它们的保守结构域或在其他生物中的同源物来猜测它们的功能。实验结果为我们研究植物抗干旱机制提供了新线索和新视野。 Seabuckthorn (Hippophae rhamnoides L.) is widly distributed throughtout the temperatureresiogn of Europe and Asia and sub-tropical plateau zone of Asia. H. rhamnoides can adapatseveral different environments, and can tolerant several envioronmental stresses (e.g, lowtemperature, high temperature, drought and salty). It has been widely used in forest restoration asthe pioneer species in China. In present study, we applied H.rhamnoides subsp. Sinensis asexperimental materials to study its drought-tolerant mechanism, and expected to findpopulational difference in drought-tolerant mechanism that may exist among populations, and tryto get some insight in drought-tolerant mechanism of it at morecular level through analyzing thechange of leaf protein expression. Three populations from high altitude wet zone, low altitude wet zone and low altitude arid znoe,respectively, were applied in our experiment, and were subjected to drought. Drought increasedthe root/shoot ratio(RS), special leaf area, long-term water use efficinency, activity of antioxidantenzymes, proline content and abscisic acid (ABA) content, declined the net photosynthesis rate(A), average leaf area (ALA), total biomass (TB). Both two low altitude populations were moredrought-tolerant than the high altitude population, and different population applied differentstratedgies to tolerant oxidant stress and drought stress. The results of the exprement in 2004 showed that Daofu population was more drought-sensitivethan Jiuzhai population. Under drought conditions, leaf relative water content (RWC) greatlydecreased in Daofu population, but not in Jiuzhai population. The large loss of water in Daofupopulation resulted in a limitation on A mainly caused by non-stomatal factors, severer suppression in growth rate and a significant reduction in ascorbic acid (AsA) content, comparedwith Jiuzhai population. The greater increase in content of ABA and proline in Daofu populationmay be also induced by large loss in water, so that enable plants to cope with sever drought. In the exprement of 2005, drought significantly increased RS, activities of catalase (CAT),peroxidase (POD), glutathione peroxidase (GPX) and ascorbate peroxidase (APX), and alsosignificantly increased ABA and proline contents. On the other hand, compared with Daofupopulation, drought induced larger RS and activities of CAT, GPX and APX, and higher ABAcontent in Dingxi population, whereas gas exchange traits, e.g., stomatal limitation value (LS) andintercellular CO2 concentration (Ci), were less responsive to drought in Dingxi population thanthose in Daofu population. All these factors enable Dingxi population to tolerant drought betterthan Daofu population. The leaf protein profile of seabuchthorn subjected to drought was analyzed. Altogether 319proteins were detected in well-watered sample, four proteins disappeard by drought (putativeABCtransporter ATP-binding protein, hypothetical protein XP-515578, Hslu219and aunidentified protein), four only appeared under drought (a probable nitrogen regulation protein(NtrX), a 4-hydroxyphenylpyruvate dioxygenase , an unnamed protein product and an identified protein), 32 drought down-regulated proteins, and 14 drought up-regulated proteins (nine wereidentified: three large subunits of Rubisco, a hypothetical protein DSM3645-23351, a putativeacyl-COA dehydrogenase, a nesprin-2, a J-type-co-chaperone HSC20 and two unmatchedproteins). These proteins may involve in β-oxidation, cross-membrane transport, cell division,cytoskeleton stabilization, iron-sulfur cluster assembly, nitrogen metabolism regulation andantioxidant substance biosynthesis or function as molecular chaperone or protease. Four proteins(a putative ABC transporter ATP-binging protein, NtrX, nesprin-2, Hslu) were new found in highplants, and their functions were estimated from their conserved domain or their homologues inother organism. Our results provided new clue and new insight for us to study thedrought-tolerant mechanism in plants.

A study of low-energy ion induced radiolysis of thiol-containing amino acid cysteine in the solid and aqueous solution states

The radiolysis of cysteine under plasma discharge and irradiation of low-energy Ion beam was investigated. The damage of cysteine in aqueous solution under discharge was assessed via the acid ninhydrin reagent and the yield of cystine produced from the reaction was analyzed by FTIR In addition, the generation of hydrogen sulfide was also identified The destruction of solid cysteine under low-energy ion beam irradiation was estimated via monitoring IR bands of different functional groups (-SH, -NH3, -COO-) of cysteine. and the production of cystine from ion-irradiated solid cysteine after dissolution in water was also verified These results may help us to understand the inactivation of sulphydryl enzymes under direct and indirect interaction with the low-energy ion irradiation (C) 2010 Elsevier B V All rights reserved.

多电荷态硫离子与氦原子和氢分子碰撞中的多电子转移过程研究

本工作采用反冲离子飞行时间技术和散射离子位置灵敏探测技术，实验研究了Sq+离子与He、H2碰撞中的多电子转移过程和分子离子的碎裂现象。研究了转移电离截面与单电子俘获截面比值和入射离子损失一个电子和两个电子的情况下，靶原子双重电离与单重电离的截面比值随入射离子能量和入射离子电荷态的变化规律，并对不同的碰撞体系的结果进行了比较。研究发现：在本工作研究的入射离子能区，对于HZ分子靶，双电子俘获自电离反应道：S2+H2→S(q-l)+2H++e-。是转移电离过程的主要贡献，而直接转移电离的贡献可以忽略；随着入射离子电荷态的增加，双电子俘获自电离的贡献增加，直接转移电离的贡献逐渐减小，但双电子俘获自电离的贡献的增加比理论预言的要慢。建立了蒙特卡罗程序模拟离子与分子碰撞中产生的具有不同的初始动能的离子碎片的飞行时间谱，深入分析和研究了Sq＋与H2分子碰撞中产生的H2+的解离过程和库仑爆炸过程，以及H＋碎片的能量分布。模拟结果与实验测量到的TOF谱的分析比较说明：在Sq＋与H2分子碰撞实验中，库仑爆炸是产生H+的主要反应道，而氢分子离子H2+发生解离产生H+的反应道相对很弱。

Degradation mechanism of polystyrene sulfonic acid membrane and application of its composite membranes in fuel cells

The lifetime behavior of a H-2/O-2 proton exchange membrane (PEM) fuel cell with polystyrene sulfonic acid (PSSA) membrane have been investigated in order to give an insight into the degradation mechanism of the PSSA membrane. The distribution of sulfur concentration in the cross section of the PSSA membrane was measured by energy dispersive analysis of X-ray, and the chemical composition of the PSSA membrane was characterized by infrared spectroscopy before and after the lifetime experiment. The degradation mechanism of the PSSA membrane is postulated as: the oxygen reduction at the cathode proceeds through some peroxide intermediates during the fuel cell operation, and these intermediates have strong oxidative ability and may chemically attack the tertiary hydrogen at the a carbon of the PSSA; the degradation of the PSSA membrane mainly takes place at the cathode side of the cell, and the loss of the aromatic rings and the SO3- groups simultaneously occurs from the PSSA membrane. A new kind of the PSSA-Nafion composite membrane, where the Nafion membrane is bonded with the PSSA membrane and located at the cathode of the cell, was designed to prevent oxidation degradation of the PSSA membrane in fuel cells. The performances of fuel cells with PSSA-Nafion101 and PSSA-recast Nafion composite membranes are demonstrated to be stable after 835 h and 240 h, respectively.

On the surface sites Of MOP/SiO2 catalyst under sulfiding conditions: IR spectroscopy and catalytic reactivity studies

The surface sites of MoP/SiO2 catalysts and their evolution under sulfiding conditions were characterized by IR spectroscopy using CO as the probe molecule. The HDS activities of thiophene were measured on the MoP/SiO2 catalyst that was subjected to different sulfidation and reactivation pretreatments. Cus Modelta+ (0 < delta less than or equal to 2) sites are probed on the surface of fresh MoP/SiO2 by molecularly adsorbed CO, exhibiting a characteristic IR band at 2045 cm(-1). The surface of MoP/SiO2 is gradually sulfided in HDS reactions, as revealed by the shift of the IR band at 2045 to ca. 2100 cm(-1). Although the surface of a MoP/SiO2 catalyst becomes partially sulfided, the HDS activity tests show that MoP/SiO2 is fairly stable in the initial stage of the HDS reaction, providing further evidence that molybdenum phosphide is a promising catalytic material for industrial HDS reactions. Two kinds of surface sulfur species are formed on the sulfided catalyst: reversibly and irreversibly bonded sulfur species. The MoP/SiO2 catalyst remains stable in the HDS of thiophene because most sulfur species formed under HDS conditions are reversibly bonded on the catalyst surface. A detrimental effect of presulfidation on the HDS activity is observed for the MoP/SiO2 catalyst treated by H2S/H-2 at temperatures higher than 623 K, which is ascribed to the formation of a large amount of the irreversibly bonded sulfur species. The irreversibly sulfided catalyst can be completely regenerated by an oxidation and a subsequent reduction under mild conditions. (C) 2003 Elsevier Inc. All rights reserved.

In situ IR spectroscopic studies on molybdenum nitride catalysts: active sites and surface reactions

Recent IR spectroscopic studies on the surface properties of fresh Mo2N/gamma-Al2O3 catalyst are presented in this paper. The surface sites of fresh Mo2N/gamma-Al2O3, both Modelta+ (0<δ<2) and N sites, are probed by CO adsorption. Two characteristic IR bands were observed at 2045 and 2200 cm(-1), due to linearly adsorbed CO on Mo and N sites, respectively. The surface N sites are highly reactive and can react with adsorbed CO to form NCO species. Unlike adsorbed CO on reduced passivated one, the adsorbed CO on fresh Mo2N/gamma-Al2O3 behaves similarly to that of group VIII metals, suggesting that fresh nitride resembles noble metals. It is found that the surface of Mo nitrides slowly transformed into sulfide under hydrotreating conditions, which could be the main reason for the activity drop of molybdenum nitride catalysts in the presence of sulfur-containing species. Some surface reactions, such as selective hydrogenation of 1,3-butadiene, isomerization of 1-butene, and hydrodesulfurization of thiophene, were studied on both fresh and reduced passivated Mo2N/gammaAl(2)O(3) catalysts using IR spectroscopy. The mechanisms of these reactions are proposed. The adsorption and reaction behaviors of these molecules on fresh molybdenum nitride also resemble those on noble metals, manifesting the unique properties of fresh molybdenum nitride catalysts. Mo and N sites are found to play different roles in the adsorption and catalytic reactions on the fresh Mo2N/gammaAl(2)O(3) catalyst. Generally, Mo sites are the main active sites for the adsorption and reactions of adsorbates; N sites are not directly involved in catalytic reactions but they modify the electronic properties of Mo sites.

Improvement of Pt catalyst for soot oxidation using mixed oxide as a support

Catalytic activity of Pt catalysts for soot oxidation was studied using temperature programmed reactions. The activity of Pt loaded over TiO2-SiO2 (Pt/TiO2-SiO2) showed higher activity than other Pt/MOx systems (MOx = TiO2, ZrO2, SiO2, Al2O3. TiO2-ZrO2. TiO2-Al2O3, ZrO2-SiO2, ZrO2-Al2O3, SiO2-Al2O3). The activity was highest when the molar ratio of TiO2/(TiO2 + SiO2) ranged from 0.4 to 0.7. The effect of pretreatment with a gas containing low SO2 concentrations on the activity was compared for Pt/SiO2, Pt/TiO2 and Pt/TiO2-SiO2. In the case of Pt/TiO2-SiO2, the activity was markedly promoted by the pretreatment whereas no variation in the activity was observed for Pt/SiO2. The difference in the behavior towards the SO, pretreatment was attributed to property difference in the supports for sulfate accumulation. The high activity of Pt/TiO2-SiO2 was also confirmed under practical conditions with a diesel engine exhaust using a catalyst-supported diesel particulate filter (DPF). (C) 2003 Elsevier Science B.V. All rights reserved.

Application of static and dynamic enclosures for determining dimethyl sulfide and carbonyl sulfide exchange in Sphagnum peatlands: Implications for the magnitude and direction of flux

A static enclosure method was applied to determine the exchange of dimethyl sulfide (DMS) and carbonyl sulfide (OCS) between the surface of Sphagnum peatlands and the atmosphere. Measurements were performed concurrently with dynamic (flow through) enclosure measurements with sulfur-free air used as sweep gas. This latter technique has been used to acquire the majority of available data on the exchange of S gases between the atmosphere and the continental surfaces and has been criticized because it is thought to overestimate the true flux of gases by disrupting natural S gas gradients. DMS emission rates determined by both methods were not statistically different between 4 and >400 nmol m−2 h−1, indicating that previous data on emissions of at least DMS are probably valid. However, the increase in DMS in static enclosures was not linear, indicating the potential for a negative feedback of enclosure DMS concentrations on efflux. The dynamic enclosure method measured positive OCS flux rates (emission) at all sites, while data using static enclosures indicated that OCS was consumed from the atmosphere at these same sites at rates of 3.7 to 55 nmol m−2 h−1. Measurements using both enclosure techniques at a site devoid of vegetation showed that peat was a source of both DMS and OCS. However, the rate of OCS efflux from decomposing peat was more than counterbalanced by OCS consumption by vegetation, including Sphagnum mosses, and net OCS uptake occurred at all sites. We propose that all wetlands are net sinks for OCS.

Iron and inorganic carbon in Liaodong Gulf sediments of Bohai Sea in China

Iron in seawater is an essential trace metal for phytoplankton that plays an important role in the marine carbon cycle. But most studies focused on oceanic iron fertilization in high nutrient low chlorophyll (HNLC) seawaters. A study of inorganic carbon (IC) forms and its influencing factors was presented in Liaodong Gulf sediments, and especially the influence of iron was discussed in detail. Inorganic carbon in Liaodong Gulf sediments was divided into five forms: NaCl, NH3·H2O, NaOH, NH2OH·HCl and HCl. The concentration of NaCl and NaOH forms were similar and they only occupied the minority of total inorganic carbon (TIC). However, NH3·H2O, NH2OH·HCl and HCl forms were the principal forms of TIC and accounted for more than 80% of TIC. Especially, the percentage of NH3·H2O form was much higher than that in the Changjiang River Estuary and Jiaozhou Bay sediments. All forms of inorganic carbon were influenced by organic carbon,pore water, iron, pH, redox potential(Eh) and sulfur potential(Es) in sediments, moreover, the influences had different characteristics for different IC forms. However, the redox reactions of iron affected mainly active IC forms. Iron had little effect on NH2OH·HCl and HCl forms of IC which were influenced mainly by pH. Iron had a stronger influence on NaCl, NaOH and NH3·H2O forms of IC; the influence of Fe2+ was higher than Fe3+ and its effect on NH3·H2O form was stronger than on NaCl and NaOH forms.

A QUANTITATIVE ASSESSMENT OF SOURCE CONTRIBUTIONS TO INHALABLE PARTICULATE MATTER POLLUTION IN METROPOLITAN BOSTON

In this paper, source apportionment techniques are employed to identify and quantify the major particle pollution source classes affecting a monitoring site in metropolitan Boston, MA. A Principal Component Analysis (PCA) of paniculate elemental data allows the estimation of mass contributions for five fine mass panicle source classes (soil, motor vehicle, coal related, oil and salt aerosols), and six coarse panicle source classes (soil, motor vehicle, refuse incineration, residual oil, salt and sulfate aerosols). Also derived are the elemental characteristics of those source aerosols and their contributions to the total recorded elemental concentrations (i.e. an elemental mass balance). These are estimated by applying a new approach to apportioning mass among various PCA source components: the calculation of Absolute Principal Component Scores, and the subsequent regression of daily mass and elemental concentrations on these scores.

Combustion synthesis and luminescent properties of the Eu3+-doped yttrium oxysulfide nanocrystalline

Nanocrystallinc Y2O2S:Eu3+ was successfully prepared with a combustion synthesis method, the corresponding bulk Y2O2S:Eu3+ was synthesized by conventional sulfur flux method. The results of XRD indicated that both bulk and nanocrystalline Y2O2S:Eu3+ have Pure hexagonal phases. The crystallite size was calculated to be about 20 nm according to Scherrer formula, which was consistent with the size as indicated by transmission electron microscopy (TEM).