Development of polymorphic EST markers in Penaeus monodon: applications in penaeid genetics

The objective of this study was to develop type I markers for genome mapping and other genetic studies of Penaeus monodon. Primers were designed based on expressed sequence tags (ESTs) from a P monodon cephalothorax cDNA library to amplify 100-300 bp products. 34 of the primer pairs successfully amplified PCR products from genomic DNA. Single-strand conformation polymorphism analysis showed that similar to 30% of the ESTs tested exhibit polymorphism in a test panel of P monodon individuals. Mendelian inheritance of the EST-derived markers has been established in two international reference mapping families of P monodon, and mapping of these markers is in progress. Some ESTs were successfully amplified from other Penaeus species (P. chinensis, P japonicus and P vannamei), indicating that the markers are applicable in cross-species comparison. Two populations of P. japonicus could be differentiated using one of the ESTS. In conclusion, the polymorphic EST markers developed in this study are applicable in genome mapping and population genetic studies of penaeid shrimp. (C) 2002 Elsevier Science B.V. All rights reserved.

Impact of eutrophication on biodiversity of the macrozoobenthos community in a Chinese shallow lake

Comparative studies on community structure and biodiversity of macrozoobenthos were carried out in three lake areas with different states of eutrophication (mesotrophic, eutrophic and hypereutrophic) in a shallow subtropic lake (Lake Donghu) in the middle basin of the Yangtze River of China. Thirty-three taxa (including six Mollusca, five Oligochaeta, 15 Chironomidae and seven other zoobenthos) were found during February 1998 to April 1999. The results show that the more eutrophic the lake water, the lower the macrozoobenthos species diversity (measured as species number, diversity index, and K-dominant curves). Abundance of the oligochaete Limnodrilus hoffmeisteri was positively correlated with the degree of eutrophication, which was due mainly to its ability to tolerate low dissolved oxygen. The chironomid Tanypus chinensis also thrived in the hypereutrophic environment; however, it was less endurable to eutrophication than L. hoffmeisteri.

Sequences of cytochrome b gene for primitive cyprinid fishes in East Asia and their phylogenetic concerning

1140 bp of cytochrome b gene were amplified and sequenced from 14 species of primitive cyprinid fishes in East Asia. Aligned with other ten cytochrome b gene sequences of cyprinid fish from Europe and North America retrieved from Gene bank, we obtained a matrix of 24 DNA sequences. A cladogram was generated by the method of Maximum likelihood for the primitive cyprinid fishes. The result indicated that subfamily Leuciscinae and Danioninae do not form a monophyletic group. In the subfamily Danioninae, Opsariichthys biden and Zacco platypus are very primitive and form a natural group and located at the root. But the genera in subfamily Danioninae are included in different groups and have not direct relationship. Among them, Aphyocypris chinensis and Yaoshanicus arcus form a monophyletic group. Tanichthys albonubes and Gobiocypris rarus have a close relation to Gobioninae. The genus Danio is far from other genera in Danioninae, In our cladogram, the genera in Leuciscinae were divided into two groups that have no direct relationship. The genera in Leuciscinae distributed in Europe, Sibera and North America, including Leuciscus, Rutilus, Phoxinus, N. crysole, Opsopoeodus emilae, form a monophyletic group. And the Leuciscinae in southern China including Ctenopharyngodon idellus, Mylopharyngodon piceus, Squalibarbus and Ochetobius elongatus have a common origination.

Bioaccumulation of polychlorinated dibenzo-p-dioxins and dibenzofurans in the foodweb of Ya-Er Lake area, China

Bioaccumulation of PCDD/F in the foodweb was investigated in the Ya-Er Lake area, which was heavily polluted by PCDD/F. The high concentrations of PCDD/F in sediment can be transferred and bioaccumulated by aquatic organisms and humans through various pathways. Benthonic invertebrate animals and aquatic plants with a lot of fibers in the root can accumulate PCDD/F from sediment and water. Snail (Bellamya aeruginosa), shrimp (Macrobranchium sp.) and freshwater mussel (Acuticosta chinensis (Lea)) took up PCDD/F from the water and maintained the emission patterns, whereas fish tended to selectively accumulate 2,3,7,8-substituted isomers. The tissues of fish-eating bird and duck (Anas platyrhynchos) were very highly contaminated by PCDD/F due to ingestion of fish and other aquatic organisms from sediment. The residual concentration in breast milk depended on the original concentration of PCDD/F in the food. A resident in Ya-Er Lake area showed a daily intake of PCDD/ F of about 9.14 pg TEQ/kg body weight/day. This is higher than the tolerable daily intake (TDI) for PCDD/F (1 pg TEQ/kg body weight/day), which was recommended by the World Health Organization (WHO). (C) 2001 Elsevier Science Ltd. All rights reserved.

安达市碱性草地土壤颗粒的分维特征

运用土壤颗粒质量分形模型计算松嫩平原低平地安达试区植被分布区和碱斑区样点土壤颗粒的分形维数,并建立分形维数与土壤颗粒不同粒级间的回归关系,以探讨土地碱化后土壤粒径分布的分形特征及其与土壤物理性状的关系。结果表明:安达试区土壤颗粒分形维数较高,平均分别仅有48.7×10-5cm/s(Pit A)和4.30×10-6cm/s(Pit B),反映了该区土壤细颗粒含量高、土壤大孔隙数量少、土壤饱和导水率低的特征;土壤颗粒分形维数与黏粒含量呈对数正相关关系,而与粉粒和砂粒含量相关性不显著,说明在安达试区,影响土壤颗粒分形维数的主要因素是黏粒含量;羊草地土壤颗粒分形维数在土壤垂直剖面上的变异较大,说明植被生长促进了土壤质地的变异;碱斑地土壤颗粒分形维数明显大于羊草地,细颗粒含量高,饱和导水率低,说明碱斑的形成恶化了土壤物理性质;土壤颗粒分形维数可以反映安达市土壤物理性质的好坏,能作为土壤退化和生态环境恶化的评价指标。研究结果可为安达市以及松嫩平原盐碱地生态环境的修复和治理提供科学依据。

西夫韦肽和盐肤木提取物体外抗HIV 活性及机制研究

本论文主要由3 个相对独立的部分组成: 西夫韦肽抗HIV 活性机制研究及其 联合用药和耐药性研究，盐肤木提取物及其化合物抗HIV 活性机制研究和精子 顶体反应抑制剂AGB 抗HIV 活性及机制研究。 HIV 侵入抑制剂是抗HIV 药物研发的热点。该类抑制剂靶定在病毒复制周 期早期，为HARRT 疗法提供了更多新的药物组合，且该类抑制剂对临床中已产 生的耐药毒株也有较好的抑制作用。目前FDA 批准上市的侵入抑制剂仅有T-20， 急需开发新一类的HIV-1 侵入抑制剂。西夫韦肽是由36 个氨基酸组成的多肽， 我们对西夫韦肽进行了一系列体外抗HIV 药效学的实验来研究西夫韦肽体外抗 HIV 活性以及作用机制。实验结果表明西夫韦肽对多种HIV 宿主细胞毒性小， 可以有效抑制HIV-1IIIB 诱导的C8166 细胞的病变效应，EC50 值仅为7.8ng/ml ， TI 值大于 384,615;在不同的检测方法中，西夫韦肽均表现出了比T-20 更好的抑 制活性，EC50 值低了13-42 倍。在对HIV-1 临床分离株、耐药株、两株HIV-2 毒株和SIVmac239 的抑制活性研究表明西夫韦肽也可以很好地抑制 HIV-1 临床株 HIV-1KM018 的复制，EC50 值低至4.4ng/ml，对耐药株HIV-174V、HIV-2 和 SIVmac239 的复制也均有较好的抑制作用。 在机制研究中，我们发现西夫韦肽极有效地抑制HIV-1慢性感染H9细胞与 正常C8166细胞间的融合作用，EC50 低至0.4ng/ml，表明西夫韦肽可以以极低的 浓度有效抑制HIV进入宿主细胞。用GST-pull down 实验进一步验证了西夫韦肽 和T-20可以很好地与HR1结合而不能与HR2结合，作用机制就是特异地与gp41的 HR1结合从而抑制了6-Helix的形成，阻断了HIV的融合过程。由于HIV的高变异 性，单一药物治疗容易产生耐药性，最终导致治疗失败。因此在新药开发中进行 药物与作用靶不同的已上市药物体外联合用药和耐药性研究是非常必要的，将对 临床应用有指导意义，我们的实验结果表明西夫韦肽与AZT联合用药体外抗HIV-1作用较单独用药好，不同检测方法联合用药比单独使用西夫韦肽的效果好 8.3-9.4倍；耐药性研究表明其体外诱导耐药性产生的时间与T-20相仿，与T-20有 交叉耐药。 我国传统的中医药是个巨大的宝库，有丰富的临床经验，中医药治疗艾滋病 有着一定的潜力。从我国国情出发，利用中医药的独特性及经济性，开发传统的 具有我国特色的艾滋病治疗天然药物成为AIDS 防治工作的当务之急。盐肤木是 中国的本土植物，在我国民间用作传统医药有着悠久的历史。盐肤木茎提取物尤 其是石油醚提取部分RC-1 具有较好的抗HIV 活性，且作用于病毒复制周期的后 期，从中分离得到的化合物1、2、4、5 和6 都是RC-1 的活性成份；盐肤木茎 提取物乙酸乙酯提取物RC-2 中也有较好的抗HIV 作用，其中的化合物8、9、 10 和13 是抗HIV 的活性成分，且作用机制各不相同，这些有效化合物的抗HIV 机制值得进一步的研究。 杀微生物剂是一种局部用药于阴道或宫颈的药物制剂，由女性自主控制防止 性传播疾病病原体包括HIV 的感染，是近年来的研究热点之一。AGB(4`-乙酰胺 苯基 4-胍基苯甲酸酯)是顶体酶的抑制剂，我们的实验表明AGB 有很好的杀精 子作用，还具有体外抗HIV-1 的作用，作用机制主要是阻断HIV-1 进入细胞。

栽培措施与文冠果结实性关系的研究

文冠果（Xanthoceras sorbifolia Bunge���是我国北方特有的木本油料树种，已被列为生物质能源树种之一。现阶段产量低是限制其发展的重要因素。结合目前文冠果生产发展的实际需要，有必要通过实践探讨出一套经济、简单易行的栽培措施，指导文冠果的高产优质栽培管理。本文以内蒙古翁牛特旗文冠果经济林场内40年生文冠果树为研究对象，对文冠果生物学、生态学特性进行定位观测和试验，研究了文冠果物候与积温的关系；野外布置不同管理措施的正交试验设计，测定了不同管理措施下文冠果产量和年生长周期内叶片矿质元素（N、P、K、Mg、Fe、Zn、Cu、Mn）的含量变化趋势，并对数据进行了分析。初步确定了文冠果物候与矿质养分动态变化、栽培管理措施与产量以及叶片养分含量之间的关系，为提高文冠果的产量、制定合理栽培管理措施提供一定理论依据，也为文冠果的进一步开发利用打下基础。 主要结论： 1.文冠果年生长周期内物候变化与积温呈显著对数函数关系，可以预测文冠果年生长发育期有效积温的理论值，也可以根据实际有效积温预测文冠果物候变化进程； 2.文冠果叶片中矿质元素变化与物候期有密切联系，大量元素间存在协同关系，水肥管理对叶片中矿质元素的变化有很大影响； 3.施肥和浇水均可提高文冠果果实产量，水肥耦合措施提高产量尤为显著：与对照相比，5月初一次浇水、施肥270g/株可使产量提高303%。

文冠果壳苷提取工艺的研究

文冠果为我国独有的油料树种，研究发现，文冠果壳乙醇提取物具有良好的促智作用，文冠果壳苷（Xanthoceracide）为齐墩果烷型五环三萜皂苷，是文冠果壳乙醇提取物中活性最强的物质。大量的药效学实验证明其具有显著改善多种记忆障碍模型小鼠的学习能力，提高大脑的缺氧耐受能力，预防并治疗多发性栓塞引起的记忆保持障碍，降低谷氨酸造成的PC12细胞死亡数量，增强对神经细胞的保护作用；数例临床试验中，一些记忆力严重减退导致生活不能自理的老年人以及智力低下的儿童服用文冠果壳提取物或文冠果壳苷后，也有明显的康复效果或治疗效果；而急性毒性试验进一步证实，这些天然产物毒性低，仅仅1/50的LD50用量就能表现出明显的改善记忆障碍作用，这预示着文冠果壳乙醇提取物将有可能开发成有效的老年痴呆症特效药。 本研究采用薄层层析法（TLC）对文冠果壳提取物中的文冠果壳苷进行定性鉴别。显色剂为10%硫酸乙醇，操作方法简便，斑点清晰，Rf适中，重现性良好。采用分光光度法对文冠果壳总皂苷含量进行测定。用文冠果壳苷作为对照品，测定波长为546 nm，文冠果壳苷含量在0.004 mg·mL-1~0.02 mg·mL-1（r=0.9998）呈现良好的线性关系，方法检验合格。采用高效液相色谱法（HPLC）对文冠果壳苷进行含量测定，文冠果壳苷含量在0.02 mg·mL-1~0.2 mg·mL-1（r=0.9992）范围内线性关系良好，方法检验合格。 采用溶剂提取法，大孔吸附树脂分离，有机溶剂萃取，硅胶柱层析分离，结晶与重结晶等方法相结合，得到纯度达98.5%的文冠果壳苷，可作标准品为开发利用文冠果壳提供依据。 以出膏率和文冠果壳苷含量为考察指标，分别考察提取次数、提取温度、乙醇浓度、料液比以及提取时间等因素对提取效果的影响。采用正交试验优化文冠果壳苷的溶剂提取工艺，最终确定提取工艺为：用70%乙醇提取，料液比为1/7（W／V），提取时间为5 h，提取温度为60 ℃，提取次数为2次。采用中试放大试验验证该工艺条件，结果表明，该提取工艺简单，易操作，提取效果较好，具有较好的应用前景。

水肥措施对文冠果叶片、果实养分分配的影响

文冠果（Xanthoceras sorbifolia Bunge���是我国北方特有的木本油料树种，已被列为生物质能源树种之一。现阶段产量低是限制其发展的重要因素。本文以内蒙古赤峰市翁牛特旗文冠果经济林场内50年生文冠果树为研究对象，对文冠果生物学、生态学特性进行定位观测和试验，分析了文冠果发育节律与积温的关系。野外布置水肥措施试验，测定了文冠果产量和叶片、果实养分元素（N、P、K、Mg、Ca、Fe、Zn、Cu、Mn）的含量，分析了水肥措施对叶片、果实养分分配的影响。 主要结论： 1.文冠果物候期与积温呈显著的直线函数关系。能够用来预测文冠果年生长发育期有效积温的理论值，为经营管理提供依据。 2.文冠果叶片中主要元素含量随物候期的变化呈下降趋势，但微量元素呈增加趋势。养分重吸收水平由大到小的顺序是：Mg>N>K>P，一定的水肥措施提高了文冠果养分再吸收水平，并降低了功能叶中的N/P、C/N。 3.文冠果果实中Ca和Fe随物候期的变化呈下降趋势，其他元素呈先减后增趋势。水肥措施降低了果实对养分元素的分配和收获系数；提高了养分利用效率。 4.合理的水肥措施能显著提高种子产量，增加含油率。最佳水肥措施为：5月初灌水50mm，施肥量为N 63 g•株-1、P2O5 83.3 g•株-1，于5、7月初分2次等量施入。 5.水分、P是文冠果生长的主要限制性因子，限制性因子具有阶段性。水分对肥效有“激发效应”。

矿山废弃物中重金属污染行为及其对白菜品质的影响

综合运用环境地球化学、土壤学、生态学和统计学的方法开展了矿山废弃物尾矿石的堆置对土壤造成的污染以及矿区土壤重金属污染对白菜（Brassica Chinensis）的卫生品质和营养品质的影响研究。静态培养试验和动力学淋溶试验证明：在环境因素（如降水）的作用下，尾矿石的堆置将对下伏土壤的重金属含量产生长久影响，随着降水量以及酸度的增加，土壤中Cu、Cd、Pb、Zn的全量和有效态含量都有所升高并且土层间的差异减小。有机酸能不同程度地促进土壤中的重金属转化为有效态，增强重金属的生物有效性；多元梭酸比一元梭酸的转化率高。另外，有机酸对重金属的转化作用随其浓度的增大而增强，在低温范围内，随温度的升高而增加。通过网室盆栽试验，研究了土壤受Cu、Cd、Pb，Zn单一重金属污染后对大白菜吸收Cu、Cd、Pb、Zn以及植物体内粗纤维和维生素C含量的影响；同时还进一步对Cd-P交互作用影响大白菜吸收Cd、P以及植物体内粗纤维和维生素C含量的情况进行了研究，结果表明大白菜从污染的土壤吸收重金属与土壤中目标元素的含量呈极显著正相关，对非目标重金属的吸收情况复杂，因不同元素间的相互作用机制而异；白菜体内粗纤维和抗坏血酸含量随土壤重金属浓度的增加而降低，它们之间呈极显著的负相关关系。在采用大白菜进行高等植物毒性试验的基础上，首次运用典型相关分析方法对Cu、Cd、Pb、Zn复合污染土壤的生态毒性进行了评价，分别建立了由4元素共同组成的复合污染土壤毒性的线性组合函数和由芽生物量抑制率和根伸长抑制率组成的植物学表征的线性组合函数，并计算了其第一典型变量Toxl（土壤毒性）和BloL（植物学表征）之间的相关系数为0.94（p＜0.01）。典型相关分析是介质毒性评价的有效方法之一。

文冠果（Xanthoceras sorbifolia）

本文对中国特有植物文冠果（Xanthoceras sorbifolia Bunge���种仁油中脂肪酸的组成及结构进行了深入的探讨。应用远端化学修饰新技术，将混合脂肪酸制备成2－烃基取代－4，4－二甲基噁唑啉衍生物的混合物，用气相色谱－质谱－计算机联用仪，结合尿素包合法对混合脂肪酸衍生物进行分析，鉴定了十二种脂肪酸组分的结构，确定了不饱和酸的双键位置。基保三种脂肪酸首次被确存在于文冠果油中，即甘碳二烯－11，14－酸，甘四碳烯－15－酸和甘四碳烷酸（木焦油酸）。本文对衍生物制备的分馏催化法和简单回流法进行了比较，肯定了后者的可行性可靠性。

中药射干的化学成分研究

本学位论文由4章组成。第一章是论文的主体，报道了中药射干的化学成分研究。第二章是中药射干代用品川射干的化学成分研究，并附带报道了西番莲化学成分的研究结果。第三章是射干、川射干及西番莲提取物化学成分串联质谱分析的报道。第四章为综述，概述了射干及鸢尾属植物的化学成分和药理研究进展。 在第一和二章中分别报道了射干（Belamcanda chinensis (L.) DC.）、川射干 （Iris tectorum Maxim.）及西番莲（Passiflora incarnate L.）化学成分的分离纯化与结构鉴定。采用正、反相硅胶柱层析、凝胶柱层析、薄层制备及HPLC等各种分离方法，从三种药用植物中共分离出68个不同的化合物，其中61个的结构得 得以鉴定，另外4个化合物的结构正在鉴定中，3个由于量少且有点杂质未作进一步的鉴定。 中药射干（Rhizoma Belamcandae）为射干植物的干燥根茎，从中共分离出53个化合物，通过红外、质谱及核磁共振等波谱方法鉴定了包括12个新化合物在内的48个，结构类型分别属于iridal型三萜及其新颖的二聚体、异黄酮、黄酮及黄酮醇、香豆素、甾体、芳香酸和脂肪酸及其甘油酯等。新化合物中有两个异黄酮类化合物，其结构分别鉴定为5,7,8,4′-四羟基-6-甲氧基异黄酮和5,6-二羟基-4′-甲氧基异黄酮-7-O-β-D-吡喃葡萄糖苷；八个新的iridal型三萜化合物分别鉴定为鸢尾烯（L）、16-甲氧基鸢尾烯、16-去羟基鸢尾烯、2-(E)-16-去羟基鸢尾烯、16-去羟基鸢尾烯B、3-乙酰基-16-去羟基鸢尾烯、iristectoroneL和iristectoroneM；两个结构骨架新颖的双三萜，分别命名为射干素A和射干素B，其分离纯化的困难以及结构的新颖和复杂突显出该论文的科学意义。除这些新化合物外，还有9个已知化合物为首次从中药射干中分离得到。此外，从中药射干的代用品川射干中分离得到7个已知化合物，主要是黄酮类成分及iridal型三萜化合物，其中1个三萜化合物为从射干中分离鉴定的新成分。另外还从西番莲中分离出8个化合物，鉴定了其中的6个，主要为黄酮碳苷。 第三章是关于射干、川射干及西番莲提取物化学成分的ESI-MS-MS分析，在初步探讨了从这些植物中分离鉴定出的一些异黄酮及黄酮碳苷的质谱裂解规律基础上，通过质谱和串联质谱分析，定性和半定量地检测了射干和川射干中主要的异黄酮成分以及西番莲中的黄酮碳苷成分，为这些药材品质的快速鉴定提供了一种简便方法。 第四章概述了射干及鸢尾属药用植物的化学和药理研究进展，特别是对其中异黄酮及三萜类成分的研究进展进行了深入系统的综述。 This dissertation is composed by four chapters. The first and second chapter reports the phytochemical investigation of three medicine plants, Belamcanda chinensis (L.)DC., Iris tectorum Maxim. and Passiflora incarnate L. Sixty eight different compounds were isolated and sixty one of them were identified. The third chapter described rapid ESI-MS-MS analysis of B. chinensis, I. tectorum, and P. incarnate. The forth part is a review about the progress of studies on the chemical constituents from Belamcanda chinensis and Iris species. Fifty-three compounds were isolated from Rhizoma Belamcandae, the rhizomes of B. chinensis by the methods of column chromatography (normal and reversed phase silica gel, Sephadex LH-20), preparative TLC and HPLC. On the basis of spectroscopic methods including IR, ESI-MS, 1-D and 2-D NMR, forty eight of them were identified as seventeen flavonoids, seventeen tritepenoids, one cumarin, five steroids and some benzene derivative etc. Among them, the structures of twelve new compounds were elucidated as 6-methoxy-5,7,8,4′-tetrahydryoxyisoflavoe, 4′-methoxy-5,6-dihydroxyisoflavone-7-O-β-D-glucopyranoside, iristectorene L, 16-methoxyisoiridogermanal, 16-dehydroxyisoiridogermanal, 2-(E)-16-dehydroxy isoiridogermanal, 16-dehydroxyiristectorene B, 3-acetyl-16-dehydroxyisoiridoger- manal, iristectorone L, iristectorone M, belamcandene A and belamcandene B. Last two new compounds are dimer of triterpenoids with a novel carbon skeleton. Beside the new compounds, nine known ones were isolated from this plant for the first time. Isolation of I. tectorum yielded seven compounds. On the basis of spectroscopic methods including ESI-MS, NMR and the comparison with authentic samples, three of them were determined as isoflavone, two of them were triterpenoids, and other two were β-sitosterol and apocynin. All of them are known compounds except one of iridal type triterpenoid, 16-dehydroxyiristectorene B, which also obtained from B. chinensis as a new compound. Isolation of P. incarnate yielded eight compounds. Six of them were determined on the basis of spectroscopic methods including ESI-MS, NMR and the comparison with authentic samples. Four of them are flavone-C-gluconside, and two are steroids. The third chapter describes the tandem mass spectrometry (ESI-MS-MS) analysis of the isoflavonoids from B. chinensis and I. tectorum, as well as C-glycosyl-flavonoide from P. incarnate, in order to explore the rapid methodology of validating the quality of the herbs. In addition, the fractionation rules of some iosflavonoids and C-glycosyl-flavonoids were discussed. The fourth chapter summarizes the research development on chemistry and pharmacology of medicine plants of B.chinensis and Iris species.

低极性化合物的电喷雾质谱检测方法及几种中藏药材的活性成分研究

本学位论文首先报道了为解决低极性化合物的电喷雾质谱（ESI-MS）分析难题而建立的一种衍生化分析方法；然后从色谱-质谱联用分析、分离纯化和结构鉴定等方面分别报道了几种中藏药材的活性成分研究。论文由下述六章组成： 第一章报道了盐酸羟胺衍生化方法在电喷雾质谱 (ESI-MS) 分析中的应用。该方法利用盐酸羟胺和羰基成肟的快速反应，建立了针对三萜酮等含酮或醛羰基低极性化合物的ESI-MS 信号增强技术。此方法不仅可应用于增强羰基化合物的ESI-MS 质谱信号，还可检测化合物中羰基的个数以及辨别涉及羰基官能团的同分异构体。此外，通过简单的氧化反应，还可将该方法拓展到三萜醇、甾醇等含羟基的低极性化合物，增强它们的ESI-MS 信号。对比已报道的相关ESI-MS 增强质谱信号的衍生化方法，此方法有经济、实用、快速和简便的显著特点。 第二章是关于野生羌活及其栽培品种化学成分的色谱-质谱联用分析。对不同产地野生羌活生长过程中活性成分的动态变化、野生羌活不同形态部位和人工栽培羌活中的活性成分含量进行了HPLC 定量分析。结果表明主要活性成分羌活醇和异欧前胡素都随生长期存在规律性变化，羌活不同形态部位中的活性成分含量也有明显不同。这些实验结果有些较好地印证了传统中医的用药理论，有些也对羌活的传统使用方法提出了新的建议。 第三章介绍了几种传统中藏药材的色谱-质谱联用及串联质谱分析。通过GC-MS 方法，从藏药材长花党参挥发油中共分离鉴定出45 个化合物；利用HPLC方法测定了该藏药材中的主要化学成分——木犀草素的含量（0.7%）；利用串联质谱技术，对西番莲和射干中的主要成分进行了快速鉴定，从西番莲中鉴定了4个黄酮碳苷；从不同产地的射干和川射干中鉴定了8 个主要异黄酮成分，其中包括一个未见报道的化合物。 第四章的内容为藏药材石莲叶点地梅的活性成分研究。从植物石莲叶点地梅(Androsace integra (Maxim.) Hand.-Mazz.) 乙醇提取物的正丁醇萃取部分共分离和鉴定了6 个化合物，利用MS 和NMR 等现代波谱学技术阐明了它们的结构：其中包括4 个三萜类化合物：分别是androsacin (1)、 ardisiacrispin A (2) 、saxifragifolin A (3) 和20(29)-lupen-3-one (4)；一个神经酰胺：4-羟基-Δ8,9(Z)-鞘氨醇-2'-羟基正二十四碳酸酰胺(5)；一个甾体类化合物：胡萝卜苷(6)。化合物1为新的13,28-epoxy-oleanane 型三萜皂苷，在其结构表征的过程中，采用LC-MS 进行糖分析，获得了值得推广的好结果。通过活性筛选发现化合物1~3 对HepG2肝癌细胞表现出不同程度的抑制活性，其中化合物2 活性最好，其IG50 为1.65μg/mL。 第五章是关于一些传统中藏药材的农药活性筛选。利用Syngenta 公司的活性筛选平台对68 种传统中藏药材醇提物进行了抗菌和除草的生物源农药活性筛选。结果表明所筛选的68 种植物提取物中，共有14 种样品表现出明显的除草/杀虫活性，其中水母雪莲花、松萝和茯神木等植物提取物还具有多种生物活性。活性成分还有待进一步追踪分离、纯化和结构鉴定。 第六章为文献综述，概述了羌活药材的研究进展。对羌活属及药用羌活植物从分类学、本草学、品质评价、人工栽培、化学成分及药理作用等方面进行了文献归纳和总结。 In this dissertation, an electrospray ionization mass spectrometry (ESI-MS) signal enhancement method, as well as the work of bioactive components study, HPLC-MS/MS application, bioassay screening, chromatograph separation and structure identification of the metabolites in several medicinal herbs have been reported. First chapter expounded a rapid, simple ESI-MS sensitivity enhancement method for detecting carbonyl groups in natural products has been developed by using hydroxylamine hydrochloride (NH2OH·HCl) as a derivatization reagent. We use the oxime formed during the derivatization reactions and its Beckmann rearrangement intermediates as a means of detecting the carbonyl groups originally present in these triterpenoids. In comparison with other derivatization methods in the literature, this method is simple, specific and can be used to detect carbonyl groups in triterpenoids which have low polarity and are poorly or non-ionizable. Moreover, it can also be used to detect hydroxyl groups by using the Dess-Martin periodinane (DMP) to convert primary and secondary hydroxyls into carbonyl groups. Chapter 2 reported an HPLC-MS method for analyzing the main bioactive compounds in both wild and cultured Notopterygium incisum. The results indicated that the main bioactive compounds varied through different seasons regularly, and in different commercial parts of this herb the content of these compounds also differed from each other. The quantitative analysis results showed that in the traditional commercial parts, the content of main chemical constitutes in Silkworm Notopterygium, Bamboo Notopterygium and Irregular-nodal Notopterygium are higher than that in Striped Notopterygium. This result is tally with the traditionally concept that the quality of Notopterygium, Bamboo Notopterygium and Irregular-nodal Notopterygium are better than that of Striped Notopterygium, which means that the quality of rhizomes is better than main roots. The chemical constituents of cultured N. incisum is reported for the first time in this dissertation and the analysis results showed some growth curves of chemical constituents in this plant, but still left some questions unanswered. Chapter 3 discussed the GC/LC-MS analysis of the traditional Chinese medicines Codonopsis thalictrifolis, Passiflora incarnate, Belamcanda chinensis and Passiflora incarnate. The main constituent, luteolin was isolated and identified from the traditional Tibet medicine of C. thalictrifolis. The quantitative analysis by HPLC has revealed that the content of luteolin in this herb is 0.7%. GC-MS was employed to analyzed chemical constituents of the essential oil from the flower of C. thalictrifolis. More than 60 peaks were detected and 45 of them were identified by comparing their spectra with that of the standards in the database and literatures. ESI-MS/MS was used to analyze the n-butanol extract of Passiflora incarnate. Based on the information of pseudo molecular ions and fragment ions of the glycosides, four major flavone-C-glycosides have been detected and identified as 7-methoxyluteolin-6-C-β-D-glucopyranoside, vitexin, swertisin and orientin. The isoflavone compounds in theextracts of three samples of B. chinensis collected in Gansu, Sichuan and Hunan, and the extract of Iris tectorum collected in Sichuan were analyzed by using TOF-HRMS and IT-MS. From the extracts of these herbs, a new isoflavone, identified as 5’,5,6,7-tetrahydroxy-3’4’-dimethoxyl isoflavon, and 7 known ones have been identified by analyzing the fragmentation patterns and their molecular formulas given by HRMS and the tandem mass spectrometry acquired by IT-MS. Chapter 4 elucidated the isolation and identification of a new triterpene saponin, androsacin (1), along with five known compounds (2-6) were isolated from the whole plants of Androsace integra (Maxim.) Hand.-Mazz., an herb used in traditional Chinese and Tibetan medicine. The chemical structure of the new compound was established as 3β-O-{β-D-glucopyranosyl-(1→4)-O-β-D-xylopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→4)-[O-β-D-glucopyranosyl-(1→2)]-α-L-arabinopyranosyl}-16α-hydroxy-13β,28-epoxy-olean-30-al by analyzing its MS, 1D- and 2D-NMR spectra. Compound 2 was cytotoxic toward HepG2 cancer cell with the GI50 value of 1.65 μg/mL. Chapter 5 described the biogenic pesticide activity screening of 68 traditional Chinese and Tibetan medicine extractions. The intention of this study is to explore bioactive natural compounds from these traditional medicinal herbs for biogenic insecticides use. Based on Syngenta’s bioassay, 14 extractions of these traditional medicines showed pesticide activities, and some of them had multi-activities on antibacterial and insecticidal. Chapter 6 is a review on the chemical and bioactivity research progress of Notopterygium incisum and N. forbesii.

贡嘎山麦吊杉(Plcea brachytyly)群落的结构研究

群落结构的确定是群落研究的基础。本文立足于个体、种群、群落三级水平，从空间和时间两种概念出发，对贡嘎山麦吊杉(Picea brachytyla)群落的结构进行了研究。经初步统计，在麦吊杉林分布的主要地段共有维管束植物93科、263属、约560种，其中种子植物71科、220属、479种。通过种子植物区系地理成份的分析表明，麦吊杉群落的区系组成具有区系成分复杂、特有现象明显和成分古老的特点。在生活型组成上，麦吊杉群落的高位芽植物所占比例最大，其中小高位芽植物最多，而在叶级谱中小叶类型占有较明显的优势。通过种—面积曲线和重要值—面积曲线两种方法求得的麦吊杉群落的最小面积在800-900平方米之间，实际应用时可选用1000平方米的保守面积。在物种的综合特征分析中，群落表现出优势种明显的特征，而存在度II占有最大的比例。物种多样性的研究表明，贡嘎山麦吊杉群落的Shannon-Wiener指数在2.33-3.26之间，Simpson指数在0.43-0.70之间，种间相遇(PIE)在0.42-0.79之间，而以Shannon-Wiener指数为基础的均匀度在34.45%-62.90%之间。麦吊杉群落的垂直结构较为复杂，分层明显。一般可划分出乔木层、灌木层、草本层和活地被层四个基本层次，乔木层通常还可划分为2-3个亚层，由高海拔至低海拔成层现象有逐渐复杂的趋势。构成麦吊杉群落的层片类型较多，其中以常绿针叶大高位芽植物为其建群层片。麦吊杉林中的层外植物较丰富，种子植物、蕨类植物、以及苔藓植物和地衣植物皆可构成其空中层片，其中苔藓植物生长繁茂，反映出群落结构的完整性。麦吊杉群落结构的水平分化较明显，经研究，其密度值在0.32-0.41之间，平均距离在1.56-1.41之间。应用Raunkiaer的频度指数方法对整个麦吊杉群落的种群频度进行调查后表明，其频度分布情况与Raunkiaer的频度定律基本上是符合的，即A>B> Cchinensis)呈Poisson分布外，其余优势种多呈集群分布。在种间联结测定时，本文选用了联结系数方法对62种乔木和60种灌木的联结特征进行了测定，结果表明，该群落可以细划为四个结构组分，各组分都有其不同的种群生态组合。本文以立木级结构代替种群年龄结构，对麦吊杉种群的动态进行了分析，结合各种群存活曲线的分析，表明麦吊杉种群正界于相对稳定与衰退时期，由于补充率小于死亡率，衰退的趋势已明显表现出来。本文还从区系成分、生活型组成、生境特点、层片结构、结构分化等方面对麦吊杉群落的特点进行了分析和讨论，并对麦吊杉种群分布格局的成因进行了探讨。