Study of physicochemical, microbial and sensory properties of surimi produced from black mouth croaker (Atrobucca nibe) at freezing condition (-18ºC)

Black mouth croaker (Atrobucca nibe) is considered as a new valuable fish stock in the Oman Sea. In this study, surimi was manufactured from nonmarket size of the fish, manually and different cryoprotectant agents were added to the surimi. Finally changes in physiochemical, microbiological and sensory quality, characteristics of the surimi and kamaboko gel samples were assessed during 6 months at freezing storage (-18ºC). Surimi samples with the addition of Iranian tragacanth gum (TG), xanthan gum (XG), chitosan (CS) and whey protein concentrate (WPC) at 1% (w/w) were prepared to evaluate their impacts as a cryoprotectant on the surimi, individually. The results showed that the whiteness and lightness indexes in all surimi samples were gradually decreased during frozen storage. This trend of decreasing was more intensity in the control sample from 61.08±0.131 to 54.21±0.067 was recorded (p<0.05). Water holding capacity (WHC) in all treatments was decreased during 6 months. The lowest WHC (g/g) was obtained in the surimi without cryoprotectants and maximum WHC was measured in Tcs and Twpc samples, respectively (p<0.05). The lowest breaking force was calculated in Txg (166.00±22.627 g) and Tc (271.50±263.16 g) during 6 months at frozen storage, respectively (p<0.05), while Twpc treatment with slight variations showed the highest breaking force (p<0.05). Also, the lowest gel strength was obtained in Txg (68.22±6.740 g.cm) after 6 month of frozen storage (p<0.05). All Kamaboko surimi gels texture profile analysis parameters decreaced with increasing shelf life. This decreasing trend in the control sample was more severe. Floding results were reduced in all samples during storage (p<0.05). The best protective results probably were obtained in WPC, chitosan and commercial cryoprotectant agents, respectively due to protein stabilization of myofibrillar proteins and the protein-protein network structure, leading to the formation of surimi gel with strong textural properties during frozen conditions. The average number of surimi polygonal structures were significantly decreased (number per mm2) and their area were significantly increased (μm2) in all treatments (p<0.05). With increasing storage time, moisture, protein contents and pH were decreaced. Maximun TVB-N index was calculated in Tc (7.93±0.400 mg/100g) and Txg (7.88±0.477), respectively (p<0.05). TBRAs index was increased in all treatments during frozen storage, while this trend was reached in maximum value in Tc (p<0.05). Sensory evaluation of the fish finger quality characteristics (color, odor, texture and overall acceptability) preapare from frozen black mouth croaker surimi was decreaced during 6 month frozen storage. After the period of frozen storage the highest quality scores were measured in Twpc, Tcs and Tcc samples, respectively (p<0.05). In this study, coliform bacteria were not found in all treatments during frozen storage. The surimi sample containing chitosan showed lower mesophilic and psychrotropic bacteria (log cfu/g) than other treatments during frozen storage (p<0.05). Salt-soluble proteins extractions of all treatments were decreased during frozen storage. This decreacing trend was highest in Tcs (45.74±0.176%) and lowest in Tc treatments after 6 month of frozen storage (29.92±0.224%) (p<0.05). Although commercial cryoprotectant agents were successful in limiting the denaturation of proteins but sugar contents were not accepted for diabetics or those who disagree with the sweet taste and high calorie food. Hence, commercial cryoprotectant agents can be replaced with whey protein concentrate and chitosan at 1% level (w/w) consider that they were showed proper protection of the surimi myofibrillar proteins during storage.

The study of biological characteristics and determination of population dynamic parameters of sawtooth barracuda (Sphyraena putnamae) in the Persian Gulf (Hormozgun Province waters)

The biological characteristics and population dynamisms of Sphyraena putnamae, were studied in the northern Persian Gulf and Oman Sea restricted to Hormuzgan province waters within 13 months period, from November 2006 up to November 2007. Biometrical and anatomical measurements were carried out, and biological surveys were conducted on 486 specimens. On the other hand, the growth and mortality parameters were estimated by using 3096 samples. These samples were collected from 3 landings, namely Bandar Abbas, Bandar Lengeh and Bandar Jask. The measurements of the minimum and maximum Fork lengths and weights were 11.7 to 8.03 cm and 135.0 to 4140.0 g, respectively. The results indicated that this species, having the Relative Length of Gut, RLG=0.34±0.002, is strongly carnivorous (often fish-eater), proven by the fact that more than 98% of its stomach contents were fish pieces. Examining the changes in the index of stomach emptiness by the percentage of CV = 0.47% indicates that this fish is Moderate feeder. The level of feeding increased in March, before spawning and decreased in June and September, simultaneously with the spawning season. There are 2 peaks of reproduction or spawning seasons during the months of April-May and September, of which the prior is assumed as the main spawning. The sex ratio (M:F) was calculated 0.5:1.0(X2 =2.11), which did not show a significant difference with expected level of 1:1 (P>0.05). The average absolute and relative reproduction rates of Sphyraena putnamae is respectively as follows: 1866827.1±255448.9 and 1097.7±94.3. The highest and the lowest diameter of matured egg are from 200 to 750 μ, and its average diameter is 402.10 ± 0.190 μ. A parameter for Saw-tooth barracuda length measurement, Lm50, based on the Fork-length, was calculated as 54.01 cm. In other words, as far as the fisheries management is concerned, the fish whose lengths are less than 54.01 cm should not be caught. The calculated level of (R2) (correlations of total length & weight), indicated strong correlations between length and weight of this fish, and the obtained formula included W =0.007100 FL 2.9295 and reinforced this assumption. The “K” Index for this fish in 3 above mentioned landings (Jask, Bandar-Abbas and Bandar-Length) were 1.24, 0.37 and 0.46 per year, respectively and the FL index for the same landings were estimated as 129, 110 and 134 cm, respectively. The growth coefficient (MONRO) for the above mentioned regions were calculated as 3.601, 3.647 and 3.917, respectively; and in the surveyed regions there were no significant differences in populations. The Total mortality coefficient (Z) was calculated 0.76, 1.12 and 1.07 per year, the Natural mortality coefficient was 0.46, 0.63 and 0.70, and the Fishing mortality coefficient (rate) (F) was found to be 0.30, 0.49 and 0.37 per year. The value of the exploitation rate (E) is equal to 0.39 per year, indicating that this species is an under-exploited resource, and there is no excessive fishing pressure on the fish supply of this species in the afore-said regions. The highest level of exploitation was found for ‘Bandar Abbas’ fishing region and the lowest level of exploitation is in ‘Bandar Lengeh’ waters.

High-order DES and zonal LES of a labyrinth seal

Hybrid numerical large eddy simulation (NLES) and detached eddy simulation (DES) methods are assessed on a labyrinth seal geometry. A high sixth order discretization scheme is used and is validated using a test case of a two dimensional vortex. The hybrid approach adopts a new blending function and along with DES is initially validated using a simple cavity flow. The NLES method is also validated outside of RANS zones. It is found that there is very little resolved turbulence in the cavity for the DES simulation. For the labyrinth seal calculations the DES approach is problematic giving virtually no resolved turbulence content. It is seen that over the tooth tips the extent of the LES region is small and is likely to be a strong contributor to excessive flow damping in these regions. On the other hand the zonal Hamilton-Jacobi approach did not suffer from this trait. In both cases the meshes used are considered to be hybrid RANS-LES adequate. Fortunately (or perhaps unfortunately) the DES profiles are in agreement with the time mean experimental measurements. It is concluded that for an inexperienced CFD practitioner this could have wider implications particularly if transient results such as unsteady loading are desired. Copyright © 2012 by the American Institute of Aeronautics and Astronautics, Inc.

Extraction of genomic DNA using a new amino silica monolithic column

A new amino silica monolithic column was developed for DNA extraction in a miniaturized format. The monolithic column was prepared in situ by polymerization of tetraethoxysilane (TEOS) and N-(beta-aminoethyl)-gamma-aminopropylmethyldimethoxysilane (AEAPMDMS). DNA was loaded in 50 mM tris(hydroxylmethyl)aminomethane-EDTA buffer at pH 7.0 and eluted with 300 mM potassium phosphate solution at pH 10.0. Under optimal condition, a 6.0-cm monolithic column provided a capacity of 56 ng DNA with an extraction efficiency of 71 +/- 5.2% (X +/- RSD). When the amino silica monolithic column was applied to extract genomic DNA from the whole blood of crucian carp, an extraction efficiency of 52 +/- 5.6% (X +/- SD) was obtained by three extractions. Since the chaotropic-based sample loading and organic solvent wash steps were avoided in this procedure, the purified DNA was suitable for downstream processes such as PCR. This amino silica monolithic column was demonstrated to allow rapid and efficient DNA purification in microscale.

Description of a new genus and two new species of labeonine fishes from South China (Teleostei : Cyprinidae)

A new genus and two new species are described from the Pearl River drainage in Guangxi Province, South China. Hongshuia, new genus, can be distinguished from all other Asian genera of the Labeonini by having a lower lip with its median lobe modified into a round, fleshy plate peripherally greatly protruded so as to form a ring-like fold that is posteromedially continuous with the mental region, and centrally sunken so as to form a round, flat, fleshy pad. This genus is distinct from all other Asian labeonine genera of the Garrina except for one newly described species of Parasinilabeo ( P. longibarbus), Pseudocrossocheilus, and Sinocrossocheilus, in the presence of well-developed maxillary barbels. Hongshuia differs from the above three genera in the lower lip morphology, and further from both Pseudocrossocheilus and Qianlabeo in the number of pharyngeal tooth rows and from Sinocrossocheilus in the colour pattern. Two new species, H. banmo and H. paoli, differ in the distribution density and degree of development of papillae on the rostral fold, depth of indentations on the distal edge of the rostral fold, presence or absence of papillae on the lower lip, size and shape of tubercles on the tip of the snout and anterior portion of the lachrymal, length, position and colour pattern of the dorsal fin, and snout length.

Redescription of Nais inflata Liang (Annelida : Oligochaeta) from China

Nais inflata Liang 1963 is redescribed on the basis of fully mature specimens collected from several localities of the Yangtze River and Yellow River. The observations and redescription resulting from the study of mature specimens supports N. inflata as a valid species. This species is characterized by a thickened body, the ventral crotchets all of the same type with distal tooth usually longer than proximal one, a greater number of dorsal chaetae per bundle, stout dorsal needles with equal minute bifid teeth (<1 mu m), presence of penial chaetae, and prostate glands on vasa deferentia instead of atria. The species is most closely related to N. communis Piguet, 1906 and N. variabilis Piguet, 1906. A table comparing allied species is provided.

Optimization of an effective extraction procedure for the analysis of microcystins in soils and lake sediments

Microcystin analysis in sediments and soils is considered very difficult due to low recovery for extraction. This is the primary limiting factor for understanding the fate of toxins in the interface between water and sediment in both the aquatic ecosystem as well as in soils. In the present study, a wide range of extraction solvents were evaluated over a wide range of pH, extraction approaches and equilibration time to optimize an effective extraction procedure for the analysis of microcystins in soils and lake sediments. The number of extractions required and acids in extraction solutions were also studied. In this procedure, EDTA-sodium pyrophosphate solution was selected as an extraction solvent based on the adsorption mechanism study. The optimized procedure proved to be highly efficient and achieved over 90% recovery. Finally, the developed procedure was applied to field soil and sediment sample collected from Chinese lakes during bloom seasons and microcystins were determined in six of ten samples. (c) 2005 Elsevier Ltd. All rights reserved.

Linichthys: A new genus of Chinese cyprinid fishes (Teleostei : Cypriniformes)

yLinichthys, new genus, is proposed for Barbodes laticeps Lin and Zhang, 1986. It can be separated from all other genera of Asian Barbinae in having a shallow depression extending along the median portion of the upper lip. Linichthys is superficially very similar to the type species of a monotypic Chinese cyprinid cavefish genus, Typhlobarbus nudiventris, in having an elongate and somewhat compressed body form, a truncate distal margin of the dorsal fin, and a soft and smooth last unbranched dorsal-fin ray, all of them being atypical for most of Asian barbins. The two genera differ in size of the eye and the number of rows of pharyngeal tooth. Although the same body form and/or structure of the dorsal fin are also shared with some species currently placed in other genera of Asian barbins, the recognition of Linichthys is warranted because of their marked differences in other characters. A redescription is also provided for the type species of the monotypic genus, Linichthys laticeps. It is known only from the upper and middle Zhu Jiang basin and the upper Chang Jiang basin in the city of Guiyang, Guizhou Province, South China.

Redescription of Difflugia tuberspinifera Hu, Shen, Gu et Gong, 1997 (Protozoa : Rhizopoda : Arcellinida : Difflugiidae) from China

The freshwater testate amoeba Difflugia tuberspinifera Hu et al. 1997 collected from pond and lake in China, is investigated by light and scanning electron microscopy. This little known taxon is redescribed and its morphology, biometry and ecology are supplied. After carefully comparison with other six similar species including Difflugia bartosi Stepanek, D. corona Wallich, D. corona cashi Deflandre, D. corona tuberculata Vucetich, D. muriformis Gauthier-Lievre et Thomas and Netzelia tuberculata (Wallich) Netzal we believe that the sub-spherical to spherical shell, the mulberry-shaped appearance, the 7-10 apertural tooth-like structures, the short collar and the conical spines numbering from 4 to 8 at the upper equatorial region in D. tuberspinifera set it apart from other species. Besides, statistical analysis indicates that D. tuberspinifera is a size-monomorphic species characterized by a main-size class and a small size range and the shell height is significant correlated with other morphometric characters at p < 0.05 excepting the number of aperture tooth-like structures and the number of spines. Moreover, D. tuberspinifera inhabits not only lotic but also lentic environment.

Microorganisms linked to Neoproterozoic microspar carbonate sedimentation in the Jilin-Liaoning area

Molar-tooth carbonate refers to a sort of rock that has ptygmatical folded structure comparable to the ivory. This kind of carbonate exists in a special time range (from Middle to Neoproterozoic). Its origin and the possibility to use it in stratigraphic correlation of the paleocontinent is the key task of the IGCP447, a project on Proterozoic molar tooth carbonates and the evolution of the earth (2001-2005). The importance lies in that the molar-tooth structure is the key to solving problems related to Precambrian biological and global geochemical events. The molar-tooth structure is associated with microorganisms. Development and recession of such carbonates have relations with the evolution process of early lives and abrupt changes in sea carbonate geochemistry. In recent years, based on researches on petrology, geochemistry and Sr isotope of molar-tooth carbonate in the Jilin-Liaoning and Xuzhou-Huaiyang area, the authors hold that it can be used as a marker for stratigraphic sequence and sedimentary facies analyses.

Coupling coefficients of gratings with graded refractive index waveguides

In this paper, the effective coupling coefficient k(eff) and the self-coupling coefficient zeta(1) are introduced to describe the characteristic of gratings in a resonant situation when the effects of radiation and other partial waves coupling are considered. The dependence of these two coupling coefficients on grating tooth shapes and depths and the dimensions of graded refractive index (GRIN) waveguides is numerically analysed. The results show that the gratings with linear GRIN waveguides have the largest \k(eff)\. The possibility of realizing a complex-coupled DFB laser, even a pure gain or loss coupled DFB laser, employing only a real refractive index coupled grating is also discussed.

Microorganisms linked to Neoproterozoic microspar carbonate sedimentation in the Jilin-Liaoning area

Molar-tooth carbonate refers to a sort of rock that has ptygmatical folded structure comparable to the ivory. This kind of carbonate exists in a special time range (from Middle to Neoproterozoic). Its origin and the possibility to use it in stratigraphic correlation of the paleocontinent is the key task of the IGCP447, a project on Proterozoic molar tooth carbonates and the evolution of the earth (2001-2005). The importance lies in that the molar-tooth structure is the key to solving problems related to Precambrian biological and global geochemical events. The molar-tooth structure is associated with microorganisms. Development and recession of such carbonates have relations with the evolution process of early lives and abrupt changes in sea carbonate geochemistry. In recent years, based on researches on petrology, geochemistry and Sr isotope of molar-tooth carbonate in the Jilin-Liaoning and Xuzhou-Huaiyang area, the authors hold that it can be used as a marker for stratigraphic sequence and sedimentary facies analyses.

余甘子、细叶草乌及土荆皮化学成分研究

本论文由三部分共6 章组成。第一部分报道了余甘子、细叶草乌和土荆皮等三种药用植物的化学成分研究成果；第二部分报道了细叶草乌和土荆皮中分离得到的化合物的活性测试，以及这两种植物的质谱分析；第三部分概述了土荆皮的研究现状。第一部分包括1－3 章。在第1 章、第2 章和第3 章中分别报道了余甘子(Phyllanthus emblica L.) 、细叶草乌(Aconitum richardsonianum var.pseudosessiliflorum) 和土荆皮( pseudolarix kaempferi) 的化学成分。采用正、反相硅胶柱层析等各种分离方法，从余甘子中共分离出10 个化合物，其中1 个为新化合物，另外还有2 个为首次从该植物中分离得到。细叶草乌的化学成分研究尚未见报道，我们从该植物中共分离出15 个化合物，其中6 个为二萜生物碱，9个为非生物碱成分。从土荆皮中分离得到16 个化合物，其中8 个二萜、1 个三萜和7 个其它类型化合物，其中有4 个化合物为首次在该植物中分离得到；从土荆皮挥发油中分离鉴定出了22 个化合物，占挥发油总量的90%。第二部分包括4－5 章。第4 章报道了从细叶草乌和土荆皮中分离得到的13个化合物的药理活性研究，结果显示，展花乌头宁和土荆乙酸葡萄糖苷等表现出较高的组织蛋白酶K 抑制活性；土荆乙酸葡萄糖苷表现出较高的组织蛋白酶B抑制活性；8-去乙酰滇乌碱表现出较高的蛋白质酪氨酸磷酸酶抑制活性。第5 章报道了细叶草乌和土荆皮总浸膏的质谱( ESI-MS ) 分析，研究结果表明，ESI-MS 法可以简单快速地检测这两种植物的主要成分；通过ESI-MS2 分析初步探讨了一些化合物的裂解规律，尝试质谱在其结构测定中的具体应用。第三部分为第6 章。从化学成分、药理、构效关系、主要成分的定量分析、及其合成研究等方面概述了土荆皮的研究进展。 This dissertation consists of three parts. The first part elaborate thephytochemical investigation of three medicinal plants, Phyllanthus emblica L.,Aconitum richardsonianum var. pseudosessiliflorum and pseudolarix kaempferi. Thesecond part reported the bioassay of 13 constituents from Aconitum richardsonianumvar. pseudosessiliflorum and pseudolarix kaempferi, and ESI-MS analysis of these twoplant . The third part is a review on the research progress of pseudolarix kaempferi.The first part is composed of three chapters. Chapters 1-3 focus on the isolationand identification of chemical constituents from Phyllanthus emblica L., Aconitumrichardsonianum var. pseudosessiliflorum and pseudolarix kaempferi. 10 compoundsincluding a new tannin were isolated from the fruits of Phyllanthus emblica by repeatcolumn chromatography over normal and reversed phase silica gel, 2 of them werefirstly reported in this plant. The chemical constituents of Aconitum richardsonianumvar. pseudosessiliflorum never reported before, 15 compounds including 6 diterpenealkaloids were isolated and identified from the roots of this plant. 16 compoundsincluding 8 diterpenes , 1 triterpene and 7 other compounds were isolated from the bark of pseudolarix kaempferi, among them, 4 compounds were firstly reported fromthe EtOH extracts of this plant, and 22 compounds were identified from its essentialoil, representing 90% of the total essential oil.The second part includes chapters 4 and 5. Chapter 4 reported thepharmacological activities of 13 compounds isolated from Aconitum richardsonianumvar. pseudosessiliflorum and pseudolarix kaempferi. Results demonstrated that chasmanine and pseudolaric acid B-β-D-glucoside exhibit relatively high anti-Cathepsin K activities; pseudolaric acid B-β-D-glucoside exhibits relatively highanti-Cathepsin B activity; 8-deacetyl-yunaconitine exhibits relatively high anti-PTP1Bactivity. Chapter 5 reported the ESI-MS analysis of extractions from Aconitumrichardsonianum var. pseudosessiliflorum and pseudolarix kaempferi, it was showedthat ESI-MS can be used as an useful tool in analyzing the major constituents of thesetwo plant much quick and easy, in addition, the fragmentation rules of somecompounds were discussed, in order to find some applications of ESI-MS2 method in their structure determination.The third part is a review on the research progress of pseudolarix kaempferi,including the chemical constituents, pharmacology, structure-activity relationship(SAP), quantitative analysis and synthesis of the major constituents.

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.

角蟾科（Megophryidae; Anura）蝌蚪的形态多样性和生态适应

角蟾科（Megophryidae）是以角蟾属（Megophrys Kuhl and Van Hasselt, 1822）为模式属而建立的，隶于无尾目（Anura），变凹型亚目（Anomocoela）。角蟾科包括2 亚科11 属142 种，分布于东洋界，从巴基斯坦、中国西部向东直到菲律宾和苏达群岛；中国有9 属75 种分布于华中和华南地区。角蟾科被认为是原始的两栖动物之一，其分类学、系统学、生态学、动物地理学的研究均深受中外科学家的瞩目。近年来，通过形态学、古生物学、细胞学、生态学、支序系统学的研究，角蟾科的分类与系统学研究取得了较大进展。与成体形态和分子系统学研究结果相比较，蝌蚪的研究存在更多的问题和挑战，尚需深入研究：（1）角蟾科蝌蚪的形态多样性分析；（2）角蟾科的系统发育关系与蝌蚪的演化，以及口漏斗的起源；（3）角蟾科蝌蚪表型分化与栖息环境和觅食行为的适应演化。针对上述问题，本文对角蟾科9 属30 种蝌蚪的形态特征，包括外部宏观形态和口器外部结构特征、口器内部显微结构、唇齿和角质颌的亚显微结构作了深入细致、多层次的比较研究；通过12s rRNA 和cytochrome b 基因构建最大简约树，采用贝叶斯系统发育进行分析，蝌蚪型的演化采用祖先性状的重建方法分析；得到如下结论：1）初步将角蟾科蝌蚪分为4 种类型；并且建立了2 种新的角蟾科蝌蚪类型。A 型：拟髭蟾型蝌蚪，该型蝌蚪包括拟髭蟾属、髭蟾属、齿蟾属和齿突蟾属的物种；B 型：新类型，掌突蟾型蝌蚪，该型蝌蚪在本文中包括掌突蟾属、小臂蟾属的物种；C 型：新类型，短腿蟾型蝌蚪，一种特化类型，该型蝌蚪在本文中仅包括短腿蟾属的物种；D 型：角蟾型蝌蚪，该型蝌蚪在本文中包括无耳蟾属、小口拟角蟾属和异角蟾属的物种。2）对角蟾科的分类进行了修订：（1）支持角蟾科两个亚科的分类系统；（2）角蟾亚科包括拟角蟾属、异角蟾属、无耳蟾属和短腿蟾属；该亚科形态差异小，系统学关系比较复杂，暂不作族级分类的再划分；（3）拟髭蟾亚科分为2 个族：拟髭蟾族，该族物种具有类型A 的蝌蚪，包括4 个属：拟髭蟾属、髭蟾属、齿蟾属、齿突蟾属；掌突蟾族，该族物种具有类型B 的蝌蚪，包括2 个属：掌突蟾属和小臂蟾属。3）结合分子系统进化关系探讨了4 种蝌蚪类型的演化。（1）角蟾科蝌蚪的最近共同祖先来自于一类具有拟髭蟾型蝌蚪性状的蝌蚪；（2）掌突蟾型蝌蚪和角蟾亚科的蝌蚪是由具有拟髭蟾型蝌蚪性状的祖先蝌蚪分别演化而来；（3）短腿蟾型蝌蚪是角蟾型蝌蚪的一种特化类型；（4）外群蝌蚪具有与拟髭蟾型蝌蚪相似的性状，进一步印证了类拟髭蟾型蝌蚪是角蟾科蝌蚪的最近共同祖先的假说；（5）具有口漏斗的蝌蚪类型是由不具口漏斗的蝌蚪类型演化而来，在角蟾科中口漏斗是一种衍生性状。4）分析了角蟾科四种蝌蚪类型与栖息环境的适应演化。（1）角蟾科蝌蚪的口部和体形的变化反映了该科蝌蚪由缓流向类似静水生境的回水凼的渐变式适应，角蟾科蝌蚪的形态显示了多方面的适应变化；（2）随着蝌蚪类型由A 向D的演化，当水速较大时，拟髭蟾型的蝌蚪营流水攀吸型生活方式；当水速递减时，掌突蟾型蝌蚪营流水附着型生活方式；当水速进一步递减时，具有较小口漏斗的短腿蟾型蝌蚪和具有大漏斗的角蟾型蝌蚪营流水浮泳型生活。角蟾科蝌蚪对于水流递减的适应演化说明蝌蚪的生态学适应是具有进化意义的；（3）蝌蚪口器内部结构的分化揭示了蝌蚪和食性的适应关系，蝌蚪以口部的唇齿与角质颌刮取或吞吸水中的物质，然后，通过口乳突有选择地过滤进入口腔中食物。拟髭蟾亚科蝌蚪的唇齿多而窄，唇齿间距宽，颌鞘粗而稀，反映了其植食性为主的特点；它们的舌前乳突一般为指状，在口腔入口处所占面积小，其机械过滤的作用很多被唇齿和角质颌分担了；而角蟾亚科的蝌蚪，其角质颌弱，其舌前乳突一般为匙状，几乎填满了口腔入口处，因此舌前乳突起了主要的机械过滤作用。The family Megophryidae is the largest and most diverse families inArchaeobatrachia, and most of its species occur in India, Pakistan, and eastward intoChina, Southeast Asia, Borneo and the Philippines to the Sunda Islands. Currently thefamily includes 142 species have been grouped into two subfamilies, Megophryinaeand Leptobrachiinae. The mountains of central and southern China are rich in speciesof Megophryidae, 75 species belong to 9 genera and two subfamilies.The family was supposed to be ideal materials of studies in many fields of biology,such as taxonomy, evolution, systematics, ecology, and biogeography. Recently, therehave a great development in taxonomy and systematics of megophryids throughstudied by morphology, paleontology, cytology, ecology, and cladistics. However,larvae of megophryids were generally unknown, although the tadpoles might be veryimportant for above studies.In this paper, we examined the evolutionary scenario of the tadpoles’ morphologyin the context of a phylogenetic framework. Our objectives are (1) to evaluate thedivergence of larval body shape and oral discs in the family Megophryidae, (2) toexplore the evolutionary trends of the larvae in megophryidae, and test if thefunnel-shaped oral disc is apomorphic, and (3) to explore the relationship of the larvalstructure, diet and microhabitat.We examined larval morphology of 30 megophryid species, the larval body shape,oral discs, the buccopharyngeal cavity, and jaw sheaths and denticles of the Chinesemegophryid frogs were re-examined. We constructed a phylogeny of the species on thebasis of published mitochondrial cytochrome b and 16S rRNA gene segments usingpartitioned Bayesian analyses. Furthermore, hypothetical changes of larval morphologywere inferred using parsimony principle on the phylogeny. The results showed that:1) Four tadpole types in Megophryidae. The larval morphological charactersseries in Chinese megophryids fall into four general categories according to the bodyshape and oral discs: (A) Leptobrachiini type, species from genera Leptobrachium,Oreolalax, Scutiger and, Vibrissaphora share this type of tadpoles. (B) Leptolalax type,species of genus Leptolalax have this type of tadpoles. (C) Brachytarsophrys type,species of the genus Brachytarsophrys have this type of tadpoles. (D) Megophryinitype, species of the genera Atympanophrys, Ophryophryne, and Xenophrys share this type of tadpoles. Of which B and C are two novel types.2）Taxonomic implications. The present study leads us to reconsider the generalclassification of tribes attributed to members of Megophryidae. More specifically,concerning the phylogenetic relationships and the two novel tadpole types describedherein, we propose a provisional taxonomy for the family but suggest that further taxasampling of other megophryids be performed to confirm this taxonomic change. TheMegophryidae is composed of two subfamilies (Leptobrachiinae and Megophryinae).The Leptobrachiinae was recogonized the two tribes: (1) tribe Leptobrachiini sensuDubois, corresponding to the tadpole of type A, including four genera, i.e.,Leptobrachium, Oreolalax, Scutiger and, Vibrissaphora; (2) tribe Leptolalaxini,corresponding to the tadpole of novel type B, including two genera, i.e., Leptolalaxand Leptobrachella. However, the relationships among the genera of Megophryinaewere largely unresolved, they recognized no monophyletic groups above the generalevel. A more thorough sampling will likely foster a better taxonomic solution.3) The larval evolutionary scenario in Megophryidae.Type A is characteristicof normal-mouthed with multiple tooth rows, representing the tadpole type of theMRCA of Chinese megophryids. Type B is characteristic of normal-mouthed withreduced tooth rows, prolonging labium, and integumetary glands. Type C ischaracteristic of no labial teeth and smaller umbeliform oral disc. Type D ischaracteristic of no labial teeth, enlarged umbeliform oral disc, representing the tadpoleof the MRCA of subfamily Megophryinae. A previous hypothesis, referring tofunnel-shaped oral discs as an apomorphy, is supported.4) The larval adaptation to habitats in Megophryidae. Tadpoles generallyadhere to substrates using their mouths, and the microhabitat that the tadpoles occupyreflects the degree of adhesion and oral complexity. The morphological changes inmegophryid tadpoles virtually allow a progressive adaptation to a changing habitatfrom faster water to slower water. Within the tadpoles of Type A to type D, the TOTbecomes smaller and smaller, and the oral disc orientates from anteroventral toumbelliform upturned, and eye position orientates from dorsal to lateral, and the trunkis more and more depressed and tail becomes relatively longer and slender. Within therunning water, the normal-mouthed with multiple tooth rows of Leptobrachiini tadpoles are correlated with lotic-suctorial, benthic feeders with anteroventral oraldisc and the largest body. With the water’s velocity decreasing, the lotic-adherentfeeders of Leptolalax tadpoles have tube-shaped labium with reduced tooth rows andintegumetary glands. And then, the smaller umbeliform in Brachytarsophrys tadpolesand the enlarged umbeliform oral disc in the Megophryini tadpoles are inhabitmicrohabitats of non-flowing backwaters of rivers, indicative of adaptive traits oflotic-neustonic surface feeders. The scheme of megophryid tadpoles andmicrohabitats provided the first clear evidence which congruent with the hypothesis ofAltig and Johnston (1989). The ecological divergence plays a general role in thedivergence and evolution of megophrid larvae. There is a definite correlation amongthe buccopharyngeal cavity, diet and feeding mechanisms, the tadpole graze orswallow the food particles, then through papillae which like a sieve and sort out foodparticles to the oesophagus. The tadpole of Leptobrachiinae possess multiple toothrows, wide intertooth distance as well as thick and sparse jaw sheath, these tadpolesinhabit bottom of the streams and graze on epiphyton or major detritus of organicmatter on the substrates, their prelingual papillae like single finger, the mechanicalpurpose of papillae served share in by tooth and jaw. The tadpoles of Megophryinaeoccur near the water surface of small streams and are the filter feeder, their dietincludes plankton and organic debris floating on the water surface, those tadpolepossess weak jaw, their prelingual papillae like spoon, the mechanical purpose ofpapillae served mostly for sieve.