中药穿龙薯蓣、黄山药和盾叶薯蓣的分子鉴别研究

为建立对中药穿龙薯蓣、黄山药和盾叶薯蓣分子鉴别的方法，我们首先研究薯蓣叶绿体psbA-trnH片段遗传多样性，探讨该片段用于中药穿龙薯蓣、黄山药和盾叶薯蓣种间分子鉴别和系统学研究中的意义。在对不同类群薯蓣种的叶绿体psbA-trnH基因间区进行ＰＣＲ扩增并测序，获得了该区间的完整序列的基础上，将所得序列用软件MEGA3.0进行相关分析。穿龙薯蓣的psbA-trnH片段全长274bp，黄山药全长279bp，盾叶薯蓣植物个体内的叶绿体DNA则有两种psbA-trnH片段，长度分别为241bp和503bp。MEGA3.0软件分析，三种薯蓣种间psbA-trnH片段序列的遗传距离（p-distance）为0.00350-0.04545，各个薯蓣种内的不同类群该序列无差异。用UPGMA法根据psbA-trnH序列的遗传距离建立系统发生树，每个种不同产地的薯蓣很明确地聚在一起，和形态分类一致。所得结果显示psbA-trnH片段序列在所研究的三种薯蓣种内保守，在种间具有明显的较大差异，而三种薯蓣及薯蓣属的系统发生关系尚须进一步研究。 以穿龙薯蓣、黄山药和盾叶薯蓣的psbA-trnH片段序列分析结果为基础，我们根据三种薯蓣在该片段上的特征序列位点设计了用于识别三种薯蓣的寡核苷酸片段并作为PCR反应的引物，将三种薯蓣的特征标记引物与psbA-trnH片段引物配合使用建立了相互识别三种薯蓣的显性和共显性两种检测方式，辅以NaOH碱法快速提取薯蓣干燥根茎总DNA，为三种薯蓣药材相互间快速准确鉴别创造了条件。 To establish molecular method for identifying Chinese medicine Dioscorea nipponica/panthaica/zingiberensis, we firstly study the Genetic diversity of psbA-trnH of Dioscorea and discuss the value of the fragment to molecular identity and systematology to three Dioscorea species (Dioscorea nipponica/panthaica/zingiberensis). The psbA-trnH fragments of different species of Dioscorea are amplified and sequenced，analyzing by software MEGA3.0. The length of Dioscorea nipponica and panthaica are 274bp and 279bp; two psbA-trnH fragments exist in Dioscorea zingiberensis individual, length of the shorter fragment is 241bp and the longer is 503bp. The interspecific Genetic distance (p-distance) of the three species is 0.00350-0.04545, and no intraspecific diversity exists. According to the Genetic distance, phylogenetic tree is built by UPGMA Method, each species of different place gather obviously, which is same to the shape classification. Results show that the psbA-trnH fragment is highly conservative at intraspecific level and obvious diversity at interspecific level, however, phylogenetic study is necessary for the Dioscorea in the future. Based on analysis to psbA-trnH fragment of Dioscorea panthaica, nipponica and zingiberensis, according to the special point of the three Yam species, primers as marker are designed for distinguishing the three species. Cooperating with primer pair psbAf-trnHr, dominant and codominant way is established for discriminating them. Moreover, assisted with NaOH method to extract total DNA of Dry stenophora of Yam, rapid and correct identity to the three species can be done.

Phosphoproteome analysis of mouse liver using immobilized metal affinity purification and linear ion trap mass spectrometry

Since protein phosphorylation is a dominant mechanism of information transfer in cells, there is a great need for methods capable of accurately elucidating sites of phosphorylation. In recent years mass spectrometry has become an increasingly viable alternative to more traditional methods of phosphorylation analysis. The present study used immobilized metal affinity chromatography (IMAC coupled with a linear ion trap mass spectrometer to analyze phosphorylated proteins in mouse liver. A total of 26 peptide sequences defining 26 sites of phosphorylation were determined. Although this number of identified phosphoproteins is not large, the approach is still of interest because a series of conservative criteria were adopted in data analysis. We note that, although the binding of non-phosphorylated peptides to the IMAC column was apparent, the improvements in high-speed scanning and quality of MS/MS spectra provided by the linear ion trap contributed to the phosphoprotein identification. Further analysis demonstrated that MS/MS/MS analysis was necessary to exclude the false-positive matches resulting from the MS/MS experiments, especially for multiphosphorylated peptides. The use of the linear ion trap considerably enabled exploitation of nanoflow-HPLC/MS/MS, and in addition MS/MS/MS has great potential in phosphoproteome research of relatively complex samples. Copyright (C) 2004 John Wiley Sons, Ltd.

Generation and analysis of 5318 expressed sequence tags from the filamentous sporophyte of Porphyra haitanensis (Rhodophyta)

Porphyra haitanensis T. J. Chang et B. F. Zheng (Bangiales, Rhodophyta) is cultivated in China and widely consumed in Asia. To gain more insight into its physiological and biochemical properties, we generated 5318 expressed sequence tags (ESTs) from the sporophyte of P. haitanensis, and upon assembling into a nonredundant set, 2535 sequences were obtained, among which only 32.2% (816) shared certain similarity with published sequences (Nr and KOG). Functional classification of such ESTs revealed that most of the transcripts were related to its conservative biological metabolism, and P. haitanensis most likely possesses cyanide-resistant respiration and a C4-like carbon-fixation pathway, both of which have never been reported in a rhodophyte before. Twenty-eight percent of the nonredundant gene clusters exhibited significant similarity to those from P. yezoensis Ueda sporophytes, and 16 genes up-regulated in P. yezoensis sporophytes were also expressed abundantly in P. haitanensis. Codon usage analysis indicated that exposure to high GC pressure might occur during evolution of P. haitanensis. These findings represent the most extensive collection of ESTs from P. haitanensis to date, and all the ESTs in this study have been submitted to GenBank (accession nos. DN604790-DN608469, EG016226-EG018540).

Reviews on The Immunological Function of Tetraspanins

Tetraspanins belongs to the transmembrane 4 superfamily(TM4SF). They can be as a bridge to connect the proteins outside or inside the cell membrane. A tetraspanins web is formed by the tetraspnins-proteins complex, and the web is believed to involve in fundamental functions of immunity system, and consequnently, signaling between cells and inside cells, regulating cell activation and adhesion, participating in the identification and infection of some virus. As a family of conservative transmembrane proteins, tetraspanins play multiplex roles in invertebrate. It was described how tetraspanin microdomains might have functions in the immune system, and how they contact with virus. In addition, the important role of tetraspanins in the innate immune system of invertebrate were discussed.

The nitrogen isotopic composition of dissolved nitrate in the Yangtze River (Changjiang) estuary, China

The nitrogen isotopic composition of dissolved nitrate (delta N-15-NO3-) in surface water of the Yangtze River estuary was determined in four seasons of 2006. delta N-15-NO3- ranged from 0.4 parts per thousand to 6.5 parts per thousand and varied with seasons and geographic regions, reflecting the dynamics of nitrogen cycling in the estuarine ecosystem. delta N-15-NO3- was markedly lower in February than in other seasons and exhibited conservative mixing, which was probably attributed to the NO3- being sourced from the atmospheric deposition and agricultural fertilizer. In the upper estuary, the influence of riverine inputs was important during all surveys. in the turbidity maximum zone, nitrification was found with nitrate depleted in N-15 in May, whereas denitrification resulting in heavy delta N-15-NO3- played an important role in August. More enriched delta N-15-NO3- values coinciding with losses of nitrate concentrations based on the conservative mixing model were found in the adjacent marine area in May, and may reflect obvious phytoplankton assimilation of dissolved nitrate. In this manner, delta N-15-NO3- may be a sensitive indicator of nitrogen sources and biogeochemical processing existing in this estuary in conjunction with the variations of dissolved nitrate and other environmental factors. (C) 2009 Elsevier Ltd. All rights reserved.

Morphological and genetic study of Prorocentrum donghaiense Lu from the East China Sea, and comparison with some related Prorocentrum species

In this paper, the detailed morphology of Prorocentrum donghaiense Lu from both field samples and cultures was examined, and a taxonomic comparison was made between P donghaiense and some related Prorocentrum spp. using morphological and molecular data and other published information. There were distinct differences among these species in morphological characteristics that historically have been presented as conservative features. The discrepancies extended beyond that of individual variations within the same species due to environmental factors. Therefore, these morphological features may not be conservative but, rather, polymorphic depending on environmental conditions. Based on this analysis, we suggest that the high-biomass bloom-forming species in the East China Sea, previously reported as Prorocentrum dentatum Stein, is P donghaiense Lu. The species reported from the East China Sea and Japanese and Korean waters appear to be the same species. Molecular data also suggest that P. dentatum (CCMP1517) and P. donghaiense are genetically identical. Therefore, the geographic distribution of P. donghaiense may be much wider than expected. (C) 2004 Elsevier B.V. All rights reserved.

Sources and transport of dissolved and particulate organic carbon in the Mississippi River estuary and adjacent coastal waters of the northern Gulf of Mexico

Dissolved organic carbon (DOC), stable carbon isotopic (delta(13)C) compositions of DOC and particulate organic carbon (POC), and elemental C/N ratios of POC were measured for samples collected from the lower Mississippi and Atchafalaya rivers and adjacent coastal waters in the northern Gulf of Mexico during the low flow season in June 2000 and high flow season in April 2001. These isotopic and C/N results combined with DOC measurements were used to assess the sources and transport of terrestrial organic matter from the Mississippi and Atchafalaya rivers to the coastal region in the northern Gulf of Mexico. delta(13)C values of both POC (-23.8parts per thousand to -26.8parts per thousand) and DOC (-25.0parts per thousand to -29.0parts per thousand) carried by the two rivers were more depleted than the values measured for the samples collected in the offshore waters. Strong seasonal variations in delta(13)C distributions were observed for both POC and DOC in the surface waters of the region. Fresh water discharge and horizontal mixing played important roles in the distribution and transport of terrestrial POC and DOC offshore. Our results indicate that both POC and DOC exhibited non-conservative behavior during the mixing especially in the mid-salinity range. Based on a simple two end-member mixing model, the comparison of the measured DOC-delta(13)C with the calculated conservative isotopic mixing curve indicated that there was a significant in situ production of marine-derived DOC in the mid- to high-salinity waters consistent with our in situ chlorophyll-a measurements. Our DOC-delta(13)C data suggest that a removal of terrestrial DOC mainly occurred in the high-salinity (>25) waters during the mixing. Our study indicates that the mid- to high- (10-30) salinity range was the most dynamic zone for organic carbon transport and cycling in the Mississippi River estuary. Variability in isotopic and elemental compositions along with variability in DOC and POC concentrations suggest that autochthonous production, bacterial utilization, and photo-oxidation could all play important roles in regulating and removing terrestrial DOC in the northern Gulf of Mexico and further study of these individual processes is warranted. (C) 2004 Elsevier B.V. All rights reserved.

强化学习算法中启发式回报函数的设计及其收敛性分析

回报函数设计的好与坏对学习系统性能有着重要作用,按回报值在状态-动作空间中的分布情况,将回报函数的构建分为两种形式:密集函数和稀疏函数,分析了密集函数和稀疏函数的特点.提出启发式回报函数的基本设计思路,利用基于保守势函数差分形式的附加回报函数,给学习系统提供更多的启发式信息,并对算法的最优策略不变性和迭代收敛性进行了证明.启发式回报函数能够引导学习,加快学习进程,从而可以实现强化学习在实际大型复杂系统应用中的实时控制和调度.

悬索桥隧道式锚碇与围岩系统的破坏模式研究

The anchorages are unparalleled structures only in a suspension bridge, and as main bearing facilities, play an important role in connecting the superstructures and the ground. The tunnel anchorage, as one alternative type of the anchorages, has more advantages over its counterpart, the gravity anchorage. With the tunnel anchorages adopted, not only can surface excavation be reduced to protect the environment, and natural condition of the rock be utilized and potential bearing capacity of surrounding rock be mobilized to save engineering cost, but also the technological predominance of auxiliary engineering measures, such as prestressed concrete, anchoring piles, rock anchors and collar beam between the two separated anchorages, can be easily cooperated to work together harmoniously under the circumstances of poor rock quality. There are plentiful high mountains and deep canyons in west part of China, and long-span bridge construction is inevitably encountered in order to realize leapfrogging development of the transportation infrastructure. Western mountainous areas usually possess the conditions for constructing tunnel anchorages, and therefore, the tunnel anchorages, which are conformed to the conception of resource conservative and sustainable society, extremely have application and popularization value in western underdeveloped region. The scientific and technological problem about the design, construction and operation of tunnel anchorages should be further investigated. Combining the engineering of western tunnel anchorages for the Balinghe Suspension Bridge, this paper probed into the survey method and in-situ test method for tunnel anchorages, scientific rock quality evaluation of surrounding rock to provide reasonable physical and mechanical parameters for design, construction and operation of tunnel anchorages, bearing capacity estimation for tunnel anchorage, deformation prediction of the anchorage-rockmass system, tunnel-anchorage slope stability analysis and the evaluation of excavation stability and degree of safety of the anchorage tunnel. The following outcomes were obtained: 1. Materials of tunnel anchorages of suspension bridge built (and in progress) at home and abroad were systematically sorted out, with the engineering geological condition and geomechanical property of surrounding rock around the anchorage tunnel, the design size of anchorages and the construction method of anchorage tunnel paid more emphasis on, to unveil the internal relationship between the engineering geological conditions of surrounding rock and the design size and axis angle of anchorages and provide references for future design, construction and study of tunnel anchorages. 2. Physical and mechanical parameters were recommended based on three domestic and foreign methods of rock quality evaluation. 3. In-situ tests, adopting the back-thrust method, of two kinds of reduced scale model, 1/30 and 1/20, for the tunnel anchorages were conducted in the declining exploration drift with rock mass at the test depth being the same as surrounding rock around real anchorages, and reliable field rockmass displacement data were acquired. Attenuation relation between the increment of distance from the anchorage and the decrement of rockmass displacement under maximum test load, and influential scope suffered by anchorage load were obtained. 4. Using similarity theory, the magnitude of real anchorage and rockmass displacement under design load and degree of safety of the anchorage system were deduced. Furthermore, inversion analysis to deformation modulus of slightly weathered dolomite rock, the surrounding rock of anchorage tunnel, was performed by the means of numerical simulation. 5. The influential law of the geometrical size to the limit bearing capacity of tunnel anchorage was studied. 6. Based on engineering geological survey data, accounting for the combination of strata layer and adverse discontinuities, the failure patterns of tunnel anchorage slope were divided into three modes: sliding of splay saddle pier slope, superficial-layer slippage, and deep-layer slippage. Using virtual work principle and taking anchorage load in account, the stability of the three kinds of failure patterns were analyzed in detail. 7. The step-by-step excavation of anchorage tunnel, the numerical overload and the staged decrement of rock strength parameters were numerically simulated to evaluate the excavation stability of surrounding rock around anchorage tunnel, the overload performance of tunnel anchorage, and the safety margin of strength parameters of the surrounding rock.