Nuclear transfer in leach (Paramisgurnus dabryanus Sauvage) by cell-to-cell electrofusion

To study nuclear transfer in the leach (Paramisgurnus dabryanus Sauvage), blastula and gastrula cells were fused with UV-inactivated oocytes by cell-to-cell electrofusion. To facilitate nuclear transfer, blastula and gastrula cells were cultured or incubated at 4 degreesC in different solutions. TC-199 medium supplemented with 20% calf serum was the best culture solution, and effectively retained the totipotence of blastula or gastrula cells for up to 10 days, It was found that gastrula cells incubated at 4 degreesC had the same totipotence as blastula cells, The optimal UV dosage for inactivation of the oocyte chromatin was 180-240 mJ cm(-2). Electrofusion was carried out in a cone-shaped fusion chamber, which permitted the recipient oocyte and the donor blastula cell to contact one another. The electrofusion procedure resulted in a 10% success rate of normal-appearing fish. Genetic analysis indicated that the nuclear material originated from the donor cell (blastomere) and the oocyte pronucleus did not take part in development.

Consensus scoring for enriching near-native structures from protein-protein docking decoys

The identification of near native protein-protein complexes among a set of decoys remains highly challenging. A stategy for improving the success rate of near native detection is to enrich near native docking decoys in a small number of top ranked decoys. Recently, we found that a combination of three scoring functions (energy, conservation, and interface propensity) can predict the location of binding interface regions with reasonable accuracy. Here, these three scoring functions are modified and combined into a consensus scoring function called ENDES for enriching near native docking decoys. We found that all individual scores result in enrichment for the majority of 28 targets in ZDOCK2.3 decoy set and the 22 targets in Benchmark 2.0. Among the three scores, the interface propensity score yields the highest enrichment in both sets of protein complexes. When these scores are combined into the ENDES consensus score, a significant increase in enrichment of near-native structures is found. For example, when 2000 dock decoys are reduced to 200 decoys by ENDES, the fraction of near-native structures in docking decoys increases by a factor of about six in average. ENDES was implemented into a computer program that is available for download at http://sparks.informatics.iupui.edu.

VEGF gene silencing by cytomegalovirus promoter driven ShRNA expression vector results in vascular development defects in zebrafish

Zebrafish has been generally considered as an excellent model in case of drug screening, disease model establishment, and vertebrate embryonic development study. In this work, the ability of human cytomegalovirus immediate early promoter (CMV promoter)-driven short hairpin RNA (shRNA) expression vector to induce shRNA against VEGF gene in zebrafish was tested, and its effect on vascular development was assed, too. Using RT-qPCR, blood vessel staining, and in situ hybridization, we confirmed certain transcriptional activity and down regulation of gene expression by the vector. In situ hybridization analysis indicated selective inhibition of NRP1 expression in the VEGF gene loss of function model, which might imply in turn that VEGF could not only activate endothelial cells directly but also could contribute to stimulating angiogenesis in vivo by a mechanism that involved up-regulation of its cognate receptor expression in zebrafish. This contributed to a better understanding of molecular mechanisms of cardiovascular development. The system improved the success rate in making inducible knockdown and widened the possibilities for better therapeutic targets in zebrafish.

全信息精细储层预测方法研究与实践-以周清庄油田沙三段油藏为例

With the continuously proceeding of petroleum exploratory development in China, exploratory development becomes more and more difficult. For increasing reserve volume and production, lithologic hydrocarbon reservoir has been the most workable, potential and universality exploration targets. In the past, Dagang Oil Field use the complicated fault reservoir theory as the guide, develop and form a suit of matching construction and instrument in prospecting complicated fault reservoir that reach top of exploration industry in China. But the research of lithologic hydrocarbon reservoir is not much, which affects the exploitation progress of lithologic hydrocarbon reservoir. In this thesis, is object, through the depth study of lithologic deposition in Shasan segment of Zhouqingzhuang Oil Field, a suit of holographic fine reservoir bed forecasting techniques is built up and finally gets following main results: 1. Applying geology, seism, drilling, logging and other information to sensitivity preferences, geological model, inversion and integrated stratum evaluation, realizing the method and flow of refined multi-information stratum forecast. 2. Built up a full three dimensional fine structural interpretation method: in view of r problem of accurately demarcating 90% inclined well, propose a inclined well air space demarcating method, make bed demarcating more exactly; in view of problem of faults demarcating and combination in seismic interpretation, propose a computational method of seismic interference based on wavelet translation, make identify the fault in different level more dependable and reasonable; for exactly identifying structural attitude, propose a velocity modeling method under multi-well restriction, make structural attitude closer to the facts. 3. Built up a high accuracy reservoir bed inversion method: in view of problem in exactly identifying reservoir and nonreservoir with conventional wave impedance inversion method in this place, propose a reservoir log response characteristic analysis and sensible log parameter inversion method. ①analysis log response of reservoir and nonreservoir in region of interest, make definite the most sensible log parameter in identifying reservoir and nonreservoir in this region; ②make sensible log parameter inversion based on wave impedance inversion, to improve inversion accuracy, the thickness of recognizable reservoir bed reach 4-5m. 4. Built up a 4-D reservoir forcasting circuit: in view of difficulty that in lithologic hydrocarbon reservoir making reservoir space characteristic clear by using structural map and reservoir forecasting techniques once only, propose a 4-D reservoir forcasting circuit. In other words, based on development conceptual design, forcast reservoir of different time, namely multiple 3D reservoir forcasting in time queue, each time the accuracy degree of reservoir forcasting is improved since apply the new well material, thereby achieve high quality and highly efficient in exploratory development. During exploratory development lithologic depositin in Shasan segment of Zhouqingzhuang Oil Field, there are thirteen wells get 100% success rate, which sufficiently proves that this suit of method is scientific and effective.

鄂尔多斯盆地上古生界低渗透砂岩有效储层识别与预测

Sulige Gasfield, with a basically proven reserve as high as one trillion cubic meters, is one giant gas field discovered in China. The major gas -bearing layers are Upper Paleozoic strata with fluvial-lacustrine sedimentary facies. Generally, gas reservoirs in this field are characteristic by "five low" properties, namely low porosity, low permeability, low formation pressure, low productivity and low gas abundance. Reservoirs in this field also feature in a large distribution area, thin single sandbody thickness, poor reservoir physical properties, thin effective reservoir thickness, sharp horizontal and/or vertical changes in reservoir properties as well as poor connectivity between different reservoirs. Although outstanding achievements have been acquired in this field, there are still several problems in the evaluation and development of the reservoirs, such as: the relation between seismic attributes and reservoir property parameters is not exclusive, which yields more than one solution in using seismic attributes to predict reservoir parameters; the wave impedance distribution ranges of sandstone and mudstone are overlapped, means it is impossible to distinguish them through the application of post-stack impedance inversion; studies on seismic petrophysics, reservoir geophysical properties, wave reflection models and AVO features have a poor foundation, makes it difficult to recognize the specific differences between tight sandstone and gas-bearing sandstone and their distribution laws. These are the main reasons causing the low well drilling success rate and poor economic returns, which usually result in ineffective development and utilization of the field. Therefore, it is of great importance to perform studies on identification and prediction of effective reservoirs in low permeable sandstone strata. Taking the 2D and 3D multiwave-multicomponent seismic exploration block in Su6-Su5 area of Sulige field as a study area and He 8 member as target bed, analysis of the target bed sedimentary characteristics and logging data properties are performed, while criteria to identify effective reservoirs are determined. Then, techniques and technologies such as pre-stack seismic information (AVO, elastic impedance, wave-let absorption attenuation) and Gamma inversion, reservoir litological and geophysical properties prediction are used to increase the precision in identifying and predicting effective reservoirs; while P-wave and S-wave impedance, ratio of P/S wave velocities, rock elastic parameters and elastic impedance are used to perform sandstone gas-bearing property identification and gas reservoir thickness prediction. Innovative achievements are summarized as follows: 1. The study of this thesis is the first time that multiwave-multicomponent seismic data are used to identify and predict non-marine classic reservoirs in China. Through the application of multiwave-multicomponents seismic data and integration of both pre-stack and post-stack seismic data, a set of workflows and methods to perform high-precision prediction of effective reservoirs in low permeable sandstone is established systematically. 2. Four key techniques to perform effective reservoir prediction including AVO analysis, pre-stack elastic wave impedance inversion, elastic parameters inversion, and absorption attenuation analysis are developed, utilizing pre-stack seismic data to the utmost and increasing the correct rate for effective reservoir prediction to 83% from the former 67% with routine methods. 3. This thesis summarizes techniques and technologies used in the identification reservoir gas-bearing properties using multiwave-multicomponent seismic data. And for the first time, quantitative analysis on reservoir fluids such as oil, gas, and/or water are carried out, and characteristic lithology prediction techniques through the integration of pre-stack and post-stack seismic prediction techniques, common seismic inversion and rock elastic parameters inversion, as well as P-wave inversion and converted wave inversion is put forward, further increasing the correct rate of effective reservoir prediction in this area to 90%. 4. Ten seismic attribute parameters are selected in the 3D multi-wave area to perform a comprehensive evaluation on effective reservoirs using weighted-factor method. The results show that the first class effective reservoir covers an area of 10.08% of the study area, while the second and the third class reservoirs take 43.8% and 46% respectively, sharply increasing the success rate for appraisal and development wells.

松辽盆地南部腰英台区块低渗透储层整体压裂改造技术研究

The Yaoyingtai Block is located within the northeastern Changling Depression of southern Songliao Basin, where the reservoir sandstones are petrophysically characterized by very low permeability, which results in the low success probability of artificial fracturing, and the low oil yield by water injection in the course of oil production. In order to improve the situations as stated above, this research aims to work out an integral fracturing technology and strategy applicable to the low permeable reservoirs in Yaoyingtai Block. Under the guidance of geological theory, reservoir engineering and technology, the subsurface occurrences of natural and hydraulic fractures in the reservoirs are expected to be delineated, and appropriate fracturing fluids and proppants are to be optimized, based on the data of drilling, well logging, laboratory and field experiments, and geological data. These approaches lay the basis of the integral fracturing technology suitable for the low permeable reservoir in the study area. Based on core sample test, in-situ stress analysis of well logging, and forward and inversion stress field modeling, as well as fluid dynamic analysis, the maximum in-situ stress field is unraveled to be extended nearly along the E-W direction (clustering along N85-135°E) as is demonstrated by the E-W trending tensional fractures. Hydraulic fractures are distributed approximately along the E-W direction as well. Faulting activities could have exerted obvious influences on the distribution of fractures, which were preferentially developed along fault zones. Based on reservoir sensitivity analysis, integrated with studies on rock mechanics, in-situ stress, natural fracture distribution and production in injection-production pilot area, the influences of primary fractures on fracturing operation are analyzed, and a diagnostic technology for primary fractures during depressurization is accordingly developed. An appropriate fracturing fluid (hydroxypropyl guar gum) and a proppant (Yixing ceramsite, with a moderate-density, 0.45-0.9mm in size) applicable to Qingshankou Formation reservoir are worked out through extensive optimization analysis. The fracturing fluid can decrease the damage to the oil reservoir, and the friction in fracturing operation, improving the effect of fracturing operation. Some problems, such as sand-out at early stage and low success rate of fracturing operations, have been effectively solved, through pre-fracturing formation evaluation, “suspension plug” fracturing, real-time monitoring and limited-flow fracturing. Through analysis of fracture-bearing tight reservoir with variable densities and dynamic analysis of influences of well patterns on fracturing by using numerical simulation, a fracturing operation scheme for the Qingshankou Formation reservoir is proposed here as being better to compress the short factures, rather than to compress the long fractures during hydraulic fracturing. It is suggested to adopt the 450m×150m inverted 9-spot well pattern in a diamond shape with wells placed parallel to fractures and a half fracture length of 60-75m.

Inhibition of egg hatching success and larvae survival of the scallop, Chlamys farreri, associated with exposure to cells and cell fragments of the dinoflagellate Alexandrium tamarense

We report an apparently novel toxic effect of the dinoflagellate Alexandrium tamarense, manifested by inhibition of the egg hatching success of the scallop, Chlamys farreri. The hatching rate of C. farreri approached only 30% of controls when its fertilised eggs were exposed for 36 h to A. tamarense cells or cellular fragments at a concentration of 100 cells/ml, and the hatching rate was just 5% after exposure to A. tamarense of 500 cells/ml. Similar exposures of the fertilised scallop eggs to two other algal species, the diatom Phaeodactylum tricornutum and the raphidophyte Heterosigma carterae, resulted in no such toxicity or inhibitory effects.. Likewise, exposure of eggs to standard STX toxin. as well as to A. tamarense cell contents (supernant of re-suspended algal cells following ultrasonication and centrifugation), did not elicit this inhibitory response. However, exposure of the scallop eggs to cell cultures, intact algal cells, or cell fragments of A. tamarense produced marked toxicity. The alga also influenced larvae at early D-shape stage of scallop. The survival rates began to decrease significantly after exposed for 6 days at concentration of 3000 cells/ml and above: no larvae could survive after 14-day exposure to A. tamarense at 10,000 cells/ml or 20-day at 5000 cells/ml. The results indicated the production of novel substances from A. tamarense which can cause adverse effects on egg hatching and survival of the scallop larvae, The experiment also found that the developmental stages before blastula was the developmental period most sensitive to the A. tamarense toxin(s) and the alga at early exponential stage had the strongest effect on egg hatching comparing with other growth phases. The adverse effect of A. tamarense on early development of scallops may cause decline of shellfish population and may have further impact on marine ecosystem. (C) 2001 Elsevier Science Ltd. All rights reserved.