Characterization of linear epitope of the beta(2)-microglobulin via combination of MALDI-TOF-MS with immunoaffinity

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF-MS), in combination with immunoaffinity provided a powerful tool for determining epitope (antigenic determinant) in protein. The linear epitope of the beta(2)-microglobulin was characterized in the paper. The method as follows: at first beta(2)-microglobulin was digested by a proteolytic enzyme to produce an appropriate set of peptide fragments, then peptide fragments containing the linear epitope were selected and separated from the pool of peptide fragments by immunoprecipitation with the monoclonal antibody. The agarose beads were collected carefully after the reaction. Unbound peptides would be washed away, while the peptides containing the epitope would remain bound to the immobilized antibody after. the beads were washed several times with appropriate buffer. At last the masses of the bound peptides were identified directly by MALDI-TOF MS. Using Endoproteinase Glu-C Endoproteinase Lys-C and Trypsin in the experiment, the linear epitope of beta(2)-microglobulin was located within peptide fragment 59-69, that is, DWSFYLLYYTE.

Endothelial cells release phenotypically and quantitatively distinct microparticles in activation and apoptosis

Background: Endothelial cells (EC) shed endothelial microparticles (EMP) in activation and apoptosis. Objectives: We compared the antigenic expression of EMP species released during activation as compared to apoptosis, in three cell lines. Methods: EC from renal and brain microvascular (MiVEC) and coronary macrovascular (MaVEC) origin were incubated with TNF-alpha to induce activation, or deprived of growth factors to induce apoptosis. Antigens expressed on EMP and EC were assayed flow cytometrically and included constitutive markers (CD31, CD51/61, CD105), inducible markers (CD54, CD62E and CD106), and annexin V binding. Results: It was found that in apoptosis, constitutive markers in EMP were markedly increased (CD31>CD105), with a concomitant decrease in expression in EC. Annexin V EC surface binding and annexin V+ EMP were more sharply increased in apoptosis than in activation. In contrast, in activation, inducible markers in EMP were markedly increased in both EMP and EC (CD62E>CD54>CD 106). Coronary MaVEC released significantly less EMP than MiVEC. Conclusion: EC release qualitatively and quantitatively distinct EMP during activation compared to apoptosis. Analysis of EMP phenotypic signatures may provide clinically useful information on the status of the endothelium. (C) 2003 Elsevier Science Ltd. All rights reserved.

博兴洼陷西部砂体成因与油气成藏研究

Based on achievements of thirty years of hydrocarbon exploration, this paper uses the modern theories and methods of sedimentology and oil accumulation to study the origin and distribution features of four sandbodies of Gaoqing, Fanjia, Zhenglizhuang and Jinjia from the third member to the lower second member of Shahejie Formation in detail. Various geophysical methods are also used to explain and to predict the spatial distribution of sandbodies, which further shows mechanism and the model of oil accumulation and illuminates the disciplinarians of oil enrichment and its controlling factors in the study area. The most favourable oil pools predicted by this paper have significant economic and social benefits, which has been confirmed by the exploration. The main conclusions and knowledge includes: (1) Resolving the problems, which remain unresolvable for a long time in the western area of Boxing depression, about the original environment and the spatial distribution of sandbodies of Gaoqing, Fanjia, Zhenglizhuang and Jinjia, and illuminating their relationships. It is suggested that two deltas or delta-related sandbody sediments, which include the delta sandbodies of Jinjia and Gaoqing and their frontal turbidite fan sandbody, are developed in the second and third members of Shahejie Formation. The sandbodies of Fanjia, Gaoqing and Zhenglizhuang are components of Gaoqing delta and belong to the sediments of various periods in different part of the delta. Whereas, the sandbody of Jinjia belong to the Jinjia delta or fan-delta created by the uplift of the Western Shandong and in some areas shows the features of juxtaposition, superimposed deposition and fingeration with the sandbodies of Gaoqing and Zhenglizhuang. (2)Proposing that the sandbodies of different origins in the deltas of Gaoqing and Jinjia have obvious different reservoir qualities, among which the delta frontal bedded sandbodies in the second member of Shahejie Formation in Zhenglizhuang are the best ones and the turbidite sandbody of Fanjia is relatively worse. This shows the direction of further reservoir prediction. (3) According to modern petroleum system theory and continental pool-formation theory, the author divided the western area of Boxing depression into the Jinjia—Zhenglizhuang—Fanjia nose structure belt pool-formation system and the Gaoqing fracture belt pool-formation system. The study area is predominantly located in the former belt and subdivided into pool-formation sub-systems of Zhenglizhuang-Fanjia and Jinjia, which have the source rock of mudstone and oil shale from the upper forth member and the third member of Shahejie Formation in Boxing depression. The hydrocarbon migration and accumulation are controlled by Jinjia-Zhenglizhuang-fanjia nose structure and Gaoqing fracture. (4)Proposing that compared with the best developed sandbodies and traps in the west area of Boxing, the source from the Boxing depression is not sufficient, which is the fundamental reason that the hydrocarbon resources in mid-west area is less than in the east of Boxing. (5) Under the direction of the new theory (fluid compartments theory) and new method of modern pool-formation mechanism, two kinds of pool-formation model are established in study, i.e. inner-compartment model and outer-compartment model. The former has abnormal pressure and is the antigenic source seal pool-forming mechanism, whereas the latter has normal pressure and is of the allochthonous source opening pool-formation mechanism. (6)The study shows that the four sandbodies of Gaoqing, Fanjia, Jinjia and Zhenglizhuang sand are all very benefit for pool-formation, among which the Fanjia sandbody is the best favourable one and is likely to form lithological reservoir and fault-lithological reservoir. But the main step of exploration in Gaoqing, Zhenglizhuang sandbodies should be finding out the fault block, reversed roof and stratum-lithological oil reservoir. (7)Established a set of guidelines and techniques for the research and exploration in the large scale of sandbodies. Proposing that the various traps related to reversed fault and basin-ward fault should be found in step slopes and gentle slopes respectively, and the lithological oil reservoir should mainly be found in the sandstone updip pinch out. It is also suggested that Fanjia sandbody is most favourable to form the lithological and fault-lithological and the Gaoqing, Zhenglizhuang and Jinjia sandbodies have the potential of forming fault block, reversed roof and stratum-lithological oil reservoir. (8) Interpretation and prediction the spatial distribution of main sandbodies based on various geophysical methods suggestion that Fanxi, Gao28 south and Gao27 east have better exploration potential.