Harpagoside (HAR) is believed to be a main compound in Scrophularia ningpoensis which possess a broad of biological activities.Human serum albumin (HSA) has important physiological roles in transportation, distribution and metabolism of many endogenous and exogenous substances in body.It is great significance in pharmacology to investigate the interaction mechanism of HAR and HSA.In this work, the interaction between HAR and HSA was investigated by fluorescence and ultraviolet absorption spectroscopy at different pH (pH=4.0, 7.4, and 9.0) and temperatures (297, 310 and 323 K).The experimental results showed that the HAR could cause the fluorescence quenching of HSA through a static quenching procedure, showing that the HAR regularly quenched the intrinsic fluorescence of HSA, and a decrease in the quenching constant was observed with an increase in temperature.Under different conditions, all the magnitude of binding constants (KA) was larger than 105 L/mol and the number of binding sites (n) in the binary system were approximate to 1.Base on the magnitude of enthalpy and entropy changes, the negative values of ΔG, ΔH and ΔS revealed that the binding of HAR with HSA was spontaneous and exothermic process, and the main interaction forces of the HAR with HAR were van der Waals forces and/or hydrogen bonding interaction.The binding distance (r) between the HAR and HSA was calculated to be about 4.2 nm based on the theory of F(o)rster′s nonradiation energy transfer, which indicated that the energy transfer from HSA to HAR occurred with high possibility.What was more, the synchronous florescence spectroscopy confirmed the conformational changes of HSA during the binding reaction.

Reduction capacity ( RC ) is an important index to evaluate the redox ability of dissolved organic matter. In order to determine the RC, hydrophilic organic fractions ( HyI ) isolated from dissolved organic matter extracted from the uncomposted and composted samples were used as electron donators and mediators, and three kinds of irons were chosen as electron acceptors. The results showed that, the RC values from the composted sample were 15. 88, 13. 41 and 51. 45 mmol e -/mol C for the electron acceptors Fe2(SO4)3, Fe(NO3)3 and FeCit, respectively, which were higher than the corresponding values (13. 45, 11. 77 and 43. 16 mmol e-/mol C) from the uncomposted sample. The electron acceptor type shows a dramatic influence on the RC value of HyI. The RC value determined by FeCit was obviously higher than that measured using Fe2( SO4 ) 3 and Fe( NO3 ) 3 , and the microbial reducing capacity of the HyI was lower than the corresponding native reducing capacity. By analyzing the special absorbencies ( SUVA254 and SUVA280 ) , absorbance ratios ( A2/A3 and A4/A6 ) and integrated area from UV-vis spectra, it can be found that the RC was affected by aromatic degree, unsaturated conjugated structure, and molecular weight. Excitation-emission matrix spectra coupled with regional integration analysis showed that the relative content of humic-like substances ( humic-like acids and fulvic-like acids) was the main factor influencing the RC value of HyI. The results obtained can be used to characterize the redox properties of HyI, and reveal its role in the transformation and degradation of pollutants during composting.

OBJECTIVETo construct the two-dimensional electrophoretic (2-DE) protein map of the human sperm head.

METHODSProtein extracts of the normal human sperm and sperm head were loaded on an 18 cm immobilized pH gradients (IPG) strip holder and separated with isoelectric focusing electrophoresis as the first dimension, and then with upright SDS-PAGE as the second. Different protein spots between them were compared with Image Master 3.0, and so were the maps of method I with Urea/Thiourea/Guanidine HCl and method II with Kit/Guanidine HCl.

RESULTSOf the whole-sperm proteome, 802 protein spots were obtained by method I, and 797 by method II. Among them, distribution patterns of 492 spots were the same. A sperm protein map was constructed with 1107 protein spots after the interaction of the spots obtained by both the methods. Of the sperm-head proteome, 428 protein spots were obtained, all found in the whole-sperm protein map after comparison.

CONCLUSIONBoth the methods could be complementarily used to construct a sperm protein map, and the map could serve as a model for the establishment of the sperm protein profile.

Electrophoresis, Gel, Two-Dimensional ; methods ; Humans ; Hydrogen-Ion Concentration ; Male ; Proteins ; analysis ; Proteomics ; methods ; Sperm Head ; chemistry

Objective: To investigate the role of CpG ODN (CpG oligodeoxynucleotide) adjuvant in enhancing the anti-bladder cancer response induced by MAGE-3 (melanoma antigen gene -3) antigen and its molecular mechanism. Methods: Mononuclear cells were isolated from HLA-A2 type peripheral blood of healthy donors by Ficoll method to prepare mature DC by conventional means. DC surface markers were detected by flow cytometry. MTT assay was used to detect the promotion effect of DCs sensitized by different means (MAGE-3, CpGODN, MAGE-3+CpG ODN, irrelevant control antigen) on the proliferation of T lymphocytes and the killing effect of CTL on BIU-87 tumor cells. The tumor mass of nude mice bearing BIU-87 bladder cell xenograft were examined on Day 7 and 11 after CpG ODN+MAGE-3 sensitized DC treatment. The expression of Bcl-2/Bax protein was detected by Western blotting while the proliferation level of xenograft cells was detected by MTT assay. Results: DCs sensitized by CpG ODN combined with MAGE-3 antigenic peptides could promote the proliferation of T lymphocytes and significantly enhance the killing effect of CTLon target BIU-87 cells (P< 0.05). Compared with other sensitized DCs, in vivo experiments showed that 7 and 11 days after treatment, both the tumor volume and weight were significantly reduced (all P<0.05), and the proliferation ability of xenograft tumor was decreased (P<0.05). Compared with other sensitization means, CpG ODN+MAGE-3 especially exhibited obvious inhibitive effect on tumor growth on Day 11, and significantly promoted the proliferation of splenic monocytes of tumor bearing mice (P<0.01); moreover, Bcl-2 expression in xenograft tissues significantly decreased（P<0.01）while Bax expression significantly increased（P<0.05 or P<0.01）on Day 3 after treatment. Conclusion: CpG ODN can promote the inhibitory effect of MAGE-3 sensitized DC on bladder cancer BIU-87 cells, which will provide experimental basis for clinical application of DC vaccine in bladder cancer treatment.