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21.
王征帆 《天然产物研究与开发》2013,25(9)
以罗丹明B为显色剂,采用分光光度法测定了护脾养胃类方剂中常见的当归、五味子、黄芩、红枣等四种中药水提物对·OH的清除能力.实验表明:这些中药永提取物对·OH都具有较强的清除作用,呈现明显的量效关系,其中红枣的清除能力最强.据此评价了这四种中药水提取物抗氧化能力大小顺序为:红枣>当归>黄芩>五味子. 相似文献
22.
Photosensitizers newly developed for photodynamic therapy of cancer need to be assessed using accurate methods of measuring reactive oxygen species (ROS). Little is known about the characteristics of the reaction of singlet oxygen (1O2) with spin traps, although this knowledge is necessary in electron spin resonance (ESR)/spin trapping. In the present study, we examined the effect of various reductants usually present in biological samples on the reaction of 1O2 with 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). The ESR signal of the hydroxyl radical (•OH) adduct of DMPO (DMPO-OH) resulting from 1O2-dependent generation of •OH strengthened remarkably in the presence of reduced glutathione (GSH), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), ascorbic acid, NADPH, etc. A similar increase was observed in the photosensitization of uroporphyrin (UP), rose bengal (RB) or methylene blue (MB). Use of 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide (DEPMPO) as a spin trap significantly lessened the production of its •OH adduct (DEPMPO-OH) in the presence of the reductants. The addition of DMPO to the DEPMPO-spin trapping system remarkably increased the signal intensity of DEPMPO-OH. DMPO-mediated generation of •OH was also confirmed utilizing the hydroxylation of salicylic acid (SA). These results suggest that biological reductants enhance the ESR signal of DMPO-OH produced by DMPO-mediated generation of •OH from 1O2, and that spin trap-mediated •OH generation hardly occurs with DEPMPO. 相似文献
23.
24.
1alpha,25-Dihydroxyvitamin D(3) (1alpha,25(OH)2D3) has been shown to modulate not only proliferation and differentiation, but also apoptosis in malignant cells, indicating that it could be useful for the treatment of cancer and psoriasis. However, little information has been available on the binding conformation of the 1alpha,25(OH)2D3 molecule and its analogs with the vitamin D receptor (VDR). Therefore, we synthesized 2alpha-fluorinated A-ring analogs of 19-nor-1alpha,25(OH)2D3 in order to investigate the VDR-binding conformation of the A-rings on the basis of the (19)F NMR analysis. The 2alpha-fluoro-19-nor-1alpha,25-dihydroxyvitamin D3 A-ring analog thus synthesized via a asymmetric catalytic carbonyl-ene cyclization, shows significant activity in transactivation. 相似文献
25.
26.
Andrew N. Hoofnagle Thomas J. Laha Thomas F. Donaldson 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(19):1639-1642
Background
The unmitigated rise in demand for the assessment of vitamin D status has taxed the ability of clinical mass spectrometry laboratories to preserve turn-around times. We aimed to improve the throughput of liquid–liquid extraction of plasma/serum for the assay of 25-hydroxy vitamin D.Methods
We designed and fabricated a flexible rubber gasket that seals two 96-well plates together to quantitatively transfer the contents of one plate to another. Using the transfer gasket and a dry-ice acetone bath to freeze the aqueous infranatant, we developed a novel liquid–liquid extraction workflow in a 96-well plate format. We applied the technology to the mass spectrometric quantification of 25-hydroxy vitamin D.Results
Cross-contamination between wells was ≤0.13%. The interassay imprecision over 132 days of clinical implementation was less than 10%. The method compared favorably to a standard liquid–liquid extraction in glass tubes (Deming slope = 1.018, Sx|y = 0.022). The accuracy of the assay was 102–105% as assessed with the recently released control materials from NIST.Conclusions
The development of a plate-sealing gasket permits the liquid–liquid extraction of clinical specimens in a moderate-throughput workflow and the reliable assay of vitamin D status. In the future, the gasket may also prove useful in other sample preparation techniques for HPLC or mass spectrometry. 相似文献27.
M. Bertamini L. Zulini R. Zorer K. Muthuchelian N. Nedunchezhian 《Photosynthetica》2007,45(3):426-432
Photoinhibition under irradiance of 2 000 μmol m−2 s−1 (HI) was studied in detached control (C) and water deficit (WD) leaves of grapevine (Vitis vinifera L.) plants. The degree of photoinhibition was determined by means of the ratio of variable to maximum chlorophyll (Chl) fluorescence
(Fv/Fm) and electron transport measurements. The potential efficiency of photosystem (PS) 2, Fv/Fm, marginally declined under HI in WD-leaves without significant increase of F0. In contrast, Fv/Fm ratio declined markedly with significant increase of F0 in C-leaves. In isolated thylakoids, the rate of whole chain and PS2 activity under HI were more decreased in C-than WD-leaves.
The artificial exogenous electron donors diphenyl carbazide, NH2OH, and Mn2+ failed to restore the HI-induced loss of PS2 activity in both C-and WD-leaves. Thus HI operates at the acceptor side of PS2
in both leaf types. Quantification of the PS2 reaction centre protein D1 following HI exposure of leaves showed pronounced
differences between C-and WD-leaves. The marked loss of PS2 activity under HI of C-leaves was due to the marked loss of D1
protein of the PS2 reaction centre. 相似文献
28.
Marie Garnier-Lhomme Axelle Grélard Cécile Loudet Banafshé Larijani 《生物化学与生物物理学报:生物膜》2007,1768(10):2516-2527
Membrane dynamics is an essential part of many cellular mechanisms such as intracellular trafficking, membrane fusion/fission and mitotic organelle reconstitution. The dynamics of membranes is dependent primarily on their phospholipid and cholesterol composition and how these molecules are ordered in relation to one another. To determine the physical status of membranes in whole cells or purified membranes of subcellular compartments we have developed a novel application exploiting solid-state 2H-NMR spectroscopy. We utilise this method to probe the dynamics of intact sperm and nuclear envelope precursor membranes. We show, using mass spectrometry, that either multilamellar or small unilamellar vesicles of deuterium-labelled palmitoyl-oleoylphosphatidylcholine can be used to probe the dynamics of sperm cells or nuclear envelope precursor membrane vesicles, respectively. Using 2H-NMR we determine the order parameters of sperm cells and nuclear envelope precursor membrane vesicles. We demonstrate that whole sperm membranes are more dynamic than nuclear envelope precursor membranes due to the higher cholesterol levels of the latter. Our new application can be exploited as a generic method for monitoring membrane dynamics in whole cells, various subcellular membrane compartments and membrane domains in subcellular compartments. 相似文献
29.
B. Shannan M. Seifert D.A. Boothman W. Tilgen J. Reichrath 《The Journal of steroid biochemistry and molecular biology》2007,103(3-5):721
Prostate cancer is the most commonly diagnosed cancer in the majority of western countries. Due to their antiproliferative and proapoptotic activity, vitamin D analogues have been introduced recently as an experimental therapy for prostate cancer. Clusterin (CLU) is a glycoprotein that has two known isoforms generated in human cells. A nuclear form of CLU protein (nCLU) is pro-apoptotic, and a secretory form (sCLU) is pro-survival. In this study, we analyzed whether proapoptotic and antiproliferative effects of 1,25(OH)2D3 on LNCaP prostate cancer cells are modulated by expression of sCLU. Using colony forming assay, we studied the effect of treatment with different doses of 1,25(OH)2D3 (10−6, 10−7, 10−10 M) on proliferation of LNCaP cells that were stable transfected and over-express sCLU (LNT-1) as compared to empty vector-transfected cells (LN/C). We also measured apoptosis using TUNEL assay. sCLU over-expression protected against both antiproliferative (30%) and proapoptotic (15%) effects of 1,25(OH)2D3, although this effect was statistically not significant. In conclusion, our findings demonstrate that expression of sCLU modulates growth regulatory effects of 1,25(OH)2D3 in prostate cancer indicating that CLU interferes with vitamin D signalling pathways. 相似文献
30.
Cecylia Tukaj Piotr Trzonkowski Jolanta Kubasik-Juraniec Andrzej Myliwski 《The Journal of steroid biochemistry and molecular biology》2007,103(3-5):525
Inhibitory effect of 1α,25dihydroxycholecalciferol (1,25D3 = calcitriol) in different cell type is well recognized but its promoting effect on vascular smooth muscle cells (SMCs) is poor established. Therefore, the aim of this study was to determine stimulatory effect of calcitriol on aortal SMCs proliferation in culture. We used the cell division analysis procedure based on the quantitative sequential halving of the stably incorporating fluorescent dye carboxyfluorescein diacetate succinimidyl ester (CFSE). This technique allowed the visualization of cycles of SMCs division by flow cytometry. Rat aortal SMCs were labeled with CFSE and cultured for up to 10 days with defined concentration of calcitriol in medium. Proliferative activity as the percentage of SMCs in different phases of the cell cycle using propidium iodide was determined. Apoptosis was assessed using Annexin-V/CFDA method. The results suggest that low concentrations of an active form of vitamin D—1α,25dihydroxycholecalciferol applied in supraphysiological concentration of 10 nmol/l is a mitogenic factor for aortal SMCs. None of the applied concentrations of calcitriol caused apoptosis. The findings well support our morphological (LM) and ultrastructural (TEM and SEM) observations. 相似文献