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1.
阿司匹林与人血清白蛋白的相互作用研究   总被引:7,自引:3,他引:4  
目的以光谱技术研究阿司匹林分子与人血清白蛋白(HSA)间结合作用机制。方法通过荧光光谱法确定阿司匹林对HSA的荧光猝灭机制。由Lineweaver-Burk双倒数作图法确定反应的解离常数。根据热力学方程讨论两者间主要的作用力类型。结合同步荧光技术考察阿司匹林对人血清白蛋白构象的影响。结果阿司匹林对HSA的荧光猝灭机制为静态猝灭。在37℃和25℃时阿司匹林与HSA的解离常数分别为KD37=1.44×10-3mol.L-1,KD25=1.96×10-3mol.L-1。结合反应热力学参数为ΔH=-19.73kJ.mol-1,△G=-16.21kJ.mol-1,ΔS=-11.77kJ.mol-1。结论两者结合的主要作用力类型是范德华力。阿司匹林与白蛋白结合后使蛋白质构象发生变化。  相似文献
2.
目的:建立葫芦素B人血清白蛋白纳米粒中药物含量的测定方法。方法:采用Diamonsil C18柱(200mm×4.6 mm,5μm);以乙腈-水-磷酸(45∶55∶0.1)作为流动相;流速为1 mL.min-1;检测波长为228 nm;柱温30℃。结果:在本实验条件下蛋白纳米粒中辅料对葫芦素B的测定无干扰,在1.0~20 mg.L-1浓度范围内线性关系良好(r=0.999 9,n=5),葫芦素B平均回收率为100.2%,RSD为0.8%(n=3)。结论:本法准确可靠,简便易行,可用于葫芦素B人血清白蛋白纳米粒中药物含量的测定。  相似文献
3.
异硫氰酸荧光素标记人血清白蛋白   总被引:6,自引:1,他引:5  
目的:用异硫氰酸荧光素(FTTC)对人血清白(HSA)进行荧光标记。方法:采用直接标记法,并考查不同料比时的荧光素分子与蛋白分子的结合情况,荧光标记物(FTTC-HSA)的荧光光谱及荧光寿命情况,结果:FTTC-HSA的激发波长为500nm,荧光波长为530nm;其固态和液态低温保存时荧光寿命分别为一年和37天。  相似文献
4.
Platelet derived growth factor (PDGF) is a key factor in the induction and progression of fibrotic diseases with the activated fibroblast as its target cell. Drug targeting to the PDGF-receptor is explored as a new approach to treat this disease. Therefore, we constructed a macromolecule with affinity for the PDGF-beta receptor by modification of albumin with a small peptide that recognises this PDGF-beta receptor. The binding of the peptide-modified albumin (pPB-HSA) to the PDGF-beta receptor was confirmed in competition studies with PDGF-BB using NIH/3T3-fibroblasts and activated hepatic stellate cells. Furthermore, pPB-HSA was able to reduce PDGF-BB-induced fibroblast proliferation in vitro, and proved to be devoid of proliferation-inducing activity itself. We assessed the distribution of pPB-HSA in vivo in two models of fibrosis and related the distribution of pPB-HSA to PDGF-beta receptor density. In rats with liver fibrosis (bile duct ligation model), pPB-HSA quickly accumulated in the liver in contrast to unmodified HSA (P<0.001). The major part of pPB-HSA in the fibrotic liver was localized in hepatic stellate cells. In rats with renal fibrosis (anti-Thy1.1 model), pPB-HSA also homed to the cells that expressed the PDGF-beta receptor, i.e. the mesangial cells in the glomeruli of the kidney. These results indicate that pPB-HSA may be applied as a macromolecular drug-carrier that accumulates specifically in cells expressing the PDGF-beta receptor, thus allowing a selective delivery of anti-fibrotic agents to these cells.  相似文献
5.
Release of Human Serum Albumin from Poly(lactide-co-glycolide) Microspheres   总被引:6,自引:0,他引:6  
Human serum albumin (HSA) was encapsulated in a 50:50 copolymer of DL-lactide/glycolide in the form of microspheres. These microspheres were used as a model formulation to study the feasibility of controlling the release of large proteins over a 20- to 30-day period. We show that HSA can be successfully incorporated into microspheres and released intact from these microspheres into various buffer systems at 37°C. A continuous release of the protein could be achieved in physiological buffers at 37°C over a 20- to 30-day period from microspheres with high protein loadings (11.6%). These results demonstrate the potential of poly(DL-lactide-co-glycolide) microspheres for continuous delivery of large proteins.  相似文献
6.
The plant derived flavonoid compound quercetin, possesses wide range of biological activities in the human body by interacting with nucleic acids, enzymes and other proteins. As has recently been shown this molecule of polyphenolic type extensively binds to human serum albumin (HSA), the most abundant carrier protein in the blood. Electronic absorption, circular dichroism (CD) spectroscopy and molecular modelling methods were used to characterize optical properties of the quercetin-HSA complex, and to gain information on the binding mechanism at molecular level. The red shift and hypochromism of the longest-wavelength absorption band of quercetin relative to the spectral properties in ethanol suggests that one or more phenolic OH groups of the bound ligand is ionized and that the exocyclic phenyl ring is not coplanar with the benzopyrone moiety. It was found that quercetin shows extrinsic optical activity on interaction with HSA. The induced CD spectra were utilized to calculate the association constant at 37 degrees (1.46+/-0.21 x 10(4)M(-1)) and to probe the ligand binding site. Results of the CD displacement experiments performed with palmitic acid and salicylate were interpreted together with the findings of molecular modelling calculation performed on the quercetin-HSA complex. Computational mapping of possible binding sites of quercetin revealed the molecule to be bound in the large hydrophobic cavity of subdomain IIA. The protein microenvironment of this site was found to be rich in polar (basic) amino acid residues which are able to help to stabilize the negatively charged ligand bound in non-planar conformation. Additionally, the position of quercetin within the binding pocket allows simultaneous binding of other ligands such as warfarin, or sodium salycilate.  相似文献
7.
人血白蛋白不良反应分析   总被引:4,自引:0,他引:4  
卜一珊 《天津药学》2007,19(1):35-37
目的:了解人血白蛋白文献报道中的不良反应情况。方法:检索CHKI收载的1994年1月—2006年10月国内公开发表的医药学期刊中所有关于人血白蛋白不良反应文献共计36篇,对其报道的病例进行统计分析。结果:人血白蛋白在临床存在不合理应用现象。人血白蛋白引起的不良反应多出现在药物输注过程中,不良反应类型主要为全身过敏反应和心脏损害。结论:临床应用人血白蛋白应严格遵循适应证,给药速度应适当,才能保证人血白蛋白安全、合理的应用。  相似文献
8.
9.
Olopatadine hydrochloride (olopatadine) is an anti-allergic drug that functions as a histamine H(1) antagonist and inhibits both mast cell degranulation and the release of arachidonic acid metabolites in various types of cells. In this study, we examined the ability of olopatadine to inhibit the expression of cytokine genes in vitro via high-affinity receptors for immunoglobulin E in mast cells, using a rat basophilic leukemia (RBL-2H3) cell line and an in vivo mouse model. Levels of gene expression in RBL-2H3 cells were determined by semi-quantitative RT-PCR, and serum interleukin-4 (IL-4) level in mice was quantified by ELISA. Olopatadine inhibited significantly the induction of IL-4 expression by mast cells both in vivo and in vitro. Olopatadine inhibited Ca(2+) influx through receptor-operated channels (ROC) without affecting Ca(2+) release from intracellular stores. Comparative analysis of olopatadine with other anti-allergic drugs and the ROC blocker SKF-96365 demonstrated that the potency of inhibition of Ca(2+) influx correlated with the degree of suppression of degranulation and arachidonic acid release. Inhibition of Ca(2+) influx decreased phosphorylation of p38 mitogen-activated protein kinase and c-Jun NH(2)-terminal kinase, which participate in regulation of cytokine (e.g. IL-4) gene expression. However, the rank order of inhibition of Ca(2+) influx did not correspond to reduction of IL-4 expression, suggesting that an unknown mechanism(s) of action, in addition to inhibition of Ca(2+) influx, is involved in the expression of cytokines in mast cells.  相似文献
10.
PURPOSE: Functional analysis of the three recombinant human serum albumin (rHSA) domains and their potential as stand-alone proteins for use as drug delivery protein carriers. METHODS: Protein structure was examined by fluorescence and CD spectroscopy. Ligand binding was estimated by ultrafiltration. Antioxidant activity was estimated by measuring the quenching of dihydrorhodamine 123. Esterase-like activity and enolase-like activity were estimated from the rate of hydrolysis of p-nitrophenyl acetate and conversion of dihydrotestosterone from the 3-keto to 3-enol form, respectively. The domains of human serum albumin (HSA) were radiolabeled with 111In to evaluate their pharmacokinetics. RESULTS: The ligand binding ability of subsites Ia and Ib could not be detected in domain II. However, the binding of ligands to subsite Ic and site II were preserved in domain II and domain III, respectively. Domain III retained about 45% of its esterase-like activity, and weaker esterase-like activity was also observed in domain I. All domains showed low enolase-like activity in a pH 7.4 phosphate buffer, but domain II had higher activity in a pH 9.2 carbonate buffer. Domain I exhibited antioxidant activity comparable to that of rHSA. All three of the 111In-radiolabeled domains were rapidly eliminated from HSA, with high accumulation in the kidneys. CONCLUSION: Domain I of HSA has great potential for further development as a drug delivery protein carrier, due to its favorable properties and the presence of a free cysteine residue.  相似文献
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