全文获取类型
收费全文 | 5915篇 |
免费 | 461篇 |
国内免费 | 2篇 |
出版年
2024年 | 5篇 |
2023年 | 18篇 |
2022年 | 50篇 |
2021年 | 123篇 |
2020年 | 88篇 |
2019年 | 99篇 |
2018年 | 176篇 |
2017年 | 147篇 |
2016年 | 234篇 |
2015年 | 336篇 |
2014年 | 407篇 |
2013年 | 461篇 |
2012年 | 556篇 |
2011年 | 529篇 |
2010年 | 346篇 |
2009年 | 294篇 |
2008年 | 377篇 |
2007年 | 331篇 |
2006年 | 310篇 |
2005年 | 266篇 |
2004年 | 289篇 |
2003年 | 194篇 |
2002年 | 179篇 |
2001年 | 135篇 |
2000年 | 91篇 |
1999年 | 76篇 |
1998年 | 32篇 |
1997年 | 34篇 |
1996年 | 23篇 |
1995年 | 17篇 |
1994年 | 11篇 |
1993年 | 14篇 |
1992年 | 21篇 |
1991年 | 18篇 |
1990年 | 22篇 |
1989年 | 15篇 |
1988年 | 7篇 |
1987年 | 9篇 |
1986年 | 3篇 |
1985年 | 8篇 |
1984年 | 2篇 |
1983年 | 2篇 |
1982年 | 3篇 |
1979年 | 2篇 |
1978年 | 3篇 |
1977年 | 3篇 |
1975年 | 2篇 |
1974年 | 3篇 |
1967年 | 2篇 |
1961年 | 1篇 |
排序方式: 共有6378条查询结果,搜索用时 31 毫秒
961.
Novel mechanistic class of fatty acid amide hydrolase inhibitors with remarkable selectivity 总被引:5,自引:0,他引:5
Ahn K Johnson DS Fitzgerald LR Liimatta M Arendse A Stevenson T Lund ET Nugent RA Nomanbhoy TK Alexander JP Cravatt BF 《Biochemistry》2007,46(45):13019-13030
Fatty acid amide hydrolase (FAAH) is an integral membrane enzyme that degrades the fatty acid amide family of signaling lipids, including the endocannabinoid anandamide. Genetic or pharmacological inactivation of FAAH leads to analgesic, anti-inflammatory, anxiolytic, and antidepressant phenotypes in rodents without showing the undesirable side effects observed with direct cannabinoid receptor agonists, indicating that FAAH may represent an attractive therapeutic target for treatment of pain, inflammation, and other central nervous system disorders. However, the FAAH inhibitors reported to date lack drug-like pharmacokinetic properties and/or selectivity. Herein we describe piperidine/piperazine ureas represented by N-phenyl-4-(quinolin-3-ylmethyl)piperidine-1-carboxamide (PF-750) and N-phenyl-4-(quinolin-2-ylmethyl)piperazine-1-carboxamide (PF-622) as a novel mechanistic class of FAAH inhibitors. PF-750 and PF-622 show higher in vitro potencies than previously established classes of FAAH inhibitors. Rather unexpectedly based on the high chemical stability of the urea functional group, PF-750 and PF-622 were found to inhibit FAAH in a time-dependent manner by covalently modifying the enzyme's active site serine nucleophile. Activity-based proteomic profiling revealed that PF-750 and PF-622 were completely selective for FAAH relative to other mammalian serine hydrolases. We hypothesize that this remarkable specificity derives, at least in part, from FAAH's special ability to function as a C(O)-N bond hydrolase, which distinguishes it from the vast majority of metabolic serine hydrolases in mammals that are restricted to hydrolyzing esters and/or thioesters. The piperidine/piperazine urea may thus represent a privileged chemical scaffold for the synthesis of FAAH inhibitors that display an unprecedented combination of potency and selectivity for use as potential analgesic and anxiolytic/antidepressant agents. 相似文献
962.
963.
Katheryn A. Resing Natalie G. Ahn 《Progress in biophysics and molecular biology》1999,71(3-4):501-523
Advances in mass spectrometry instrumentation, protocols for sample handling, and computational methods provide powerful new approaches to solving problems in analytical biochemistry. This review summarizes recent work illustrating ways in which mass spectrometry has been used to address questions relevant to signal transduction. Rather than encompass all of the instruments or methodologies that might be brought to bear on these problems, we present an overview of commonly used techniques, promising new methodologies, and some applications. 相似文献
964.
J Klein-Seetharaman J Hwa K Cai C Altenbach W L Hubbell H G Khorana 《Biochemistry》1999,38(25):7938-7944
Cysteines were introduced, one at a time, at amino acid positions 55-75 in the cytoplasmic region connecting helices I and II in rhodopsin. In each of the 21 cysteine mutants, the reactive native cysteine residues (C140 and C316) were replaced by serine. Except for N55C, all mutants formed rhodopsin-like chromophores and had normal photobleaching characteristics. The efficiency of GT activation was reduced only for K66C, K67C, L68C, and P71C. The reactivity of the substituted cysteine in each mutant toward 4, 4'-dithiodipyridine (4-PDS) was investigated in the dark. The mutants F56C to L59C and I75C were unreactive to 4-PDS under the conditions used, suggesting that they are embedded in the micelle or protein interior. The mutants V63C, H65C-T70C, and N73C reacted rapidly, while the remainder of the mutants reacted more slowly, and varied in reactivity with sequence position. For the mutants derivatized with 4-PDS, the rate of release of thiopyridone from the resulting thiopyridinyl-cysteine disulfide bond by dithiothreitol was investigated in the dark and in the light. Marked changes in the rates of thiopyridone release in the light were found at specific sites. Collectively, the data reveal tertiary interactions of the residues in the sequence investigated and demonstrate structural changes due to photoactivation. 相似文献
965.
Twenty-one single-cysteine substitution mutants were prepared in the sequence 56-75 between transmembrane helices I and II at the cytoplasmic surface of bovine rhodopsin. Each mutant was reacted with a sulfhydryl-specific reagent to produce a nitroxide side chain. The electron paramagnetic resonance of the labeled proteins in dodecyl maltoside solution was analyzed to provide the relative mobility and accessibility of the nitroxide side chain to both polar and nonpolar paramagnetic reagents. The results indicate that the hydrophobic-water interface of the micelle intersects helices I and II near residues 64 and 71, respectively. Thus, the sequence 64-71 is in the aqueous phase, while 56-63 and 72-75 lie in the transmembrane helices I and II, respectively. The lipid-facing surfaces on transmembrane helices I and II near the cytoplasmic surface correspond to approximately 180 degrees and 90 degrees of arc on the helical surfaces, respectively. Photoactivation of rhodopsin produced changes in structure in the region investigated, primarily around helix II. However, these changes are much smaller than those noted by spin labels in helix VI (Altenbach, C., Yang, K., Farrens, D., Farahbakhsh, Z., Khorana, H. G., and Hubbell, W. L. (1996) Biochemistry 35, 12470). 相似文献
966.
In order to develop a serum-free (SF) medium for the production of erythropoietin (EPO) by suspension culture of recombinant Chinese hamster ovary (rCHO) cells, a statistical optimization approach based on a Plackett-Burman design was adopted. A basal medium was prepared by supplementing Iscove's modified Dulbecco's medium (IMDM) with Fe(NO3)3.9H2O, CuCl2 and ZnSO4.7H2O which are generally contained in SF medium formulations. Insulin, transferrin and ethanolamine were also supplemented to the basal medium to determine their optimal concentrations. From this statistical analysis, glutamate, serine, methionine, phosphatidylcholine, hydrocortisone and pluronic F68 were identified as positive determinants for cell growth. The SF medium was formulated by supplementing the basal medium with components showing positive effects on cell growth in suspension culture. An EPO titer in this optimized SF medium was 79% of that in IMDM supplemented with 5% dialyzed fetal bovine serum (dFBS). Furthermore, the in vitro and in vivo biological activities of EPO produced in the SF medium were comparable to those produced in the serum-supplemented medium. Taken together, the results obtained here show that a Plackett-Burman design facilitates the development of SF media for the production of EPO by suspension culture of rCHO cells. 相似文献
967.
S Y Choi F Kwok J H Bahn S G Jeon Y K Ahn B H Yoon B R Lee K S Choi G Z Gao 《BioFactors (Oxford, England)》1999,10(1):35-42
Six monoclonal antibodies that recognize porcine brain pyridoxal kinase have been selected and designated as PK67, PK86, PK91, PK144, PK252 and PK275. A total of six monoclonal antibodies recognizing different epitopes of the enzyme were obtained, of which four inhibited the enzyme activity. When total proteins of porcine brain homogenate separated by SDS-PAGE were subjected to monoclonal antibodies, a single reactive protein band of molecular weight 39 kDa which comigrated with purified porcine pyridoxal kinase was detected. Using the anti-pyridoxal kinase antibodies as probes, the cross reactivities of brain pyridoxal kinase from human and other mammalian tissues and from avian sources were also investigated. Among human and all animal tissues tested, immunoreactive bands on Western blots appeared to have the same molecular mass of 39 kDa. These results indicate that mammalian brains contain only one major type of immunologically similar pyridoxal kinase, although some properties of the enzymes reported previously differed from one another. 相似文献
968.
Reexamining the classification proposed by Hurusawa, the phylogeny of Far EasternEuphorbia subgenusEsula was analyzed using thirteen morphological and seventeen phenolic compound data. These data were analyzed independently and
in combination using PAUP under the assumptions of Fitch parsimony. Ten species, comprised of three sections and five subsections
within Far EasternEuphorbia subg.Esula and one outgroup from subg.Chamaesyce, were used as terminal taxa. The phylogenetic results did not support the sectional classifications within subg.Esula proposed by Hurusawa. SectionDecussatae was nested in the paraphyletic sectEsula in all of the analyses, and the relationship of sectHelioscopiae was equivocal among data sets. The disagreement of data sets over the placement ofEuphorbia ebracteolata is probably due to a hybrid origin of the species and missing phenolic data forE. pallasii. A sister-group relationship of the Korean endemicE. fauriei with the widespreadE. pekinensis was strongly supported by the morphological and phenolic data. 相似文献
969.
The mixing properties of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) were examined in liquid-crystalline phase using fluorescent probes incorporated into lipid bilayers. The excimer to monomer (E/M) fluorescence ratio of 1-hexadecanoyl-2-(1-pyrenedecanoyl)-sn-glycero-3-phosphocholine (PPC) versus PPC concentration was higher for binary mixtures containing phosphatidylcholine (PC)/phosphatidylethanolamine (PE) (1:1) compared to PC matrix. When POPC was gradually replaced with POPE, the E/M ratio also increased suggesting the enhanced lateral mobility or the lateral enrichment of PPC into domains or both. Evidences for the PE-induced domain formation were further provided by resonance energy transfer between 2-(4, 4-difluoro-5-methyl-4-boro-3a, 4a-diaza-s-indacene-3-dodecanoyl)-1-hexadecanoyl-sn-glycero- 3-phospho choline and PPC, which was enhanced as a function of PE concentration, and by the polarization of 1,6-diphenyl-1,3, 5-hexatriene. In addition, PE reduced free volume and polarity of lipid bilayers as measured by the emission fluorescence of 1,2-bis PPC and 6-lauroyl-2-dimethylaminonaphthalene. When POPE analogs with a methylated head group instead of normal POPE were used, the diminished effect on the domain formation was shown in the order N-methyl PE > N,N-dimethyl PE. The results suggest that the mixing properties of POPE and POPC are not random but that lipid domains of phospholipids are formed. 相似文献
970.
Inhibition of Mitogen-Activated Kinase Signaling Sensitizes HeLa Cells to Fas Receptor-Mediated Apoptosis
下载免费PDF全文
![点击此处可从《Molecular and cellular biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Tim H. Holmstrm Stefanie E. F. Tran Victoria L. Johnson Natalie G. Ahn Sek C. Chow John E. Eriksson 《Molecular and cellular biology》1999,19(9):5991-6002
The Fas receptor (FasR) is an important physiological mediator of apoptosis in various tissues and cells. However, there are also many FasR-expressing cell types that are normally resistant to apoptotic signaling through this receptor. The mitogen-activated protein kinase (MAPK) signaling cascade has, apart from being a growth-stimulating factor, lately received attention as an inhibitory factor in apoptosis. In this study, we examined whether MAPK signaling could be involved in protecting FasR-insensitive cells. To this end, we used different approaches to inhibit MAPK signaling in HeLa cells, including treatment with the MAPK kinase inhibitor PD 98059, serum withdrawal, and expression of dominant-interfering MAPK kinase mutant protein. All of these treatments were effective in sensitizing the cells to FasR-induced apoptosis, demonstrating that MAPK indeed is involved in the control of FasR responses. The MAPK-mediated control seemed to occur at or upstream of caspase 8, the initiator caspase in apoptotic FasR responses. Transfection with the constitutively active MAPK kinase abrogated FasR-induced apoptosis also in the presence of cycloheximide, indicating that the MAPK-generated suppression of FasR-mediated apoptotic signaling is protein synthesis independent. In cells insensitive to FasR-induced apoptosis, stimulation of the FasR with an agonistic antibody resulted in significant MAPK activation, which was inhibited by PD 98059. When different cell types were compared, the FasR-mediated MAPK activation seemed proportional to the degree of FasR insensitivity. These results suggest that the FasR insensitivity is likely to be a consequence of FasR-induced MAPK activation, which in turn interferes with caspase activation. 相似文献