全文获取类型
收费全文 | 1168篇 |
免费 | 69篇 |
国内免费 | 1篇 |
专业分类
1238篇 |
出版年
2022年 | 11篇 |
2021年 | 10篇 |
2020年 | 9篇 |
2019年 | 14篇 |
2018年 | 21篇 |
2017年 | 29篇 |
2016年 | 33篇 |
2015年 | 35篇 |
2014年 | 50篇 |
2013年 | 69篇 |
2012年 | 76篇 |
2011年 | 75篇 |
2010年 | 48篇 |
2009年 | 44篇 |
2008年 | 50篇 |
2007年 | 65篇 |
2006年 | 63篇 |
2005年 | 58篇 |
2004年 | 61篇 |
2003年 | 40篇 |
2002年 | 42篇 |
2001年 | 24篇 |
2000年 | 31篇 |
1999年 | 25篇 |
1998年 | 18篇 |
1997年 | 17篇 |
1996年 | 10篇 |
1995年 | 5篇 |
1994年 | 7篇 |
1993年 | 6篇 |
1992年 | 15篇 |
1991年 | 11篇 |
1990年 | 7篇 |
1989年 | 14篇 |
1988年 | 9篇 |
1987年 | 12篇 |
1986年 | 9篇 |
1985年 | 12篇 |
1984年 | 9篇 |
1983年 | 7篇 |
1982年 | 6篇 |
1981年 | 5篇 |
1980年 | 6篇 |
1979年 | 10篇 |
1978年 | 5篇 |
1976年 | 10篇 |
1975年 | 9篇 |
1974年 | 9篇 |
1973年 | 5篇 |
1967年 | 5篇 |
排序方式: 共有1238条查询结果,搜索用时 15 毫秒
61.
S T Christian J A Monti W H Finley 《Biochemical and biophysical research communications》1977,79(3):966-972
We have observed distinct differences in the polarization of fluorescence and temperature dependent emission intensity of the highly fluorescent phospholipid derivative (1-acyl-2-(N-4-nitrobenzo-2-oxa-1,3-diazole)--aminocaproyl phosphatidylcholine (NBD-PC), when incorporated in the plasma membranes of normal and cystic fibrosis fibroblasts. Fluorescence polarization measurements indicate that the fluorochrome has a much higher degree of rotational mobility in cystic fibrosis fibroblasts as compared with normal cells. Temperature dependent transitions in the emission intensity of NBD-PC incorporated in normal fibroblasts are indicated at 17.7 and 21.2° C while the abnormal cell membranes apparently undergo transitions at 8.7 and 13.5° C. These differences might be due to changes in plasma membrane composition and/or organization, in the case of the cystic fibrosis cells. 相似文献
62.
Ilari A Latella MC Ceci P Ribacchi F Su M Giangiacomo L Stefanini S Chasteen ND Chiancone E 《Biochemistry》2005,44(15):5579-5587
The role of the ferroxidase center in iron uptake and hydrogen peroxide detoxification was investigated in Listeria innocua Dps by substituting the iron ligands His31, His43, and Asp58 with glycine or alanine residues either individually or in combination. The X-ray crystal structures of the variants reveal only small alterations in the ferroxidase center region compared to the native protein. Quenching of the protein fluorescence was exploited to assess stoichiometry and affinity of metal binding. Substitution of either His31 or His43 decreases Fe(II) affinity significantly with respect to wt L. innocua Dps (K approximately 10(5) vs approximately 10(7) M(-)(1)) but does not alter the binding stoichiometry [12 Fe(II)/dodecamer]. In the H31G-H43G and H31G-H43G-D58A variants, binding of Fe(II) does not take place with measurable affinity. Oxidation of protein-bound Fe(II) increases the binding stoichiometry to 24 Fe(III)/dodecamer. However, the extent of fluorescence quenching upon Fe(III) binding decreases, and the end point near 24 Fe(III)/dodecamer becomes less distinct with increase in the number of mutated residues. In the presence of dioxygen, the mutations have little or no effect on the kinetics of iron uptake and in the formation of micelles inside the protein shell. In contrast, in the presence of hydrogen peroxide, with increase in the number of substitutions the rate of iron oxidation and the capacity to inhibit Fenton chemistry, thereby protecting DNA from oxidative damage, appear increasingly compromised, a further indication of the role of ferroxidation in conferring peroxide tolerance to the bacterium. 相似文献
63.
Margherita Sosio Giuseppe Amati Carmela Cappellano Edoardo Sarubbi Federica Monti Stefano Donadio 《Molecular microbiology》1996,22(1):43-51
SecA protein, the ATPase promoting translocation of proteins across the Escherichia coli inner membrane, contains two ATP-binding domains that differ greatly in their affinity for bound nucleotide. In order to define more precisely the location of the high-affinity nucleotide-binding site, oligonucleotide-directed mutagenesis was used to introduce cysteine residues into the SecA sequence, and a cysteine-specific cleavage reagent was employed to generate defined peptides of SecA protein after photocross-linking with [α-32P]-ATP. This analysis revealed that the nucleotide was cross-linked between amino acid residues 75 and 97 of SecA protein. The biochemical function of the high affinity ATP-binding domain was explored by subcellular fractionation studies which demonstrated that SecA proteins defective in this region were found almost exclusively in their integral membrane form, while SecA proteins with defects in the low-affinity ATP-domain showed a normal distribution of cytosolic, peripheral and integral membrane forms. Interestingly, the SecA51(Ts) protein that has a Leu to Pro substitution at amino acid residue 43 bound ATP with high affinity, but its fractionation pattern and translocation ATPase activity were similar to those of proteins with defects in the high-affinity ATP-binding site. These results delimit more precisely the high-affinity ATP-binding domain of SecA, indicate the importance of the early amino-terminal region of SecA protein in the functioning of this domain, and demonstrate the role of this domain in regulating penetration of SecA protein into the inner membrane. Our results lead to a simple model for the regulation of a cycle of SecA insertion into, and de-insertion from, the inner membrane by the activity of the high-affinity ATP-binding domain. 相似文献
64.
F. Demay S. Geffroy C. Tiffoche M. de Monti M. L. Thieulant 《Cell biology and toxicology》1996,12(4-6):317-324
Analysis of the mechanism of action of estrogen receptor shows protein and mRNA polymorphism within distinct pituitary receptor-positive cells. The lactotropes exhibit unique properties in these mechanisms that distinguish them from gonadotropes. Therefore, this cell type constitutes an especially interesting model in the male as well as in the female for estrogen receptor studies.Abbreviations PRL
prolactin
- E2
estradiol
- ER
estrogen receptor
- GnRH
gonadotropin releasing hormone
- PMSG
pugnant mare serum gonadotropin 相似文献
65.
Mesnard F Azaroual N Marty D Fliniaux MA Robins RJ Vermeersch G Monti JP 《Planta》2000,210(3):446-453
Nitrogen metabolism was monitored in suspension cultured cells of Nicotiana plumbaginifolia Viv. using nuclear magnetic resonance (NMR) spectroscopy following the feeding of (15NH4)2SO4 and K15NO3. By using two-dimensional 15N-1H NMR with heteronuclear single-quantum-coherence spectroscopy and heteronuclear multiple-bond-coherence spectroscopy sequences,
an enhanced resolution of the incorporation of 15N label into a range of compounds could be detected. Thus, in addition to the amino acids normally observed in one-dimensional
15N NMR (glutamine, aspartate, alanine), several other amino acids could be resolved, notably serine, glycine and proline. Furthermore,
it was found that the peak normally assigned to the non-protein amino-acid γ-aminobutyric acid in the one-dimensional 15N NMR spectrum was resolved into a several components. A peak of N-acetylated compounds was resolved, probably composed of the intermediates in arginine biosynthesis, N-acetylglutamate and N-acetylornithine and, possibly, the intermediate of putrescine degradation into γ-aminobutyric acid, N-acetylputrescine. The occurrence of 15N-label in agmatine and the low detection of labelled putrescine indicate that crucial intermediates of the pathway from glutamate
to polyamines and/or the tobacco alkaloids could be monitored. For the first time, labelling of the peptide glutathione and
of the nucleotide uridine could be seen.
Received: 29 March 1999 / Accepted: 15 July 1999 相似文献
66.
François-Yves Dupradeau Guillaume Le Flem Jean-Michel Wieruszeski Manuel Dauchez Alain Alix Véronique Larreta-Garde Jean-Pierre Monti 《Letters in Peptide Science》1997,4(4-6):489-495
H-NMR studies of the bovine insulin S-sulfonatedB-chain are reported in H2O/D2O (9/1) and inglycerol-d5 (5 M) using two-dimensional NMRspectroscopy. The first results show that the oxidizedinsulin B-chain secondary structure differs from thatof native insulin by a loss of the -helixbetween the two disulfide bridges and that theglycerol favours the structuring of the peptide. 相似文献
67.
François-Yves Dupradeau Guillaume Le Flem Jean-Michel Wieruszeski Manuel Dauchez Alain Alix Véronique Larreta-Garde Jean-Pierre Monti 《International journal of peptide research and therapeutics》1997,4(4-6):489-495
Summary
1H-NMR studies of the bovine insulin S-sulfonated B-chain are reported in H2O/D2O (9/1) and in glycerol-d
5 (5 M) using two-dimensional NMR spectroscopy. The first results show that the oxidized insulin B-chain secondary structure
differs from that of native insulin by a loss of the α-helix between the two disulfide bridges and that the glycerol favours
the structuring of the peptide. 相似文献
68.
K Flanagan K Fitzgerald J Baker K Regnstrom S Gardai F Bard S Mocci P Seto M You C Larochelle A Prat S Chow L Li C Vandevert W Zago C Lorenzana C Nishioka J Hoffman R Botelho C Willits K Tanaka J Johnston T Yednock 《PloS one》2012,7(7):e40443
TH17 cells enter tissues to facilitate pathogenic autoimmune responses, including multiple sclerosis (MS). However, the adhesion molecules involved in the unique migratory capacity of TH17 cells, into both inflamed and uninflamed tissues remain unclear. Herein, we characterize MCAM (CD146) as an adhesion molecule that defines human TH17 cells in the circulation; following in vitro restimulation of human memory T cells, nearly all of the capacity to secrete IL-17 is contained within the population of cells expressing MCAM. Furthermore, we identify the MCAM ligand as laminin 411, an isoform of laminin expressed within the vascular endothelial basement membranes under inflammatory as well as homeotstatic conditions. Purified MCAM-Fc binds to laminin 411 with an affinity of 27 nM, and recognizes vascular basement membranes in mouse and human tissue. MCAM-Fc binding was undetectable in tissue from mice with targeted deletion of laminin 411, indicating that laminin 411 is a major tissue ligand for MCAM. An anti-MCAM monoclonal antibody, selected for inhibition of laminin binding, as well as soluble MCAM-Fc, inhibited T cell adhesion to laminin 411 in vitro. When administered in vivo, the antibody reduced TH17 cell infiltration into the CNS and ameliorated disease in an animal model of MS. Our data suggest that MCAM and laminin 411 interact to facilitate TH17 cell entry into tissues and promote inflammation. 相似文献
69.
E Villa R Vukotic C Cammà S Petta A Di Leo S Gitto E Turola A Karampatou L Losi V Bernabucci A Cenci S Tagliavini E Baraldi N De Maria R Gelmini E Bertolini M Rendina A Francavilla 《PloS one》2012,7(9):e44624
Introduction
Chronic hepatitis C is the main cause of death in patients with end-stage liver disease. Prognosis depends on the increase of fibrosis, whose progression is twice as rapid in men as in women. Aim of the study was to evaluate the effects of reproductive stage on fibrosis severity in women and to compare these findings with age-matched men.Materials and Methods
A retrospective study of 710 consecutive patients with biopsy-proven chronic hepatitis C was conducted, using data from a clinical database of two tertiary Italian care centers. Four age-matched groups of men served as controls. Data about demographics, biochemistry, liver biopsy and ultrasonography were analyzed. Contributing factors were assessed by multivariate logistic regression analysis.Results
Liver fibrosis was more advanced in the early menopausal than in the fully reproductive (P<0.0001) or premenopausal (P = 0.042) group. Late menopausal women had higher liver fibrosis compared with the other groups (fully reproductive, P<0.0001; premenopausal, P = <0.0001; early menopausal, P = 0.052). Multivariate analyses showed that male sex was independently associated with more severe fibrosis in the groups corresponding to premenopausal (P = 0.048) and early menopausal (P = 0.004) but not late menopausal pairs. In women, estradiol/testosterone ratio decreased markedly in early (vs. reproductive age: P = 0.002 and vs. premenopausal: P<0.0001) and late menopause (vs. reproductive age: P = 0.001; vs. premenopausal: P<0.0001). In men age-matched with menopausal women, estradiol/testosterone ratio instead increased (reproductive age group vs. early: P = 0.002 and vs. late M: P = 0.001).Conclusions
The severity of fibrosis in women worsens in parallel with increasing estrogen deprivation and estradiol/testosterone ratio decrease. Our data provide evidence why fibrosis progression is discontinuous in women and more linear and severe in men, in whom aging-associated estradiol/testosterone ratio increase occurs too late to noticeably influence the inflammatory process leading to fibrosis. 相似文献70.
This work presents a detailed kinetic study that shows the coupling between the E2→E1 transition and Rb(+) deocclusion stimulated by Na(+) in pig-kidney purified Na,K-ATPase. Using rapid mixing techniques, we measured in parallel experiments the decrease in concentration of occluded Rb(+) and the increase in eosin fluorescence (the formation of E1) as a function of time. The E2→E1 transition and Rb(+) deocclusion are described by the sum of two exponential functions with equal amplitudes, whose rate coefficients decreased with increasing [Rb(+)]. The rate coefficient values of the E2→E1 transition were very similar to those of Rb(+)-deocclusion, indicating that both processes are simultaneous. Our results suggest that, when ATP is absent, the mechanism of Na(+)-stimulated Rb(+) deocclusion would require the release of at least one Rb(+) ion through the extracellular access prior to the E2→E1 transition. Using vanadate to stabilize E2, we measured occluded Rb(+) in equilibrium conditions. Results show that, while Mg(2+) decreases the affinity for Rb(+), addition of vanadate offsets this effect, increasing the affinity for Rb(+). In transient experiments, we investigated the exchange of Rb(+) between the E2-vanadate complex and the medium. Results show that, in the absence of ATP, vanadate prevents the E2→E1 transition caused by Na(+) without significantly affecting the rate of Rb(+) deocclusion. On the other hand, we found the first evidence of a very low rate of Rb(+) occlusion in the enzyme-vanadate complex, suggesting that this complex would require a change to an open conformation in order to bind and occlude Rb(+). 相似文献