全文获取类型
收费全文 | 972篇 |
免费 | 91篇 |
出版年
2021年 | 6篇 |
2020年 | 7篇 |
2019年 | 10篇 |
2018年 | 9篇 |
2017年 | 11篇 |
2016年 | 9篇 |
2015年 | 32篇 |
2014年 | 31篇 |
2013年 | 37篇 |
2012年 | 41篇 |
2011年 | 47篇 |
2010年 | 37篇 |
2009年 | 34篇 |
2008年 | 57篇 |
2007年 | 48篇 |
2006年 | 40篇 |
2005年 | 44篇 |
2004年 | 52篇 |
2003年 | 34篇 |
2002年 | 32篇 |
2001年 | 30篇 |
2000年 | 36篇 |
1999年 | 21篇 |
1998年 | 9篇 |
1997年 | 13篇 |
1996年 | 11篇 |
1995年 | 7篇 |
1994年 | 13篇 |
1993年 | 7篇 |
1992年 | 16篇 |
1991年 | 12篇 |
1990年 | 21篇 |
1989年 | 27篇 |
1988年 | 14篇 |
1987年 | 15篇 |
1986年 | 19篇 |
1985年 | 25篇 |
1984年 | 16篇 |
1983年 | 11篇 |
1982年 | 7篇 |
1981年 | 7篇 |
1980年 | 6篇 |
1979年 | 7篇 |
1978年 | 8篇 |
1977年 | 6篇 |
1975年 | 8篇 |
1973年 | 6篇 |
1971年 | 8篇 |
1967年 | 5篇 |
1960年 | 8篇 |
排序方式: 共有1063条查询结果,搜索用时 31 毫秒
921.
Wang J Della Penna K Wang H Karczewski J Connolly TM Koblan KS Bennett PB Salata JJ 《American journal of physiology. Heart and circulatory physiology》2003,284(1):H256-H267
We established HEK-293 cell lines that stably express functional canine ether-à-go-go-related gene (cERG) K(+) channels and examined their biophysical and pharmacological properties with whole cell patch clamp and (35)S-labeled MK-499 ([(35)S]MK-499) binding displacement. Functionally, cERG current had the hallmarks of cardiac delayed rectifier K(+) current (I(Kr)). Channel opening was time- and voltage dependent with threshold near -40 mV. The half-maximum activation voltage was -7.8 +/- 2.4 mV at 23 degrees C, shifting to -31.9 +/- 1.2 mV at 36 degrees C. Channels activated with a time constant of 13 +/- 1 ms at +20 mV, showed prominent inward rectification at depolarized potentials, were highly K(+) selective (Na(+)-to-K(+) permeability ratio = 0.007), and were potently inhibited by I(Kr) blockers. Astemizole, terfenadine, cisapride, and MK-499 inhibited cERG and human ERG (hERG) currents with IC(50) values of 1.3, 13, 19, and 15 nM and 1.2, 9, 14, and 21 nM, respectively, and competitively displaced [(35)S]MK-499 binding from cERG and hERG with IC(50) values of 0.4, 12, 35, and 0.6 nM and 0.8, 5, 47, and 0.7 nM, respectively. cERG channels had biophysical properties appropriate for canine action potential repolarization and were pharmacologically sensitive to agents known to prolong QT. A novel MK-499 binding assay provides a new tool to detect agents affecting ERG channels. 相似文献
922.
Kevin?WhittingstallEmail author Gerhard?Stroink Larry?Gates JF?Connolly Allen?Finley 《Biomedical engineering online》2003,2(1):14
Background
The electroencephalogram (EEG) reflects the electrical activity in the brain on the surface of scalp. A major challenge in this field is the localization of sources in the brain responsible for eliciting the EEG signal measured at the scalp. In order to estimate the location of these sources, one must correctly model the sources, i.e., dipoles, as well as the volume conductor in which the resulting currents flow. In this study, we investigate the effects of dipole depth and orientation on source localization with varying sets of simulated random noise in 4 realistic head models. 相似文献923.
Although many treatments have been proposed for the prevention of intestinal adhesions, none has been completely effective. For bowel obstruction due to adhesions the initial approach should be conservative. If operation becomes necessary, the best results depend on avoidance of trauma and infection, division of adhesions with cautery, use of mesothelial grafts, instillation of intraperitoneal hyaluronidase and stimulation of early postoperative peristalsis. In the event of massive adhesions or failure of other treatment, intestinal plication is the treatment of choice. 相似文献
924.
It is generally considered that tracks of cell adhesiveness are important in controlling cell migration during the development and regeneration of many tissues. In order to investigate this experimentally, a number of techniques have in the past been employed to make patterns of differential adhesiveness for in vitro studies. However, practical limitations on patterning resolution and the introduction of residual topography to the experimental substrata have restricted their usefulness. Here we describe a simplified photolithographic technique for patterning cell adhesiveness which allows a high degree of flexibility and precision. We have quantified, using adhesion and spreading characteristics of BHK cells, the differential adhesiveness that can be created on patterned surfaces, how this alters with the duration of exposure to serum proteins, and how this, in turn, relates to the persistence of cell patterning despite increases in cell density. We believe that this technique will prove extremely useful for the detailed in vitro examination of the mechanisms controlling cell behavior as it offers a degree of precision and ease of fabrication that has previously been unavailable. 相似文献
925.
Mason AB Halbrooks PJ James NG Connolly SA Larouche JR Smith VC MacGillivray RT Chasteen ND 《Biochemistry》2005,44(22):8013-8021
Each homologous lobe of human serum transferrin (hTF) has one Fe(3+) ion bound by an aspartic acid, a histidine, two tyrosine residues, and two oxygens from the synergistic anion, carbonate. Extensive characterization of these ligands in the N-terminal lobe has been carried out. Despite sharing the same set of ligands, there is a substantial amount of evidence that the N- and C-lobes are inequivalent. Studies of full-length hTF have shown that iron release from each lobe is kinetically distinguishable. To simplify the assessment of mutations in the C-lobe, we have created mutant hTF molecules in which the N-lobe binds iron with high affinity or not at all. Mutations targeting the C-lobe liganding residues have been introduced into these hTF constructs. UV-visible spectral, kinetic, and EPR studies have been undertaken to assess the effects of each mutation and to allow direct comparison to the N-lobe. As found for the N-lobe, the presence of Y517 in the C-lobe (equivalent to Y188 in the N-lobe) is absolutely essential for the binding of iron. Unlike the N-lobe, however, mutation of Y426 (equivalent to Y95) does not produce a stable complex with iron. For the mutants that retain the ability to bind iron (D392S and H585A), the rates of release are considerably slower than those measured for equivalent mutations in the N-lobe at both pH 7.4 and pH 5.6. Equilibrium binding experiments with HeLa S(3) cells indicate that recombinant hTF, in which Y426 or H585 is mutated, favor a closed or nearly closed conformation while those with mutations of the D392 or Y517 ligands appear to promote an open conformation. The differences in the effects of mutating the liganding residues in the two lobes and the subtle indications of cooperativity between lobes point to the importance of the transferrin receptor in effecting iron release from the C-lobe. Significantly, the equilibrium binding experiments also indicate that, regardless of which lobe contains the iron, the free energy of binding is equivalent and not additive; each monoferric hTF has a free energy of binding that is 82% of diferric hTF. 相似文献
926.
927.
928.
The current studies describe a new, robust cell-based functional assay useful to characterize L-type voltage-dependent calcium channels and their antagonists. The basis of this assay is measurement in plate format of Ca2+ influx through the L-type Ca2+ channel complex (alpha1C, alpha2delta, and beta2a subunits) in response to potassium-mediated depolarization; EC(50)=11 mM [K+](o). The Ca2+ influx was inhibited by the L-type Ca2+ channel antagonist, nimodipine; IC(50)=59 nM. These cells were also transfected with the Kir2.3 inward rectifier K(+) channel, which allows for changing the cell membrane potential by modulation of extracellular [K](o); -65 mV in physiological [K](o) and -28 mV in 30 mM [K](o) containing buffer. The conformational state of the voltage-sensitive Ca2+ channel is altered under these different conditions. Under the depolarized condition, nimodipine was a more potent antagonist, inhibiting Ca2+ influx with an IC(50) value of 3 nM. The results demonstrate that the interaction of nimodipine and other antagonists with the channel is modulated by changes in membrane potential and thus the state of the channel. Overall, this novel assay can be used to identify state-dependent calcium channel antagonists and should be useful for evaluating state-dependent inhibitory potency of a large number of samples. 相似文献
929.
Application of 2-D free-flow electrophoresis/RP-HPLC for proteomic analysis of human plasma depleted of multi high-abundance proteins 总被引:1,自引:0,他引:1
Moritz RL Clippingdale AB Kapp EA Eddes JS Ji H Gilbert S Connolly LM Simpson RJ 《Proteomics》2005,5(13):3402-3413
Free-flow electrophoresis (FFE) and rapid (6 min) RP-HPLC was used to fractionate human citrate-treated plasma. Prior to analysis, the six most abundant proteins in plasma were removed by immunoaffinity chromatography; both depleted plasma and the fraction containing the six abundant proteins depleted were taken for MS-based analysis. Fractionated proteins were digested with trypsin and the generated peptides were subjected to MS-based peptide sequencing. To date, 78 plasma proteins have been unambiguously identified by manual validation from 16% (15/96 FFE total fractions) of the collected FFE pools; 55 identifications were based on > or = 2 tryptic peptides and 23 using single peptides. The molecular weight range of proteins and peptides isolated by this method ranged from approximately 190 K (e.g., Complement C3 and C4) to approximately 4-6 K (e.g., CRISPP and Apolipoprotein C1). This FFE/RP-HPLC approach reveals low-abundance proteins and peptides (e.g., L-Selectin approximately 17 ng/mL and the cancer-associated SCM-recognition, immunodefense suppression, and serine protease protection peptide (CRISPP) at approximately 0.5-1 ng/mL), where CRISPP was found in association with alpha-1-antitrypsin as a non-covalent complex, in the fraction containing the depleted high-abundance proteins. In contrast to shotgun proteomic approaches, the FFE/RP-HPLC method described here allows the identification of potentially interesting peptides to be traced back to their protein of origin, and for the first time, has confirmed the "protein sponge" hypothesis where the 35 residue CRISPP polypeptide is non-covalently complexed with the major circulating plasma protein alpha-1-antitrypsin. 相似文献
930.
Lignans from the roots of Echinops giganteus 总被引:2,自引:0,他引:2
Two new lignans, (+)-4-(3-methylbutanoyl)-2,6-di(3,4-dimethoxy)phenyl-3,7-dioxabicyclo[3.3.0]octane and (+)-4-hydroxy-2,6-di(3,4-dimethoxy)phenyl-3,7-dioxabicyclo[3.3.0]octane, together with the known lupeol and sitosteryl beta-D-glucopyranoside, have been isolated from the roots of Echinops giganteus var. lelyi C. D. Adams (Compositae). The structures were elucidated on the basis of spectral studies and comparison with published data. 相似文献