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101.
102.
ATPase and Proton-Translocating Activities in a Plasma Membrane-Enriched Fraction from Cotyledons of Ricinus communis 总被引:1,自引:0,他引:1
Sucrose gradient centrifugation was used to separate the microsomalmembranes and purify the plasma membrane ATPase from Ricinuscotyledons. The pellet from a three-step (30, 34, 38%) sucrosegradient was enriched in plasma membrane as determined by acombination of marker assays. The partially purified plasma membrane ATPase was magnesium-dependentand had a pH optimum of 6.5. It showed high sensitivity to vanadate,erythrosin B, SW 26, DCCD and PCMBS but low sensitivity to azide,nitrate and NEM. Substitution of calcium for magnesium resultedin low activity, and in the presence of magnesium, calcium wasinhibitory. KCl stimulation was low (less than 50%) and of thepotassium salts tested all were stimulatory except which was inhibitory. Specificity for nucleotide triphosphateswas high, greatest activity occurring with ATP. Proton-pumping activity measured using quinacrine fluorescencequenching was inhibited by vanadate and erythrosin B but notby nitrate and oligomycin indicating that activity was mainlydue to the plasma membrane ATPase. Key words: ATPase, cotyledons, plasma membrane, proton pumping, Ricinus communis 相似文献
103.
The organization of microtubules (MTs) in the cortex of cells at interphase is an important element in morphogenesis. Mechanisms
controlling the initiation of MTs and their spatial ordering, however, are largely unknown. Our recent study concerning the
generation of a radial array of MTs in stomatal guard cells inAllium showed that the MTs initiate in a cortical MT-organizing zone adjacent to the ventral wall separating the two young guard
cells (Marc, Mineyuki and Palevitz, 1989, Planta179, 516, 530). In an attempt to detect MT-ordering mechanisms separate from the sites of MT initiation, we now employ various
drugs to manipulate the geometry and integrity of the ventral wall and thereby also the associated MT-organizing zone. In
the presence of cytochalasin D the ventral wall is tilted away from its normal mid-longitudinal anticlinal alignment, while
treatments with the herbicide chloroisopropyl-N-phenylcarbamate (CIPC) induce the formation of a branched ventral wall. Nonetheless, in either case the MTs still
form a radial array, although this is asymmetric as it is centered in accordance with the misaligned or branched ventral wall.
Since the MTs maintain their original course undisturbed as they extend beyond the abnormal ventral wall, there is no evidence
for the presence of an inherent MT-ordering mechanism at locations remote from MT-initiation sites. Following treatments with
caffeine, which abolishes the formation of the ventral wall, the MTs revert to a transversely oriented cylindrical array as
in normal epidermal cells. Thus the presence of the ventral wall, and presumably also the associated MT-organizing zone, is
essential for the establishment of the radial array. The MT-organizing zone is therefore involved not only in the initiation
of MTs, but also in determining their spatial order throughout the cell cortex.
We thank Drs. Richard J. Cyr and Yoshi Mineyuki for providing valueable suggestions during the course of this work, and Ms.
Elizabeth Bruce printing some of the figures. This research was supported by Funds from the National Science Foundation grants
DCB-8703292 to B.A.P. and DCB-8803286 to B.A.P. and J.M. 相似文献
104.
Cora-Jean S. Edgell Jill E. Haizlip C. Robert Bagnell Joan P. Packenham Paul Harrison Barry Wilbourn Victoria J. Madden 《In vitro cellular & developmental biology. Plant》1990,26(12):1167-1172
Summary Weibel-Palade bodies are ultrastructurally defined organelles found only in vascular endothelial cells. Because endothelium
in corpo is very dispersed, isolation and further characterization of this organelle has been dependent on increasing the
number of cells in culture. However, primary isolates of endothelial cells have a limited replication potential and tend to
senesce in culture. In this report, EA.hy926, a continuously replicating cell line derived from human endothelium, is shown
to contain Weibel-Palade bodies. Electron micrographs demonstrate the ultrastructural characteristics of these tissue-specific
organelles and their cytoplasmic distribution in EA.hy926 cells. Von Willebrand factor, which has been shown to exist in Weibel
Palade bodies, is demonstrated by immunofluorescence in discrete rod-shaped organelles whose size, shape, and distribution
are consistent with that of Weibel-Palade bodies in primary endothelial cell cultures. Rapid release of von Willebrand factor
can be induced by calcium ionophore, and large multimeric forms of the protein are found in EA.hy926 cells. These two properties
are consistent with the function currently ascribed to Weibel Palade bodies: storage of multimerized von Willebrand factor.
Thus ultrastructural, immunologic, and functional data establish the existence of this as yet poorly understood tissue-specific
organelle in a continuous, vigorously replicating human cell line. 相似文献
105.
How to Characterize a Biological Antioxidant 总被引:15,自引:0,他引:15
Barry Halliwell 《Free radical research》1990,9(1):1-32
An antioxidant is a substance that, when present at low concentrations compared to those of an oxidizable substrate, significantly delays or prevents oxidation of that substrate. Many substances have been suggested to act as antioxidants in vivo, but few have been proved to do so. The present review addresses the criteria necessary to evaluate a proposed antioxidant activity. Simple methods for assessing the possibility of physiologically-feasible scavenging of important biological oxidants (superoxide, hydrogen peroxide, hydroxyl radical, hypochlorous acid, haem-associated ferryl species, radicals derived from activated phagocytes, and peroxyl radicals, both lipid-soluble and water-soluble) are presented, and the appropriate control experiments are described. Methods that may be used to gain evidence that a compound actually does function as an antioxidant in vivo are discussed. A review of the pro-oxidant and anti-oxidant properties of ascorbic acid that have been reported in the literature leads to the conclusion that this compound acts as an antioxidant in vivo under most circumstances. 相似文献
106.
107.
The calcium-dependent modulation of the affinity of the cyclic nucleotide-gated (CNG) channels for adenosine 3′,5′-cyclic
monophosphate (cAMP) was studied in enzymatically dissociated rat olfactory receptor neurons, by recording macroscopic cAMP-activated
currents from inside-out patches excised from their dendritic knobs. Upon intracellular addition of 0.2 mm Ca2+ (0.2 Ca) the concentration of cAMP required for the activation of half-maximal current (EC50) was reversibly increased from 3 μm to about 30 μm. This Ca2+-induced affinity shift was insensitive to the calmodulin antagonist, mastoparan, was abolished irreversibly by a 2-min exposure
to 3 mm Mg2++ 2 mm EGTA (Mg + EGTA), and was not restored by the application of calmodulin (CAM). Addition of CAM plus 0.2 mm Ca2+ (0.2 Ca + CAM), further reversibly shifted the cAMP affinity from 30 μm to about 200 μm. This affinity shift was not affected by Mg + EGTA exposure, but was reversed by mastoparan. Thus, the former Ca2+-only effect must be mediated by an unknown endogenous factor, distinct from CAM. Removal of this factor also increased the
affinity of the channel for CAM. The affinity shift induced by Ca2+-only was maintained in the presence of the nonhydrolyzable cAMP analogue, 8-bromo-cAMP and the phosphatase inhibitor, microcystin-LR,
ruling out modulation by phosphodiesterases or phosphatases. Our results indicate that the olfactory CNG channels are modulated
by an as yet unidentified factor distinct from CAM.
Received: 26 December 1995/Revised: 14 March 1996 相似文献
108.
109.
ALWYN WILLIAMS MAGGIE CUSACK 《Lethaia: An International Journal of Palaeontology and Stratigraphy》1996,29(4):349-360
Organophosphatic shells of the brachiopod Lingula squarniformis , collected from Scottish Lower Carboniferous shales and mudstones of intertidal to sublittoral provenance, have been studied to ascertain chemico-structural changes resulting from fossilization. Enough original shell has been preserved at ultrastructural and molecular levels to confirm that Carboniferous and Recent integuments are homologous with stratiform successions of apatitic to organic laminae forming rhythmic sets. One of the main organic constituents, the acidic, hydrophilic gel glycosaminoglycans (GAGs), is the dominant component towards the tops of rhythms. During fossilization of the Carboniferous shells, GAGs degraded incrementally without disturbing apatitic ultrastructures, and the spaces so created became partly filled with sheets of recrystal-lized apatite with some kaolinite or with books and plates of kaolinite. The kaolinite in the shells contrasts with the illite of the entombing sediments and suggests that degrading acidic GAGs mediated in clay formation in situ . The sediments also contain framboidal pyrite, which is virtually absent from the shells themselves but is usually even more abundant, with a greater range of trace metals, in the sedimentary fills of complete shells. This imbalance suggests mediation by another gel, the glycocalyx, secreted by the inner epithelium of the brachiopod mantle. The glycocalyx would have lined the shell interior and could have served as a sorption film for dissolved metals precipitated as compounds on decomposition of body tissue. 相似文献
110.
A note on optimality in lattice square designs 总被引:1,自引:0,他引:1