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Janet G. Salisbury John M. Graham Charles A. Pasternak 《Journal of biochemical and biophysical methods》1979,1(6):341-347
Thymocytes from rats and mice have been separated into large and small cell populations in high yield and purity by isopycnic centrifugation in a discontinuous (three-layered) gradient of Percoll. Cells are mixed with the middle layer and during low-speed centrifugation the small, denser cells sediment to the bottom interface whilst the large, less dense cells float to the top interface. Red blood cells, and small debris particles separate into the bottom and top layers respectively. Studies on the uptake of [3H]TdR show that the small cells are non-dividing while more than half of the large cells are capable of division. 相似文献
445.
Following transposon Tn5 mutagenesis of the plant growth-promoting rhizobacterium Pseudomonas putida GR12-2, mutants that were able to grow in the presence of the tryptophan analog 5-fluorotryptophan were selected. Seven of
the 50 5-fluorotryptophan-resistant mutants overproduced the phytohormone indoleacetic acid (IAA). Of these seven mutants,
the highest level of IAA was observed with strain P. putida GR12-2/aux1, which produced four times the amount of indoleacetic acid synthesized by the wild-type strain. Strain P. putida GR12-2/aux1, in contrast to the wild type, lost the ability to stimulate the elongation of the roots of canola seedlings under gnotobiotic
conditions. The growth rate, siderophore production, and 1-aminocyclopropane-1-carboxylate deaminase activity of mutant strain
P. putida GR12-2/aux1 were identical to those of the wild-type strain. The role of IAA in the mechanism of plant growth stimulation by P. putida GR12-2 and other plant growth-promoting rhizobacteria is discussed. 相似文献
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C A Pasternak 《Indian journal of biochemistry & biophysics》1990,27(6):363-364
Many disease affect cell behaviour by an effect at the cell surface, often leading to altered communication across the plasma membrane. Two examples of this from our own work are presented. The first concerns the induction of pores, leading to a breach of insulating properties of the cell membrane, by agents as diverse as certain viruses, bacterial and animal toxins, or immune molecules. In each case, membrane damage can be prevented by divalent cations such as Ca2+ or Zn2+. The second example concerns the effect of stress stimuli on the ability of cells to take up glucose. Different stresses, such as hyperthermia, toxic chemicals or infection by certain viruses, cause cells to increase glucose uptake. As with insulin-stimulated glucose uptake, the mechanism is by translocation of the glucose transporter protein from an intracellular (inactive) site to the plasma membrane. 相似文献
449.
Opiates and enkephalins: a common binding site mediates their analgesic actions in rats 总被引:3,自引:0,他引:3
3H-Labelled opiate and enkephalin ligands appear to bind with highest affinity to a single site responsible for their analgesic properties. Administered , naloxazone, an irreversible opiate, selectively inhibits for over 24 hours the high affinity binding of 3H-labelled mu, and kappa opiates and enkephalins. This inhibition of binding gradually resolves over 3 days, perhaps correlating with receptor turnover. Naloxazone treatment also abolishes morphine, D-ala2-met5-enkephalinamide and betah-endorphin analgesia. Although morphine and D-ala2-met5-enkephalinamide bind with similar potencies to the high affinity site, morphine's potency for the low affinity D-ala2-met5-enkephalinamide site is far less than the enkephalin analog. These results imply that all 3H-ligands examined bind with highest affinity to a mu-like receptor while low affinity D-ala2-met5-enkephalinamide binding, with a KD of 6 nM, represents a delta-like receptor. 相似文献
450.
Naloxazone irreversibly inhibits the high affinity binding of [125I]D-ala2-D-leu5-enkephalin 总被引:2,自引:0,他引:2
Scatchard analyses of [125I]D-ala2-D-leu5-enkephalin binding in rat brain membranes are curvilinear, suggesting low and high affinity sites. Treating the membranes with naloxazone abolishes the high affinity binding with slight effect on low affinity binding. Displacement of [125I]-D-ala2-D-leu5-enkephalin binding by morphine in untreated membranes is biphasic. Displacement by morphine in naloxazone-treated tissue is monophasic, with no inhibition by low concentrations of morphine. Naloxazone treatment has little effect on displacements by unlabeled D-ala2-D-leu5-enkephalin. Binding in N4TG1 neuroblastoma cells, which demonstrates a linear Scatchard plot with single affinity constant similar to that of the low affinity binding in brain, is less sensitive to naloxazone's actions. Naloxazone treatment inhibits D-ala2-D-leu5-enkephalin analgesia. 相似文献