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We used water-soluble styryl pyridinium dyes that fluoresce at the membrane-water interface to study vesicle traffic in endothelial
cells. Cultured endothelial cells derived from bovine and human pulmonary microvessels were incubated in styryl probes, washed
to remove dye from the plasmalemmal outer face, and observed by digital fluorescence microscopy. Vesicles that derived from
plasmalemma by endocytosis were filled with the styryl dye. These vesicles were distributed throughout the cytosol as numerous
particles of heterogeneous diameter and brightness. Vesicle formation was activated 2-fold following addition of extracellular
albumin whereas a control protein, immunoglobulin G, had no effect. Dye uptake was abrogated by labeling at low temperatures
and inhibitors of phosphoinositide-3-kinase (PI 3-kinase). Tyrosine kinase inhibitors (genistein and herbimycin A) prevented
the albumin-induced vesicle formation. Cytochalasin B prevented vesicle redistribution indicating involvement of actin filaments
in translocation of endosomes away from sites of vesicle formation. Styryl dye was lost from cells by exocytosis as evident
by the disappearance of discrete fluorescent particles. N-ethylmaleimide and botulinum toxin types A and B caused cells to accumulate increased number of vesicles suggesting that exocytosis was regulated by NSF-dependent
SNARE mechanism. The results suggest that phosphoinositide metabolism regulates endocytosis in endothelial cells and that
extracellular albumin activates endocytosis by a mechanism involving tyrosine phosphorylation, whereas exocytosis is a distinct
process regulated by the SNARE machinery. The results support the hypothesis that albumin regulates its internalization and
release in vascular endothelial cells via activation of specific endocytic and exocytic pathways. 相似文献
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Ryan J. Haasl Bret A. Payseur 《Evolution; international journal of organic evolution》2014,68(9):2737-2744
Long dinucleotide repeats found in exons present a substantial mutational hazard: mutations at these loci occur often and generate frameshifts. Here, we provide clear and compelling evidence that exonic dinucleotides experience strong selective constraint. In humans, only 18 exonic dinucleotides have repeat lengths greater than six, which contrasts sharply with the genome‐wide distribution of dinucleotides. We genotyped each of these dinucleotides in 200 humans from eight 1000 Genomes Project populations and found a near‐absence of polymorphism. More remarkably, divergence data demonstrate that repeat lengths have been conserved across the primate phylogeny in spite of what is likely considerable mutational pressure. Coalescent simulations show that even a very low mutation rate at these loci fails to explain the anomalous patterns of polymorphism and divergence. Our data support two related selective constraints on the evolution of exonic dinucleotides: a short‐term intolerance for any change to repeat length and a long‐term prevention of increases to repeat length. In general, our results implicate purifying selection as the force that eliminates new, deleterious mutants at exonic dinucleotides. We briefly discuss the evolution of the longest exonic dinucleotide in the human genome—a 10 x CA repeat in fibroblast growth factor receptor‐like 1 (FGFRL1)—that should possess a considerably greater mutation rate than any other exonic dinucleotide and therefore generate a large number of deleterious variants. 相似文献
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L. G. Firbank 《Oecologia》1993,94(3):351-355
The changing populations of weeds during 13 years of the Broadbalk continuous wheat experiment were analysed to investigate the extent of differences in shortterm variability of cover between species. The data were from two sections of the experiment where winter wheat was grown continuously under herbicide treatment for 13 and 6 years respectively. Logistic regressions were fitted to the data. Equisetum arvense showed significant long-term increases on both sections; long-term trends were also detected in the longer data run for Agrostis stolonifera, Cirsium arvensa, Poa trivialis, Ranunculus arvensis and Vicia sativa, and for Medicago lupulina on the shorter data run. Variation around long-term trends was low in the case of Equisetum, and, in the longer data run, for Cirsium and Tussilago farfara, and high for Poa spp. and Vicia. Cover values on the two sections were positively correlated for Alopecurus myosuroides, Equisetum, Poa annua and Tripleurospermum inodorum. There was a weak correlation between C-S-R strategy and short-term variability; the more competitive species displayed less variability than the ruderal species. Furthermore, species regenerating from persistent seed banks were more variable in the short term than those regenerating from short-lived seed or bud banks. This can be explained by differences in response to year-to-year variation in environmental conditions, those species with persistent seed banks being typically more sensitive to annual fluctuations in the environment than those without. 相似文献
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Mehdi Razzaghi 《Biometrical journal. Biometrische Zeitschrift》1991,33(7):775-779
The problem of testing the equality of means of two normal populations is considered when independent random samples of random sizes are given with the total number of observations from both populations being a fixed number. An application in forestry is discussed. 相似文献