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31.
32.
Gerald M. Kidder Douglas J. Barron Joanna B. Olmsted 《Development genes and evolution》1988,197(2):110-114
Summary We have examined the persistence of midbody channels during the second, third, and fourth cleavage cycles of the mouse using immunofluorescence to map the distribution of midbody microtubule bundles in intact embryos. Electron microscopy showed these bundles to be a characteristic feature of midbodies throughout the interphase period. In recently-divided embryos at each cleavage stage the number of midbodies was half the number of blastomeres, and declined towards zero as the next cleavage approached. This indicated to us that the only midbodies present in each stage were those which had arisen in the immediately-preceding division. Of those blastomeres which were in mitosis at the time of fixation, less than 4% were connected via a midbody to another blastomere, demonstrating that persistence of midbodies beyond a single cleavage cycle is a rare event. We conclude that midbody channels in our embryos are likely to connect only pairs of sister blastomeres because midbodies do not persist through multiple cleavage cycles. Midbody channels cannot, therefore, be regarded as providing extensive cell coupling in advance of the onset of gap junctional communication. 相似文献
33.
34.
Gerald A. Merrill David Miller John Chirgwin Paul M. Horowitz 《Journal of Protein Chemistry》1992,11(2):193-199
Rhodanese has been utilized as a model enzyme for the study of protein structure-function relationships. The enzyme has recently been cloned and the recombinant enzyme is now available for investigation. However, prior to use in structure-function studies, the recombinant enzyme must be shown to have the same structure and activity as the bovine liver enzyme used in the previous studies. An immunological study of the conformations of these enzyme conformers is described. Three antibodies (two monoclonal and one polyclonal, site-directed antibody) were shown to detect distinct and nonoverlapping epitopes. The epitopes of the monoclonal antirhodanese antibodies (R207 and MAB11) were mapped to the same CNBr digest fragment of the amino terminal domain of rhodanese, and the epitope of the site-directed antibody prepared against the interdomain tether sequence of rhodanese (PAT-T1) was mapped to that region of rhodanese (residues 142–156). The rhodanese conformers were studied by monitoring the accessibility of the epitopes recognized by each antibody in each conformer using an indirect ELISA. None of the antibodies could detect its epitope on the purified liver enzyme. Two of the antibodies (R207 and PAT-T1) could also not detect their epitopes on the recombinant enzyme. However, MAB11 did detect a conformational difference between the natural and recombinant rhodanese conformers, indicating the conformational difference is localized in the first 73 amino acids of rhodanese. This difference presumably reflects the difference in the histories of the two enzymes and may be due to differences in enzyme folding, differences in the purification procedures, and differences in storage conditions—all of which could influence the final conformation of the enzyme. 相似文献
35.
Toshihiro Mitaka Gerald L. Sattler Henry C. Pitot Yohichi Mochizuki 《Virchows Archiv. B, Cell pathology including molecular pathology》1992,62(1):329-335
Phenotypes of the cells developing into small colonies after days of primary culture of adult rat hepatocytes in serum-free
modified Dulbecco Modified Eagles’ medium containing 10 mM nicotinamide and 10 ng/ml epidermal growth factor were analyzed
immunocytochemically, cytochemically and ultrastructurally. Albumin, cytokeratin 8 and 18 were seen by immunocytochemical
techniques in the cells of the small colonies at Day 6. Transferrin, α-antitrypsin, ceruloplasmin, and haptoglobin, proteins
secreted by mature hepatocytes, were faintly stained in these cells as was α-fetoprotein. These proteins were secreted into
the culture medium as evidenced by immunoblot analysis. γ-Glutamyltransferase, alkaline phosphatase and glucose 6-phosphatase
were not present in the cells of the small colonies as well as the surrounding hepatocytes at Day 6 of culture. In addition,
ultrastructural examinations of the cells in the small colonies indicated that these cells not only had many characteristic
mitochondria and desmosomes, but also a few small peroxisomes. Such cells, even after 20 days in culture were proliferating,
as evidenced by the intranuclear presence of the proliferating cell nuclear antigen. The potential relation of these cells
to hepatocytes which may serve as the principal reserve for replicating hepatocytes is discussed. 相似文献
36.
Gerald W. Hart 《Current opinion in cell biology》1992,4(6):1017-1023
Protein glycosylation is more abundant and structurally diverse than all other types of post-translational modifications combined. Protein-bound saccharides range from dynamic monosaccharides on nuclear and cytoplasmic proteins, to enormously complex 'recognition' molecules on extracellular N- or O-linked glycoproteins or proteoglycans. Recent elucidation of a few of the myriad functions of these saccharides has finally opened a crack in the door to one the last great frontiers of biochemistry. 相似文献
37.
Jagjivan R. Mehta Kyle G. Braund Gerald A. Hegreberg Vijay Thukral 《Neurochemical research》1991,16(2):129-135
Erythrocyte membranes and their liposomes were prepared from clinically normal dogs and Labrador retrievers with hereditary muscular dystrophy. The static and dynamic components of fluidity of each membrane were then assessed by steady-state fluorescence polarization techniques using limiting hindered fluorescence anisotropy and order parameter values of 1,6-diphenyl-1,3,5-hexatriene (DPH) and fluorescence anisotropy values ofdl-2-(9-anthroyl)-stearic acid anddl-12-(9-anthroyl)-stearic acid, respectively. Membrane lipids were extracted and analyzed by thin-layer chromatography and gas chromatography. The results of these studies demonstrated that the lipid fluidity of erythrocyte membranes, and their liposomes, prepared from dystrophic dogs were found to possess significantly lower static and dynamic components of fluidity than control counterparts. Analysis of the composition of membranes from dystrophic dogs revealed a higher ratio of saturated fatty acyl chain/unsaturated chains (w/w) and lower double-bond index. Alterations in the fatty acid composition such as decrease in levels of linoleic (18:2) and arachidonic (20:4) acids and increase in palmitic (16:0) and stearic (18:0) acids were also observed in the membranes of dystrophic animals. These associated fatty acyl alterations could explain, at least in part, the differences in membrane fluidity between dystrophic and control dogs. 相似文献
38.
39.
Hans-B?rje Jansson A. Jeyaprakash Gerald C. Coles Nahum Marban-Mendoza Bert M. Zuckerman 《Journal of nematology》1986,18(4):570-574
Caenorhabditis elegans and Panagrellus redivivus were investigated for surface carbohydrates using fluorescent-labelled and ferritin-labelled lectins. Rhodamine-labelled Concanavalin A was specifically located in the cephalic region of both species. Rhodamine-labelled wheat germ agglutinin was located over the entire cuticle of P. redivivus but was absent on C. elegans. Rhodamine-labelled peanut agglutinin and Limax flavus agglutinin did not label nematodes of either species. Galactose and sialic acid were not detected on either species, whereas mannose-glucose residues were specifically localized in the head areas of both species. No detectable N-acetylglucosamine occurred on C. elegans, but it was evenly distributed over the cuticle surface of P. redivivus. 相似文献
40.
Francis Chaouloff Guy A. Kennett Bernard Serrurrier Daniele Merino Gerald Curzon 《Journal of neurochemistry》1986,46(5):1647-1650
Rats were trained to run on a horizontal treadmill for 2 h at 20 m/min. This activity considerably increased plasma free tryptophan (TRP) (+70%) but did not alter plasma total TRP levels and had little or no effect on plasma concentrations of the other large neutral amino acids (LNAAs) that compete with TRP for entry into the brain. Brain TRP levels increased by 80%. The only other brain LNAA to be affected by exercise was threonine, which rose moderately. The results indicate that increased plasma free TRP was specifically responsible for the increase of brain TRP after 2 h of exercise. Brain lysine was also increased whereas glycine, alanine, and gamma-aminobutyric acid were decreased. The differences between the present findings and those previously obtained following 2 h immobilization stress are discussed. 相似文献