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41.
Michele Solem Angela Helmrich Paul Collodi David Barnes 《Molecular and cellular biochemistry》1991,100(2):141-149
Summary Human S-protein is a serum glycoprotein that binds and inhibits the activated complement complex, mediates coagulation through interaction with antithrombin III and plasminogen activator inhibitor I, and also functions as a cell adhesion protein through interactions with extracellular matrix and cell plasma membranes. A full length cDNA clone for human S-protein was isolated from a lambda gt11 cDNA library of mRNA from the HepG2 hepatocellular carcinoma cell line using mixed oligonucleotide sequences predicted from the amino-terminal amino acid sequence of human S-protein. The cDNA clone in lambda was subcloned into pUC18 for Southern and Northern blot experiments. Hybridization with radiolabeled human S-protein cDNA revealed a single copy gene encoding S-protein in human and mouse genomic DNA. In addition, the S-protein gene was detected in monkey, rat, dog, cow and rabbit genomic DNA. A 1.7 Kb mRNA for S-protein was detected in RNA from human liver and from the PLC/PRF5 human hepatoma cell line. No S-protein mRNA was detected in mRNA from human lung, placenta, or leukocytes or in total RNA from cultured human embryonal rhabdomyosarcoma (RD cell line) or cultured human fibroblasts from embryonic lung (IMR90 cell line) and neonatal foreskin. A 1.6 Kb mRNA for S-protein was detected in mRNA from mouse liver and brain. No S-protein mRNA was detected in mRNA from mouse skeletal muscle, kidney, heart or testis. 相似文献
42.
The activating anions are found to induce an unexpectedly high (up to 8-fold for sulphite) increase of ATPase activity in intact rat liver mitochondria. This effect is not determined by the observed changes in Km and Ki (ADP) values. The stimulation seems to be caused by dissociation of the inactive complex of ATPase with Mg·ADP. The quantity of this complex formed in the course of ATP hydrolysis is approx. 90% of the total ATPase content in intact mitochondria. The data on toluene-permeabilized mitochondria suggest that the high content of the complex is a result of the stabilizing effect of some matrix macromolecules. 相似文献
43.
The components of magnesium efflux in squid axons have been studied under internal dialysis and voltage clamp conditions. The present report rules out the existence of an ATP-dependent, Na0- and Mg0-independent Mg2+ efflux (ATP-dependent Mg2+ pump) leaving the Mg2+---Na+ exchange system as the only mechanism for Mg2+ extrusion. The main features of the Mg2+ efflux are: (1) The efflux is completely dependent on ATP. (2) The efflux can be activated either by external Na+ (forward Mg2+---Na+ exchange) or external Mg2+ (Mg2+---Mg2+ exchange). (3) The mobility of the Mg2+ exchanger in the Na0+-loaded form is greater than that in the Mg2+-loaded one. (4) In variance with the Na+---Ca2+ exchange mechanism, Mg2+---Mg2+ exchange is not activated by external monovalent cations. (5) ATPγS replaces ATP in activating Mg2+---Na+ exchange suggesting that a phosphorylation/dephosphorylation process regulates this transport mechanism. 相似文献
44.
Claudio Nicolini Andrew S. Belmont Antonietta Martelli 《Cell biochemistry and biophysics》1986,8(2):103-117
Using HeLa S-3 cells synchronized by selective detachment, in this paper we report a parallel study of nuclear morphology
and autoradiography grain patterns between middle G1 and middle S phases: Our results show two distinct [3H]-thymidine labeling patterns. The first “peripheral” labeling pattern has a characteristic nuclear size distribution, in
contrast to the heterogeneous and varying size distributions of Feulgen-stained nuclei, and apparently is characteristic of
very early S phase. The sizes of the second labeling pattern—homogeneous or inhomogeneous grain distribution throughout the
nucleus—are equal or larger than the first and vary with S phase progression. Together, the corresponding nuclear sizes of
the labeled nuclei represent the larger extreme of nuclear areas, and the labeling index closely parallels the fraction of
nuclei with areas larger than the minimum size of the labeled nuclei. These results suggest a characteristic nuclear size
(reflecting unique intranuclear DNA distribution) as a necessary, if not sufficient, requirement for S phase initiation. Parallel
experimentation with rat liver cells—synchronized in vivo by partial hepatectomy and analyzed by thin section autoradiography—confirms
the existence of a peripheral labeling pattern in both the very early part and the very late part of S phase, which reconciles
our data with previous results and points to the fact that both initiation and termination sites for DNA replication are near
the nuclear periphery. 相似文献
45.
Kaoru Miyazaki Keisuke Mashima Nobuhiko Yamashita Jinpei Yamashita Takekazu Horio 《In vitro cellular & developmental biology. Plant》1985,21(1):62-66
Summary We have previously reported the transformation by Rous sarcoma virus of a cloned epithelial cell line (BRL) established from
Buffalo rat liver by H. Coon. The nontransformed (BRL) and transformed (RSV-BRL) cells grew at comparable rates in culture,
whereas only the transformed cells were tumorigenic in vivo. We report here on the existence in rat and mouse sera of a growth
inhibitor for the nontransformed BRL cells. The transformed BRL cells (RSV-BRL) were insensitive to this inhibitor. The inhibitory
activity was not prominent in sera from other species of animals tested except for rabbit; this serum inhibited the growth
of RSV-BRL cells more strongly than that of BRL cells. The growth inhibitor was partially purified from rat serum. It is a
protein free of lipid and has a molecular weight of about 220 000. The inhibitor could be separated into three components
of pI 4.6, 5.2 (major) and 5.6 by isoelectric electrophoresis.
EDITOR'S STATEMENT Although compelling theoretical arguments sometimes can be made for the likely existence of growth-inhibitory
substances of physical relevance in the control of cell proliferation, experiments aimed at identifying and studying such
factors often are difficult to design and interpret, and little strong data exists to suggest that growth-inhibitory substances
are important regulatorsin vivo. The information presented in this paper represents a start toward developing a useful system for studying growth-inhibitory
factor. David W. Barnes 相似文献
46.
Structural evidence for ligand-induced sequential conformational changes in glyceraldehyde 3-phosphate dehydrogenase 总被引:2,自引:0,他引:2
Glyceraldehyde 3-phosphate dehydrogenase is a tetramer of four chemically identical subunits which requires the cofactor nicotinamide adenine dinucleotide (NAD) for activity. The structure of the holo-enzyme from Bacillus stearothermophilus has recently been refined using X-ray data to 2.4 A resolution. This has facilitated the structure determination of both the apo-enzyme and the enzyme with one molecule of NAD bound to the tetramer. These structures have been refined at 4 A resolution using the constrained-restrained parameter structure factor least-squares refinement program CORELS. When combined with individual atomic temperature factors from the holo-enzyme, these refined models give crystallographic R factors of 30.2% and 30.4%, respectively, for data to 3 A resolution. The apo-enzyme has 222 molecular symmetry, and the subunit structure is related to that of the holo-enzyme by an approximate rigid-body rotation of the coenzyme binding domain by 4.3 degrees with respect to the catalytic domains, which form the core of the tetramer. The effect of this rotation is to shield the coenzyme and active site from solvent in the holo-enzyme. In addition to the rigid-body rotation, there is a rearrangement of several residues involved in NAD binding. The structure of the 1 NAD enzyme is asymmetric. The subunit which contains the bound NAD adopts a conformation very similar to that of a holo-enzyme subunit, while the other three unliganded subunits are very similar to the apo-enzyme conformation. This result provides unambiguous evidence for ligand-induced sequential conformational changes in B. stearothermophilus glyceraldehyde 3-phosphate dehydrogenase. 相似文献
47.
Cathepsin B, H, L and D activities in liver lysosomes were compared between species. Although cathepsin B and D were detected in bovine, pig, chicken and rat liver, striking species differences were evident for cathepsin H and L. Cathepsin L activity was particularly high in chicken lysosomal extracts, but could not be detected in bovine and pig extracts. Whereas there was no significant cathepsin H activity in bovine extracts, rat liver lysosomal extracts contained large amounts of cathepsin H activity. 相似文献
48.
Summary A monoclonal antibody against pea-leaf calmodulin was used to localise this calcium-binding protein on frozen sections of compound eyes of several arthropod species and on nitrocellulose replicas of electrophoretically separated peptides of isolated photoreceptor membrane from crayfish, fly, and squid. We report the presence of immunochemically detectable amounts of calmodulin specifically associated with the photoreceptor microvilli of rhabdomeral photoreceptors. A weak immunofluorescent signal was also observed in the cytoplasm of retinula cells. The presence of calmodulin in rhabdomeral microvilli is discussed in view of its possible implication in phototransduction and/or involvement in cytoskeletal structures associated with photoreceptor membranes in invertebrates. 相似文献
49.
Ralf Morgenstern Claes Guthenberg Bengt Mannervik Joseph W. DePierre 《FEBS letters》1983,160(1-2):264-268
The amount and nature of glutathione transferases in rat liver microsomes were determined using immunological techniques. It was shown that cytosolic glutathione transferase subunits A plus C, and B plus L were present at levels of 2.4 ± 0.6 and 1.5 ± 0.1 μg/mg microsomal protein, respectively. These levels are 10-times higher than those for non-specific binding of cytosolic components judging from the distribution of lactate dehydrogenase, a cytosolic marker. The possibility that a portion of these glutathione transferases is functionally localized on the endoplasmic reticulum is discussed. A previously described microsomal glutathione transferase which is distinct from the cytosolic enzymes is present in an amount of 31 ± 6 μg/mg microsomal protein. 相似文献
50.
The herbicide glyphosate (N-phosphonomethyl glycine) is a potent reversible inhibitor of the 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase activity of the purified arom multienzyme complex from Neurospora crassa. Inhibition of the EPSP synthase reaction by glyphosate is competitive with respect to phosphoenolpyruvate, with K(i) 1.1 microM, and uncompetitive with respect to shikimate-3-phosphate. The kinetic patterns are consistent with a compulsory order sequential mechanism in which either PEP or glyphosate can bind to an enzyme: shikimate-3-phosphate complex. 相似文献