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961.
Adenylate cyclase activity measured in membranes of cultured normal rat kidney (NRK) fibroblasts was markedly increased by prior treatment of the intact cells with trypsin. Cell population density influenced the extent of activation observed. Trypsin treatment of sparse cells significantly enhanced adenylate cyclase activity, whereas similar treatment of confluent cells caused only a slight increase in adenylate cyclase activity. The degree of activation noted after trypsin treatment also varied depending on the adenylate cyclase function measured. Activity determined in the presence of GTP alone showed the greatest increase after trypsin treatment. Similar enhancement of adenylate cyclase activity of a washed cell membrane preparation was achieved by the addition of low concentrations of trypsin directly to the adenylate cyclase reaction mixture. The membranes of confluent NRK fibroblasts initially exhibited higher adenylate cyclase activity than did membranes of sparse cells. The present results suggest that this change in adenylate cyclase activity at cell confluence is not due to an increase in the amount of adenylate cyclase in the cell membrane but rather to a change in membrane components that regulate its activity. Proteolytic activation of adenylate cyclase appears to result from degradation of cell membrane proteins that modulate the activity of this enzyme. 相似文献
962.
Characterization of an immune complex kinase in immunoprecipitates of avian sarcoma virus-transformed fibroblasts. 总被引:5,自引:0,他引:5
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Kinase activity detected in immune complexes containing the src gene product of the avian sarcoma virus has been reported. To further characterize this immune complex kinase, we developed a routine quantitative assay involving trichloroacetic acid precipitation on filters. The enzyme reaction required either Mg2+ or Mn2+, but was inactive with Ca2+. The kinetics of the phosphorylation reaction indicated a transient enzyme activity limited by rapid substrate-dependent inactivation of the enzyme. A variety of nucleoside and deoxyribonucleoside triphosphates (dATP, ATP, GTP, CTP, dGTP, TTP, dCTP) served as phosphoryl donors. The phosphorylation of immunoglobulin G was inhibited by the presence of nucleoside diphosphates. Deoxyribonucleoside diphosphates can either stimulate or inhibit the kinase reaction depending upon the concentration used. The unusual enzymatic properties of the immune complex kinase raise the possibility that the enzyme does not function as a protein kinase in vivo, but rather belongs to a different class of kinases (nucleotide kinases) which adventitiously phosphorylates immunoglobulin G when immunoprecipitated with immune serum. 相似文献
963.
Quantitation of a transformation-sensitive, adhesive cell surface glycoprotein. Decrease of several untransformed permanent cell lines
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We have quantitated the transformation-sensitive, cell surface LETS glycoprotein on many untransformed cell types. By SDS-polyacrylamide gel electrophoresis, this trypsin-sensitive iodinatable glycoprotein comprises 1-3% of total cellular protein of the seven early passage cell types tested. In contrast, it constitutes less than 0.15% of the protein in four of six continuous cell lines. This decrease is reflected in alterations both in [14C]glucosamine labeling and in the immunofluorescent staining of early passage vs. these four permanent cell lines. These results help to clarify previous experiments in which CSP, a purified LETS protein, partially restored a fibroblastic phenotype to cells transformed by tumor viruses. These findings also indicate that a major decrease in this cell surface glycoprotein can occur in the establishment of a continuous cell line without resulting in cellular transformation. 相似文献
964.
Cyclic AMP increases the concentration of insulin receptors in cultured fibroblasts and lymphocytes.
P Thomopoulos F C Kosmakos I Pastan E Lovelace 《Biochemical and biophysical research communications》1977,75(2):246-252
Incubation of SV40 transformed fibroblasts with dibutyryl cyclic AMP, 8-bromo-cyclic AMP, or 1-methyl-3-isobutylxanthine (MIX), a phosphodiesterase inhibitor, produced a two-fold increase in insulin receptor concentration without an effect on receptor affinity. The increase was dose-dependent, was observed after 8 hrs of treatment, and reached a maximum level by 12 to 24 hours. Upon removal of the nucleotide, receptor number decreased towards basal level.Incubation of cultured human lymphocytes (IM-9 line) with cyclic AMP derivatives or MIX also increased the number of insulin receptors without an alteration in receptor affinity. This effect was partially blocked by inhibition of protein synthesis and was independent of changes in cell cycle. The increase in insulin receptors was a specific response to cyclic AMP as the number of receptors for human growth hormone was unaltered. Incubation with 8-bromo-cyclic GMP did not alter the level of insulin binding. 相似文献
965.
A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting. 总被引:1,自引:0,他引:1
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Immunotoxoin-mediated cell targeting (IMCT) is a technique for conditionally ablating specific cell types based on the cytotoxic activity of a recombinant immunotoxin anti-Tac (Fv)-PE40. To examine the feasibility of this technique for the negative selection in mouse embryonic stem (ES) cells, we investigated the responsiveness of cells expressing human interleukin-2 receptor alpha subunit to anti-Tac(Fv)-PE40. The immunotoxin treatment efficiently eliminated only ES cells bearing the receptor as a consequence of the target specificity of anti-Tac(Fv)-PE40, indicating that IMCT can be used as a novel strategy for positive and negative selection to enrich ES cell clones with a targeted mutation. 相似文献
966.
The dynamics of three mutually coupled cortical neurons with time delays in the coupling are explored numerically and analytically.
The neurons are coupled in a line, with the middle neuron sending a somewhat stronger projection to the outer neurons than
the feedback it receives, to model for instance the relay of a signal from primary to higher cortical areas. For a given coupling
architecture, the delays introduce correlations in the time series at the time-scale of the delay. It was found that the middle
neuron leads the outer ones by the delay time, while the outer neurons are synchronized with zero lag times. Synchronization
is found to be highly dependent on the synaptic time constant, with faster synapses increasing both the degree of synchronization
and the firing rate. Analysis shows that pre-synaptic input during the inter-spike interval stabilizes the synchronous state,
even for arbitrarily weak coupling, and independent of the initial phase. The finding may be of significance to synchronization
of large groups of cells in the cortex that are spatially distanced from each other. 相似文献
967.
968.
969.
The elusive and enigmatic origin of AIDS-associated Kaposi's sarcoma (AIDS-KS) makes it a complex tumor and therefore difficult to treat. Here we demonstrate that AIDS-KS cells express surface interleukin-4 (IL-4) receptors, and that IL-4 toxin (IL-4(38-37)-PE38KDEL) is specifically cytotoxic to these cells. Intratumoral, intraperitoneal and intravenous administration of IL-4 toxin in nude mice with established subcutaneous AIDS-KS tumors caused considerable anti-tumor activity in a dose-dependent manner, with highest dose producing durable complete responses. Metabolic changes, including cachexia and lymphopenia, induced by KS tumors were prevented by IL-4 toxin treatment. This report establishes IL-4(38-37)-PE38KDEL as an experimental therapeutic agent for the treatment of AIDS-KS. 相似文献
970.
Wallace D. Dawson S. Robert Young Zongren Wang Linda W. Liu Ira F. Greenbaum Lisa M. Davis Bryan K. Hall 《Mammalian genome》1999,10(7):730-733
Fluorescence-labeled DNA probes constructed from three whole house mouse (Mus domesticus) chromosomes were hybridized to metaphase spreads from deer mouse (Peromyscus maniculatus) to identify homologies between the species. Mus Chr 7 probe hybridized strongly to the ad-centromeric two-thirds of Peromyscus Chr 1q. Most of Mus 3 probe hybridized principally to two disjunct segments of Peromyscus Chr 3. Mus Chr 9 probe hybridized entirely to the whole Peromyscus Chr 7. Three Peromyscus linkage groups were assigned to chromosomes, based on linkage homology with Mus. The data also are useful in interpretation of chromosomal evolutionary history in myomorphic rodents.
Received: 1 December 1998 / Accepted: 17 February 1999 相似文献