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41.
42.
A new method for cell permeabilization reveals a cytosolic protein requirement for Ca2+ -activated secretion in GH3 pituitary cells 总被引:7,自引:0,他引:7
Ca2+ is a major regulator of exocytosis in secretory cells, however, the biochemical mechanisms underlying regulation remain to be identified. To render the secretory apparatus accessible for biochemical studies, we have developed a cell permeabilization method (cell cracking) which utilizes mechanical shear. GH3 pituitary cells subjected to cracking were permeable to macromolecules but retained a normal cytoplasmic ultrastructure including secretory granules. Incubation of the permeable cells at 30-37 degrees C with 0.1-1.0 microM Ca2+ and millimolar MgATP resulted in the release of the secretory proteins, prolactin (PRL) and a proteoglycan, but not lysosomal enzymes. Extensively washed permeable cells were incapable of releasing PRL in response to Ca2+ and MgATP addition. However, addition of cytosol was found to restore Ca2+-activated, MgATP-dependent PRL release. The cytosolic factor responsible for activity was thermolabile and protease sensitive. The protein was partially purified, and its molecular mass was estimated to be equivalent to that of a globular protein of 200-350 kDa by molecular sieve chromatography. Inhibitors of calmodulin or protein kinase C (trifluroperazine, calmidazolium, H-7) failed to inhibit Ca2+-activated PRL release, and the required cytosolic protein could not be replaced by purified calmodulin, calmodulin-dependent protein kinase II, protein kinase C, or calpactin I. Further purification and characterization of the cytosolic protein should reveal the nature of biochemical events involved in regulated secretory exocytosis. 相似文献
43.
A. Rigotto S.R. Cotta A.C.F. Dias J.L.N. Carvalho F.D. Andreote 《Letters in applied microbiology》2020,71(5):444-450
Sugarcane cultivation supports Brazil as one of the largest world sugar and ethanol producer. In order to understand the impact of changing sugarcane harvest from manual to mechanized harvest, we studied the effect of machinery traffic on soil and consequently soil compaction upon soil microbial communities involved in nitrogen cycling. The impact of sugarcane harvest was dependent on soil depth and texture. At deeper soil layers, mechanized harvesting increases the abundance of nitrogen fixers and denitrifying communities (specifically nosZ clade I and II) while manual harvesting increases the abundance of ammonia oxidizers (specifically AOA) and increases denitrifying communities (nosZ clade I and II) on top and at intermediate depth. The effect of change on the harvest system is more evident on sandy soil than on clay soil, where soil indicators of compaction (bulk density and penetration resistance) were negatively correlated with soil microorganisms associated with the nitrogen cycle. Our results point to connections between soil compaction and N transformations in sugarcane fields, besides naming biological variables to be used as proxies for alterations in soil structure. 相似文献
44.
Patterns generated from ecological surveys are rarely tested in similar habitats to assess the accuracy of predictions. Testing empirically derived predictions provides a strong tool for establishing the consistency of general patterns in ecology. We test the consistency of beetle community associations with habitat complexity in open canopy forests and make both community and morphospecies-level comparisons with results from a previous study. We use Normalized Difference Vegetation Indices (NDVIs) from remote sensing as a surrogate for habitat complexity. The positive relationships between NDVIs and site-based beetle species richness and abundance were consistent in open canopy forests both south and north of Sydney, Australia. NDVIs were also useful for predicting differences in beetle composition in open canopy forests. Taxon-specific responses to NDVI differences in 'southern forests' were very similar to responses in 'northern forests', most likely reflecting beetle trophic roles. This study shows that NDVIs can be used as rapid biodiversity indicators, when integrated with identified faunal responses to vegetation structure, provided that the lower vegetation strata may be measured by remote sensing. 相似文献
45.
46.
Anne F Mannion Stephane Kämpfen Urs Munzinger Ines Kramers-de Quervain 《Arthritis research & therapy》2009,11(5):R139-13
Introduction
Patient's expectations are variably reported to influence self-rated outcome and satisfaction after medical treatment; this prospective study examined which of the following was the most important unique determinant of global outcome/satisfaction after total knee arthroplasty (TKA): baseline expectations; fulfilment of expectations; or current symptoms and function. 相似文献47.
Tamar F Brionez Shervin Assassi John D Reveille Thomas J Learch Laura Diekman Michael M Ward John C Davis Jr Michael H Weisman Perry Nicassio 《Arthritis research & therapy》2009,11(6):R182-9
Introduction
Functional status is an integral component of health-related quality of life in patients with ankylosing spondylitis (AS). The purpose of this study was to investigate the role of psychological variables in self-reported functional limitation in patients with AS, while controlling for demographic and medical variables. 相似文献48.
49.
F. L. Laksmana P. J. A. Hartman Kok H. Vromans H. W. Frijlink K. Van der Voort Maarschalk 《AAPS PharmSciTech》2009,10(3):732-742
Next to the coating formulation, process conditions play important roles in determining coating quality. This study aims to
develop an operational window that separates layering from agglomeration regimes and, furthermore, the one that leads to the
best coating quality in a fluidized bed coater. The bed relative humidity and the droplet size of the coating aerosol were
predicted using a set of engineering models. The coating quality was characterized using a quantitative image analysis method,
which measures the coating thickness distribution, the total porosity, and the pore size in the coating. The layering regime
can be achieved by performing the coating process at a certain excess of the viscous Stokes number (ΔSt
v). This excess is dependent on the given bed relative humidity and droplet size. The higher the bed relative humidity, the
higher is the ΔSt
v required to keep the process in the layering regime. Further, it is shown that using bed relative humidity and droplet size
alone is not enough to obtain constant coating quality. The changes in bed relative humidity and droplet size have been identified
to correlate to the fractional area of particles sprayed per unit of time. This parameter can effectively serve as an additional
parameter to be considered for a better control on the coating quality. High coating quality is shown to be achieved by performing
the process close to saturation and spraying droplets small enough to obtain high spraying rate, but not too small to cause
incomplete coverage of the core particles. 相似文献
50.
David A. Dik Daniel R. Marous Jed F. Fisher 《Critical reviews in biochemistry and molecular biology》2017,52(5):503-542
The lytic transglycosylases (LTs) are bacterial enzymes that catalyze the non-hydrolytic cleavage of the peptidoglycan structures of the bacterial cell wall. They are not catalysts of glycan synthesis as might be surmised from their name. Notwithstanding the seemingly mundane reaction catalyzed by the LTs, their lytic reactions serve bacteria for a series of astonishingly diverse purposes. These purposes include cell-wall synthesis, remodeling, and degradation; for the detection of cell-wall-acting antibiotics; for the expression of the mechanism of cell-wall-acting antibiotics; for the insertion of secretion systems and flagellar assemblies into the cell wall; as a virulence mechanism during infection by certain Gram-negative bacteria; and in the sporulation and germination of Gram-positive spores. Significant advances in the mechanistic understanding of each of these processes have coincided with the successive discovery of new LTs structures. In this review, we provide a systematic perspective on what is known on the structure–function correlations for the LTs, while simultaneously identifying numerous opportunities for the future study of these enigmatic enzymes. 相似文献