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61.
62.
Inhibitors of Urokinase and Thrombin in Cultured Neural Cells 总被引:2,自引:1,他引:1
Steven L. Wagner Alice L. Lau Ann Nguyen Jun Mimuro David J. Loskutoff Paul J. Isackson† Dennis D. Cunningham 《Journal of neurochemistry》1991,56(1):234-242
Recent studies have suggested important roles for certain proteases and protease inhibitors in the growth and development of the CNS. In the present studies, inhibitors of urokinase or thrombin in cultured neural cells and serum-free medium from the cells were identified by screening for components that formed sodium dodecyl sulfate-stable complexes with 125I-urokinase or 125I-thrombin. Rinsed glioblastoma possessed two components that complexed 125I-urokinase. One was type 1 plasminogen activator inhibitor (PAI-1), because the 125I-urokinase-containing complexes were immunoprecipitated with anti-PAI-1 antibodies. The other component formed complexes with 125I-urokinase that were not recognized by antibodies to PAI-1 or protease nexin-1 (PN-1). Its identity is unknown. In addition to these cell-bound components, the glioblastoma cells also secreted two inhibitors that formed complexes with 125I-urokinase; one was PAI-1, and the other was PN-1. The secreted PN-1 also formed complexes with 125I-thrombin. It was the only thrombin inhibitor detected in these studies. Human neuroblastoma cells did not contain components that formed detectable complexes with either 125I-urokinase or 125I-thrombin. However, human neuroblastoma cells did contain very low levels of PN-1 mRNA and PN-1 protein. Added PN-1 bound to the surface of both glioblastoma and neuroblastoma cells. This interaction accelerated the inhibition of thrombin by PN-1 and blocked the ability of PN-1 to form complexes with 125I-urokinase. Thus, cell-bound PN-1 was a specific thrombin inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
63.
M J Carrier S N Chatfield G Dougan U T Nowicka D O'Callaghan J E Beesley S Milano E Cillari F Y Liew 《Journal of immunology (Baltimore, Md. : 1950)》1992,148(4):1176-1181
The feasibility of using Salmonella typhimurium aroA mutant (SL3261) to deliver protein therapeutic agents was investigated in a murine model system. We have constructed an Escherichia coli expression plasmid designed to express the human protein IL-1 beta. This plasmid expresses IL-1 beta to high levels (greater than 30% total cell protein) in E. coli. In Salmonella the IL-1 beta is expressed constitutively to about 10% total cell protein, as verified by Western blotting analysis using polyclonal rabbit anti-IL-1 beta antibody. The protein is produced in a soluble and biologically active form. BALB/c mice administered orally or i.v. with S. typhimurium aroA mutants carrying the plasmid produced highly significant antibody responses against human IL-1 beta as determined by a solid-phase RIA. Furthermore, mice injected with the construct were significantly protected against lethal gamma-irradiation (850 rad). This study therefore demonstrates that the vaccine strain of Salmonella mutants can also be used effectively to deliver therapeutic proteins in vivo. 相似文献
64.
Danielle-Julie Carrier Nathalie Chauret Michael Mancini Pierre Coulombe Ronald Neufeld Martin Weber Jean Archambault 《Plant cell reports》1991,10(5):256-259
Ginkgo biloba cells were cultured in two 500 mL shake flasks and in 2 L and 6 L immobilization bioreactors using MS medium supplemented with 1 mg.L–1 NAA, 0.1 mg.L–1 K and 30 g.L–1 sucrose. Specific growth rates were 0.06 d–1, 0.11 d–1 and 0.07 d–1 for the 2 L and 6 L bioreactors and shake flask cultures, respectively. Extracellular phosphate, nitrate, ammonium and carbohydrate uptake rates of the bio reactor cultures were approximately 17 to 39% slower than those of shake flask cultures. The specific oxygen uptake and carbon dioxide transfer rates of immobilized Ginkgo biloba cells ranged from 0.027 to 0.041 mmol O2.g–1.d.w.hr–1 (maximum uptake at 14 days) and 0.020 to 0.057 mmol CO2g. –1.d.w.hr–1 (maximum production at 14 days). Extracts from the biomass of the two immobilized and shake flask suspension cultures were analysed for ginkgolide A by GC-MS. Yields of 7, 17, 19 and 7 ng.g. –1d.w. of ginkgolide A were determined for shake flask 1, shake flask 2 and the 2 L and 6 L immobilized cultures, respectively. Traces of ginkgolide B were detected with the signal to noise ratio, however, being too low for positive confirmation of this last product.Abbreviations CTR
Carbon dioxide transfer rate
- DO
Dissolved oxygen
- g.d.w.
Gram dry weight
- GA
Ginkgolide A
- GB
Ginkgolide B
- GC
Gas chromatography
- GC-MS
Gas chromatography-mass spectrometry
- HPLC
High performance liquid chromatography
- K
Kinetin
- MS
Murashige and Skoog salt medium
- N1K1MS
Complete Murashige and Skoog medium supplemented with 1 mg.L–1 NAA, 0.1 mg.L–1 K and 30g.L–1 sucrose
- NAA
Naphthaleneacetic acid
- OTR
Oxygen transfer rate
- PAF
Platelet Aggregating Factor
- qCO2
Specific carbon dioxide production rate
- qO2
Specific oxygen uptake rate
- u
Specific growth rate 相似文献
65.
Bruce A. Macheri William M. F. Lee Mary Alice Westrick 《Molecular and cellular biochemistry》1982,47(2):81-95
Summary We have reviewed the studies on neutral glycosphingolipids and gangliosides of normal and leukemia human leukocytes. In this review, we examine (a) the glycosphingolipid composition of various leukocyte populations, (b) the differences in glycosphingolipids found among subsets of these cells, (c) the possible use of these compounds as markers of differentiation, and (d) the changes in glycosphingolipid composition that occur with leukemogenesis. 相似文献
66.
Alice Bourke Hayes 《American journal of botany》1981,68(6):733-740
Auxin treatment results in hyponastic curvature of the primary leaves of Phaseolus vulgaris L. var pinto. Ethylene production by hyponastic leaves is detected within 1 hr after treatment with IAA in concentrations at or above 1 μm. The amount of ethylene detected is proportional to the concentration of auxin applied. Untreated control leaves and leaves treated with 2,3,5-tri-iodobenzoic acid or gibberellic acid did not produce ethylene detectable by our equipment. The hyponastic curvature induced by auxin treatment can be inhibited by exogenous application of ethylene or ethylene-generating compounds, and these treatments produce epinasty in auxin-treated leaves. Treatment with inhibitors of ethylene synthesis or action, such as aminoethoxy-vinylglycine, carbon dioxide, or heat treatment, prolong hyponasty. The planar form, therefore, appears to be affected by both hyponastic auxin effect and an epinastic ethylene effect. 相似文献
67.
Benzodiazepines are known to facilitate GABA-ergic transmission at synaptic sites, while sodium valproate is an anticonvulsant drug which is reported to elevate GABA levels in the brain. In order to determine whether these two drugs interact functionally at GABA receptor sites, graded doses of chlordiazepoxide (CDZ) and sodium valproate were injected bilaterally into the nucleus accumbens and their effect on the dopamine (DA)-induced stimulation of motor activity was studied. Both of these compounds, as well as GABA, produced an inhibition of the hyperactivity induced by the bilateral injection of DA into the nucleus accumbens. Bicuculline, the GABA receptor antagonist, blocked the effect of CDZ on the DA-induced hyperactivity. A low dose of CDZ (2 μg), which by itself did not significantly inhibit the effect of DA, potentiated the inhibition of the hyperactivity produced by valproate. These results suggest that CDZ and sodium valproate can interact functionally at GABA-ergic sites in the central nervous system. 相似文献
68.
Alice S. Tempel 《Oecologia》1981,51(1):97-106
Summary The acceptance of secondary plant metabolites as herbivore deterrents rests primarily on their deleterious effects on herbivores. Efforts to demonstrate differential fitness in natural plant populations with varying concentrations of tannin have failed, since coevolved plant predators may physiologically or behaviorally circumvent the defense, which results in apparently equal amounts of damage to defended and undefended individuals. In this study, two approaches were used to overcome this difficulty. 1) Theoretically, more energy should be allocated to the defense of parts which contribute more heavily to the plant's fitness. Bracken fern clones produce fronds throughout the growing season. Fronds which are produced early should be more heavily defended than late-emerging fronds which will return less photosynthate per unit cost of production. The results of this study do not support this prediction; it appears that the production of tannin is more closely linked to environmental factors such as water stress than to date of frond emergence. Fronds which emerged in August contained as much tannin as fronds which emerged in May. 2) By recording the temporal occurrence of herbivore damage in bracken ferns, it was found that in fronds which escaped attack until after reaching maturity there was a significant negative correlation between tannin concentration in the frond and the amount of damage experienced. This result supports the generally accepted assumption that herbivory has been a selective force in the evolution of tannin as a defensive substance. 相似文献
69.
70.
Plant tumor reversal associated with the loss of foreign DNA 总被引:1,自引:0,他引:1
Fun-Mei Yang Alice L. Montoya Eugene W. Nester Milton P. Gordon 《In vitro cellular & developmental biology. Plant》1980,16(1):87-92
Summary Transformation of plant tissues into crown gall tumors has been associated with the transfer of a portion of a tumor-inducing
plasmid (Ti-plasmid) into plant DNA. Various laboratories have regenerated normal-appearing plants from a number of crown
gall tumors. This study investigates the fate of the foreign DNA in a series of tissues derived from various parts of a plant
regenerated from the tumor BT-37 by Braun and his coworkers. It was found that all the foreign DNA sequences were lost from
tissues that had lost all their tumorous traits; whereas the plasmid DNA sequences were still present in tissues that appeared
normal but still exhibited tumorous traits when returned to tissue culture media. From these studies it would appear that
the presence of the Ti-plasmid sequences in the plant DNA is required for the maintenance of the transformed state.
Presented in the Symposium on Gene Transfer, Differentiation and Neoplasia in Plant and Animal Cells at the 30th Annual Meeting
of the Tissue Culture Association, Seattle, Washington, June 10–14, 1979. This symposium was supported in part by Grant CA
26748 from the National Cancer Institute, DHEW, and Grant RD-67 from the American Cancer Society. 相似文献