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141.
The processing of polypeptide neurotrophins in the nervous system is poorly understood. In this paper, we provide information on the effects of C-terminal processing of nerve growth factor. Three forms of recombinant mouse beta-nerve growth factor (rNGF) were produced and isolated from insect cells infected with a recombinant baculovirus. The three purified forms of rNGF exhibited distinct biological activities and differed in their abilities to compete with high affinity binding of mouse beta-nerve growth factor (mNGF). However, they were chemically and structurally indistinguishable from each other. All three forms of rNGF differed from mature mNGF from mouse submaxillary gland in that the C-terminal Arg-Gly dipeptide had not been proteolytically removed. Removal of the C-terminal dipeptide by gamma-NGF peptidase treatment converted the three forms into a single form identical with mature mNGF. The above results demonstrate that a single polypeptide of rNGF, due to the presence of a C-terminal dipeptide, exhibits three stable dimeric protein conformations with distinct biological activities. The apparent lack of gamma-NGF peptidase in the nervous system raises the possibility that the biologically significant form of NGF may differ from mature mNGF; such a difference may be of physiological relevance. 相似文献
142.
A study on the constitutive equation of blood. 总被引:1,自引:0,他引:1
143.
The mean δD value of petiole water of Pterocarpus indicus Willd (δD = −9.0 ± 2.5‰, n = 3) was not significantly different from the mean value of stem water (−8.3 ± 2.8‰, n = 3). δD values of main vein water ranged from −11.1 to + 12.0‰ (n = 14) and increased along the main vein from petiole to the tip of leaves. Mesophyll water was highly enriched in deuterium (mean δD = +32.0 ± 2.0‰, n = 19) when compared with stem, petiole, and vein water. δD values of mesophyll water for different areas of the lamina, however, were not homogenous and could differ by as much as 20‰. 相似文献
144.
Alteration of Components of Leaf Water Potential and Water Content in Velvetleaf under the Effects of Long-Term Humidity Difference 总被引:1,自引:0,他引:1 下载免费PDF全文
Velvetleaf (Abutilon theophrasti Medik.) was grown in growth chambers set at 45 or 85% relative humidity at 30°C, CO2 350 microliters per liter and 1000 micromoles per square meter per second of photosynthetically active radiation. Soil water potential was maintained at −0.05 megapascal by subirrigation with half strength Hoagland solution. The third, fourth, and fifth leaves from the base of 21- and 25-day-old plants were used for pressure-volume measurements. Components of leaf water status including water potential (osmotic and potential associated with the apoplast), leaf water content (apoplasmic and symplasmic water), and elastic modulus of leaf tissue were determined. Results indicate: (a) persistent dry air generated leaves with lower water potential at a given relative water content than did humid air; (b) the higher total leaf water content in plants grown in dry air was related to an increase in apoplasmic water, whereas symplasmic water remained similar in both humidity treatments; (c) difference in leaf water potential between low and high humidity treatments was related to decreased potential associated with the apoplast but not to a change in cell wall elasticity. 相似文献
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C F Semenkovich C C Luo M K Nakanishi S H Chen L C Smith L Chan 《The Journal of biological chemistry》1990,265(10):5429-5433
Detailed structure-function information about human lipoprotein lipase (LPL) is unavailable because it is difficult to purify large amounts of the enzyme for study. To circumvent this problem, we constructed an in vitro LPL expression vector. Human LPL cDNA was cloned and inserted into the expression vector p91023(B). After transfection of COS M-6 cells with the human LPL cDNA construct, LPL enzyme activity was detected in cell extracts and culture medium. Purified human apolipoprotein C-II caused a 5-fold stimulation of the recombinant human LPL expressed in vitro. Using site-specific mutagenesis, Ala residues were substituted for Asn residues at two potential N-linked glycosylation sites (positions 43 and 359) and at a third unrelated Asn (position 257) in the LPL cDNA. RNA blot analysis demonstrated the presence of a single mRNA species in COS cells transfected with wild-type and mutant LPL expression vectors. Intracellular and secreted LPL activity was absent in the construct containing an Ala for Asn mutation at position 43, whereas the same substitutions at positions 257 and 359 did not appreciably affect activity. LPL activity was also absent in another construct containing a Gln for Asn mutation at position 43. Quantitation of LPL protein mass concomitant with measurement of enzyme activity showed that substitution of Ala or Gln for Asn at position 43 resulted in the production of an enzymatically inactive protein which accumulated intracellularly but was not secreted into the culture medium. Our report represents an initial documentation of the expression of cloned human LPL in vitro and of the importance of Asn-43 for both enzyme activity and secretion. 相似文献
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Many insects have evolved resistance to abamectin but the mechanisms involved in this resistance have not been well characterized. P-glycoprotein (P-gp), an ATP-dependent drug-efflux pump transmembrane protein, may be involved in abamectin resistance. We investigated the role of P-gp in abamectin (ABM) resistance in Drosophila using an ABM-resistant strain developed in the laboratory. A toxicity assay, Western blotting analysis and a vanadate-sensitive ATPase activity assay all demonstrated the existence of a direct relationship between P-gp expression and ABM resistance in these flies. Our observations indicate that P-gp levels in flies' heads were higher than in their thorax and abdomen, and that both P-gp levels and LC50 values were higher in resistant than in susceptible and P-gp-deficient strains. In addition, P-gp levels in the blood–brain barrier (BBB) of resistant flies were higher than in susceptible and P-gp-deficient flies, which is further evidence that a high level of P-gp in the BBB is related to ABM resistance. Furthermore, we found greater expression of Drosophila EGFR (dEGFR) in the resistant strain than in the susceptible strain, and that the level of Drosophila Akt (dAkt) was much higher in resistant than in susceptible flies, whereas that in P-gp-deficient flies was very low. Compared to susceptible flies, P-gp levels in the resistant strain were markedly suppressed by the dEGFR and dAkt inhibitors lapatinib and wortmannin. These results suggest that the increased P-gp in resistant flies was regulated by the dEGFR and dAkt pathways and that increased expression of P-gp is an important component of ABM resistance in insects. 相似文献