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231.
Summary The toxicity of chromium and tin on growth, photosynthetic carbon-fixation, oxygen evolution, heterocyst differentiation and nitrogenase activity ofAnabaena doliolum and its interaction with bivalent cations has been studied. Some interacting cations, viz. Ca2+, Mg2+ and Mn2+, substantially antagonised the toxic effects of chromium and tin with reference to growth, heterocyst differentiation and nitrogenase activity in the following hierarchal sequence: Ca2+ > Mg2+ > Mn2+. However, the sequence of hierarchy was Mg2+ > Ca2+ > Mn2+ for carbon fixation and Mn2+ > Mg2+ > Ca2+ for photosynthetic oxygen evolution. Synergistically inhibitory patterns were noticed for all the parameters, viz. growth,14CO2 uptake, oxygen evolution, heterocyst differentiation and nitrogenase activity ofA. doliolum when Ni2+, Co2+ and Zn2+ were combined with the test metals in the growth medium. These cations followed the following sequence of synergistic inhibition: Ni2+ > Co2+ > Zn2+. Among all the interacting cations, Ca2+, Mg2+ and Mn2+ exhibited antagonistic effects which relieved the test cyanobacterium from metal toxicity. In contrast to this, Ni2+, CO2+ and Zn2+ showed synergistic inhibition which potentiating the toxicity of test metals in the N2-fixing cyanobacteriumA. doliolum. It is evident from the present study that bivalent cations, viz. Ca2+, Mg2+, Mn2+, Ni2+, Co2+ and Zn2+, may appreciably regulate the toxicity of heavy metals in N2-fixing cyanobacteria if present in aquatic media. 相似文献
232.
We have studied the mechanisms of breakdown of 2'-5' oligoadenylates. We monitored the time-courses of degradation of ppp(A2'p5')nA (dimer to tetramer) and of 5'OH-(A2'p5')nA (dimer to pentamer) in unfractionated L1210 cell extract. The 5' triphosphorylated 2'-5' oligoadenylates are converted by a phosphatase activity. However, 2'-5' oligoadenylates are degraded mainly by phosphodiesterase activity which splits the 2'-5' phosphodiester bond sequentially at the 2' end to yield 5' AMP and one-unit-shorter oligomers. The nonlinear least-squares curve-fitting program CONSAM was used to fit these kinetics and to determine the degradation rate constant of each oligomer. Trimers and tetramers, whether 5' triphosphorylated or not, are degraded at the same rate, whereas 5' triphosphorylated dimer is rapidly hydrolyzed and 5'-OH dimer is the most stable oligomer. The interaction between degradation enzymes and the substrate strongly depends on the presence of a 5' phosphate group in the vicinity of the phosphodiester bond to be hydrolyzed; indeed, when this 5' phosphate group is present, as in pp/pA2'p5'A/or A2'/p5'A2'p5'A/, affinity is high and maximal velocity is low. Such a degradation pattern can control the concentration of 2'-5' oligoadenylates active on RNAse L either by limiting their synthesis (5' triphosphorylated dimer is the primer necessary for the formation of longer oligomers) and/or by converting them into inhibitory (e.g., monophosphorylated trimer) or inactive (e.g., nonphosphorylated oligomers) molecules. 相似文献
233.
Summary The two high affinity calcium binding sites of the cardiac (Ca2+ + Mg2+)-ATPase have been identified with the use of Eu3+. Eu3+ competes for the two high affinity calcium sites on the enzyme. With the use of laser-pulsed fluorescent spectroscopy, the environment of the two sites appear to be heterogeneous and contain different numbers of H2O molecules coordinated to the ion. The ion appears to be occluded even further in the presence of ATP. Using non-radiative energy transfer studies, we were able to estimate the distance between the two Ca2+ sites to be between 9.4 to 10.2 A in the presence of ATP. Finally, from the assumption that the calcium site must contain four carboxylic side chains to provide the 6–8 ligands needed to coordinate calcium, and based on our recently published data, we predict the peptidic backbone of the two sites. 相似文献
234.
Angelo N. Belcastro Wade Parkhouse Goeff Dobson James S. Gilchrist 《Molecular and cellular biochemistry》1988,83(1):27-36
The purpose of this study was to examine the Ca2+-Mg2+ myofibrillar ATPase and protein composition of cardiac and skeletal muscle following strenuous activity to voluntary exhaustion. Sprague-Dawley rats (200 g) were assigned to a control and exercised group, with the run group completing 25 m·min–1 and 8% grade for 1 hour. Following activity, the myocardial Ca2+–Mg2+ myofibrillar ATPase activity -pCa relationship had undergone a rightward shift in the curve. Electrophoretic analysis revealed a change in the pattern of cardiac myofibrillar protein bands, particularly in the 38–42 Kdalton region. Enzymatic analysis of myofibrillar proteins from plantaris muscle, revealed no change in Ca2+ regulation following exercise. Electronmicrographic and electrophoretic analysis revealed extensively disrupted sarcomeric structure and a change in the ratio of several plantaris myofibrillar proteins. No difference was observed for myosin: Actin: tropomyosin ratios; however a dramatic reduction in 58 and 95 Kdalton proteins were evident. The results indicate that prolonged running is associated with similar responses in cardiac and skeletal muscle myofibrillar protein compositions. The abnormalities in myofibrillar ultrastructure may implicate force transmission failure as a factor in exercised-induced muscle damage and/or fatigue. 相似文献
235.
Terrence L. Scott 《Molecular and cellular biochemistry》1988,82(1-2):51-54
Summary The Ca2+ binding site region of the Ca2+ — ATPase of skeletal muscle sarcoplasmic reticulum was labeled with several fluorescent analogs of dicyclohexylcarbodiimide. As has been shown by Chadwick and Thomas [1, 2], in the absence of Ca2+ in the medium, labeling with the naphthyl carbodiimide results in the inhibition of enzyme activity. Further, Ca2+ occupancy of the high affinity sites of the enzyme protects against incorporation into the site(s). The fluorescent carbodiimide has been used to determine the depth of the site of label incorporation relative to the aqueous-bilayer interfaces by quenching studies using spin-labeled fatty acid derivatives. The series of quenchers used have their spin-label moiety located at different positions along the fatty acid chain. It was found that after suitable correction for differences in partitioning of the various derivatives, the order of quenching efficiency was 16 - > 12- > 10- > 7- > 5-NS, indicating that the naphthyl moiety is near the center of the bilayer. In contrast, quenching with the aqueous-restricted I– indicated that the label is accessible from the external milieu, likewise for a presumed aqueous quencher, acrylamide. The aqueous quenchers accessibilities were altered upon Ca2+ binding to the ATPase. Quenching of the intrinsic fluorescence with the x-NS derivatives indicates that the ATPase tryptophan residues are primarily localized at the aqueous-membrane interfaces, with the order of quenching being 5- > 7- > 10- > 12- > 16-NS. The trp residue(s) which changes its fluorescence upon Ca2+ binding is shown to be near the membrane surface. 相似文献
236.
C. J. Duncan 《Cell and tissue research》1988,253(2):457-462
Summary This study compares the action of inhibitors of the eicosanoid cascade on calcium-induced myofilament damage in cardiac muscle of the perfused frog heart and incubated frog ventricle slices, and in skeletal muscle of incubated mammalian diaphragm and isolated and saponin-skinned amphibian pectoris cutaneous muscle. Mepacrine (10-5M) and indomethacin (3×10-6M) protected completely against myofilament damage induced by entry of calcium in the calcium-paradox in frog heart. However, inhibition of phospholipase A2 (PLA2) (with chlorpromazine, 2×10-4M, or mepacrine, 10-5M, 5x10-5M), of cyclo-oxygenase enzymes (with indomethacin, 3x10-6M to 10-5M or BW755C, 3.8x10-4M), or of lipoxygenase enzymes (with BW755C, 3.8x10-4M or nordihydroguaiaretic acid, 2x10-6M or 5x10-6M) all failed in intact cardiac or skeletal muscle cells to prevent the myofilament damage that is rapidly triggered by 10-2M caffeine, 6x10-6M ruthenium red, 10-4M DNP or 5 g ml-1 A23187. These agents also failed completely to protect against myofilament damage in saponin-skinned amphibian skeletal muscle when [Ca]i was raised to 8x10-6M. Thus, inhibition of PLA2 does not protect the myofilament apparatus against calcium released intracellularly, and it is suggested that mepacrine and indomethacin can block entry of calcium in the calcium-paradox in the amphibian heart. Chlorpromazine (2x10-4M) and mepacrine (10-3M) at zero [Ca] caused severe myofilament damage in skinned muscle, possibly due to an effect on membranes. Since inhibitors of PLA2 and of lipoxygenases prevent efflux of creatine kinase and sarcolemma damage in mammalian skeletal muscle, it is evident that experimentally-induced rises in [Ca]i (by caffeine or A23187) can trigger two separate pathways: (i) PLA2 and the arachidonic acid cascade which culminate in membrane damage, and (ii) a different, Ca-activated system that causes rapid damage of myofilaments. 相似文献
237.
Barbara J. Javor 《Archives of microbiology》1988,149(5):433-440
Seven strains of extremely halophilic bacteria (Halobacterium spp., Halococcus spp., and Haloarcula sp.) fixed CO2 under light and dark conditions. Light enhanced CO2 fixation in Halobacterium halobium but inhibited it in Halobacterium volcanii and Haloarcula strain GN-1. Propionate stimulated 14CO2 incorporation in some strains, but inhibited it in others. Semi-starvation in basal salts plus glycerol induced enhanced CO2 fixation rates. 14CO2 fixation in semi-starved cells was stimulated by NH
4
+
or pyruvate and inhibited by succinate and acetate in most strains. No possible reductant was found. In cell-free extracts of H. halobium, NH
4
+
but not propionate stimulated 14CO2 fixation. No RuBP carboxylase activity was detected. The main 14C-labeled -keto acid detected after a 2-min incubation with 14CO2 and pyruvate was pyruvate. Little or no -ketobutyrate was detected among the early products of propionate-stimulated CO2 fixation. Glycine was the major amino acid synthesized during a 2-min incubation with NH
4
+
, propionate, and 14CO2. Propionate-stimulated CO2 fixation was sensitive to trimethoprim and insensitive to avidin. A novel pathway for non-reductive CO2 fixation involving a glycine synthase reaction with CO2, NH
4
+
, and a methyl carbon derived from the -carbon cleavage of propionate is tentatively proposed.Abbreviations used BBS
buffered basal salts
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- MOPS
3-(N-morpholino)propanesulfonic acid
- DNPH
2,4-dinitrophenylhydrazine
- DNP
dinitrophenyl
- TLC
thin-layer chromatography
- FH4
tetrahydrofolate
This work was supported by National Science Foundation grant PCM-8116330 and Petroleum Research Fund grant PRF 13704-AC2 相似文献
238.
Anaerobic bacteria degrading 2-methoxyethanol were enriched from freshwater sediments, and three strains were isolated in pure culture. Two of them were Grampositive non-spore-forming rods and grew strictly anaerobically by acetogenic fermentation. Optimal growth occurred at 30°C, initial pH 7.5–8.0. 2-Methoxyethanol and 2-ethoxyethanol were fermented to acetate and corresponding alcohols. Hydrogen plus carbon dioxide, formate, acetoin, l-malate, lactate, pyruvate, fructose, and methoxyl groups of 3,4,5-trimethoxybenzoate and 3,4,5-trimethoxycinnamate were fermented to acetate. 1,2-Propanediol was fermented to acetate, propionate, and propanol. Strain MuME1 was described as a new species, Actetobacterium malicum. It had a DNA base composition of 44.1 mol% guanine plus cytosine. The third strain, which was identified as Pelobacter venetianus, fermented 2-methoxyethanol to methanol, ethanol, and acetate. 相似文献
239.
Abstract Nif− mutants of Rhodobacter capsulatus carrying mutations either in the nifR4 regulatory gene or in the nifH structural gene both outgrew the wild-type strain B10 in mixed chemostat cultures under conditions favouring nitrogenase-mediated H2 production by the wild-type (ammonia as limiting nutrient, inert argon atmosphere, light as energy source), whereas under aerobic conditions in the dark, or in batch culture, the growth of Nif− mutants was not favoured. Nitrogenase-mediated H2 production therefore appears to be detrimental to the growth of R. capsulatus in nitrogen-limited continuous culture, as may also be the case for other nitrogen fixers. 相似文献
240.
A synchronization treatment was initiated when each of 1227 heifers (four trials) was tailpainted. The tailpaint was sprayed with an aerosol raddle at the end of the treatment period. The heifers were in herds of 20 to 279 animals. Each herd was observed for estrus at selected post treatment intervals. A heifer was considered to be (or to have been) in estrus when the raddle was rubbed off. In three of the trials, animals which had the raddle removed were inseminated at 48h following the end of the synchronization treatment. The tailpaint of an inseminated animal was scored from 0 (less than 10% of the paint remained) to 5 (more than 90% of the paint remained) and was then reraddled with a second color. The detection-insemination sequence was always repeated at 72 and 96h, and sometimes at 120h. Animals which had been previously inseminated, but then had paint scores reduced by at least 2 units were reinseminated 24h later. Over the four trials, 94.5% of the heifers were detected in estrus through the use of the tailpaint and raddle system. The remaining 67 animals included only 10 (0.8%) which had ovulated without being detected in estrus. The reinsemination rate on consecutive days was 11.3% and was highest among animals that had a tailpaint score of 4 or 5 at 48h. The proportion of animals detected in estrus at selected posttreatment intervals varied with the different synchronization treatments used within one herd, or with the same treatment used in different herds. The combination of tailpaint, raddling, tailpaint scoring and reraddling is a simple sequence which can be effectively used to detect estrus among heifers synchronized in research or commercial herds. 相似文献