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41.
Wendell L. Combest Timothy J. Bloom Lawrence I. Gilbert 《Journal of neurochemistry》1988,51(5):1581-1591
The effects of the naturally occurring polyamines spermine and spermidine on phosphorylation promoted by cyclic AMP (cAMP)-dependent protein kinase (PK) (cAMP-PK; EC 2.7.1.37) were studied using the brain of the tobacco hornworm, Manduca sexta. Four particulate-associated peptides (280, 34, 21, and 19 kilodaltons) in day 1 pupal brains are endogenous substrates for a particulate type II cAMP-PK. These phosphoproteins are present in brain synaptosomal, as well as microsomal, particulate fractions but are not present in the cytosol. They are distributed throughout the CNS and PNS and are present in several nonneuronal tissues as well. Phosphorylation of these proteins via cAMP-PK was inhibited markedly by micromolar concentrations of spermine and spermidine. Other particulate-associated peptides phosphorylated via a Ca2+/calmodulin-PK or Ca2+ and cAMP-independent PKs were unaffected by polyamines, whereas the phosphorylation of a 260-kilodalton peptide was markedly enhanced. Spermine did not exert its inhibitory effect indirectly by enhancement of cAMP or ATP hydrolysis or via proteolysis, but its action appears to involve a substrate-directed inhibition of cAMP-PK-promoted phosphorylation as well as enhanced dephosphorylation. Although addition of spermine resulted in marked ribosome aggregation in synaptosomal and microsomal particulate fractions, this phenomenon was not involved in the inhibition of cAMP-PK-promoted phosphorylation. 相似文献
42.
Summary The complete physical map of the mitochondrial genome of the Owen cytoplasm of sugar beet has been determined from overlapping cosmid clones. The genome is 386 kb in size and has a multicircular organisation generated by homologous recombination across repeated DNA elements. The location of the rRNA genes and several polypeptide genes has been determined. In addition the mitochondrial genome was found to contain a sequence of chloroplast DNA including part of the 16 S rRNA gene. 相似文献
43.
Mahan James R.; Sherman Timothy D.; Funkhouser Edward A. 《Plant & cell physiology》1988,29(4):735-737
The thermal dependence of two of the reactions catalyzed bythe nitrate reductase from Chlorella vulgaris was determined.The activation energies for NADH:nitrate oxidoreductase (EC1.6.6.1
[EC]
) and NADH:Cytochrome c oxidoreductase (EC 1.6.99.3
[EC]
)are 42.1 kJ?mol1 and 21.5 kJ?mol1, respectively.Since the thermal dependency of the two enzymes is different,ratios of the activities will vary with temperature. The importanceof both rigorous thermal control during nitrate reductase assaysas well as the need to specify the temperature at which theratio of activities for the enzyme are clearly established.
1Present Address: Cropping Systems Research Laboratory, USDA-ARS,Route 3, Box 215, Lubbock, TX 79401, U.S.A. (Received November 25, 1987; Accepted March 2, 1988) 相似文献
44.
Shmuel Muallem Stephen J. Pandol Timothy G. Beeker 《The Journal of membrane biology》1988,106(1):57-69
Summary
45Ca fluxes and free-cytosolic Ca2+ ([Ca2+]
i
) measurements were used to study the effect of Ca2+-mobilizing hormones on plasma membrane Ca2+ permeability and the plasma membrane Ca2+ pump of pancreatic acinar cells. We showed before (Pandol, S.J., et al., 1987.J. Biol. Chem.
262:16963–16968) that hormone stimulation of pancreatic acinar cells activated a plasma membrane Ca2+ entry pathway, which remains activated for as long as the intracellular stores are not loaded with Ca2+. In the present study, we show that activation of this pathway increases the plasma membrane Ca2+ permeability by approximately sevenfold. Despite that, the cells reduce [Ca2+]i back to near resting levels. To compensate for the increased plasma membrane Ca2+ permeability, a plasma membrane Ca2+ efflux mechanism is also activated by the hormones. This mechanism is likely to be the plasma membrane Ca2+ pump. Activation of the plasma membrane Ca2+ pump by the hormones is time dependent and 1.5–2 min of cell stimulation are required for maximal Ca2+ pump activation. From the effect of protein kinase inhibitors on hormone-mediated activation of the pump and the effect of the phorbol ester 12-0-tetradecanoyl phorbol, 13-acetate (TPA) on plasma membrane Ca+ efflux, it is suggested that stimulation of protein kinase C is required for the hormone-dependent activation of the plasma membrane Ca2+ pump. 相似文献
45.
The stem-galling sawfly Euura lasiolepisuses one or more plant wound compounds resulting from oviposition scars as cues in host discrimination (avoiding sites occupied by conspecifics). Four experiments were conducted to test hypotheses about how Euurapartitions resources. Experiment 1 demonstrated that Euuraavoids ovipositing on nodes with scars from previous ovipositions. Experiment 2 showed no evidence that the sawfly uses oviposition-deterring pheromones and indicated there is a time lag following oviposition before the oviposition scar becomes a deterrent. Experiment 3 showed that sawflies avoid artificially formed scars, demonstrating that a plant cue alone can lead to host discrimination. Experiment 4 showed that visual or tactile cues are not necessary for host discrimination and indicated that a plant wound compound functions as an oviposition deterrent. Both experimental results and field surveys showed that Euuraoviposition scars were more uniformly distributed than expected if sawflies were ignoring previous ovipositions. 相似文献
46.
一、前言哈巴德布鲁克实验林(Hubbard Brook Experimental Forest)位于美国东北部新罕布什尔州中部的白山国家森林中。该地区位于典型温带湿润气候区内,年平均降水量为129.5cm,全年月平均降水量变化不大,冬雪夏雨。蒸发蒸腾量以每年6—9月为最大(Likens等,1977;Bormann等,1979)。该实验林为北美温带落叶阔叶林,属红果云杉(Picea rubens)-阔叶林。Hubbard Brook Ecosystem Study(HBES)是开始最 相似文献
47.
A rapid fluorometric DNA assay for the measurement of cell density and proliferation in vitro 总被引:4,自引:0,他引:4
Timothy A. McCaffrey Lily A. Agarwal Babette B. Weksler 《In vitro cellular & developmental biology. Plant》1988,24(3):247-252
Summary Many research efforts require the accurate determination of cell density in vitro. However, physical cell counting is inaccurate,
time-intensive and requires removal of the cells from their growth environment, thereby introducing a host of potential artifacts.
The current studies document a very simple method of determining cell density in microtiter wells via DNA-enhanced fluorescence.
Fixed cells are stained with the A-T intercalating DNA stains DAPI or Hoechst 33342 and then fluorescence is quantified in
a plate fluorometer. Fluorescence is shown to be linearly related to cell density as determined by two physical counting methods.
The validity of the method is established in determining serum-stimulated growth of smooth muscle cells and in mitogen-induced
growth of endothelial cells. The fixed cells can be stored for prolonged periods, thus allowing time-course proliferation
assays without interassay variations. The fixed cells are also suitable for determinations of antigens of interest by ELISA.
This method is potentially valuable in many in vitro systems where the quantification of cell density and proliferation is
necessary.
This work supported in part by NIH Cardiovascular Training Grant HL07423 and a grant from the American Federation for Aging
Research to T. M. and HL35724 to B. W.
EDITOR’S STATEMENT The technique described in this paper represents an approach to quantifying cell density in adherent monolayers
of cultured cells in microtiter wells that is rapid and simple and does not require radioisotopes or removal of cells. 相似文献
48.
Indu S. Ambudkar Timothy Lockwich Yukiharu Hiramatsu Bruce J. Baum 《Molecular and cellular biochemistry》1992,114(1-2):73-77
Conclusions While it is generally accepted that Ca2+ plays an important regulatory role in the physiology of a number of non-excitable cells, the mechanisms which regulate intracellular [Ca2+ are far from well established. Ca2+ transporting mechanisms which distribute Ca2+ intracellularly as well as those which allow influx of extracellular Ca2+ are involved in mediating intracellular Ca2+ homestasis. In this paper we have described recent studies on the regulation of the Ca2+ influx system in the data, it appears that the process of Ca2+ entry is extremely complex and may involve several levels of regulation. Understanding the molecular basis of these regulatory mechanisms presents a challeging problem for future studies. 相似文献
49.
Debra L. Forthman Suzanne D. Elder Roger Bakeman Timothy W. Kurkowski Constance C. Noble Samuel W. Winslow 《Zoo biology》1992,11(3):187-195
Bears are extremely popular among the zoo-going public, yet while many zoo exhibits have undergone dramatic design changes in recent years, most bears continue to be housed in moated grottos constructed largely of gunite. In these traditional exhibits they frequently demonstrate stereotypic locomotor patterns and are often encouraged by the public to beg. Thus, the manner in which most captive bears are exhibited does not facilitate conservation education. It is possible, however, to provide bears with opportunities to demonstrate species-typical feeding and foraging behaviors, even in standard exhibits. Subjects were four individuals of three bear species. Feeding enrichment was provided to one bear per week during three mornings during the summers of 1989 and 1990. Overall, animals were more active, less passive and less often engaged in abnormal behaviors during sessions with enrichment. Effects showed individual variation and were more profound during the second year of the study, when a greater variety of enrichment items was presented. These results suggest that simple and inexpensive methods of enrichment may have a significant, positive influence on the behavior of captive bears. © 1992 Wiley-Liss, Inc. 相似文献
50.
In vivo ionizing irradiations produce deletions in the hprt gene of human T-lymphocytes 总被引:6,自引:0,他引:6
Janice A. Nicklas J. Patrick O'Neill Timothy C. Hunter Michael T. Falta Malcolm J. Lippert David Jacobson-Kram Jerry R. Williams Richard J. Albertini 《Mutation research》1991,250(1-2):383-396
The hprt T-lymphocyte cloning assay, which detects mutations occurring in vivo in humans, has been used to examine mutants induced in patients receiving radioimmunoglobulin therapy (RIT) for cancer. Samples from 13 patients before treatment (controls) and 15 samples from 12 patients after treatment were studied for both mutant frequencies and molecular changes in the hprt mutant T-cell clones. Patients were studied up to 48 months after treatment. Post-RIT patients showed increased mutant frequencies as compared to pre-treatment values. T-cell receptor (TCR) gene analysis of mutant T-cell clones demonstrated that 84% arose independently, both pre- and post-treatment, which is the same proportion as seen in normal individuals. However, several individuals did show large sets of mutants with the same TCR gene rearrangement patterns. Molecular analysis of mutants demonstrated a greater proportion of mutations with hprt gene changes on Southern blots after RIT treatment than before (40% versus 20%). RIT increases the proportion of mutations with total rather than partial gene deletions or other gross structural changes compared to normal individuals or pre-treatment patients. These studies are defining the spectrum for radiation-induced hprt gene mutations in vivo in human T-lymphocytes. 相似文献