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991.
992.
Hadley KC Borrelli MJ Lepock JR McLaurin J Croul SE Guha A Chakrabartty A 《Cell stress & chaperones》2011,16(5):549-561
The inability of cells to maintain protein folding homeostasis is implicated in the development of neurodegenerative diseases,
malignant transformation, and aging. We find that multiphoton fluorescence imaging of 1-anilinonaphthalene-8-sulfonate (ANS)
can be used to assess cellular responses to protein misfolding stresses. ANS is relatively nontoxic and enters live cells
and cells or tissues fixed in formalin. In an animal model of Alzheimer’s disease, ANS fluorescence imaging of brain tissue
sections reveals the binding of ANS to fibrillar deposits of amyloid peptide (Aβ) in amyloid plaques and in cerebrovascular
amyloid. ANS imaging also highlights non-amyloid deposits of glial fibrillary acidic protein in brain tumors. Cultured cells
under normal growth conditions possess a number of ANS-binding structures. High levels of ANS fluorescence are associated
with the endoplasmic reticulum (ER), Golgi, and lysosomes—regions of protein folding and degradation. Nuclei are virtually
devoid of ANS binding sites. Additional ANS binding is triggered by hyperthermia, thermal lesioning, proteasome inhibition,
and induction of ER stress. We also use multiphoton imaging of ANS binding to follow the in vivo recovery of cells from protein-damaging
insults over time. We find that ANS fluorescence tracks with the binding of the molecular chaperone Hsp70 in compartments
where Hsp70 is present. ANS highlights the sensitivity of specific cellular targets, including the nucleus and particularly
the nucleolus, to thermal stress and proteasome inhibition. Multiphoton imaging of ANS binding should be a useful probe for
monitoring protein misfolding stress in cells. 相似文献
993.
A cytoplasmic NADH oxidase (NOX) was purified from a soil bacteria, Brevibacterium sp. KU1309, which is able to grow in the medium containing 2-phenylethanol as the sole source of carbon under an aerobic condition. The enzyme catalyzed the oxidation of NADH to NAD+ involving two-electron reduction of O2 to H2O2. The molecular weight of the enzyme was estimated to be 102 kDa by gel filtration and 57 kDa by SDS-PAGE, which indicates that the NOX was a homodimer consisting of a single subunit. The enzyme was stable up to 70 degrees C at a broad range of pH from 7 to 11. The enzyme activity increased about ten-fold with the addition of ammonium salt, while it was inhibited by Zn2+ (39%), Cu2+ (41%), Hg2+ (72%) and Ag+ (37%). The enzyme acts on NADH, but not on NADPH. The regeneration of NAD+ utilizing this enzyme made selective oxidation of mandelic acid or L: -phenylalanine possible. This thermostable enzyme is expected to be applicable as a useful biocatalyst for NAD+ recycling. 相似文献
994.
Differential role of microenvironment in microencapsulation for improved cell tolerance to stress 总被引:6,自引:0,他引:6
Sun ZJ Lv GJ Li SY Yu WT Wang W Xie YB Ma X 《Applied microbiology and biotechnology》2007,75(6):1419-1427
The effect of the microenvironment in alginate–chitosan–alginate (ACA) microcapsules with liquid core (LCM) and solid core
(SCM) on the physiology and stress tolerance of Sacchromyces cerevisiae was studied. The suspended cells were used as control. Cells cultured in liquid core microcapsules showed a nearly twofold
increase in the intracellular glycerol content, trehalose content, and the superoxide dismutase (SOD) activity, which are
stress tolerance substances, while SCM did not cause the significant physiological variation. In accordance with the physiological
modification after being challenged with osmotic stress (NaCl), oxidative stress (H2O2), ethanol stress, and heat shock stress, the cell survival in LCM was increased. However, SCM can only protect the cells
from damaging under ethanol stress. Cells released from LCM were more resistant to hyperosmotic stress, oxidative stress,
and heat shock stress than cells liberated from SCM. Based on reasonable analysis, a method was established to estimate the
effect of microenvironment of LCM and SCM on the protection of cells against stress factors. It was found that the resistance
of LCM to hyperosmotic stress, oxidative stress, and heat shock stress mainly depend on the domestication effect of LCM’s
microenvironment. The physical barrier of LCM constituted by alginate–chitosan membrane and liquid alginate matrix separated
the cells from the damage of oxidative stress and ethanol stress. The significant tolerance against ethanol stress of SCM
attributed to the physical barrier consists of solid alginate–calcium matrix and alginate–chitosan membrane. 相似文献
995.
The biological effects of rare-earth ions on the organism have been studied using Pr3+ as a probe ion and Escherichia coli cell as a target. Atomic force microscopy (AFM) observation of the surface of E. coli cells shows that the presence of Pr3+ substantially changes the structure of the outer membrane. By induced coupled plasma-mass spectrometry (ICP-MS), more Cu2+ was found in the cells grown in the presence of Pr3+, indicating changes of cell permeability. Using energy dispersive X-ray spectroscopy (EDX), Ca2+ is found on the outer surface of the original cell. It is proposed that Pr3+ can replace Ca2+ from the binding sites because of their close ionic radii and similar ligand speciality. 相似文献
996.
Geister TL Lorenz MW Hoffmann KH Fischer K 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》2009,179(1):87-98
Phenotypic plasticity may allow an organism to adjust its phenotype to environmental needs. However, little is known about
environmental effects on offspring biochemical composition and turnover rates, including energy budgets and developmental
costs. Using the tropical butterfly Bicyclus anynana and employing a full-factorial design with two oviposition and two developmental temperatures, we explore the consequences
of temperature variation on egg and hatchling composition, and the associated use and turnover of energy and egg compounds.
At the lower temperature, larger but fewer eggs were produced. Larger egg sizes were achieved by provisioning these eggs with
larger quantities of all compounds investigated (and thus more energy), whilst relative egg composition was rather similar
to that of smaller eggs laid at the higher temperature. Turnover rates during embryonic development differed across developmental
temperatures, suggesting an emphasis on hatchling quality (i.e. protein content) at the more stressful lower temperature,
but on storage reserves (i.e. lipids) at the higher temperature. These differences may represent adaptive maternal effects.
Embryonic development was much more efficient at the lower temperature, providing a possible mechanism underlying the temperature-size
rule. 相似文献
997.
Small eukaryotes, cells with a diameter of less than 5 mum, are fundamental components of lacustrine planktonic systems. In this study, small-eukaryote diversity was determined by sequencing cloned 18S rRNA genes in three libraries from lakes of differing trophic status in the Massif Central, France: the oligotrophic Lake Godivelle, the oligomesotrophic Lake Pavin, and the eutrophic Lake Aydat. This analysis shows that the least diversified library was in the eutrophic lake (12 operational taxonomic units [OTUs]) and the most diversified was in the oligomesotrophic lake (26 OTUs). Certain groups were present in at least two ecosystems, while the others were specific to one lake on the sampling date. Cryptophyta, Chrysophyceae, and the strictly heterotrophic eukaryotes, Ciliophora and fungi, were identified in the three libraries. Among the small eukaryotes found only in two lakes, Choanoflagellida and environmental sequences (LKM11) were not detected in the eutrophic system whereas Cercozoa were confined to the oligomesotrophic and eutrophic lakes. Three OTUs, linked to the Perkinsozoa, were detected only in the Aydat library, where they represented 60% of the clones of the library. Chlorophyta and Haptophyta lineages were represented by a single clone and were present only in Godivelle and Pavin, respectively. Of the 127 clones studied, classical pigmented organisms (autotrophs and mixotrophs) represented only a low proportion regardless of the library's origin. This study shows that the small-eukaryote community composition may differ as a function of trophic status; certain lineages could be detected only in a single ecosystem. 相似文献
998.
Galaud JP Carrière M Pauly N Canut H Chalon P Caput D Pont-Lezica RF 《The Plant journal : for cell and molecular biology》1999,17(1):111-118
We constructed a high-efficiency expression library from Arabidopsis cDNA clones by introducing a poly (dC) stretch at the 5' end of the clones. This library enables the synthesis of proteins from all the cDNA clones present. We have screened the high-efficiency expression library with antibodies raised against total proteins from Arabidopsis plasmalemma and tonoplast. With the positive clones, we have constructed two cDNA ordered libraries enriched in genes encoding plasmalemma (522 clones) and tonoplast proteins (594 clones). Partial sequencing of both libraries shows that a high proportion (47%) of the clones encoded putative membrane proteins, or membrane-associated proteins. When sequenced, 55% of the cDNAs were new EST sequences for Arabidopsis, 26% were similar to genes present in other plants or organisms, and 29% were not referenced in any databank. Immunoscreening of the two cDNA ordered libraries with antibodies raised against proteins from Arabidopsis cells submitted to osmotic stress allows the selection of genes over- and under-expressed in stress conditions. 相似文献
999.
P Codogno C Bauvy A P Sève M Hubert E Ogier-Denis M Aubery J Hubert 《Journal of cellular biochemistry》1992,50(1):93-102
Nonhistone proteins were extracted in 0.4 M NaCl from membrane-depleted nuclei of HeLa cells grown in the presence or the absence of [5,6-3H]fucose. Control experiments strongly suggest that most extracted proteins were indeed nuclear components. Several proteins, present in the 0.4 M NaCl nuclear extract, with M(r) ranging from 35,000 to 115,000 were identified on Western blots as fucosylated glycoproteins owing to their binding to the fucose-specific lectin, Ulex europeus agglutinin I. Results of experiments involving mild alkaline treatment and peptide N-glycosidase F digestion showed that the carbohydrate moieties of these fucosylated nuclear glycoproteins were N-linked to the polypeptide backbone. Analysis of the N-glycans revealed the presence of two populations of sialylated oligosaccharides on the basis of their relative molecular masses. The sensitivity of the high-M(r) oligosaccharides to endo-beta-galactosidase and their incorporation of [3H]glucosamine suggest that they could contain repeating N-acetyllactosamine units. [3H]Fucose incorporated into nuclei was confined to the nucleoli, as judged by autoradiography of sections cut through cells grown in the presence of [3H]fucose. Electron microscopy autoradiography showed that the fibrillar centers were never labeled, while silver grains were observed on the dense and the granular components of nucleoli. Taking into account of these data most nuclear fucosylated glycoproteins extracted in 0.4 M NaCl might be nucleolar ribonucleoproteins. 相似文献
1000.
Molecular cloning of a widely distributed microsatellite core sequence from the cultivated mushroom Agaricus bisporus 总被引:3,自引:0,他引:3
Barroso G Sonnenberg AS Van Griensven LJ Labarère J 《Fungal genetics and biology : FG & B》2000,31(2):115-123
An Agaricus bisporus microsatellite with the tetranucleotide motif TATG tandemly repeated was isolated from an A. bisporus library enriched in repeated sequences. The use of the 16-mer oligonucleotide (TATG)4 indicates that many loci contain nearby copies of the microsatellite in opposite orientations. The wide distribution of the microsatellite in the A. bisporus genome was assessed (i) by polyacrylamide gel electrophoresis of the products generated by directed amplification of microsatellite-region DNA (DAMD) and (ii) by hybridization of these products with A. bisporus chromosomes separated by pulsed-field gel electrophoresis. This is, to our knowledge, the first microsatellite reported in the cultivated edible mushrooms. DAMD-PCR products were generated using DNA of three Pleurotus species (P. pulmonarius, P. sajor-caju, and P. florida), indicating that (TATG)4 repeats are also present in these cultivated species. The variability found within closely related strains indicates that such microsatellites are useful in fingerprinting and studying genetic variability in wild and commercial mushrooms. 相似文献