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21.
22.
Frank W. Crow Kenneth B. Tomer Michael L. Gross James A. McCloskey Donald E. Bergstrom 《Analytical biochemistry》1984,139(1):243-262
The positive and negative ion fast atom bombardment (FAB) mass spectra and fast atom bombardment collisionally activated decomposition (CAD) spectra of a series of nucleosides and two dinucleotides are reported. The nucleosides studied are substituted forms of guanosine, adenosine, nebularine, tubercidin, uridine, and related pyrimidines. The FAB and CAD data both contain similar information. The CAD spectra are found to provide some structural information not found in the FAB mass spectra. Tandem mass spectrometry also allows emphasis to be put on weak fragments which are either not observed in the FAB mass spectrum or are lost in the matrix ion signals. 相似文献
23.
Alfred Berteloot Christiane Malo Sylvie Breton Michel Brunette 《The Journal of membrane biology》1991,122(2):111-125
Summary Kinetic data in (brush-border) membrane vesicles which rely on the validity of the initial rate assumption for their interpretation and depend on tracer flux studies using the rapid filtration technique for their experimental measurement have been limited to some extent by the absence of techniques that would allow for real-time data analysis. In this paper, we report on our successful design of a fast sampling, rapid filtration apparatus (FSRFA) which seems to fill up this technical gap since showing the following characteristics: (i) rapid injection (5 msec) and mixing (less than 100 msec) of small amounts of vesicles (10–40 l) with an incubation medium (0.2–1.0 ml); (ii) fast (20 to 80 msec depending on the sample volume) and multiple (up to 18 samples at a maximal rate of 4/sec) sampling of the uptake mixture followed by rapid quenching in the stop solution (approximately 5 msec) according to a predetermined time schedule (any time combination from 0.25 to 9999 sec); and (iii) fast, automated, and sampling-synchronized filtration and washings of the quenched uptake medium (only 15–20 sec are necessary for the first filtration followed by two washings and extra filtrations). As demonstrated using adult human jejunal brush-border membrane vesicles and Na+-d-glucose cotransport as models, the FSRFA accurately reproduces the manual aspects of the rapid filtration technique while allowing for very precise initial rate determinations. Moreover, the FSRFA has also been designed to provide as much versatility as possible and, in its present version, allows for a very precise control of the incubation temperature and also permits a few efflux protocols to be performed. Finally, its modular design, which separates the fast sampling unit from the rapid filtration device, should help in extending its use to fields other than transport measurement. 相似文献
24.
We analyzed the fiber-type composition of the soleus muscle in rats and mice to determine whether the adult proportion of fiber types is fixed soon after birth or whether it changes during postnatal maturation. We examined muscles from animals varying in age from 1 week to 1 year using monoclonal antibodies that distinguish between fast and slow isoforms of myosin heavy chains. In cross sections of unfixed muscle containing profiles of all myofibers in the muscle, we counted the fibers that stained with antibodies to fast myosin, and in adjacent sections, those that stained positive with an antibody to slow myosin. We also counted the total number of fibers in each section. Rat soleus contained about 2500 myofibers, and mouse about 1000 at all ages studied, suggesting that myogenesis ceases in soleus by 1 week after birth or sooner. In mouse soleus, the relative proportions of fibers staining positive with fast and slow myosin antibodies were similar at all ages studied, about 60%–70% being fast and 30%–40% slow. In rat soleus, however, the proportions of fast antibody-positive and slow antibody-positive fibers changed dramatically during postnatal maturation. At 1 week after birth, about 50% of rat soleus fibers stained with fast myosin antibodies, whereas between 1 and 2 months this value fell to about 10%. In mouse, about 10% of fibers at 1 week, but none at 1 year, reacted with both fast and slow antibodies, whereas in rat, fewer than 3% bound both antibodies to a significant degree at 1 week. It is puzzling why, in rat soleus, the majority of apparently fast fibers present at 1 week is converted to a slow phenotype, whereas in mouse soleus the predominant change appears to be the suppression of fast myosin expression in a subset of fibers that expresses both myosin types at 1 week. It is possible that this may be related to differences in size and the amount of body growth between these two species. 相似文献
25.
26.
Myosin light chain kinase binding to plastic 总被引:3,自引:0,他引:3
Methionine-81 and/or -8 of the transmembrane sialoglycoprotein, glycophorin A, have been specifically alkylated with 13CH3I to produce the sulfonium ion derivatives [S-[13C]methylmethionine-8]glycophorin A and [S-[13C]methylmethionine-8 and -81]glycophorin A. 13C NMR spectra of these species show that the resonances of the methyl groups of the modified glycophorins occur at 26.1 ppm downfield from Me4Si. A spin-lattice relaxation time of 0.4 was observed for the 13C-enriched methyl resonances of the sulfonium ion derivatives of Met-8 and -81, which corresponds to an effective correlation time of < 2× 10?10 s. Demethylation of the 2 glycophorin A sulfonium ion species with 2-mercaptoethanol produces native glycophorin A which now has the ε-carbon of the methionine residue(s) 45% isotopically enriched. The ε-carbon of Met-8 was found to occur at 15.7 ppm downfield from Me4Si whereas the ε-carbon of Met-81 exhibited an unusual chemical shift of 2.0 ppm downfield from Me4Si. The spin-lattice relaxation time of both resonances was found to be ~0.3 s. 相似文献
27.
Paul Jensén 《Physiologia plantarum》1981,52(4):437-441
Uptake of Rb+ from a complete nutrient solution with 2.0 mM Rb+ was studied in roots of spring wheat seedlings ( Triticum aestivum L. cv. Svenno) with different K+ levels. The relationship between Rb+ uptake and concentration of K+ in the roots indicated a negative feedback mechanism operating through allosteric control. The Rb+ uptake process in root cells was divided into two steps: (1) binding of the ion in the free space, and (ii) transmembrane transport into the cytoplasm. Metabolic and non-metabolic components of uptake were separated by addition of the metabolic inhibitor 2,4-dinitrophenol (DNP) to the nutrient solution. It is suggested that metabolic Rb+ uptake requires energy in two uptake steps (for binding to the carrier entity in the free space and for transmembrane transport) or in one step only (for transmembrane transport), dependent on the K+ status of the roots. The change from metabolic to non-metabolic binding in the free space is accomplished by changing the conformational state of the carrier (slow/fast transitions). There may be a hysteretic effect on metabolic Rb+ uptake through a slow transition between carrier states. This is superimposed on the negative cooperativity, strengthening further cooperativity at intermediate K+ levels in the roots. Non-metabolic Rb+ uptake probably consists of two components, a carrier-mediated (facilitated diffusion) and a parallel diffusive component. 相似文献
28.
David C. Heimbrook Stephen G. Sligar 《Biochemical and biophysical research communications》1981,99(2):530-535
We have examined the 5--hydroxylation of camphor by cytochrome P450 in [18O] water/buffer solution. In the reaction of the reconstituted multienzyme system, no 18O-label is observed in the product alcohol. Similarly, in the m-chloroperbenzoic acid or cumene hydroperoxide supported reactions with ferric P450, solvent oxygen is not incorporated into hydroxycamphor. When the analagous reaction is carried out using iodosobenzene as the exogenous oxidant, however, the alcoholic oxygen of the product is derived entirely from the solvent. These results cannot be explained by equilibration of the iodosobenzene oxygen with solvent water before reacting with P450, and suggest a unique mechanism for iodosobenzene-supported P450 oxygenations. We propose two distinct mechanistic activities for cytochrome P450: a hydroxylase, and an oxene transferase, with the former encompassing the classic oxygenase as well as “peroxygenase” reactions. 相似文献
29.
The chemical reaction between (±)-styrene oxide and N-acetylcysteine produces both positional isomers ( and ) as a mixture of diastereoisomers with a preference for the benzylic thioether isomer (2 : 1). Synthesis of the mercapturic acid conjugates from either (+)- or (?)-styrene oxide produces only two of the four possible stereoisomers. The single diastereoisomers of and were separated by high pressure liquid chromatography (HPLC) and identified by 1H- and 13C-nuclear magnetic resonance (NMR). The relative stereochemistry at the benzylic carbon center of the mercapturic acid conjugates was assigned on the basis of the established chemical correlation between optically pure styrene oxide and its precursor mandelic acid, and considerations on the mechanism of ring opening of epoxides by sulfur nucleophiles. The stereochemical definition of the isomers – should prove useful in investigations of the biotransformation of the glutathione (GSH) conjugates of styrene oxide. 相似文献
30.
R. B. Cumming Marva F. Walton J. C. Fuscoe B. A. Taylor J. E. Womack F. H. Gaertner 《Biochemical genetics》1979,17(5-6):415-431
A single formamidase, which is different from the formamidases found in other tissues, occurs in the brains of mice. This enzyme is here called formamidase-5 and the gene symbol is designated For-5. Two alleles are recognized on the basis of their differential heat sensitivity: For-5
b is relatively heat stable and is present in strain C57BL/6J, while For-5
d is relatively heat sensitive and is present in strain DBA/2J. The heat sensitivity of formamidase-5 in 44 other inbred strains and substrains was tested and found to resemble that of C57BL/6J or DBA/2J. Thirty-six recombinant inbred strains derived from progenitors that differed at For-5 were studied to test for single-gene inheritance and linkage with other loci. Complete concordance was found with the esterase-10 locus (Es-10), indicating close linkage. The 99% upper confidence limit of the distance between For-5 and Es-10 is 3.7 centimorgans (cM). Es-10 is located on chromosome 14 about 19 cM from the centromere. An independent demonstration of linkage of For-5 with Es-10 and another chromosome 14 marker, hairless (hr), is provided by the finding that the HRS/J strain, which has been sibmated for 60 generations with forced heterozygosity at the hr locus, is cosegregating at For-5 and Es-10. A survey of 32 inbred strains and substrains revealed that the For-5
d allele is associated with the Es-10
b allele, and that the For-5
b allele is associated with Es-10
a and Es-10
c. Formamidase-5 segregates as expected in the F2 generation of crosses between strains bearing For-5
b and For-5
d alleles. It is possible that this unique formamidase of the brain is involved in the metabolism of a neurotransmitter substance.This research was sponsored in part by the Department of Energy under contract with the Union Carbide Corporation and in part by NIH Research Grant GM-18684 from the National Institute of General Medical Sciences. J. C. F. is a predoctoral Fellow supported by Grant CA 09104 from the National Cancer Institute. The Biology Division of Oak Ridge National Laboratory and the Jackson Laboratory are fully accredited by the American Association for Accreditation of Laboratory Animal Care. 相似文献