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151.
The effect of acetaldehyde concentrations on the relative rates of formation of acetaldehyde-modified hemoglobins 总被引:1,自引:0,他引:1
L B Nguyen C M Peterson 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1984,177(2):226-233
The effect of various concentrations of acetaldehyde (0, 0.05, 0.1, 0.25, 0.5, 1.0, and 5.0 mM) on the relative rates of formation of hemoglobin acetaldehyde adducts detected in fractions eluted from cation exchange high-pressure liquid chromatography (HPLC) was investigated. When the hemoglobin and acetaldehyde mixtures were incubated at 37 degrees C for various time intervals up to 24 hr, increased amounts of HbA1c could be observed after 2 hr incubation with 1 mM or greater concentrations of acetaldehyde, or after 4 hr incubation with at least 0.5 mM acetaldehyde. An increase in the HbA1a + b fraction was not observed with 4 hr incubation time until the acetaldehyde level reached 1 mM. The HPLC method detected no difference in minor hemoglobins from alcoholic and normal subjects. Incubation of red blood cells at 37 degrees C for 1 hr with six consecutive pulses of 0.05 mM [14C]acetaldehyde showed no differences in the amounts of minor hemoglobins determined chromatographically at various pulse intervals. However, the measure of the 14C-label incorporation into hemoglobin showed that adducts eluting in the HbA1a+b fraction were formed at a faster rate than those eluting in the HbA1c or HbA0 fraction, respectively. The specific activities of the HbA1a+b fractions at 2, 4, and 6 pulses were 34, 128, and 949 cpm/mg hemoglobin; those of the HbA1c fraction were 15, 58, and 174 cpm/mg hemoglobin. This evidence of modification of hemoglobin by physiological levels of acetaldehyde from 14C-label incorporation suggests that an assay more sensitive than chromatographic separation of adducts might be clinically useful in detecting alcoholism or monitoring alcohol detoxification programs. 相似文献
152.
Human genes for glutathione S-transferases 总被引:11,自引:2,他引:9
The tissue distribution of different glutathione S-transferases (GST) is analysed by electrophoresis. The existence of GST"e" (erythrocyte), GST3, GST1, and GST2 is confirmed. GST"e" the fastest and most thermolabile of different GST analysed is observed only in erythrocyte cells. GST3 which migrates more slowly than GST"e" is present in all tissues and cells analysed, excepted for erythrocyte cells in which only GST"e" is observed. GST1 presents a polymorphism with four phenotypes, 1, 1/2, 2, and 0 controlled by three alleles 1, 2, and 0 (null). With the sample of 56 livers analysed the different frequencies obtained are 9%, 5%, 43%, 43% for the phenotypes 1, 1/2, 2, and 0 respectively and 0.074 (p), 0.279 (q), 0.647 (r) for the alleles, 1, 2, and 0 (null). GST2 presents variant patterns due probably, in the majority of cases, to post-synthetic modifications rather than allelic variation. Two new GST are described, GST4 and GST5. GST4 abundant in muscle tissue is a dimeric protein. GST4 forms with GST1 a heterodimeric band. GST5 is observed in brain homogenates. For the chromosome localization the results obtained by man (leucocytes)-mouse somatic cell hybrid analysis indicate that the gene for leucocytes GST is on chromosome 11. This gene is the structural GST3 gene. 相似文献
153.
B Descamps-Latscha M N Feuillet-Fieux A Baruchel C Patereau A T Nguyen 《Comptes rendus de l'Académie des sciences. Série III, Sciences de la vie》1984,298(15):419-422
We have previously shown that monoclonal anti-T cell antibodies bound to their specific targets can trigger the activation of monocyte/macrophage oxidative metabolism through an Fc receptor-mediated interaction. The present study demonstrates that IgG coated platelets from patients with thrombocytopenia-associated diseases can induce a similar respiratory burst activation in polymorphonuclear and mononuclear phagocytes from normal individuals. The intensity of the oxidative reaction as measured by luminol-dependent chemiluminescence is in close correlation with the level of surface-bound IgG molecules as determined by a radioactive anti-immunoglobulin assay. This new methodology to evaluating IgG fixed on human platelets by their capacity to trigger the generation of highly reactive oxygen species by granulocytes and monocytes has also suggested a new mechanism in the genesis of thrombocytopenia associated with autoimmune diseases. 相似文献
154.
An anaerobic modification of conventional polyscrylamide-gel electrophoretic equipment is described. The modified apparatus has been applied to the separation of Azotobacter vinelandii nitrogenase components and should prove useful in the analysis of other O2-sensitive proteins. Electrophoresis in reducing gels can be followed with a dithionite-resistant tracking dye, potassium gualazulene-1-sulfonate. 相似文献
155.
Use of Adenosine 5′-Triphosphate as an Indicator of the Microbiota Biomass in Rumen Contents
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A number of techniques were tested for their efficiency in extracting adenosine 5′-triphosphate (ATP) from strained rumen fluid (SRF). Extraction with 0.6 N H2SO4, using a modification of the procedure described by Lee et al. (1971), was the most efficient and was better suited for extracting particulate samples. Neutralized extracts could not be stored frozen before assaying for ATP because large losses were incurred. The inclusion of internal standards was necessary to correct for incomplete recovery of ATP. The ATP concentration in rumen contents from a cow receiving a ration of dried roughage (mainly alfalfa hay) ranged from 31 to 56 μg of ATP per g of contents. Approximately 75% of the ATP was associated with the particulate material. The ATP was primarily of microbial origin, since only traces of ATP were present in the feed and none was found in “cell-free” rumen fluid. Fractionation of the bacterial and protozoal populations in SRF resulted in the isolation of an enriched protozoal fraction with a 10-fold higher ATP concentration than that of the separated rumen bacteria. The ATP pool sizes of nine functionally important rumen bacteria during the exponential phase of growth ranged from 1.1 to 17.6 μg of ATP per mg of dry weight. This information indicates that using ATP as a measure of microbial biomass in rumen contents must be done with caution because of possible variations in the efficiency of extraction of ATP from rumen contents and differences in the concentration of ATP in rumen microbes. 相似文献
156.
R.A. Gravel K.F. Lam D. Mahuran A. Kronis 《Archives of biochemistry and biophysics》1980,202(2):669-670
A procedure is described for a simple two-step purification of human liver propionyl-CoA carboxylase. The method is based on acid and carbon tetrachloride extraction to remove other biotin carboxylases followed by an 800-fold purification through biotin-pretreated, monomeric avidin-Sepharose 4B-CL with elution of active enzyme using a biotin gradient. The enzyme had a sedimentation coefficient of 17.4 S and polyacrylamide gel electrophoresis after reduction and alkylation revealed two nonidentical polypeptide chains of 75,000 and 60,000 Mr. The heavier chain was identified as the biotin-containing subunit by electrophoresis after avidin binding. 相似文献
157.
Structural Analysis of the dur Loci in S. CEREVISIAE: Two Domains of a Single Multifunctional Gene 总被引:13,自引:1,他引:12
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In Saccharomyces cerevisiae, the degradation of urea to carbon dioxide and ammonia is catalyzed by urea carboxylase and allophanate hydrolase. The loci coding for these enzymes (dur1 and dur2) are very tightly linked on the right arm of chromosome II between pet11 and met8. Pleiotropic mutations that fail to complement mutations in either of the dur loci were found to be predominantly located in or near the dur2 locus. We interpret these data as suggesting that the two dur loci might in reality be domains of a single gene that codes for a multifunctional polypeptide. In view of this conclusion, we have renamed the dur loci as the dur1,2 locus. 相似文献
158.
C. P. Georgopoulos B. Lam A. Lundquist-Heil C. F. Rudolph J. Yochem M. Feiss 《Molecular & general genetics : MGG》1979,172(2):143-149
Summary The E. coli dnaK (groPC756) gene product is essential for bacteriophage DNA replication. Bacterial DNA segments carrying this gene have been cloned onto a bacteriophage vector. The product of the dnaK gene has been identified on SDS polyacrylamide gels after infection of UV-irradiated E. coli cells. The dnaK gene codes for a polypeptide with an apparent molecular weight of 93,000-Mr. Transducing phages carrying amber mutations in the dnaK gene fail to induce the synthesis of the 93,000-Mr polypeptide chain upon infection of sup
+ bacteria, but do so upon infection of supF bacteria. E. coli carrying the dnaK756 mutation are, in addition, temperature sensitive for growth at 43° C. It is shown that the dnaK756 mutation results in an overproduction of the dnaK gene product at that temperature. 相似文献
159.
Graphical methods have traditionally been the principal means for estimation of parameters (e.g., affinity constants, cooperativity parameters, and concentrations of receptor sites) in enzymology and ligand-binding problems. The present report provides a review of these methods as well as new results, as applied to three coordinate systems popularly used in ligand-binding studies: vs [Bound]. vs [Free], and vs [Total]. We consider two extremely general models, the statistical mechanical model and the Adair model for equilibrium ligand binding. We also consider a very specialized case of receptor interaction wherein the equilibrium constannt of dissociation is linearly related to receptor occupancy. We collect previously described equations and derive new ones, to enable the user to estimate the parameters of the models in terms of relatively easily measurable graphical characteristics. We have evaluated the performance of these methods in representative cases using Monte Carlo studies. The results indicate the kind of precision and accuracy which can be obtained with typical experimental designs. Depending upon the magnitude of experimental error, the graphical methods can provide dependable values for the binding parameters. However, in general, the results obtained by the graphical methods should be regarded as reasonable initial estimates for further refinement by weighted nonlinear least-squares curve fitting. 相似文献
160.
Dominique Weil Nguyen Van-Cong Catherine Finaz R. Rebourcet Chantal Cochet J. de Grouchy J. Frézal 《Human genetics》1977,36(2):205-211
Summary 22 independent man-hamster (HGPRT–) hybrids using male human cells with balanced reciprocal translocation t(X;2)(p22;q32) were analysed for human genes localized on chromosome 2 (IDHS, MDHS), on chromosome X (PGK, GAL, G6PD) and for the different chromosomes in relation with the balanced reciprocal translocation (chr.2, chr.2q–, chr.Xp+).The following results were obtained:The chromosomes 2 and 2q– are absent in the 22 hybrids.In 9 hybrids, the absence of MDHS in spite of the presence of the chromosome Xp+ indicates that the gene for MDHS is not localized on this chromosome (or that the gene for MDHS is not on the segment 2q32-2qter translocated on X).In 14 hybrids, the three markers of X (PGK, GAL, G6PD) and IDHS are expressed in the presence of the chromosome Xp+. This result indicates that the genes for these markers are on Xp+ or that the genes PGK, GAL, G6PD are on X without the Xp22-Xter segment, translocated on the chr.2, and that the gene for IDHS is on the 2q32-2qter segment translocated on X.In 8 hybrids, in the absence of the intact chromosome Xp+, the higher percentage of the presence of G6PD (7 hybrids) and the lower percentage of the presence of IDHS (3 hybrids) are explained by the fact that these hybrids selected in HAT medium had to retain a segment of Xp+ bearing the human gene HGPRT. G6PD appeared very close to HGPRT and IDHS very distant from HGPRT.The study of the different correlations between the presence and the absence of these four markers on Xp+ in the different hybrids indicates the following order on the chromosome Xp+ from p to q: IDHs — PGK — GAL — G6PD.
Groupe INSERM: Directeur J. Frézal
Groupe CNRS, ER, 149: Directeur J. de Grouchy 相似文献
Groupe INSERM: Directeur J. Frézal
Groupe CNRS, ER, 149: Directeur J. de Grouchy 相似文献