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91.
Dynein ATPase is inhibited selectively in vitro by erythro-9-[3-2-(hydroxynonyl)]adenine 总被引:8,自引:0,他引:8
S M Penningroth A Cheung P Bouchard C Gagnon C W Bardin 《Biochemical and biophysical research communications》1982,104(1):234-240
Dynein (ATP phosphohydrolase, EC 3.6.1.3) extracted from sea urchin sperm tails was inhibited by erythro-9-[3-2-(hydrosynonyl)]adenine in a dosedependent fashion; at the 50% inhibitory concentration, 0.23 mM, twelve other ATP-metabolizing enzymes were notsignificantly affected. Actomyosin and myosin ATPase activities were enhanced 1.5- to 2-fold by millimolar concentrations of erythro-9-[3-2-(hydroxynonyl)]adenine. Enzyme kinetic analysis supported a model of linear mixed-type inhibition, which suggests that the binding site for erythro-9-[3-2-(hydroxynonyl)]adenine on dynein is remote from the ATPase active site. As a selective inhibitor , erythro-9-[3-2-(hydroxynonyl)]adenine appears to offer a biochemical criterion for identifying dynein isozymes in tissue extracts. 相似文献
92.
Metabolism and activation of benzo[a]pyrene by mouse and rat skin in short-term organ culture and in vivo 总被引:1,自引:0,他引:1
The metabolic activation of BP was examined in mouse and rat skin in vivo and in short-term organ culture. In mouse skin, larger quantities of ether- and water-soluble metabolites were formed and more BP became bound covalently to DNA and protein than in rat skin. Qualitative differences in the formation of dihydrodiol metabolites and of BP-deoxyribonucleoside adducts between mouse and rat skin were also observed. Organ culture techniques may not provide a true model of metabolic activation in vivo because it was found that the covalent binding of BP to DNA and protein was reduced in skin maintained in culture despite an accumulation of dihydrodiol and other ether-soluble metabolites. In addition, the proportions of the syn- and anti-isomers of BP-7,8-diol 9,10-oxide involved in the formation of adducts with deoxyguanosine differed between skin treated in organ culture and in vivo. 相似文献
93.
Elena C. McCoy G.David McCoy Herbert S. Rosenkranz 《Biochemical and biophysical research communications》1982,108(3):1362-1367
Characterization of a mutant strain (TA98/1,8-DNP6) resistant to the mutagenicity of nitrated polycyclic aromatic hydrocarbons (nitroarenes) revealed that it was also non-responsive to the mutagenic action of nitroso- and N-hydroxylaminoarenes. The mutant strain was fully sensitive to the mutagenic action of the corresponding hydroxamic acid ester. These results suggest that TA98/1,8-DNP6 is deficient in a specific esterifying enzyme and that esterification of the penultimate mutagenic metabolites of nitro- and aminoarenes (, arylhydroxylamines) to form potent electrophiles is controlled by a specific gene. 相似文献
94.
A case of somatic instability affecting aleurone colour in a strain of maize from India with flint background was analysed.
The somatic instability is localized to theC
1 (Inhibitor) allele ofC locus on the short arm of chromosome 9. Molecular tests indicated thatAc is not present in the Indian stock and the evidence is consistent with the involvement of theEn (Spm) transposable element in the instability. The presence of theEn (Spm)-like element in the stock would suggest that these elements have been present in the maize genome for a long time. A new
allele ofshrunken (sh1) gene with a somewhat unorthodox breeding behaviour is also described. 相似文献
95.
Interaction of a spin-labeled 9-aminoacridine with DNA was studied by electron spin resonance spectroscopy. Accurate determination of the binding parameters, equilibrium dissociation constant (KD), and total number of ligand-binding sites was obtained using Scatchard and Lineweaver-Burk plots. The competition between 9-aminoacridine and its spin-labeled derivative was examined by a similar analysis of the spin-label signals. The practical interest of this method lies in the fact that the precision and the simplicity of the measurements allow the quantitative determination of the binding capacity of any intercalative drug which interacts specifically with adenine/thymine bases of DNA. 相似文献
96.
The genetic effects of MNNG, 4NQO and ICR-170 have been compared on 5 different UV-sensitive strains and a standard wild-type strain of Neurospora crassa with regard to inactivation and the induction of forward-mutations at the ad-3A and ad-3B loci. Whereas all UV-sensitive strains (upr-1, uvs-2, uvs-3, uvs-5 and uvs-6) are more sensitive to inactivation by MNNG and ICR-170 than wild-type, only uvs-5 shows survival comparable to wild-type after 4NQO treatment, all other strains are more sensitive to 4NQO. In contrast to the effects on inactivation, a wide variety of effects were found for the induction of ad-3A and ad-3B mutations: higher forward-mutation frequencies than were found in wild-type were obtained after treatment with MNNG or 4NQO for upr-1 and uvs-2, no significant increase over the spontaneous mutation frequency was found with uvs-3 after MNNG, 4NQO or ICR-170 treatment; mutation frequencies comparable to that found in wild-type were obtained with uvs-6 after MNNG, 4NQO or ICR-170 treatment and with upr-1 after ICR-170 treatment. Lower forward-mutation frequencies than were found in wild-type were obtained with uvs-2 after ICR-170 treatment and with uvs-5 after MNNG, 4NQO or ICR-170 treatment. These data clearly show that the process of forward-mutation at the ad-3A and ad-3B loci is under genetic control by mutations at other loci (e.g. upr-1, uvs-2, uvs-3, uvs-5 and uvs-6) and that the effect is markedly mutagen-dependent. 相似文献
97.
Steroid acid esters, synthesized by modifying the 17-ketol side chain of prednisolone, were tested for their in vitro ability to stabilize heavy mitochondrial lysosomes prepared from rat liver. Membrane stabilization was determined by assessing capability of steroids to decrease extrusion of the marker enzymes (acid phosphatase, beta-glucuronidase and aryl sulfatase) from lysosomes incubated in hypo-osmotic sucrose-Tris acetate buffer. Results indicated that prednisolone (1) significantly inhibited the lysosomal release of acid phosphatase as did the new anti-inflammatory steroid, methyl 20-dihydroprednisolonate. Methyl prednisolonate exhibited weak membrane stabilization capacities and 20-dihydroprednisolonic acid, a metabolic product of methyl 20-dihydroprednisolonate, showed virtually no membrane stabilization. 相似文献
98.
Robert S.U. Baker Antonio M. Bonin Ieva Stupans Gerald M. Holder 《Mutation research》1980,71(1):43-52
A highly significant enhancement of mutagenicity occurs with 11 polycyclic aromatic hydrocarbons when 3-methylcholanthrene-induced guinea pig liver S9 is substituted for Aroclor-induced rat liver S9 in the Ames test. The use of MC-induced guinea pig liver S9 is particularly valuable for detecting the weak mutagenicity of benz[c]acridine, which is barely positive in a standard Ames assay. However, anthracene and phenanthrene, which are generally considered not to be carcinogens, remain non-mutagenic for strain TA100. This enhancement of mutagenicity does not correlate with arylhydrocarbon hydroxylase activities of the various liver preparations and does not apply to certain other non-PAH mutagens, including β-naphthylamine, aflatoxin B1 and 4-dimethylaminoazobenzene. 相似文献
99.
Asynchronous 9L cells were separated into relatively homogeneously-sized populations using centrifugal elutriation with both
a conventional collection method and a long collection method. A substantial increase in the homogeneity of the volume distributions
and in the degree of synchrony of the separated fractions was obtained using the long collection method. Autoradiographic
data indicated that fractions containing ≥97% G1 cells, ≥80% S cells, and 70–75% G2 cells could be routinely recovered with this procedure. Recovery in these fractions varied from 5 to 8% of the total number
of cells elutriated. The colony forming efficiency (CFE) of cells from fractions representing each phase of the cell cycle
was a constant 60–70%, which was comparable to the 60–80% usually found for asynchronous 9L cells. The percentage of cells
in the G1, S, and G2 phases in the elutriated fractions was more accurately determined from the volume distribution than from computer fits of
the DNA histogram obtained from flow cytometry. In general, the degree of synchrony was related to the coefficient of variation
(CV) of the volume distributions of the elutriated fractions. The CV was about 14% for all elutriated fractions. When the
≥97% G1 population was allowed to progress to S and G2, the CVs were about 17 and 20.2%, respectively. Thus, the best nonperturbing method for obtaining synchronous 9L cells in
the S or G2 phases was direct elutriation with the long collection method. 相似文献
100.
Christine De Paillerets Jacques Gallay Michel Vincent Monique Rogard Annette Alfsen 《生物化学与生物物理学报:生物膜》1981,644(1):134-142
The lipid dynamics of the adrenocortical microsomal membranes was studied by monitoring the fluorescence anisotropy and excited state lifetime of a set of anthroyloxy fatty acid probes (2-, 7-, 9- and 12-(9-anthroyloxy)-stearic acid (AP) and 16-(9-anthroyloxy)palmitic acid (AS). It was found that a decreasing polarity gradient from the aqueous membrane interface to the membrane interior, was present. This gradient was not modified by the proteins, as evidenced by comparison of complete membranes and derived liposomes, suggesting that the anthroyloxy probes were not in close contact with the proteins. An important change of the value of the mean rotational relaxation time as a function of the position of the anthroyl ring along the acyl chain was evidenced. In the complete membranes, a relatively more fluid medium was evidenced in the C16 as compared to the C2 region, while the rotational motion appeared to be the most hindered at the C7–C9 level. In the derived liposomes, a similar trend was observed but the mobility was higher at all levels. The decrease of the mean rotational relaxation time was more important for 12-AS and 16-AP. Temperature dependence of the mean rotational relaxation time of 2-AS, 12-AS and 16-AP in the complete membranes revealed the existence of a lipid reorganization occurring around 27°C and concerning mainly the C16 region. The extent to which the acyl chain reacted to this perturbation at the C12 level depended on pH. The presence of proteins increased the apparent magnitude of this reorganization and also modified the critical temperature from approx. 23°C in the derived liposomes to approx. 27°C in the complete membranes. Thermal dependence of the maximum velocity of the , the second enzyme in the enzymatic sequence, responsible for the biosynthesis of the in the adrenal cortex microsomes, was studied. The activation energy of the catalyzed reaction was found to be low and constant () in the temperature range 16–40°C at pH 7.5, 8.5 and 9, corresponding to the minimum, intermediate and maximum rate, respectively. A drastic increase of the activation energy () was observed at temperature below 16°C at pH 7.5. A correlated change of the as function of temperature was detected; at 36°C while at 13°C the value shifted to 8.7. The pH range of the group ionization was narrower at 13°C. In contrast with the behaviour of the , the was apparently unaffected by the lipid reorganization at 27°C. It is suggested that this enzyme possesses a different and more fluid lipid environment than the bulk lipids. 相似文献