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61.
Investigations of the uptake of ammonium (NH
4
+
) by Rhodopseudomonas capsulata B100 supported the presence of an NH
4
+
transport system. Experimentally NH
4
+
was determined by electrode or indophenol assay and saturation kinetics were observed with two apparent K
m's of 1.7 M and 11.1 M (pH 6.8, 30°) and a V
max at saturation of 50–60 nmol/min·mg protein. The optimum pH and temperature were 7.0 and 33° C, respectively. The Q10 quotient was calculated to be 1.9 at 100 M NH
4
+
, indicating enzymatic involvement. In contrast to the wild type, B100, excretion of NH
4
+
, not uptake, was observed in a glutamine auxotroph, R. capsulata G29, which is derepressed for nitrogenase and lacks glutamine synthetase activity. G29R1, a revertant of G29, also took up NH
4
+
at the same rate as wild type and had fully restored glutamine synthetase activity. Partially restored derivatives, G29R5 and G29R6, grew more slowly than wild type on NH
4
+
as the nitrogen source, remained derepressed for nitrogenase in the presence of NH
4
+
, and displayed rates of NH
4
+
uptake in proportion to their glutamine synthetase activity. Ammonium uptake and glutamine synthetase activity were also restored in R. capsulata G29 exconjugants which had received the plasmid pPS25, containing the R. capsulata glutamine synthetase structural gene. These data suggest that NH
4
+
transport is tightly coupled to assimilation.Abbreviations used CHES
cyclohexylaminoethanesulfonic acid
- GS
glutamine synthetase
- SDS
sodium dodecylsulfate 相似文献
62.
Barbara Siemieniako Ewa M. Rakowicz-Szulczyńska Antoni Horst 《Molecular and cellular biochemistry》1985,65(2):131-141
Non-histone chromatin proteins synthesized during chicken embryonic liver development were labeled with [3H]tryptophan and [3H]methionine and characterized by electrophoresis. During embryonic development protein/DNA ratio in chromatin was low (1.30-1.62) but synthesis of non-histone protein was high. Especially one characteristic fraction K (MW 18 000), tightly bound with DNA was preferentially associated with DNAase II sensitive, active transcribed sequences. In 7-day old and adult chicken synthesis of all non-histone proteins was low, fraction K was absent or synthesized only in small amounts in association with non-active sequences, however protein/DNA ratio in chromatin was high (2.30-2.33). 相似文献
63.
Glutathione (GSH) dissolved in Eagle's MEM and added to cultures o of V79-E cells in concentrations between 2.5 × 10–4 and 10–3 moles/l for 1 h induces a dose-dependent cell cycle delay, sister chromatid exchanges and clastogenic damage. 7–8% of the metaphases showed endoreduplication at a recovery phase of 25 and 30 h after treatment with 10–3 molesll GSH. Higher concentrations were lethal. The highest tolerated dose corresponds to the intracellular GSH level in V79-E cells. In the same range of concentrations, glutathione disulfide was inactive. Endoreduplication induction by GSH is G2-phase specific and endoreduplication metaphases show a reduced occurrence of single SCEs when extrapolated to the diploid complement. The adverse effects of GSH are independent of the presence of serum in the culture fluid but completely abolished when the treatment is performed in Hank's solution instead of MEM. The mechanism of genotoxicity of exogenous GSH is discussed but, at present, no pertinent explanation can be given.Abbreviations BUdR
5-bromodeoxyuridine
- GSH
glutathione
- GSSG
glutathione disulfide
- SCE
sister chromatid exchange 相似文献
64.
65.
Barbara A. Brennessel Kathleen J. Keyes 《In vitro cellular & developmental biology. Plant》1985,21(7):402-408
Summary The artificial sweetener saccharin inhibits binding of epidermal growth factor (EGF) to cultured rat pituitary tumor cells
(GH4C1 cells). Saccharin also causes morphological alterations in these cells, resulting in pronounced elongation, stretching, and
firmer attachment of cells to the culture dishes. These alterations in cell shape are similar to those observed after treatment
of GH4C1 cells with EGF and with thyrotropin-releasing hormone (TRH), both of which enhance prolactin (PRL) production in these cells.
After assaying for PRL in saccharin-treated cultures, it was observed that this sweetener is also capable of stimulating PRL
production two-to sixfold in a dose-dependent manner. Enhancement of PRL production can be observed at 0.5 mM saccharin, yet this is 10 times less than the saccharin concentration required to alter cell shape. These effects of saccharin
on cell morphology and on PRL production are reversible in GH4C1 cell cultures. When added to cultures along with maximal concentrations of EGF or TRH, the effects of saccharin on PRL production
are additive, suggesting that the actions of saccharin are mediated by a somewhat different pathway from that of the peptide
hormones. Pulse labeling studies indicate that the enhancement of PRL production is highly specific inasmuch as saccharin
was found to decrease the overall rate of protein synthesis in these cells. Saccharin also causes a decrease in the rate of
DNA synthesis under these treatment conditions. Mitomycin C, which similarly inhibited DNA synthesis, had no effect on cell
morphology or PRL production.
This investigation was supported by a Faculty Research Grant from Wheaton College 相似文献
66.
Giorgio M. Hanozet Barbara Giordana Paolo Parenti Andrea Guerritore 《The Journal of membrane biology》1984,81(3):233-240
Summary In brush border membrane vesicles from the midgut ofPhilosamia cynthia larvae (Lepidoptera) thel- andd-alanine uptake is dependent on a potassium gradient and on transmembrane electrical potential difference. Each isomer inhibits the uptake of the other form: inhibition ofl-alanine uptake byd-alanine is competitive, whereas inhibition ofd-alanine uptake byl-alanine is noncompetitive. Transstimulation experiments as well as the different pattern of specificity to cations suggest the existence of two transport systems. Kinetic parameters for the two transporters have been calculated both when Kout>Kin and Kout=Kin.d-alanine is actively transported also by the whole midgut, but it is not metabolized by the intestinal tissue. 相似文献
67.
A method is described for the extraction of enkephalin-like peptides from peripheral nerve using chloroform and acidic methanol to facilitate a differential extraction of peptides and lipid. Porcine splanchnic nerve contains enkephalin-like peptides in low amounts compared to porcine adrenal medulla and striatum. Gel filtration chromatography reveals the presence of enkephalin-like peptides in both processed and cryptic forms. This is the first reported isolation and partial characterization of these peptides in splanchnic nerve. The presence of these peptides in this nerve provides support for the contention that the splanchnic nerve can modulate catecholamine release from the adrenal medulla through an effect on opiate receptors located on chromaffin cells. 相似文献
68.
Barbara Stay Lynda S. Ostedgaard Stephen S. Tobe Alain Strambi Eugene Spaziani 《Journal of insect physiology》1984,30(8):643-651
The free (non-conjugated) ecdysteroid in the ovaries during the first gonadotrophic cycle of Diploptera punctata was identified as 20-hydroxyecdysone. The hormone, quantified by radioimmunoassay and by ultraviolet absorbance, was detectable in the ovary toward the end of vitellogenesis; the quantity increased rapidly during chorion formation. Ovaries with chorionated eggs contained 67 μg of 20-hydroxyecdysone per g fresh weight. The haemolymph free-ecdysteroid, not identified physicochemically, was quantified by radioimmunoassays. The highest concentration was observed at adult emergence; the titre declined between days 1–3 and then remained at a relatively constant level through oviposition (which occurs between day 7 and 8); titres in pregnant females were higher. Ovariectomized females exhibited the same pattern of ecdysteroid titres in the haemolymph as the sham operated controls throughout the period corresponding to the first gonadotrophic cycle. Thus the ovary may not be the only source of haemolymph ecdysteroid related to reproduction in adult females. 相似文献
69.
Dynamics of orange peel fermentation during ensilage 总被引:2,自引:0,他引:2
G. Ashbell G. Pahlow Barbara Dinter Z. G. Weinberg 《Journal of applied microbiology》1987,63(4):275-279
The dynamics of fermentation during ensilage were studied on orange peel (variety Shamouti) ensiled in 50 plastic containers, 10 kg in each, with an outlet for seepage. At predetermined intervals the containers were weighed and samples were taken from three of them for chemical and microbial analysis. Fermentation losses amounted to up to ca one-third of the fresh peel dry matter (DM) content. Most losses occurred within 10 d of commencement of fermentation, and were attributable to gas release. The major fermentation products were ethanol, lactic and acetic acids (16, 3 and 3% of DM, respectively). The dominant microbial populations were lactobacilli and yeasts. Tests are in progress to inhibit the yeasts and thereby reduce fermentation losses. 相似文献
70.
Barbara A. Israel Warren I. Schaeffer 《In vitro cellular & developmental biology. Plant》1987,23(9):627-632
Summary Using both normal and transformed rat liver epithelial cells to prepare cytoplasmic hybrids (cybrids) we have found evidence
to support the theory that the cytoplasm from a normal cell can suppress tumorigenicity. A unique aspect of this study is
that all of the cells utilized, both normal and malignantly transformed, were derived from an original cloned cell. We found
that fusing cytoplasts from normal cells to malignantly transformed whole cells resulted in cybrid clones which, when injected
into newborn rat pups, isogenic with those from which the cell culture was initiated, yielted tumors in 51% of the animals
injected compared to 92% of the animals injected with the tumorigenic parent. Those animals that did develop tumors from the
cybrid cells survived longer than those injected with cells from the tumorigenic parent. Thus, the cybrid, formed of cytoplasm
from both parents, was less tumorigenic than the malignantly transformed parent cell. When reconstituted cells were prepared
by fusing cytoplasts from normal cells with karyoplasts from malignantly transformed cells, a situation in which essentially
all of the cytoplasm of the reconstituted cell is derived from normal cells, the tumorigenic phenotype was extinguished.
This work was supported in part by United States Public Health Service grant CA12056, and grant CA09100 from the National
Cancer Institute, Bethesda, MD. This work is partial fulfillment for the degree of Doctor of Philosophy for B.A.I. 相似文献