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31.
Summary Human umbilical vein endothelial cells (HUV-EC) grew rapidly in vitro in medium supplemented with epidermal growth factor,
fetal bovine serum (FBS) and human diploid fibroblast-conditioned medium. The effect of FBS could be replaced partially by
bovine serum albumin, cholesterol, and vitamin E, and completely by further addition of serum dialysate or refeeding every
other day. Among these components, fibroblast-conditioned medium is essential for HUV-EC growth. The HUV-EC were cultured
serially for over 50 population doublings in the 10% FBS containing fibroblast-conditioned medium and for over 40 population
doublings in the serum-free medium. Mitogenic factor(s) present in the medium conditioned by fibroblasts may be related to
endothelial cell growth factor and play an important role angiogenesis and regeneration of vascular endothelium in vitro. 相似文献
32.
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
33.
Molecular cloning and sequence analysis of cDNA for human transferrin 总被引:10,自引:0,他引:10
G Uzan M Frain I Park C Besmond G Maessen J S Trépat M M Zakin A Kahn 《Biochemical and biophysical research communications》1984,119(1):273-281
A cDNA clone for human transferrin was identified from a human liver cDNA library by pre-screening with different ss-cDNA probes against length-fractionated liver mRNAs, positive hybridization-selection and nucleotide sequence analysis. The insert was of 1 kb, encoding human transferrin from aminoacid 403 through the COOH terminus, with a 3' non coding region of 166 nucleotides. This insert hybridized with a single major mRNA species of about 2.4 kb and several genomic DNA restriction fragments. Hybridization of the Southern blots with different parts of the transferrin insert and at different stringences suggest that the various bands observed correspond to splice sites inside one gene rather than to hybridization to several related genes. Finally, a single or a low number of transferrin gene copies seem to exist in the human genome. 相似文献
34.
N E Owen 《Biochemical and biophysical research communications》1984,125(2):500-508
The regulation of Na/K/Cl cotransport was investigated in vascular smooth muscle cells. That a Na/K/Cl cotransport system exists was established by the finding that the ouabain insensitive K influx was sensitive to the "loop" diuretic bumetanide. Furthermore, bumetanide sensitive K influx was dependent upon the presence of both Na and Cl in the extracellular milieu. Bumetanide sensitive K influx was inhibited by agents which elevate cellular cyclic AMP levels, and to a lesser extent by agents which elevate cellular cyclic GMP levels. When serum, EGF or TPA was added, bumetanide sensitive K influx was enhanced. These results suggest that vascular smooth muscle cells have a ouabain insensitive, bumetanide sensitive Na/K/Cl cotransport system which is stimulated by serum, EGF or TPA and inhibited by cAMP or cGMP. 相似文献
35.
36.
C. A. Lapp M. E. Stachura J. M. Tyler Y. S. Lee 《In vitro cellular & developmental biology. Plant》1987,23(10):686-690
Summary GH3 cell secretory activity was studied in long-term perifusion to define previously reported spontaneous increases in growth
hormone (GH) and prolactin production (PRL). Mechanically harvested cells (1×107/column) were perifused at 4 ml/h for 72 h. A basal period of variable duration (8 to 12 h), during which hormone secretion
was stable, was followed by steadily increasing secretion rates. Changes in cell number were not sufficient to acount for
increased jormone secretion rates: a) there was no significant change in cell count after 72 h (0.97±0.03×107;n=18); b) mean cell column DNA content increased 25.5% above the base value, whereas GH secretion rose 385% and PRL rose 178%
(n=5). Observed differences in the duration of the basal secretion period, the basal secretory rate, and the magnitude of secretory
rate increase were associated with several variables: a) variablility within a subline was a function of passage number: GH
secretion decreased and PRL secretion increased with subculture number; b) cells with identical lot and freeze numbers, but
received at different times, behaved differently; c) the presence of an antifungal agent (nystatin) altered hormone secretion
reproducibly. Conclusions: a) rates of GH and PRL secretion rise spontaneously in perifusion without a proportional increase
in GH3 cell number; b) fluctuations in the rate of GH3 cell secretion of GH and PRL are not entirely random but are determined by several definable variables.
Supported by a grant to MES from the National Institutes of Health (AM33388) and in part by the Medical Research Service of
the Veterans Administration. 相似文献
37.
Jan Kopecký Josef Houštěk Eva Szarska Zdeněk Drahota 《Journal of bioenergetics and biomembranes》1986,18(6):507-519
The proteolipid subunit of H+-ATPase was labeled by [14C]N,N-dicyclohexylcarbodiimide in bovine heart mitochondria. The radioactive labeling was followed using various systems of sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). When using discontinuous SDS-PAGE (Laemmli, U.K., 1970,Nature (London)227, 680–685) a monomeric (Mr 7600±1500) and a dimeric form (Mr 17,800±1200) of the proteolipid were detected, while only the monomeric form was found on urea (8 M) containing gels (SDS-PAGE according to Laemmli; or Swank, R. T., and Munkers, K. D., 1971,Anal. Biochem.
39, 462–477). When using SDS-PAGE with Na-Pi buffer (Weber, K., and Osborn, M., 1969,J. Biol. Chem.
244, 4406–4442), only a dimeric form of the proteolipid (Mr 15,000±1000) was detected. Experimental data indicate that the different patterns of proteolipid separation are related to the presence of the two distinct proteolipid conformations in the SDS solution. 相似文献
38.
Akemichi Ueno Naokatu Arakaki Toshiya Oribe Yoshiro Takeda 《Molecular and cellular biochemistry》1986,70(2):121-130
Summary A two-chain polypeptide, which corresponds to amino acid residues 115–143 and 144–184(185) of bovine serum albumin, connected to each other by a disulfide bridge, potentiated the effects of insulin on glucose transport and glucose metabolism in isolated rat adipocytes. Although the peptide alone had little activity, it shifted the concentration-response curves of insulin-stimulated D-[I-14C]glucose oxidation, 2-deoxyglucose transport, and lipid synthesis from D-[U-14C]glucose to lower insulin concentrations. It also increased the maximal responses of these parameters to insulin. However, it did not affect insulin binding to adipocytes. The peptide protected insulin considerably from degradation, but this effect alone cannot account for its effect in increasing the maximal responses to the hormone, and even when degradation of a submaximal concentration of insulin was suppressed by bacitracin, the peptide still had an enhancing effect. These results suggest not only that the peptide influences a step distal to receptor-mediated insulin binding but also that inhibition of insulin degradation alone cannot explain its total effect.The peptide lost its insulin-stimulating activity completely when it was further digested with V8 or lysinespecific endopeptidase, or when it was reduced and then carboxamidomethylated or oxidized with performic acid. Similar active tryptic fragments were obtained from human and rat albumins.Insulin-stimulating peptides should be useful in studies on the mechanisms of insulin action including both the sensitivities and responsiveness of target cells to the hormone.Abbreviations ISP
insulin-stimulating peptide
- HEPES
N-(2-hydroxyethyl)piperazine-N-2-ethanesulfonic acid
- HPLC
high-performance liquid chromatography
- SDS
sodium dodecyl sulfate 相似文献
39.
Concerted stimulation of PI-turnover, Ca2+-influx and histamine release in antigen-activated rat mast cells 总被引:2,自引:0,他引:2
Phospholipid metabolism in rat mast cells activated by antigen was examined with reference to phosphatidylinositol (PI) turnover. Upon antigen stimulation, histamine release from passively sensitized mast cells with IgE was potentiated by adding phosphatidylserine (PS). The addition of antigen to [3H]glycerol-prelabeled and sensitized mast cells induced a marked loss of radioactivity of PI and a concurrent accumulation of 1,2-diacylglycerol (DG) and phosphatidic acid (PA) within 5 to 60 sec. Furthermore, this antigen-induced PI breakdown was enhanced in the presence of Mg2+. Histamine release occurred in parallel with PI breakdown. On the other hand, the transient Ca2+ influx into mast cells, as measured by uptake of 45Ca2+, was found to occur quickly after cells were activated by antigen, which was concerted with PI breakdown. These results suggest that enhanced PI turnover may be an important step in the biochemical sequence of events leading to release of histamine, and that not only Ca2+ but also Mg2+ appears to take a part in stimulus-response coupling in rat mast cells. 相似文献
40.
An antiserum to pure glutamate decarboxylase (GAD) when incubated with rat cortical synaptosomes in the presence of complement caused release of 33-53% of lactate dehydrogenase (LDH) and 22-41% of total GAD. In addition most of the gamma-aminobutyrate (GABA) present was released. Anti-GAD antiserum alone, or complement alone, were without action. The antiserum plus complement had no effect on noradrenaline or choline uptake, and did not release choline acetylase (ChAT). Anti-ChAT serum plus complement released 30-37% of ChAT and 10-13% of LDH. It prevented choline uptake. This serum did not produce GAD release or prevent GABA, choline or noradrenaline uptake. When cortical synaptosomes were exposed to both antisera plus complement, their actions were strictly additive. The data indicate specific lysis of GABAergic and cholinergic synaptosomal sub-populations. 相似文献