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1.
Papers on the mechanisms of translation initiation in mammals studied by reconstruction of initiation complexes from individual components are reviewed. The author points to the constraints of this approach and to the pitfalls ignoring which one might come to erroneous conclusions and even artifacts. In addition, some methods employed in the field as well as some technical problems are discussed in the paper, together with the means of obviating them. The review could be a guidebook for newcomers into this quite labor-consuming field. 相似文献
2.
A mutant strain of Penicillium citrinum grown in a chemically-defined production medium, yielded 145 mg compactin l–1. The medium also facilitated spectrophotometric analysis of compactin. Addition of KH2PO4in the production medium did not increase the compactin production, while addition of a surfactant, Tween 80, increased compactin to 175 mg l–1. Inoculation with 107 spores ml–1 and initial pH of 6.5–7 were the most suitable for compactin production. 相似文献
3.
Protein Tyrosine Kinase Activity and Its Endogenous Substrates in Rat Brain: A Subcellular and Regional Survey 总被引:21,自引:11,他引:10
The rat CNS contains high levels of tyrosine-specific protein kinases that specifically phosphorylate the tyrosine-containing synthetic peptide poly(Glu80,Tyr20). The phosphorylation of this peptide is rapid and occurs with normal Michaelis-Menten kinetics. Using this peptide to assay for enzyme activity, we have measured the protein tyrosine kinase activity in homogenates from various regions of rat CNS. A marked regional distribution pattern was observed, with high activity present in cerebellum, hippocampus, olfactory bulb, and pyriform cortex, and low activity in the pons/medulla and spinal cord. The distribution of protein tyrosine kinase activity was examined in various subcellular fractions of rat forebrain. The majority of the activity was associated with the particulate fractions, with enrichment in the crude microsomal (P3) and crude synaptic vesicle (LP2) fractions. Moreover, the subcellular distribution of pp60csrc, a well-characterized protein tyrosine kinase, was examined by immunoblot analysis using an affinity-purified antibody specific for pp60csrc. The subcellular distribution of pp60csrc paralleled the overall protein tyrosine kinase activity. In addition, using an antibody specific for phosphotyrosine, endogenous substrates for protein tyrosine kinases were demonstrated on immunoblots of homogenates from the various regions and the subcellular fractions. The immunoblots revealed numerous phosphotyrosine-containing proteins that were present in many of the CNS regions examined and were associated with specific subcellular fractions. The differences in tyrosine-specific protein kinase activity, and in phosphotyrosine-containing proteins, observed in various regional areas and subcellular fractions may reflect specific functional roles for protein tyrosine kinase activity in mammalian brain. 相似文献
4.
Charles G. Crispens Philip S. Porter John R. J. Sorenson 《Biological trace element research》1986,10(1):19-24
Plasma copper and zinc levels were measured in SJL/J mice, an inbred strain characterized by a high, spontaneous incidence
of reticulum cell sarcoma (RCS). The changes with age in mean concentrations of these metals were consistent with a physiological
response that is required for remission of neoplasia. Treatment of SJL/J mice with a copper complex, Cu(II)(3,4-diisopropylsalicylate)2 (Cu 3,5-DIPS), dissolved in a 10% Tween 80-saline solution revealed a decrease in survival and decline in the incidence of
RCS at 52 wk of age. The toxic effects of Cu 3,5-DIPS therapy appeared to be related to the intraperitoneal route of administration
and to extracellular deposition of collagen. The inhibitory effect on tumor development was not related to Cu 3,5-DIPS. Rather,
Tween 80 was found to be the factor of importance. 相似文献
5.
F. Morishita A. Shimada Y. Takeda M. Fujimoto H. Katayama K. Yamada 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1996,166(8):467-472
To investigate the functions of GTP-binding protein(s) in the melanosome-aggregating response in fish melanophores, the effects
of activators of G-proteins, namely, mastoparan and compound 48/80, were examined in cultured melanophores of the balck-moor
goldfish, Carassius auratus. Both mastoparan and compound 48/80 induced an approximately 40% increase in the GTP-hydrolyzing activity in the melanophore
membranes compared to the basal level. In intact melanophores, these compounds inhibited the effect of 3-isobutyl-1-methylxanthine,
which induced the accumulation of intracellular cAMP. Pretreatment of melanophores with pertussis toxin at 1 μg ⋅ ml-1 for 15 h attenuated the inhibitory effect of mastoparan on the accumulation of cAMP. However, pretreatment with the toxin
only slightly attenuated the inhibitory effect of compound 48/80 on the accumulation of cAMP. In addition, compound 48/80
at 1 mg ⋅ ml-1 induced full aggregation of the melanosomes in melanophores, though mastoparan at 5 μmol ⋅ l-1 induced only 10–20% aggregation of melanophores. These results suggest that mastoparan and compound 48/80 can each activate
the inhibitory G-protein in goldfish melanophores, which results in inhibition of adenylate cyclase activity. This signal-transduction
pathway is involved in the aggregation of melanosomes in these cells.
Accepted: 3 June 1996 相似文献
6.
E. Thies T. Jenkins F. Stutzenberger 《World journal of microbiology & biotechnology》1994,10(6):657-663
Tween 80 (0.1%, v/v) added to Thermomonospora curvata growing in minimal medium caused a transient lowering of the dry cell mass, decreased the optimal growth temperature of the thermophile from 62 to 54°C, and increased extracellular esterase activity. Cells grown in the presence of Tween 80 had decreased concentrations of branched chain fatty acids and increased concentrations of oleic acid. The detergent removed surface protuberances from mycelia and increased the liberation of enzymes active against crystalline cellulose, but did not stimulate liberation of enzymes active against carboxymethylcellulose, starch or pectin. 相似文献
7.
Mortimer M. Civan Jonathan Robbins Simon Broad Enrique Rozengurt David A. Brown 《The Journal of membrane biology》1993,133(1):51-59
Summary Differentiated neuroblastoma cells exhibit both the delayed rectifier potassium current (I
K) and the M-current (I
M). The present study was designed to determine the roles of protein kinase C (PKC) and of the calmodulin-binding protein 80K/MARCKS, a prominent substrate for PKC and possible regulator of these currents. Neuroblastoma x glioma (NG108-15) hybrid cells transfected with m1 muscarinic receptors were grown with 1% fetal bovine serum (FBS) without the prostaglandin E1 (PGE1) and isobutylmethylxanthine (IBMX) usually added in preparation for electrophysiological studies. Under these conditions, the usual pleomorphism was largely abolished, leaving two populations of small cells with stellate and spherically symmetrical geometries. Whole-cell patch clamping indicated that the two cell types had identical electrophysiological properties, displaying: I
k, a small current through a T-like Ca2+ channel, and no M-current.Stimulation with carbachol shifted the distribution of cells to a more stellate morphology within 24 hr and later (after 48 hr) reduced the PKC substrate 80K/MARCKS by 22±7%. In contrast to the stimulation of I
k observed with cardiac cells, PKC activation produced only a small inhibition of I
k, which was independent of carbachol pretreatment. Thus, PKC and 80K/MARCKS can be dissociated from the regulation of I
k in neuroblastoma cells.Supported in part by research grants from the National Institutes of Health (DK-40145 and EY-08343) and from the U.K. Medical Research Council.We thank Dr. Peter J. Parker for his generous gift of PKC, and Yvonne Vallis for her skillful assistance with the cultures and harvesting of the NG108-15 transfected cells. 相似文献
8.
Kazuyoshi Masuda Shunji Nagata Koichiro Hirano Yasushi Takagishi Hidematsu Hirai 《Microbiology and immunology》1993,37(2):165-167
This paper describes an attempt to effectively induce antibody-dependent cell-mediated cytotoxicity (ADCC) in nude mice. A monoclonal antibody against α-fetoprotein, 80G, coadministered with spleen cells from other nude mice bearing HuH-7N (xenograft of human hepatoma cell line, HuH-7) significantly suppressed the growth of HuH-7N as compared to treatment with 80G alone. 80G with spleen cells from normal nude mice also had some suppressive effect. In contrast, no effect was observed with each spleen cells alone as well as 80G alone. These results suggest that further supply of effector cells could enhance ADCC activity in nude mice. 相似文献
9.
Eva Pocsik Rudolf Mihalik Maria Penzes Hansruedi Loetscher Harald Gallati Bharat B. Aggarwal 《Journal of cellular biochemistry》1995,59(3):303-316
The cell cycle has been shown to regulate the biological effects of human tumor necrosis factor (TNF), but to what extent that regulation is due to the modulation of TNF receptors is not clear. In the present report we investigated the effect of the cell cycle on the expression of surface and soluble TNF receptors in human histiocytic lymphoma U-937. Exposure to hydroxyurea, thymidine, etoposide, bisbensimide, and democolcine lead to accumulation of cells primarily in G1/S, S, S/G2/M, G2/M, and M stages of the cell cycle, respectively. Whilie no significant change in TNF receptors occurred in cells arrested in G1/S or S/G2 stages, about a 50% decrease was observed in cells at M phase of the cycle. Scatchard analysis showed a reduction in receptor number rather than affinity. In contrast, cells arrested at S phase (thymidine) showed an 80% increase in receptor number. The decrease in the TNF receptors was not due to changes in cell size or protein synthesis. The increase in receptors, however, correlated with an increase in total protein synthesis (to 3.8-fold of the control levels). A proportional change was observed in the p60 and p80 forms of the TNF receptors. A decrease in the surface receptors in cells arrested in M phase correlated with an increase in the amount of soluble receptors. The cellular response to TNF increased to 8- and 2-fold in cells arrested in G1 and S phase, respectively; but cells at G2/M phase showed about 6-fold decrease in response. In conclusion, our results demonstrate that the cell cycle plays an important role in regulation of cell-surface and soluble TNF receptors and also in the modulation of cellular response. © 1995 Wiley-Liss, Inc. 相似文献
10.
K. Norrby S. Bergström P. Druvefors 《In vitro cellular & developmental biology. Plant》1984,20(8):607-614
Summary The intact membranous rat mesentery was cultured in Eagle's minimum essential medium containing no serum or only low concentrations
of serum. The procedure is in some important respects superior to previous organ culture techniques. To estimate the extent
of disturbance of homeostasis of the tissue in culture, the spontaneous mast-cell histamine release was quantitated after
preculture preparation of the specimens and after different intervals in culture. Also, the proliferation of fibroblasts and
mesothelial cells that predominate in the mesentery was assessed at 48 h by cytofluorometric quantitation of DNA in single-tissue
cells.
Spontaneous histamine release was time dependent during cultivation, amounting to ca. 50% at 48 h, and was affected by the
medium used for moistening the tissue before cultivation. Culturing also brought about great spontaneous increase in the proliferation
of fibroblasts and mesothelial cells, the rate being related to the concentration of serum. Addition of the mast-cell secretagogues
48/80 or polymyxin B at 1 h caused rapid release of 50 to 60% of the histamine and was followed by augmented proliferation
in the serum-containing media.
The spontaneous increase of cell proliferation in tissue culture may be causally related to mast-cell secretion. Further studies
are needed to define factors influencing the spontaneous mast-cell secretion and the mast-cell-dependent mitogenesis in normal
tissue cells
Supported by grants from the Swedish Medical Research Council (Project 5942) and State Board for Animal Experiments. 相似文献