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1.
A number of different membrane associated isoforms of the neural cell adhesion molecule (N-CAM) have previously been identified. Here the structure of a novel secreted isoform of N-CAM is established by analysis of a cDNA corresponding to an N-CAM mRNA from human skeletal muscle. The mRNA incorporates a novel sequence block into the extracellular domain, which introduces an in-frame stop codon and thus prematurely terminates the coding sequence, generating a truncated N-CAM polypeptide. Analysis of genomic clones indicates that the inserted sequence is present as a discrete exon within the human N-CAM gene, and Northern analysis shows it to be associated specifically with a 5.2 kb mRNA species from skeletal muscle and brain. Stable transfectants expressing the secreted isoform accumulate it in the cytoplasm and release it to the culture medium. In contrast, cells transfected with cDNA encoding lipid-tailed N-CAM express it predominantly at the cell surface. The existence of a secreted isoform may further expand the spectrum of N-CAM function beyond its known involvement in intercellular adhesion to extracellular matrix interactions. 相似文献
2.
The metabolism of pregnenolone in subcellular fractions of the testes of the macaque (Macaca fascicularis) has been studied using capillary gas chromatography to characterize and quantify the metabolites, after their conversion into the O-methyloxime and/or trimethylsilyl ether derivatives. The microsomal incubations yielded the greatest quantities of metabolites, with lesser amounts in the mitochondrial fraction. The cytosolic fraction contained no significant quantity of metabolites after incubation, except for 5alpha-androst-16-en-3 beta-ol. This, and other odorous androst-16-enes, found in the microsomal fraction, are of particular interest in the context of animal communication because of their possible pheromonal role. Pregnenolone was converted into androst-5-ene-3 beta,17 beta-diol, androst-4-ene-3,17-dione and testosterone, suggesting that both classical pathways for testosterone synthesis were operating. Testosterone was further converted into 5 alpha-reduced androstanediols, especially in the microsomal fraction. 相似文献
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A method for the determination of desferrioxamine-available iron in tissue fractions is described which involves incubation with desferrioxamine, extraction of desferrioxamine and its iron-bound form, ferrioxamine, and quantitation of these two forms of the drug by reversed-phase hplc analysis. Chelatable iron levels in the 1-10µMolar region could be accurately and reproducibly measured using this technique.
The desferrioxamine-available iron levels in both the cortex and medulla of rabbit kidneys were significantly elevated (up to 2-fold) after the organs had been subjected to 2 hours warm ischaemia or 24 hours cold storage at 0°C In hypertonic citrate solution. There was no change in the total iron content of the tissues under these circumstances and thus a redistribution of intracellular iron to more available pools had presumably taken place as a result of ischaemia. This redistribution of iron may be an important factor in the initiation of peroxidative damage to cell membranes upon reperfusion of the organ with oxygen. 相似文献
The desferrioxamine-available iron levels in both the cortex and medulla of rabbit kidneys were significantly elevated (up to 2-fold) after the organs had been subjected to 2 hours warm ischaemia or 24 hours cold storage at 0°C In hypertonic citrate solution. There was no change in the total iron content of the tissues under these circumstances and thus a redistribution of intracellular iron to more available pools had presumably taken place as a result of ischaemia. This redistribution of iron may be an important factor in the initiation of peroxidative damage to cell membranes upon reperfusion of the organ with oxygen. 相似文献
6.
Neuronal process outgrowth of human sensory neurons on monolayers of cells transfected with cDNAs for five human N-CAM isoforms 总被引:14,自引:7,他引:7 下载免费PDF全文
P Doherty C H Barton G Dickson P Seaton L H Rowett S E Moore H J Gower F S Walsh 《The Journal of cell biology》1989,109(2):789-798
Full length cDNAs for a variety of human N-CAM isoforms have been transfected into mouse L-cells and/or 3T3 cells. Three independent clones of each cell line that were shown to express human N-CAM were tested for their ability to support the morphological differentiation of sensory neurons. The cell surface expression of N-CAM isoforms, linked to the membrane directly by an integral transmembrane spanning domain or indirectly via covalent attachment to a glycosyl-phosphatidylinositol moiety, were consistently found to be associated with a significant increase in the morphological differentiation of both human and rat dorsal root ganglion neurons. Modification of the extracellular structure of both classes of N-CAM, consequent to the expression of a glycosylated 37-amino acid sequence normally found expressed exclusively in muscle N-CAM isoforms did not obviously affect the ability of transfected cells to support increased neuronal differentiation. 3T3 cells that were transfected with a full length cDNA encoding a secreted N-CAM isoform, and that have previously been shown to secrete N-CAM into the growth media rather than link it to the membrane did not significantly differ from control cells in their ability to support neuronal differentiation. These data provide direct evidence for both transmembrane and lipid-linked N-CAM isoforms being components of the regulatory machinery that determines neuronal morphology and process outgrowth. 相似文献
7.
William R. Gower Jr. Robert M. Risch Constantine V. Godellas Peter J. Fabri 《In vitro cellular & developmental biology. Animal》1994,30(3):151-161
Summary A new human pancreatic cancer (HPAC) cell line was established from a nude mouse xenograft (CAP) of a primary human pancreatic
ductal adenocarcinoma. In culture, HPAC cells form monolayers of morphologically heterogenous, polar epithelial cells, which
synthesize carcinoembryonic antigen, CA 19-9, CA-125, cytokeratins, antigens for DU-PAN-2, HMFG1, and AUA1, but do not express
chromogranin A or vimentin indicative of their pancreatic ductal epithelial cell character. In the presence of serum, HPAC
cell DNA synthesis was stimulated by insulin, insulin growth factor-I, epidermal growth factor, and TGF-α but inhibited
by physiologic concentrations of hydrocortisone and dexamethasone. Dose-dependent inhibition of DNA synthesis was limited
to steroids with glucocorticoid activity. The inhibitory effect of dexamethasone was abolished by the glucocorticoid antagonist
RU 38486. Binding of [3H]dexamethasone to cytosolic proteins was specific and saturable at 4° C. Scatchard analysis of binding data demonstrated
a single class of high-affinity binding sites (Kd=3.8±0.9 nM; Bmax=523±128 fmol/mg protein). Western blot analysis revealed a major protein band that migrated at a Mr of 96 kDa. Northern blot analysis identified an mRNA of approximately 7 kilobases which hybridized with a specific glucocorticoid
receptor complementary DNA probe (OB7). These findings support a role for glucocorticoids in the regulation of human malignant
pancreatic cell function. 相似文献
8.
David J. Gower Jens B. Rasmussen Simon P. Loader Mark Wilkinson 《African Journal of Ecology》2004,42(2):83-87
A report is given of an adult caecilian, Scolecomorphus kirkii, found in the gut of a specimen of the snake Atractaspis aterrima from the Udzungwa Mountains, Tanzania. Both predator and prey are largely fossorial in soil, and their ecology is poorly known, such that this is the first reported predator of any scolecomorphid caecilian. The caecilian was ingested head first and much of the flesh from the anterior of the specimen had been digested. The prey/predator mass ratio is 0.48. This value is substantially higher than reported for A. aterrima from West Africa, and refutes the notion that this species feeds only on small prey. Most reported predators of caecilians are snakes, and a brief review is presented. 相似文献
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Commentary: Carbon Metabolism of the Terrestrial Biosphere: A Multitechnique Approach for Improved Understanding 总被引:2,自引:0,他引:2
J. G. Canadell H. A. Mooney D. D. Baldocchi J. A. Berry J. R. Ehleringer C. B. Field S. T. Gower D. Y. Hollinger J. E. Hunt R. B. Jackson S. W. Running G. R. Shaver W. Steffen S. E. Trumbore R. Valentini B. Y. Bond 《Ecosystems》2000,3(2):115-130
Understanding terrestrial carbon metabolism is critical because terrestrial ecosystems play a major role in the global carbon
cycle. Furthermore, humans have severely disrupted the carbon cycle in ways that will alter the climate system and directly
affect terrestrial metabolism. Changes in terrestrial metabolism may well be as important an indicator of global change as
the changing temperature signal. Improving our understanding of the carbon cycle at various spatial and temporal scales will
require the integration of multiple, complementary and independent methods that are used by different research communities.
Tools such as air sampling networks, inverse numerical methods, and satellite data (top-down approaches) allow us to study
the strength and location of the global- and continental-scale carbon sources and sinks. Bottom-up studies provide estimates
of carbon fluxes at finer spatial scales and examine the mechanisms that control fluxes at the ecosystem, landscape, and regional
scales. Bottom-up approaches include comparative and process studies (for example, ecosystem manipulative experiments) that
provide the necessary mechanistic information to develop and validate terrestrial biospheric models. An iteration and reiteration
of top-down and bottom-up approaches will be necessary to help constrain measurements at various scales. We propose a major
international effort to coordinate and lead research programs of global scope of the carbon cycle.
Received 7 May 1999; accepted 28 September 1999. 相似文献