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Molecular cloning and characterization of an alpha1,3 fucosyltransferase, CEFT-1, from Caenorhabditis elegans 总被引:1,自引:1,他引:1
We report on the identification, molecular cloning, and characterization of
an alpha1,3 fucosyltransferase (alpha1,3FT) expressed by the nematode,
Caenorhabditis elegans . Although C. elegans glycoconjugates do not express
the Lewis x antigen Galbeta1-- >4[Fucalpha1-->3]GlcNAcbeta-->R,
detergent extracts of adult C.elegans contain an alpha1,3FT that can
fucosylate both nonsialylated and sialylated acceptor glycans to generate
the Lexand sialyl Lexantigens, as well as the lacdiNAc-containing acceptor
GalNAcbeta1-->4GlcNAcbeta1-- >R to generate GalNAcbeta1-->4
[Fucalpha1-->3]GlcNAcbeta1-->R. A search of the C.elegans genome
database revealed the existence of a gene with 20-23% overall identity to
all five cloned human alpha1,3FTs. The putative cDNA for the C.elegans
alpha1,3FT (CEFT-1) was amplified by PCR from a cDNA lambdaZAP library,
cloned, and sequenced. COS7 cells transiently transfected with cDNA
encoding CEFT-1 express the Lex, but not sLexantigen. The CEFT-1 in the
transfected cell extracts can synthesize Lex, but not sialyl Lex, using
exogenous acceptors. A second fucosyltransferase activity was detected in
extracts of C. elegans that transfers Fuc in alpha1,2 linkage to Gal
specifically on type-1 chains. The discovery of alpha-fucosyltransferases
in C. elegans opens the possibility of using this well-characterized
nematode as a model system for studying the role of fucosylated glycans in
the development and survival of C.elegans and possibly other helminths.
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Paul C. Schroy Anil K. Rustgi Ekaterini Ikonomu Xiu P. Liu Joseph Polito Chris Andry J. Connor O'Keane 《Journal of cellular physiology》1994,161(1):111-123
The polar-planar compound hexamethylene bisacetamide (HMBA) can inhibit HT29 colon carcinoma cell growth and induce a more benign phenotype, as defined by decreased anchorage-independent clonogenicity, loss of a cell surface malignancy marker, and decreased in vivo tumorigenicity. The principle aim of this study was to determine whether HMBA's effects on HT29 cell growth and biologic behavior correlate with effects on intestinal differentiation. Parallel studies were performed with sodium butyrate (NaBT), a potent inducer of intestinal differentiation HT29 cell growth, proliferation, and markers of intestinal differentiation were assayed after short- and long-term treatment with HMBA, NaBT, or the combination. Both 5 mM HMBA and 5 mM NaBT were potent inhibitors of monolayer growth; in combination their effects were nearly additive. Inhibition of DNA synthesis was detectable within 6 h of treatment and was preceded by down-regulation of c-myc expression. Soft agar clonogenicity was also decreased by 90%, > 99%, and > 99% by HMBA, NaBT, and the combination, respectively. Despite these parallel effects on growth and in vitro markers of a benign phenotype, effects on intestinal differentiation were discordant. NaBT induced significant increases in membrane-associated alkaline phosphatase activity, cytosolic mucin content, PAS+/diastase-resistant cells, and ultrastructural evidence of intestinal cell differentiation. HMBA not only failed to induce markers of intestinal differentiation, but attenuated NaBT's effects when used in combination. These data suggest that growth and intestinal differentiation may be independently regulated in HT29 cells. They also suggest that expression of intestinal markers of differentiation is not a prerequisite for the acquisition of a more benign phenotype. © 1994 Wiley-Liss, Inc. 相似文献
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A study of molecular dynamics and freezing phase transition in tissues by proton spin relaxation
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Muscle, spleen, and kidney tissues from 4-wk-old C57 black mice were studied by proton magnetic resonance. Spin-lattice relaxation times at high fields and in the rotating frame, as well as the spin-spin relaxation times, are reported as a function of temperature in the liquid and frozen phase. Motions of large molecules and of water molecules and their changes at the freezing phase transition are studied. The shortcomings of the two-state fast-exchange relaxation model are discussed. 相似文献