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1.
Regulation of collagenase and collagenase mRNA production in early- and late-passage human diploid fibroblasts 总被引:6,自引:0,他引:6
The levels of collagenase and collagenase mRNA produced by early-passage (less than 40% of lifespan completed) and late-passage (greater than 80% of lifespan completed) cultures of human fibroblasts were analyzed. The constitutive levels of collagenase and collagenase mRNA produced by the late-passage cultures were 10-30 x greater than the levels observed in similarly treated early-passage cultures. Immunofluorescence analysis established that the percentage of collagenase-positive cells was also greater (77% vs. 4%) in the late-passage cultures. To determine whether the difference in collagenase production resulted from cell-derived regulatory factors, collagenase production was examined in cultures plated onto substrates coated with fibroblast extracellular matrix (ECM). Collagenase and collagenase mRNA production was enhanced in both types of cultures, although amounts produced by ECM-induced early-passage cultures was significantly less than that produced by similarly treated late-passage cultures. Collagen-coated substrates also induced collagenase synthesis. 相似文献
2.
Lars Iversen †Eileen Mulvihill †Betty Haldeman ‡Nils Henrik Diemer Frank Kaiser Malcolm Sheardown Peter Kristensen 《Journal of neurochemistry》1994,63(2):625-633
Abstract: Metabotropic glutamate receptors mediate their intracellular response by coupling to G proteins and may be divided into three subfamilies: mGluR1 and mGluR5, which stimulate phosphatidylinositol hydrolysis; mGluR2 and mGluR3, which are negatively coupled to cyclic AMP formation; and mGluR4 and mGluR6, which also inhibit forskolin-stimulated cyclic AMP formation. The mGluR4 subtypes may represent l -2-amino-4-phosphonobutyrate-sensitive presynaptic autoreceptors, and two alternatively spliced variants of the mGluR4 coding for two receptors with different C termini have been identified. Using in situ hybridization, we measured the levels of mGluR1–mGluR5 mRNA in regions of the rat brain 24 h after transient global ischemia, a time point when no neuronal damage can yet be observed morphologically. In the hippocampus, the mRNA levels for mGluR1, mGluR2, and mGluR5 were decreased, mGluR3 mRNA levels were unchanged, and the mGluR4 mRNA levels were strongly increased. The strongest increase appeared to be in the mRNA encoding mGluR4b. The mGluR4 mRNA was also increased in the parietal cortex, whereas the ventral posteromedial thalamic nucleus showed a small decrease in its mRNA content. These results suggest that vulnerable neurons react to an increased extracellular glutamate concentration by differential regulation of the mRNA for pre- and postsynaptically located metabotropic glutamate receptors. 相似文献
3.
Summary The present study investigates the effects of phenylsuccinate (PS), an inhibitor of the mitochondrial ketodicarboxylate carrier (KCC), on release of-aminobutyric acid (GABA), glutamate (Glu), glutamine (Gln), and glycine (Gly), induced by potassium chloride (KCl) and by cardiac arrest caused by a halothane overdose. Microdialysates were collected from the hippocampus of anaesthetized rats, and analyzed by HPLC. Continuous perfusion of 50 mM PS through the dialysis probe, reduces release of GABA induced by KCl (50 mM for 10 min through the dialysis probe) by up to 72%. In addition, PS abolished KCl-induced release of Glu. Release of GABA during cardiac arrest was not reduced by PS, whereas PS reduced release of Glu in the early stage of cardiac arrest. PS furthermore increased the basal level of Gln, and reversed a decrease of Gln induced by cardiac arrest.It is proposed that the KCC is present in GABA'ergic neurons of the rat hippocampus, and that GABA, released by KCl, can be synthesized in a KCC dependent manner. It is also suggested that ischemia-induced release of GABA, to some extent, has a non-transmitter origin. The results furthermore indicate that uptake of Gln into GABA'ergic and Glu'ergic neurons is not regulated by simple demand mechanisms.Abbreviations PS
phenylsuccinate
- KCC
ketodicarboxylate carrier
- GABA
-aminobutyric acid
- Glu
glutamate
- Gln
glutamine
- Gly
glycine
-
-KG
-ketoglutarate
- Mal
malate
- KRB-buffer
Krebs-Ringer bicarbonate-buffer
- HPLC
high pressure liquid chromatography 相似文献
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In many bird species, there is a floating population of females that are excluded from breeding because of competition for limited breeding resources. Female floaters may enhance their reproductive success by engaging in intraspecific brood parasitism. We studied female floaters in a population of European starlings, Sturnus vulgaris, in order to determine their identity and potential parasitic behaviour. Females were caught after being attracted to nestboxes with artificial nests during 1993-1995. None of the females was known to have a nest of her own at capture but 47% of the females either laid an egg in the nest or carried a fully developed egg within the reproductive tract, indicating that they were intraspecific brood parasites. The floating females were significantly younger and smaller than breeding females. Of 13 females equipped with radiotransmitters and followed daily, all but one started a breeding attempt of their own after 3-8 days and the majority settled as secondary females or mated with males where the original female had disappeared. This suggests that females that are unable to compete successfully for nest sites or males early in the breeding season may use intraspecific brood parasitism to enhance reproductive success during the period that they are constrained from breeding. The importance of settling rapidly because of a seasonal decline in reproductive success may also promote the evolution of intraspecific brood parasitism in the starling. The relative reproductive success of combining egg dumping with breeding compared with traditional breeding will depend on the costs of delaying breeding as well as the probability of finding a mate later in the breeding season. Copyright 1999 The Association for the Study of Animal Behaviour. 相似文献
6.
Effects of habitat fragmentation on the fitness of two common wetland species,Carex davalliana and Succisa pratensis 总被引:1,自引:0,他引:1
Small habitat size and spatial isolation may cause plant populations to suffer from genetic drift and inbreeding, leading to a reduced fitness of individual plants. We examined the germination, establishment, growth, and reproductive capacity of two characteristic species of mown fen meadows, Carex davalliana, and Succisa pratensis, common in Switzerland. Plants were grown from seeds, which were collected in 18 habitat islands, differing in size and in degree of isolation. We used both common garden and reciprocal transplant experiments to assess effects of habitat fragmentation. In the common garden, plants of Carex originating from small habitat islands yielded 35% less biomass, 30% fewer tillers, and 45% fewer flowering tillers than plants from larger ones. In contrast, plants of Succisa originating from small habitat islands yielded 19% more biomass, 14% more flower heads and 35% more flowers per flower head than plants from larger ones. Moreover, plants of Succisa from small isolated habitats yielded 32% more rosettes than did plants from small connected islands. Reciprocally transplanted plants of Succisa originating from small habitat islands produced 7% more rosettes than plants from larger ones. There was no effect of small habitat size and isolation on germination and establishment of both species in the field. Our results document genetic differences in performance attributable to habitat fragmentation in both species. We suggest that fitness loss in Carex is caused by inbreeding depression, whereas in Succisa the differences in fitness are more likely caused by genetic differentiation. Our study implies that habitat fragmentation affects common habitat-specific species, such as Carex and Succisa, as well as rare ones. 相似文献
7.
Binz PA Abdi F Affolter M Allard L Barblan J Bhardwaj S Bienvenut WV Bulet P Burgess J Carrette O Corthals G Delalande F Diemer H Favreau P Giuliano E Gueguen Y Guillaume E Hahner S Man P Michalet S Neri D Noukakis D Palagi P Paroutaud P Pimenta DC Quadroni M Resemann A Richert S Rybak J Sanchez JC Scherl A Scheurer S Schweiger Hufnagel U Siethoff C Suckau D van Dorsselaer A Wagner Redeker W Walter N Stöcklin R 《Proteomics》2003,3(8):1562-1566
After the success of the mass spectrometry (MS) round table that was held at the first Swiss Proteomics Society congress (SPS'01) in Geneva, the SPS has organized a proteomics application exercise and allocated a full session at the SPS'02 congress. The main objective was to encourage the exchange of expertise in protein identification, with a focus on the use of mass spectrometry, and to create a bridge between the users' questions and the instrument providers' solutions. Two samples were sent to fifteen interested labs, including academic groups and MS hardware providers. Participants were asked to identify and partially characterize the samples. They consisted of a complex mixture of peptide/proteins (sample A) and an almost pure recombinant peptide carrying post-translational modifications (sample B). Sample A was an extract of snake venom from the species Bothrops jararaca. Sample B was a recombinant and modified peptide derived from the shrimp Penaeus vannamei penaeidin 3a. The eight labs that returned results reported the use of a wide range of MS instrumentation and techniques. They mentioned a variety of time and manpower allocations. The origin of sample A was generally identified together with a number of database protein entries. The difficulty of the sample identification lay in the incomplete knowledge of the Bothrops species genome sequence and is discussed. Sample B was generally and correctly identified as penaeidin. However, only one group reported the full primary structure. Interestingly, the approaches were again varied and are discussed in the text. 相似文献
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10.
BelAiba RS Djordjevic T Petry A Diemer K Bonello S Banfi B Hess J Pogrebniak A Bickel C Görlach A 《Free radical biology & medicine》2007,42(4):446-459
NADPH oxidases have been identified as sources of reactive oxygen species (ROS) in vascular cells. In addition to the initially described enzyme containing gp91phox (NOX2), several homologues to NOX2 have been identified. Whereas NOX1, NOX2, and NOX4 are expressed in endothelial cells, a functional role of NOX5 containing additional N-terminal calcium-binding domains of varying sequences has not been reported in these cells. NOX5 protein was found in the endoplasmic reticulum of human microvascular endothelial cells (HMEC-1) and in the vascular wall. HMEC-1 cells expressed NOX5beta and NOX5delta as well as a variant lacking calcium-binding domains (NOX5S). NOX5beta and NOX5S increased basal ROS levels. Ionomycin exclusively enhanced NOX5beta-mediated ROS production. Although p22phox, when overexpressed, interacted with both NOX5 proteins, it was not essential for NOX5-mediated ROS production. NOX5 proteins stimulated endothelial cell proliferation and the formation of capillary-like structures whereas depletion of NOX5 by siRNA prevented these responses to thrombin. These data show that endothelial cells express different NOX5 variants including NOX5S lacking calcium-binding domains. NOX5 proteins are functional, promoting endothelial ROS production, proliferation, and the formation of capillary-like structures and contribute to the endothelial response to thrombin. These findings suggest that NOX5 variants play a novel role in controlling ROS-dependent processes in the vasculature. 相似文献