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51.
52.
BHK cells transfected with human cathepsin D (CD) cDNA normally segregate the autologous hamster cathepsin D while secreting a large proportion of the human proenzyme. In the present work, we have utilized these transfectants to examine to what extent the mannose-6-phosphate-dependent pathway for lysosomal enzyme segregation contributes to the differential sorting of human and hamster CD. We report that, in recipient control BHK cells, the rate of mannose-6-phosphate-dependent endocytosis of human procathepsin D secreted by transfected BHK cells is lower than that of hamster procathepsin D and much lower than that of human arylsulphatase A. The missorted human enzyme bears phosphorylated oligosaccharides and most of its phosphate residues are “uncovered”, like the autologous enzyme. Thus, despite both the Golgi-associated modifications of oligosaccharides, i.e. the phosphorylation of mannose and the uncovering of mannose-6-phosphate residues, which proceed on human and hamster procathepsin D with comparable efficiency, only the latter is accurately packaged into lysosomes. Ammonium chloride partially affects the lysosomal targeting of cathepsin D in control BHK cells, whereas in transfected cells, this drug strongly inhibits the maturation of human procathepsin D and slightly enhances its secretion. These data indicate that: (1) over-expression of a lysosomal protein does not saturate the Golgi-associated reactions leading to the synthesis of mannose-6-phosphate; (2) a portion of cathepsin D is targeted independently of mannose-6-phosphate receptors in the transfected BHK cells; and (3) whichever mechanism for lysosomal delivery of autologous procathepsin D is involved, this is not saturated by the high rate of expression of human cathepsin D. 相似文献
53.
Pomara Nunzio Shao Binbin Wisniewski Thomas Mehta Pankaj D. 《Neurochemical research》1998,23(12):1563-1566
This report examines plasma amyloid proteins A40 and A42 and apolipoprotein E (apoE) levels and their relationships with age in non-demented older adults with (N = 32) or without the apoE-4 allele (N = 94). A levels did not differ between the groups whereas the 4 allele was associated with a significant reduction in plasma apoE. In subjects with the 4 allele, increasing age was associated with significant reduction in plasma A40. Subjects without the 4 allele showed a significant positive correlation between A40 and A42 levels. There was also a significant correlation between plasma A40 and apoE levels in all subjects. 相似文献
54.
The sensilla ampullacea on the apical antennomere of the leaf-cutting ant Atta sexdens were investigated regarding both their responses to CO2 and their ultrastructure. By staining the sensillum during recording, we confirmed that the sensilla ampullacea are responsible for CO2 perception. We showed that the sensory neurons of the sensilla ampullacea are continuously active without adaptation during stimulation with CO2 (test duration: 1 h). This feature should enable ants to assess the absolute CO2 concentration inside their nests. Sensilla ampullacea have been found grouped mainly on the dorso-lateral side of the distal antennal segment. Scanning and transmission electron microscopic investigations revealed that the external pore opens into a chamber which connects to the ampulla via a cuticular duct. We propose protection against evaporation as a possible function of the duct. The ampulla houses a peg which is almost as long as the ampulla and shows cuticular ridges on the external wall. The ridges are separated by furrows with cuticular pores. The peg is innervated by only one sensory neuron with a large soma. Its outer dendritic segment is enveloped by a dendritic sheath up to the middle of the peg. From the middle to the tip numerous dendritic branches (up to 100) completely fill the distal half of the peg. This is the first report of a receptor cell with highly branched dendrites and which probably is tuned to CO2 exclusively. 相似文献
55.
This work was based on the analysis of digital images of histochemical profile from subcutaneous lesions in sporotrichosis
(ST) and chromoblastomycosis (CM) patients. An additional aim was the detection of carbohydrate expression using lectin histochemical
analysis of the different carbohydrates in the fungal cell wall from four different species (Sporothrix
schenckii, Fonsecaea pedrosoi, Phialophora verrucosa, and Cladophialophora carrionii) associated with diseases mentioned earlier. Slides from tissue biopsies from ST and CM positive patients (n = 10, each) were stained according to routine techniques. Slides were incubated with 25 μg/ml of Con A lectins and WGA conjugated
to peroxidase. Digital image analysis was carried out in a workstation using OPTIMAS™ software system. Routine histochemistry
results indicated that there is significantly higher collagen deposition and elastic fibers in ST characteristic lesions compared
with that found in CM cases. The ST interstitial fibrosis area was larger than in CM lesions. Comparative lectin binding showed
a positive and intense lectin staining pattern in the cell wall of S. schenckii, suggesting a higher expression of glucose/mannose and N-acetyl glucosamine in their cell surface as evidenced by Con A and WGA, respectively. However, these lectins were not effective
to recognize some carbohydrates moieties in the F. pedrosoi, P. verrucosa, and C. carrionii. Such findings contribute to additional information about specific recognition processes between fungal parasites and their
host cell targets may be mediated by the interaction of carbohydrate-binding proteins, such as lectins, on the surface of
one type of cell that combine with complementary sugars on the surface of another cells into fibro-connective tissues associated
with lesions. 相似文献
56.
57.
Carmen Martínez Ana M. Ortiz Yasmina Juarranz Amalia Lamana Iria V. Seoane Javier Leceta Rosario García-Vicu?a Rosa P. Gomariz Isidoro González-álvaro 《PloS one》2014,9(1)
Objective
Suitable biomarkers are essential for the design of therapeutic strategies in personalized medicine. Vasoactive intestinal peptide (VIP) has demonstrated immunomodulatory properties in autoimmune murine and ex vivo human models. Our aim was to study serum levels of VIP during the follow-up of an early arthritis (EA) cohort and to analyze its value as a biomarker predicting severity and therapeutic requirements.Methods
Data from 91 patients on an EA register were analyzed (76% rheumatoid arthritis (RA), 24% undifferentiated arthritis, 73% women, and median age 54 years; median disease duration at entry, 5.4 months). We collected per protocol sociodemographic, clinical, and therapeutic data. VIP levels were determined by enzyme immunoassay in sera harvested from the 91 patients (353 visits; 3.9 visit/patient) and from 100 healthy controls. VIP values below the 25th percentile of those assessed in healthy population were considered low. To determine the effect of independent variables on VIP levels, we performed a longitudinal multivariate analysis nested by patient and visit. A multivariate ordered logistic regression was modeled to determine the effect of low VIP serum levels on disease activity at the end of follow-up.Results
VIP concentrations varied considerably across EA patients. Those fulfilling the criteria for RA had the lowest values in the whole sample, although no significant differences were observed compared with healthy donors. Disease activity, which was assessed using DAS28, inversely correlated with VIP levels. After a two-year follow-up, those patients with low baseline levels of VIP displayed higher disease activity and received more intensive treatment.Conclusion
Patients who are unable to up-regulate VIP seem to have a worse clinical course despite receiving more intense treatment. Therefore, measurement of VIP levels may be suitable as a prognostic biomarker. 相似文献58.
Fernanda Genre Raquel López-Mejías Mercedes García-Bermúdez Santos Casta?eda Carlos González-Juanatey Javier Llorca Alfonso Corrales Bego?a Ubilla José A. Miranda-Filloy Trinitario Pina Carmen Gómez-Vaquero Luis Rodríguez-Rodríguez Benjamín Fernández-Gutiérrez Alejandro Balsa Dora Pascual-Salcedo Francisco J. López-Longo Patricia Carreira Ricardo Blanco Isidoro González-álvaro Javier Martín Miguel A. González-Gay 《PloS one》2014,9(9)
Introduction
Rheumatoid arthritis is an inflammatory disease with high incidence of cardiovascular disease due to accelerated atherosclerosis. Osteoprotegerin (OPG) has been associated with increased risk of atherosclerotic disease in the general population. Several polymorphisms in the OPG gene with functional effects on cardiovascular disease in non-rheumatic individuals have been described. Therefore, we aimed to analyze the effect of three of these functional OPG polymorphisms on the risk of cardiovascular disease in a large and well-characterized cohort of Spanish patients with rheumatoid arthritis.Methods
Three OPG gene variants (rs3134063, rs2073618 and rs3134069) were genotyped by TaqMan assays in 2027 Spanish patients with rheumatoid arthritis. Anti-cyclic citrullinated peptide (anti-CCP) antibody testing was positive in 997 of 1714 tested. Also, 18.3% of the whole series had experienced cardiovascular events, including 5.4% with cerebrovascular accidents. The relationship between OPG variants and cardiovascular events was assessed using Cox regression.Results
No association between OPG gene variants and cardiovascular disease was observed in the whole group of rheumatoid arthritis patients or in anti-CCP positive patients. Nevertheless, a protective effect of CGA haplotype on the risk of cardiovascular disease in general, and specifically in the risk of cerebrovascular complications after adjusting for sex, age at disease diagnosis and traditional cardiovascular risk factors was disclosed in anti-CCP negative patients (HR = 0.54; 95%CI: 0.31–0.95; p = 0.032 and HR = 0.17; 95%CI: 0.04–0.78; p = 0.022, respectively).Conclusion
Our results indicate a protective effect of the OPG CGA haplotype on cardiovascular risk, mainly due to a protective effect against cerebrovascular events in anti-CCP negative rheumatoid arthritis patients. 相似文献59.
Giulia Giannuzzi Nunzio Lobefaro Eleonora Paradies Angelo Vozza Giuseppe Punzi Carlo M. T. Marobbio 《Molecular biotechnology》2014,56(2):157-165
The Lpp2981 gene from Legionella pneumophila, the causative agent of Legionnaire’s disease, was cloned into the pMWT7 plasmid. The construct was used to express this gene in Escherichia coli. Five different bacterial strains were tested to overexpress the gene but without success. Sequence analysis revealed a cluster of four rare codons near the 5′-end of the gene. These codons were replaced with those commonly used in E. coli. The mutated Lpp2981 gene was successfully expressed in all the E. coli strains tested. The expressed protein (with an apparent molecular mass of 30 kDa) was collected in the insoluble fraction of the cell lysate, purified as inclusion bodies and functionally reconstituted into liposomes. The highest level of overexpression was obtained in E. coli C0214 after 6 h of induction with isopropyl-β-d-thiogalactopyranoside at 37 °C, yielding 74 mg of purified protein per liter of culture. We conclude that the clustering of rare codons at the 5′-end of the open-reading frame is a critical factor for the heterologous expression of Lpp2981 in E. coli. 相似文献
60.