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Xu J Reumers J Couceiro JR De Smet F Gallardo R Rudyak S Cornelis A Rozenski J Zwolinska A Marine JC Lambrechts D Suh YA Rousseau F Schymkowitz J 《Nature chemical biology》2011,7(5):285-295
Many p53 missense mutations possess dominant-negative activity and oncogenic gain of function. We report that for structurally destabilized p53 mutants, these effects result from mutant-induced coaggregation of wild-type p53 and its paralogs p63 and p73, thereby also inducing a heat-shock response. Aggregation of mutant p53 resulted from self-assembly of a conserved aggregation-nucleating sequence within the hydrophobic core of the DNA-binding domain, which becomes exposed after mutation. Suppressing the aggregation propensity of this sequence by mutagenesis abrogated gain of function and restored activity of wild-type p53 and its paralogs. In the p53 germline mutation database, tumors carrying aggregation-prone p53 mutations have a significantly lower frequency of wild-type allele loss as compared to tumors harboring nonaggregating mutations, suggesting a difference in clonal selection of aggregating mutants. Overall, our study reveals a novel disease mechanism for mutant p53 gain of function and suggests that, at least in some respects, cancer could be considered an aggregation-associated disease. 相似文献
44.
Lupberger J Zeisel MB Xiao F Thumann C Fofana I Zona L Davis C Mee CJ Turek M Gorke S Royer C Fischer B Zahid MN Lavillette D Fresquet J Cosset FL Rothenberg SM Pietschmann T Patel AH Pessaux P Doffoël M Raffelsberger W Poch O McKeating JA Brino L Baumert TF 《Nature medicine》2011,17(5):589-595
Hepatitis C virus (HCV) is a major cause of liver disease, but therapeutic options are limited and there are no prevention strategies. Viral entry is the first step of infection and requires the cooperative interaction of several host cell factors. Using a functional RNAi kinase screen, we identified epidermal growth factor receptor and ephrin receptor A2 as host cofactors for HCV entry. Blocking receptor kinase activity by approved inhibitors broadly impaired infection by all major HCV genotypes and viral escape variants in cell culture and in a human liver chimeric mouse model in vivo. The identified receptor tyrosine kinases (RTKs) mediate HCV entry by regulating CD81-claudin-1 co-receptor associations and viral glycoprotein-dependent membrane fusion. These results identify RTKs as previously unknown HCV entry cofactors and show that tyrosine kinase inhibitors have substantial antiviral activity. Inhibition of RTK function may constitute a new approach for prevention and treatment of HCV infection. 相似文献
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Crystel Bonnet M’hamed Grati Sandrine Marlin Jacqueline Levilliers Jean-Pierre Hardelin Marine Parodi Magali Niasme-Grare Diana Zelenika Marc Délépine Delphine Feldmann Laurence Jonard Aziz El-Amraoui Dominique Weil Bruno Delobel Christophe Vincent Hélène Dollfus Marie-Madeleine Eliot Albert David Catherine Calais Jacqueline Vigneron Bettina Montaut-Verient Dominique Bonneau Jacques Dubin Christel Thauvin Alain Duvillard Christine Francannet Thierry Mom Didier Lacombe Françoise Duriez Valérie Drouin-Garraud Marie-Françoise Thuillier-Obstoy Sabine Sigaudy Anne-Marie Frances Patrick Collignon Georges Challe Rémy Couderc Mark Lathrop José-Alain Sahel Jean Weissenbach Christine Petit Françoise Denoyelle 《Orphanet journal of rare diseases》2011,6(1):1-19
Background
Usher syndrome (USH) combines sensorineural deafness with blindness. It is inherited in an autosomal recessive mode. Early diagnosis is critical for adapted educational and patient management choices, and for genetic counseling. To date, nine causative genes have been identified for the three clinical subtypes (USH1, USH2 and USH3). Current diagnostic strategies make use of a genotyping microarray that is based on the previously reported mutations. The purpose of this study was to design a more accurate molecular diagnosis tool.Methods
We sequenced the 366 coding exons and flanking regions of the nine known USH genes, in 54 USH patients (27 USH1, 21 USH2 and 6 USH3).Results
Biallelic mutations were detected in 39 patients (72%) and monoallelic mutations in an additional 10 patients (18.5%). In addition to biallelic mutations in one of the USH genes, presumably pathogenic mutations in another USH gene were detected in seven patients (13%), and another patient carried monoallelic mutations in three different USH genes. Notably, none of the USH3 patients carried detectable mutations in the only known USH3 gene, whereas they all carried mutations in USH2 genes. Most importantly, the currently used microarray would have detected only 30 of the 81 different mutations that we found, of which 39 (48%) were novel.Conclusions
Based on these results, complete exon sequencing of the currently known USH genes stands as a definite improvement for molecular diagnosis of this disease, which is of utmost importance in the perspective of gene therapy. 相似文献47.
Maillard P Walic M Meuleman P Roohvand F Huby T Le Goff W Leroux-Roels G Pécheur EI Budkowska A 《PloS one》2011,6(10):e26637
A distinctive feature of HCV is that its life cycle depends on lipoprotein metabolism. Viral morphogenesis and secretion follow the very low-density lipoprotein (VLDL) biogenesis pathway and, consequently, infectious HCV in the serum is associated with triglyceride-rich lipoproteins (TRL). Lipoprotein lipase (LPL) hydrolyzes TRL within chylomicrons and VLDL but, independently of its catalytic activity, it has a bridging activity, mediating the hepatic uptake of chylomicrons and VLDL remnants. We previously showed that exogenously added LPL increases HCV binding to hepatoma cells by acting as a bridge between virus-associated lipoproteins and cell surface heparan sulfate, while simultaneously decreasing infection levels. We show here that LPL efficiently inhibits cell infection with two HCV strains produced in hepatoma cells or in primary human hepatocytes transplanted into uPA-SCID mice with fully functional human ApoB-lipoprotein profiles. Viruses produced in vitro or in vivo were separated on iodixanol gradients into low and higher density populations, and the infection of Huh 7.5 cells by both virus populations was inhibited by LPL. The effect of LPL depended on its enzymatic activity. However, the lipase inhibitor tetrahydrolipstatin restored only a minor part of HCV infectivity, suggesting an important role of the LPL bridging function in the inhibition of infection. We followed HCV cell entry by immunoelectron microscopy with anti-envelope and anti-core antibodies. These analyses demonstrated the internalization of virus particles into hepatoma cells and their presence in intracellular vesicles and associated with lipid droplets. In the presence of LPL, HCV was retained at the cell surface. We conclude that LPL efficiently inhibits HCV infection by acting on TRL associated with HCV particles through mechanisms involving its lipolytic function, but mostly its bridging function. These mechanisms lead to immobilization of the virus at the cell surface. HCV-associated lipoproteins may therefore be a promising target for the development of new therapeutic approaches. 相似文献
48.
Hillaire ML van Eijk M van Trierum SE van Riel D Saelens X Romijn RA Hemrika W Fouchier RA Kuiken T Osterhaus AD Haagsman HP Rimmelzwaan GF 《PloS one》2011,6(9):e25005
The emergence of influenza viruses resistant to existing classes of antiviral drugs raises concern and there is a need for novel antiviral agents that could be used therapeutically or prophylacticaly. Surfactant protein D (SP-D) belongs to the family of C-type lectins which are important effector molecules of the innate immune system with activity against bacteria and viruses, including influenza viruses. In the present study we evaluated the potential of recombinant porcine SP-D as an antiviral agent against influenza A viruses (IAVs) in vitro. To determine the range of antiviral activity, thirty IAVs of the subtypes H1N1, H3N2 and H5N1 that originated from birds, pigs and humans were selected and tested for their sensitivity to recombinant SP-D. Using these viruses it was shown by hemagglutination inhibition assay, that recombinant porcine SP-D was more potent than recombinant human SP-D and that especially higher order oligomeric forms of SP-D had the strongest antiviral activity. Porcine SP-D was active against a broad range of IAV strains and neutralized a variety of H1N1 and H3N2 IAVs, including 2009 pandemic H1N1 viruses. Using tissue sections of ferret and human trachea, we demonstrated that recombinant porcine SP-D prevented attachment of human seasonal H1N1 and H3N2 virus to receptors on epithelial cells of the upper respiratory tract. It was concluded that recombinant porcine SP-D holds promise as a novel antiviral agent against influenza and further development and evaluation in vivo seems warranted. 相似文献
49.
Bidet M Joubert O Lacombe B Ciantar M Nehmé R Mollat P Brétillon L Faure H Bittman R Ruat M Mus-Veteau I 《PloS one》2011,6(9):e23834
Background
Sonic hedgehog (Shh) signaling plays a crucial role in growth and patterning during embryonic development, and also in stem cell maintenance and tissue regeneration in adults. Aberrant Shh pathway activation is involved in the development of many tumors, and one of the most affected Shh signaling steps found in these tumors is the regulation of the signaling receptor Smoothened by the Shh receptor Patched. In the present work, we investigated Patched activity and the mechanism by which Patched inhibits Smoothened.Methodology/Principal Findings
Using the well-known Shh-responding cell line of mouse fibroblasts NIH 3T3, we first observed that enhancement of the intracellular cholesterol concentration induces Smoothened enrichment in the plasma membrane, which is a crucial step for the signaling activation. We found that binding of Shh protein to its receptor Patched, which involves Patched internalization, increases the intracellular concentration of cholesterol and decreases the efflux of a fluorescent cholesterol derivative (BODIPY-cholesterol) from these cells. Treatment of fibroblasts with cyclopamine, an antagonist of Shh signaling, inhibits Patched expression and reduces BODIPY-cholesterol efflux, while treatment with the Shh pathway agonist SAG enhances Patched protein expression and BODIPY-cholesterol efflux. We also show that over-expression of human Patched in the yeast S. cerevisiae results in a significant boost of BODIPY-cholesterol efflux. Furthermore, we demonstrate that purified Patched binds to cholesterol, and that the interaction of Shh with Patched inhibits the binding of Patched to cholesterol.Conclusion/Significance
Our results suggest that Patched may contribute to cholesterol efflux from cells, and to modulation of the intracellular cholesterol concentration. This activity is likely responsible for the inhibition of the enrichment of Smoothened in the plasma membrane, which is an important step in Shh pathway activation. 相似文献50.
渤海莱州湾表层沉积物中底栖有孔虫分布特征及其环境意义 总被引:2,自引:0,他引:2
对渤海莱州湾海域240个站位表层沉积物中底栖有孔虫群落进行了分析,共鉴定常见的底栖有孔虫42种。结果表明,莱州湾表层沉积物中底栖有孔虫主要以玻璃质壳为主(平均丰度达70.9%),瓷质壳含量次之,胶结壳含量最低;玻璃质壳占有孔虫全群的百分含量,随水深的增加而增加;从黄河口向外海方向,有孔虫分异度和丰度都逐渐增大。该海域底栖有孔虫平面分布的主要控制因素为盐度和底质沉积物类型,大体可分为两个组合分区,I区为Ammonia beccarii-Quinqueloculina spp.组合,代表盐度较低的近岸海陆过渡浅水环境;II区为Cribrononionsub-incertum-Protelphidium tuberculatum组合,代表盐度较高的远岸内陆架环境。 相似文献