全文获取类型
收费全文 | 14630篇 |
免费 | 1584篇 |
国内免费 | 2篇 |
出版年
2022年 | 81篇 |
2021年 | 169篇 |
2020年 | 119篇 |
2019年 | 159篇 |
2018年 | 168篇 |
2017年 | 140篇 |
2016年 | 301篇 |
2015年 | 568篇 |
2014年 | 538篇 |
2013年 | 715篇 |
2012年 | 823篇 |
2011年 | 735篇 |
2010年 | 528篇 |
2009年 | 506篇 |
2008年 | 633篇 |
2007年 | 636篇 |
2006年 | 571篇 |
2005年 | 543篇 |
2004年 | 552篇 |
2003年 | 501篇 |
2002年 | 501篇 |
2001年 | 463篇 |
2000年 | 407篇 |
1999年 | 367篇 |
1998年 | 217篇 |
1997年 | 205篇 |
1996年 | 187篇 |
1995年 | 203篇 |
1994年 | 190篇 |
1993年 | 209篇 |
1992年 | 435篇 |
1991年 | 321篇 |
1990年 | 330篇 |
1989年 | 309篇 |
1988年 | 270篇 |
1987年 | 224篇 |
1986年 | 230篇 |
1985年 | 238篇 |
1984年 | 192篇 |
1983年 | 166篇 |
1982年 | 123篇 |
1981年 | 98篇 |
1980年 | 98篇 |
1979年 | 137篇 |
1978年 | 102篇 |
1977年 | 90篇 |
1976年 | 85篇 |
1975年 | 77篇 |
1974年 | 107篇 |
1973年 | 95篇 |
排序方式: 共有10000条查询结果,搜索用时 31 毫秒
991.
Accurate and early detection of coronary artery disease (CAD) is essential, as the disease remains one of the leading causes of death in the industrialised world. For this purpose, several invasive as well as noninvasive modalities are available. The current gold standard to detect significant narrowing of the coronary arteries is invasive coronary angiography, which allows direct visualisation of the coronary arteries with high spatial and temporal resolution. In addition, if abnormalities are demonstrated, direct intervention is possible. However, it is also an invasive technique that is associated with substantial patient discomfort, costs and a small but distinct risk of potentially life-threatening complications. 相似文献
992.
Ahmed EA van der Vaart A Barten A Kal HB Chen J Lou Z Minter-Dykhouse K Bartkova J Bartek J de Boer P de Rooij DG 《DNA Repair》2007,6(9):1243-1254
In male germ cells the repair of DNA double strand breaks (DSBs) differs from that described for somatic cell lines. Irradiation induced immunofluorescent foci (IRIF's) signifying a double strand DNA breaks, were followed in spermatogenic cells up to 16 h after the insult. Foci were characterised for Mdc1, 53BP1 and Rad51 that always were expressed in conjecture with gamma-H2AX. Subsequent spermatogenic cell types were found to have different repair proteins. In early germ cells up to the start of meiotic prophase, i.e. in spermatogonia and preleptotene spermatocytes, 53BP1 and Rad51 are available but no Mdc1 is expressed in these cells before and after irradiation. The latter might explain the radiosensitivity of spermatogonia. Spermatocytes from shortly after premeiotic S-phase till pachytene in epithelial stage IV/V express Mdc1 and Rad51 but no 53BP1 which has no role in recombination involved repair during the early meiotic prophase. Mdc1 is required during this period as in Mdc1 deficient mice all spermatocytes enter apoptosis in epithelial stage IV when they should start mid-pachytene phase of the meiotic prophase. From stage IV mid pachytene spermatocytes to round spermatids, Mdc1 and 53BP1 are expressed while Rad51 is no longer expressed in the haploid round spermatids. Quantifying foci numbers of gamma-H2AX, Mdc1 and 53BP1 at various time points after irradiation revealed a 70% reduction after 16 h in pachytene and diplotene spermatocytes and round spermatids. Although the DSB repair efficiency is higher then in spermatogonia where only a 40% reduction was found, it still does not compare to somatic cell lines where a 70% reduction occurs in 2 h. Taken together, DNA DSBs repair proteins differ for the various types of spermatogenic cells, no germ cell type possessing the complete set. This likely leads to a compromised efficiency relative to somatic cell lines. From the evolutionary point of view it may be an advantage when germ cells die from DNA damage rather than risk the acquisition of transmittable errors made during the repair process. 相似文献
993.
Profiling phosphoproteins of yeast mitochondria reveals a role of phosphorylation in assembly of the ATP synthase 总被引:1,自引:0,他引:1
Reinders J Wagner K Zahedi RP Stojanovski D Eyrich B van der Laan M Rehling P Sickmann A Pfanner N Meisinger C 《Molecular & cellular proteomics : MCP》2007,6(11):1896-1906
Mitochondria are crucial for numerous cellular processes, yet the regulation of mitochondrial functions is only understood in part. Recent studies indicated that the number of mitochondrial phosphoproteins is higher than expected; however, the effect of reversible phosphorylation on mitochondrial structure and function has only been defined in a few cases. It is thus crucial to determine authentic protein phosphorylation sites from highly purified mitochondria in a genetically tractable organism. The yeast Saccharomyces cerevisiae is a major model organism for the analysis of mitochondrial functions. We isolated highly pure yeast mitochondria and performed a systematic analysis of phosphorylation sites by a combination of different enrichment strategies and mass spectrometry. We identified 80 phosphorylation sites in 48 different proteins. These mitochondrial phosphoproteins are involved in critical mitochondrial functions, including energy metabolism, protein biogenesis, fatty acid metabolism, metabolite transport, and redox regulation. By combining yeast genetics and in vitro biochemical analysis, we found that phosphorylation of a serine residue in subunit g (Atp20) regulates dimerization of the mitochondrial ATP synthase. The authentic phosphoproteome of yeast mitochondria will represent a rich source to uncover novel roles of reversible protein phosphorylation. 相似文献
994.
Like the cyst walls of other protists, the spore coat of Dictyostelium discoideum is formed de novo to protect the enclosed dormant cell from stress. Spore coat assembly is initiated by exocytosis of protein and polysaccharide precursors at the cell surface, followed by the infusion of nascent cellulose fibrils, resulting in an asymmetrical trilaminar sandwich with cellulose filling the middle layer. A molecular complex consisting of cellulose and two proteins, SP85 and SP65, is associated with the inner and middle layers and is required for proper organization of distinct proteins in the outer layer. Here we show that, unlike SP85 and other protein precursors, which are stored in prespore vesicles, SP65 is, like cellulose, synthesized just in time. By tagging the SP65 locus with green fluorescent protein, we find that SP65 is delivered to the cell surface via largely distinct vesicles, suggesting that separate delivery of components of the cellulose-SP85-SP65 complex regulates its formation at the cell surface. In support of previous in vivo studies, recombinant SP65 and SP85 are shown to interact directly. In addition, truncation of SP65 causes a defect of the outer layer permeability barrier as seen previously for SP85 mutants. These observations suggest that assembly of the cellulose-SP85-SP65 triad at the cell surface is biosynthetically regulated both temporally and spatially and that the complex contributes an essential function to outer layer architecture and function. 相似文献
995.
996.
Fina A. S Kurreeman Leonid Padyukov Rute B Marques Steven J Schrodi Maria Seddighzadeh Gerrie Stoeken-Rijsbergen Annette H. M van der Helm-van Mil Cornelia F Allaart Willem Verduyn Jeanine Houwing-Duistermaat Lars Alfredsson Ann B Begovich Lars Klareskog Tom W. J Huizinga Rene E. M Toes 《PLoS medicine》2007,4(12)
997.
Ingrid van der Pluijm George A Garinis Renata M. C Brandt Theo G. M. F Gorgels Susan W Wijnhoven Karin E. M Diderich Jan de Wit James R Mitchell Conny van Oostrom Rudolf Beems Laura J Niedernhofer Susana Velasco Errol C Friedberg Kiyoji Tanaka Harry van Steeg Jan H. J Hoeijmakers Gijsbertus T. J van der Horst 《PLoS biology》2007,5(1)
998.
Henri C van der Heyde James M Burns William P Weidanz John Horn Irene Gramaglia John P Nolan 《Cytometry. Part A》2007,71(4):242-250
BACKGROUND: Measuring antibody production in response to antigen exposure or vaccination is key to disease prevention and treatment. Our understanding of the mechanisms involved in the antibody response is limited by a lack of sensitive analysis methods. We address this limitation using multiplexed microsphere arrays for the semi -quantitative analysis of antibody production in response to malaria infection. METHODS: We used microspheres as solid supports on which to capture and analyze circulating antibodies. Antigen immobilized on beads captured antigen-specific antibodies for semi- quantitative analysis using fluorescent secondary antibodies. Anti-immunoglobulin antibodies on beads captured specific antibody isotypes for affinity estimation using fluorescent antigen. RESULTS: Antigen-mediated capture of plasma antibodies enables determination of antigen-specific antibody "titer," a semi-quantitative parameter describing a convolution of antibody abundance and avidity, as well as parameters describing numbers of antibodies bound/bead at saturation and the plasma concentration-dependent approach to saturation. Results were identical in single-plex and multiplex assays, and in qualitative agreement with similar parameters derived from ELISA-based assays. Isotype-specific antibody-mediated capture of plasma antibodies allowed the estimation of the affinity of antibody for antigen. CONCLUSION: Analysis of antibody responses using microspheres and flow cytometry offer significant advantages in speed, sample size, and quantification over standard ELISA-based titer methods. 相似文献
999.
Blodgett JA Thomas PM Li G Velasquez JE van der Donk WA Kelleher NL Metcalf WW 《Nature chemical biology》2007,3(8):480-485
Phosphinothricin tripeptide (PTT, phosphinothricylalanylalanine) is a natural-product antibiotic and potent herbicide that is produced by Streptomyces hygroscopicus ATCC 21705 (ref. 1) and Streptomyces viridochromogenes DSM 40736 (ref. 2). PTT has attracted widespread interest because of its commercial applications and unique phosphinic acid functional group. Despite intensive study since its discovery in 1972 (see ref. 3 for a comprehensive review), a number of steps early in the PTT biosynthetic pathway remain uncharacterized. Here we report a series of interdisciplinary experiments involving the construction of defined S. viridochromogenes mutants, chemical characterization of accumulated intermediates, and in vitro assay of selected enzymes to examine these critical steps in PTT biosynthesis. Our results indicate that early PTT biosynthesis involves a series of catalytic steps that to our knowledge has not been described so far, including a highly unusual reaction for carbon bond cleavage. In sum, we define a pathway for early PTT biosynthesis that is more complex than previously appreciated. 相似文献
1000.
Yang XX Hawle P Bebelman JP Meenhuis A Siderius M van der Vies SM 《FEMS yeast research》2007,7(6):796-807
Cdc37p, the p50 homolog of Saccharomyces cerevisiae, is an Hsp90 cochaperone involved in the targeting of protein kinases to Hsp90. Here we report a role for Cdc37p in osmoadaptive signalling in this yeast. The osmosensitive phenotype that is displayed by the cdc37-34 mutant strain appears not to be the consequence of deficient signalling through the high osmolarity glycerol (HOG) MAP kinase pathway. Rather, Cdc37p appears to play a role in the filamentous growth (FG) pathway, which mediates adaptation to high osmolarity parallel to the HOG pathway. The osmosensitive phenotype of the cdc37-34 mutant strain is aggravated upon the deletion of the HOG gene. We report that the hyper-osmosensitive phenotype of the cdc37-34, hog1 mutant correlates to a reduced of activity of the FG pathway. We utilized this phenotype to isolate suppressor genes such as KSS1 that encodes a MAP kinase that functions in the FG pathway. We report that Kss1p interacts physically with Cdc37p. Like Kss1p, the second suppressor that we isolated, Dse1p, is involved in cell wall biogenesis or maintenance, suggesting that Cdc37p controls osmoadapation by regulating mitogen-activated protein kinase signalling aimed at adaptive changes in cell wall organization. 相似文献