全文获取类型
收费全文 | 88603篇 |
免费 | 6036篇 |
国内免费 | 16篇 |
专业分类
94655篇 |
出版年
2023年 | 577篇 |
2022年 | 756篇 |
2021年 | 1457篇 |
2020年 | 1195篇 |
2019年 | 1365篇 |
2018年 | 2493篇 |
2017年 | 2290篇 |
2016年 | 3018篇 |
2015年 | 3976篇 |
2014年 | 4169篇 |
2013年 | 5515篇 |
2012年 | 6505篇 |
2011年 | 5966篇 |
2010年 | 3862篇 |
2009年 | 3247篇 |
2008年 | 4753篇 |
2007年 | 4568篇 |
2006年 | 4231篇 |
2005年 | 3623篇 |
2004年 | 3605篇 |
2003年 | 3225篇 |
2002年 | 2967篇 |
2001年 | 2114篇 |
2000年 | 2057篇 |
1999年 | 1645篇 |
1998年 | 838篇 |
1997年 | 623篇 |
1996年 | 614篇 |
1995年 | 576篇 |
1994年 | 487篇 |
1993年 | 453篇 |
1992年 | 883篇 |
1991年 | 796篇 |
1990年 | 723篇 |
1989年 | 704篇 |
1988年 | 655篇 |
1987年 | 623篇 |
1986年 | 564篇 |
1985年 | 554篇 |
1984年 | 508篇 |
1983年 | 407篇 |
1982年 | 350篇 |
1981年 | 274篇 |
1980年 | 273篇 |
1979年 | 359篇 |
1978年 | 321篇 |
1975年 | 315篇 |
1974年 | 351篇 |
1973年 | 353篇 |
1972年 | 295篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
991.
Anoek Friskes Lisa Koob Lenno Krenning Tesa M Severson Emma
S Koeleman Xabier Vergara Michael Schubert Jeroen van
den
Berg Bastiaan Evers Anna G Manjn Stacey Joosten Yongsoo Kim Wilbert Zwart Ren
H Medema 《Nucleic acids research》2022,50(17):9930
Cells respond to double-strand breaks (DSBs) by activating DNA damage response pathways, including cell cycle arrest. We have previously shown that a single double-strand break generated via CRISPR/Cas9 is sufficient to delay cell cycle progression and compromise cell viability. However, we also found that the cellular response to DSBs can vary, independent of the number of lesions. This implies that not all DSBs are equally toxic, and raises the question if the location of a single double-strand break could influence its toxicity. To systematically investigate if DSB-location is a determinant of toxicity we performed a CRISPR/Cas9 screen targeting 6237 single sites in the human genome. Next, we developed a data-driven framework to design CRISPR/Cas9 sgRNA (crRNA) pools targeting specific chromatin features. The chromatin context was defined using ChromHMM states, Lamin-B1 DAM-iD, DNAseI hypersensitivity, and RNA-sequencing data. We computationally designed 6 distinct crRNA pools, each containing 10 crRNAs targeting the same chromatin state. We show that the toxicity of a DSB is highly similar across the different ChromHMM states. Rather, we find that the major determinants of toxicity of a sgRNA are cutting efficiency and off-target effects. Thus, chromatin features have little to no effect on the toxicity of a single CRISPR/Cas9-induced DSB. 相似文献
992.
Ramiro S. Arrieta Darío A. Lijtmaer Pablo L. Tubaro 《Biological journal of the Linnean Society. Linnean Society of London》2013,110(3):528-542
The process of speciation is a crucial aspect of evolutionary biology. In this study, we analysed the patterns of evolution of postzygotic reproductive isolation in Galliformes using information on hybridization and genetic distance among species. Four main patterns arose: (1) hybrid inviability and sterility in F1 hybrids increase as species diverge; (2) the presence of geographical overlap does not affect the evolution of postzygotic isolation; (3) the galliforms follow Haldane's rule; (4) hybrid inviability is higher in F2 than in F1 hybrids, but does not appear to be increased in the backcrosses. This study contributes to the growing evidence suggesting that the patterns of evolution of postzygotic isolation and the process of speciation are shared among avian groups (and animals in general). In particular, our results support the notion of F2 hybrid inviability as being key for the maintenance of species genetic integrity when prezygotic isolation barriers are overcome in closely related species, in which postzygotic isolation in the F1 hybrid might still not be fully developed. To the contrary, hybrids from backcrosses did not show serious inviability problems (at least not more than F1 hybrids), demonstrating that they could generate gene flow among bird species. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 528–542. 相似文献
993.
994.
Common bean (Phaseolus vulgaris) has become a cosmopolitan crop, but was originally domesticated in the Americas and has been grown in Latin America for several thousand years. Consequently an enormous diversity of bean nodulating bacteria have developed and in the centers of origin the predominant species in bean nodules is R. etli. In some areas of Latin America, inoculation, which normally promotes nodulation and nitrogen fixation is hampered by the prevalence of native strains. Many other species in addition to R. etli have been found in bean nodules in regions where bean has been introduced. Some of these species such as R. leguminosarum bv. phaseoli, R. gallicum bv. phaseoli and R. giardinii bv. phaseoli might have arisen by acquiring the phaseoli plasmid from R. etli. Others, like R. tropici, are well adapted to acid soils and high temperatures and are good inoculants for bean under these conditions. The large number of rhizobia species capable of nodulating bean supports that bean is a promiscuous host and a diversity of bean-rhizobia interactions exists. Large ranges of dinitrogen fixing capabilities have been documented among bean cultivars and commercial beans have the lowest values among legume crops. Knowledge on bean symbiosis is still incipient but could help to improve bean biological nitrogen fixation. 相似文献
995.
996.
Summary From a callus culture which was started from explants of a single plant of Crepis capillaris, a line was isolated which could grow on auxin- and kinetin-free medium (in contrast to the other lines of the culture). The calluses of this habituated line grew on hormone-free medium as teratomas. Regenerated plants were obtained from the habituated line and from other lines of the culture as control. In the secondary callus cultures which originated from these plants, the subcultures belonging to the initially transformed line no longer had the ability to grow on hormone-free medium. Since this line was also a chromosome mutant and the same mutation was present in its regenerated plants and in the secondary culture line of them, the loss of the auxin-autotrophy cannot be explained in this case on the basis of a selection of cells with karyotypes different from those of the primary culture. 相似文献
997.
Szerman N Schroh I Rossi AL Rosso AM Krymkiewicz N Ferrarotti SA 《Bioresource technology》2007,98(15):2886-2891
Cyclodextrins (CD) are cyclic oligosaccharides with multiple applications in the food, pharmaceutical, cosmetic, agricultural and chemical industries. In this work, the conditions used to produce CD with cyclodextrin glycosyltransferase from Bacillus circulans DF 9R were optimized using experimental designs. The developed method allowed the partial purification and concentration of the enzyme from the cultural broth and, subsequently, the CD production, using the same cassava starch as enzyme adsorbent and as substrate. Heat-treatment of raw starch at 70 degrees C for 15 min in the presence of adsorbed cyclodextrin glycosyltransferase allowed the starch liquefaction without enzyme inactivation. The optimum conditions for CD production were: 5% (w/v) cassava starch, 15 U of enzyme per gram of substrate, reaction temperature of 56 degrees C and pH 6.4. After 4h, the proportion of starch converted to CD reached 66% (w/w) and the weight ratio of alpha-CD:beta-CD:gamma-CD was 1.00:0.70:0.16. 相似文献
998.
During the G1/S transition, p21 proteolysis is mediated by Skp2; however, p21 reaccumulates in G2 and is degraded again in prometaphase. How p21 degradation is controlled in mitosis remains unexplored. We found that Cdc20 (an activator of the ubiquitin ligase APC/C) binds p21 in cultured cells and identified a D box motif in p21 necessary for APC/C(Cdc20)-mediated ubiquitylation of p21. Overexpression of Cdc20 or Skp2 destabilized wild-type p21; however, only Skp2, but not Cdc20, was able to destabilize a p21(D box) mutant. Silencing of Cdc20 induced an accumulation of p21, increased the fraction of p21 bound to Cdk1, and inhibited Cdk1 activity in p21(+/+) prometaphase cells, but not in p21(-/-) cells. Thus, in prometaphase Cdc20 positively regulates Cdk1 by mediating the degradation of p21. We propose that the APC/C(Cdc20)-mediated degradation of p21 contributes to the full activation of Cdk1 necessary for mitotic events and prevents mitotic slippage during spindle checkpoint activation. 相似文献
999.
Rendal Vázquez ME Díaz Román TM Rodríguez Cabarcos M Zavanella Botta C Domenech García N González Cuesta M Sánchez Dopico MJ Pértega Díaz S Andión Núñez C 《Cell and tissue banking》2008,9(2):101-107
To analyse the influence of cold ischemic time (CIT) (2–24 h) and of cryopreservation (liquid phase) on the viability of the
valvular fibroblasts and in the presence of apoptosis. Cardiac valves from 10 pigs were evaluated by anatomo-pathological
study of the wall, muscle and leaflet. At the same time, the presence of cellular death due to apoptosis was investigated
in two ways; directly on tissue by Apodetec system and by two-colour flow cytometry assay analyzing a suspension of fibroblast
from valve leaflets using Anexina V and propidium iodure (PI). We established three groups of samples to compare different
experimental conditions: 2 h of ischemia (group 1), 24 h of ischemia (group 2), and a programme of cryopreservation (−1°C/min)
after 2 h of ischemia, followed by storage in liquid nitrogen during a week and thawing was performed (group 3). The analysis
of viabilities showed slight differences between all three groups. The results indicated CIT of 24 h undergoing more structural
affectation than CIT of 2 h. Flow cytometry analysis did not show important differences between groups; however cryopreserved
samples (group 3) slightly less viability and a higher percentage of death by apoptosis than group 1 and 2 using flow cytometry.
Apoptosis was confirmed on tissue from all valves but mainly in samples of group 2 and group 3. In summary, the viability
of the valves in the case of ischemic times of 2 h, 24 h or after cryopreservation/thawing differs slightly. The death of
the cells is mainly mediated by necrosis and not by apoptosis. 相似文献
1000.
Jean-François Fournier Yushma Bhurruth-Alcor Branislav Musicki Jérome Aubert Michèle Aurelly Claire Bouix-Peter Karinne Bouquet Laurent Chantalat Marion Delorme Bénédicte Drean Gwenaelle Duvert Nicolas Fleury-Bregeot Blanche Gauthier Karine Grisendi Craig S. Harris Laurent F. Hennequin Tatiana Isabet Florence Joly Loïc Tomas 《Bioorganic & medicinal chemistry letters》2018,28(17):2985-2992
A series of squaramide-based hydroxamic acids were designed, synthesized and evaluated against human HDAC enzyme. Squaramides were found to be potent in the Hut78 cell line, but initially suffered from low solubility. Leads with improved solubility and metabolic profiles were shown to be class I, IIB and IV selective. 相似文献