全文获取类型
收费全文 | 528篇 |
免费 | 61篇 |
国内免费 | 1篇 |
专业分类
590篇 |
出版年
2022年 | 5篇 |
2021年 | 10篇 |
2020年 | 4篇 |
2019年 | 8篇 |
2018年 | 12篇 |
2017年 | 9篇 |
2016年 | 19篇 |
2015年 | 30篇 |
2014年 | 20篇 |
2013年 | 24篇 |
2012年 | 45篇 |
2011年 | 43篇 |
2010年 | 13篇 |
2009年 | 23篇 |
2008年 | 24篇 |
2007年 | 20篇 |
2006年 | 31篇 |
2005年 | 19篇 |
2004年 | 10篇 |
2003年 | 12篇 |
2002年 | 8篇 |
2001年 | 14篇 |
2000年 | 15篇 |
1999年 | 23篇 |
1998年 | 3篇 |
1997年 | 7篇 |
1996年 | 6篇 |
1995年 | 9篇 |
1994年 | 6篇 |
1993年 | 4篇 |
1992年 | 8篇 |
1991年 | 13篇 |
1990年 | 8篇 |
1989年 | 9篇 |
1988年 | 7篇 |
1987年 | 3篇 |
1986年 | 3篇 |
1985年 | 6篇 |
1984年 | 6篇 |
1983年 | 7篇 |
1982年 | 4篇 |
1981年 | 4篇 |
1979年 | 5篇 |
1978年 | 5篇 |
1977年 | 3篇 |
1975年 | 4篇 |
1973年 | 2篇 |
1969年 | 2篇 |
1968年 | 2篇 |
1957年 | 2篇 |
排序方式: 共有590条查询结果,搜索用时 0 毫秒
21.
3' untranslated regions of alfamo- and ilar-virus RNAs fold into a series of stem-loop structures to which the coat protein binds with high affinity. This binding plays a role in initiation of infection ('genome activation') and has been thought to substitute for a tRNA-like structure that is found at the 3' termini of related plant viruses. We propose the existence of an alternative conformation of the 3' ends of alfamo- and ilar-virus RNAs, including a pseudoknot. Based on (i) phylogenetic comparisons, (ii) in vivo and in vitro functional analyses of mutants in which the pseudoknot has been disrupted or restored by compensatory mutations, (iii) competition experiments between coat protein and viral replicase, and (iv) investigation of the effect of magnesium, we demonstrate that this pseudoknot is required for replication of alfalfa mosaic virus. This conformation resembles the tRNA-like structure of the related bromo- and cucumo-viruses. A low but specific interaction with yeast CCA-adding enzyme was found. The existence of two mutually exclusive conformations for the 3' termini of alfamo- and ilar-virus RNAs could enable the virus to switch from translation to replication and vice versa. The role of coat protein in this modulation and in genome activation is discussed. 相似文献
22.
23.
24.
Quan-Xiang Wei Rainer Claus Thomas Hielscher Daniel Mertens Aparna Raval Christopher C. Oakes Stephan M. Tanner Albert de la Chapelle John C. Byrd Stephan Stilgenbauer Christoph Plass 《PloS one》2013,8(1)
We previously reported a rare germline variant (c.1-6531) that resulted in allele–specific expression (ASE) of death-associated protein kinase 1 (DAPK1) and predisposition to chronic lymphocytic leukemia (CLL). We investigated a cohort of CLL patients lacking this mutation for the presence of ASE of DAPK1. We developed a novel strategy that combines single-nucleotide primer extension (SNuPE) with MALDI-TOF mass spectrometry, and detected germline DAPK1 ASE in 17 out of 120 (14.2%) CLL patients associated with a trend towards younger age at diagnosis. ASE was absent in 63 healthy controls. Germline cells of CLL patients with ASE showed increased levels of DNA methylation in the promoter region, however, neither genetic nor further epigenetic aberrations could be identified in the DAPK1 5′ upstream regulatory region, within distinct exons or in the 3′-UTR. We identified B-lymphoid malignancy related cell line models harboring allelic imbalance and found that allele-specific methylation in DAPK1 is associated with ASE. Our data indicate that ASE at the DAPK1 gene locus is a recurrent event, mediated by epigenetic mechanisms and potentially predisposing to CLL. 相似文献
25.
Eva Veronesi Frank Antony Simon Gubbins Nick Golding Alison Blackwell Peter PC. Mertens Joe Brownlie Karin E. Darpel Philip S. Mellor Simon Carpenter 《PloS one》2013,8(8)
Background
Culicoides biting midges (Diptera: Ceratopogonidae) are the biological vectors of globally significant arboviruses of livestock including bluetongue virus (BTV), African horse sickness virus (AHSV) and the recently emerging Schmallenberg virus (SBV). From 2006–2009 outbreaks of BTV in northern Europe inflicted major disruption and economic losses to farmers and several attempts were made to implicate Palaearctic Culicoides species as vectors. Results from these studies were difficult to interpret as they used semi-quantitative RT-PCR (sqPCR) assays as the major diagnostic tool, a technique that had not been validated for use in this role. In this study we validate the use of these assays by carrying out time-series detection of BTV RNA in two colony species of Culicoides and compare the results with the more traditional isolation of infectious BTV on cell culture.Methodology/Principal Findings
A BTV serotype 1 strain mixed with horse blood was fed to several hundred individuals of Culicoides sonorensis (Wirth & Jones) and C. nubeculosus (Mg.) using a membrane-based assay and replete individuals were then incubated at 25°C. At daily intervals 25 Culicoides of each species were removed from incubation, homogenised and BTV quantified in each individual using sqPCR (Cq values) and virus isolation on a KC-C. sonorensis embryonic cell line, followed by antigen enzyme-linked immunosorbent assay (ELISA). In addition, comparisons were also drawn between the results obtained with whole C. sonorensis and with individually dissected individuals to determine the level of BTV dissemination.Conclusions/Significance
Cq values generated from time-series infection experiments in both C. sonorensis and C. nubeculosus confirmed previous studies that relied upon the isolation and detection of infectious BTV. Implications on the testing of field-collected Culicoides as potential virus vectors by PCR assays and the use of such assays as front-line tools for use in diagnostic laboratories in this role are discussed. 相似文献26.
Andrew E. Shaw Maxime Ratinier Sandro Filipe Nunes Kyriaki Nomikou Marco Caporale Matthew Golder Kathryn Allan Claude Hamers Pascal Hudelet Stéphan Zientara Emmanuel Breard Peter Mertens Massimo Palmarini 《Journal of virology》2013,87(1):543-557
Coinfection of a cell by two different strains of a segmented virus can give rise to a “reassortant” with phenotypic characteristics that might differ from those of the parental strains. Bluetongue virus (BTV) is a double-stranded RNA (dsRNA) segmented virus and the cause of bluetongue, a major infectious disease of livestock. BTV exists as at least 26 different serotypes (BTV-1 to BTV-26). Prompted by the isolation of a field reassortant between BTV-1 and BTV-8, we systematically characterized the process of BTV reassortment. Using a reverse genetics approach, our study clearly indicates that any BTV-1 or BTV-8 genome segment can be rescued in the heterologous “backbone.” To assess phenotypic variation as a result of reassortment, we examined viral growth kinetics and plaque sizes in in vitro experiments and virulence in an experimental mouse model of bluetongue disease. The monoreassortants generated had phenotypes that were very similar to those of the parental wild-type strains both in vitro and in vivo. Using a forward genetics approach in cells coinfected with BTV-1 and BTV-8, we have shown that reassortants between BTV-1 and BTV-8 are generated very readily. After only four passages in cell culture, we could not detect wild-type BTV-1 or BTV-8 in any of 140 isolated viral plaques. In addition, most of the isolated reassortants contained heterologous VP2 and VP5 structural proteins, while only 17% had homologous VP2 and VP5 proteins. Our study has shown that reassortment in BTV is very flexible, and there is no fundamental barrier to the reassortment of any genome segment. Given the propensity of BTV to reassort, it is increasingly important to have an alternative classification system for orbiviruses. 相似文献
27.
PR Marcelino MS Aoki AFS Arruda CG Freitas A Mendez-Villanueva A Moreira 《Biology of sport / Institute of Sport》2016,33(1):37-42
The purpose of this study was to investigate the effect of court size on physiological responses and physical performance of young elite basketball players. Twelve male basketball players (18.6 ± 0.5 years; 88.8 ± 14.5 kg; 192.6 ± 6.5 cm) from an under-19 team performed two small-sided games (matches) with different court areas (28x15 m and 28x9 m; 28x15 and 28x9 protocols). The number of players (3x3) was kept the same in each protocol. The players performed a repeated-sprint ability (RSA) test before and after each match. Blood lactate concentration was collected before (pre) and after (post) the matches, and the session rating of perceived exertion (session-RPE) was determined 30 minutes after the match. Best and mean time in the RSA test were not different between the 28x15 and the 28x9 match protocols (p > 0.05). A significant difference was observed for lactate concentration from pre- to post-match (p < 0.05) in both protocols (28x15 and 28x9); however, there was no significant interaction between protocols. A similar session-RPE mean score (28x15: 7.2 ± 1.4 and 28x9: 6.6 ± 1.4) was detected for both protocols (p > 0.05, ES=0.41). In summary, the results of the current study suggest that the different court areas induced similar responses. Although there was no significant difference in effort perception, players tended to perceive a greater effort in the larger court size. 相似文献
28.
Tobias Abzieher Somayeh Moghadamzadeh Fabian Schackmar Helge Eggers Florian Sutterlüti Amjad Farooq Danny Kojda Klaus Habicht Raphael Schmager Adrian Mertens Raheleh Azmi Lukas Klohr Jonas A. Schwenzer Michael Hetterich Uli Lemmer Bryce S. Richards Michael Powalla Ulrich W. Paetzold 《Liver Transplantation》2019,9(12)
29.
Recognizing cross‐ecosystem responses to changing temperatures: soil warming impacts pelagic food webs 下载免费PDF全文
Samuel B. Fey Andrew N. Mertens Lucas J. Beversdorf Katherine D. McMahon Kathryn L. Cottingham 《Oikos》2015,124(11):1473-1481
The energy and materials that move across ecosystem boundaries influence food web structure and key ecosystem functions. Despite the acknowledged importance of such ecological subsidies, surprisingly little information is available regarding the role of environmental temperature in influencing subsidy quality and the response of the recipient ecosystem. We evaluated the impacts of temperature‐mediated changes in leaves from deciduous trees, an important subsidy from terrestrial to freshwater ecosystems, on both the producer‐based and detritivore‐based components of a pelagic pond food web in a field mesocosm experiment. We hypothesized that variation in leaf chemistry driven by increased soil temperature would alter both the quality of leaf subsidies and the pond response. We collected red maple Acer rubrum leaves from heated and ambient temperature plots from the long‐term soil warming experiment at the Harvard Experimental Forest and added them to 167‐l field mesocosms containing established plankton communities, creating ‘no leaf’, ‘ambient leaf’ and ‘heated leaf’ treatments during autumn 2012. We then monitored physical, chemical, and biological responses to treatments until the mesocosms froze six weeks later. Experimental soil warming altered the chemical composition of deciduous leaves, the physical and chemical environment of the aquatic ecosystems to which leaves were added, and the pelagic pond food webs as measured by community composition. Compared to leaves from ambient‐temperature soils, leaves from warmed soils initially resulted in lower water column phosphorus and dissolved organic carbon, reducing bacterial densities. However, the diminished carbon and phosphorus resulting from soil warming also increased light availability that ultimately stimulated cladoceran zooplankton relative to ambient‐temperature leaves. Our results suggest that changes in temperature can alter ecological subsidies in unanticipated ways, and suggest that accurately predicting the potential consequences of climate change will require conducting research across ecosystem boundaries. 相似文献
30.
Pierre Soubeyran Carine Bellera Jean Goyard Damien Heitz Hervé Curé Hubert Rousselot Gilles Albrand Véronique Servent Olivier Saint Jean Isabelle van Praagh Jean-Emmanuel Kurtz Stéphane Périn Jean-Luc Verhaeghe Catherine Terret Christophe Desauw Véronique Girre Cécile Mertens Simone Mathoulin-Pélissier Muriel Rainfray 《PloS one》2014,9(12)