全文获取类型
收费全文 | 6074篇 |
免费 | 588篇 |
国内免费 | 3篇 |
专业分类
6665篇 |
出版年
2024年 | 8篇 |
2023年 | 44篇 |
2022年 | 117篇 |
2021年 | 205篇 |
2020年 | 114篇 |
2019年 | 125篇 |
2018年 | 165篇 |
2017年 | 118篇 |
2016年 | 212篇 |
2015年 | 394篇 |
2014年 | 409篇 |
2013年 | 476篇 |
2012年 | 622篇 |
2011年 | 550篇 |
2010年 | 315篇 |
2009年 | 276篇 |
2008年 | 400篇 |
2007年 | 377篇 |
2006年 | 356篇 |
2005年 | 294篇 |
2004年 | 276篇 |
2003年 | 233篇 |
2002年 | 187篇 |
2001年 | 44篇 |
2000年 | 22篇 |
1999年 | 27篇 |
1998年 | 53篇 |
1997年 | 29篇 |
1996年 | 20篇 |
1995年 | 17篇 |
1994年 | 22篇 |
1993年 | 15篇 |
1992年 | 25篇 |
1991年 | 13篇 |
1990年 | 10篇 |
1989年 | 6篇 |
1988年 | 7篇 |
1987年 | 8篇 |
1986年 | 4篇 |
1985年 | 4篇 |
1984年 | 9篇 |
1983年 | 5篇 |
1982年 | 9篇 |
1981年 | 8篇 |
1980年 | 8篇 |
1979年 | 5篇 |
1976年 | 4篇 |
1973年 | 3篇 |
1972年 | 2篇 |
1971年 | 2篇 |
排序方式: 共有6665条查询结果,搜索用时 0 毫秒
81.
82.
83.
Stephanie Lamer Peter R. Jungblut 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2001,752(2)
In theory, peptide mass fingerprinting by matrix assisted laser desorption–ionization mass spectrometry (MALDI-MS) has the potential to identify all of the proteins detected by silver staining on gels. In practice, if the genome of the organism investigated is completely sequenced, using current techniques, all proteins stained by Coomassie Brilliant Blue can be identified. This loss of identification sensitivity of ten to hundred-fold is caused by loss of peptides by surface contacts. Therefore, we performed digestion and transfer of peptides in the lower μl range and reduced the number of steps. The peptide mix obtained from in-gel or on-blot digestion was analyzed directly after digestion or after concentration on POROS R2 beads. Eight protein spots of a 2-DE gel from Mycobacterium bovis BCG were identified using these four preparation procedures for MALDI-MS. Overall, on-blot digestion was as effective as in-gel digestion. Whereas higher signal intensities resulted after concentration, hydrophilic peptides are better detected by direct measurement of the peptide mix without POROS R2 concentration. 相似文献
84.
Eva Maire Joshua Cinner Laure Velez Cindy Huchery Camilo Mora Stephanie Dagata Laurent Vigliola Laurent Wantiez Michel Kulbicki David Mouillot 《Ecology letters》2016,19(4):351-360
The depletion of natural resources has become a major issue in many parts of the world, with the most accessible resources being most at risk. In the terrestrial realm, resource depletion has classically been related to accessibility through road networks. In contrast, in the marine realm, the impact on living resources is often framed into the Malthusian theory of human density around ecosystems. Here, we develop a new framework to estimate the accessibility of global coral reefs using potential travel time from the nearest human settlement or market. We show that 58% of coral reefs are located < 30 min from the nearest human settlement. We use a case study from New Caledonia to demonstrate that travel time from the market is a strong predictor of fish biomass on coral reefs. We also highlight a relative deficit of protection on coral reef areas near people, with disproportional protection on reefs far from people. This suggests that conservation efforts are targeting low‐conflict reefs or places that may already be receiving de facto protection due to their isolation. Our global assessment of accessibility in the marine realm is a critical step to better understand the interplay between humans and resources. 相似文献
85.
Dylan Alexander Carlin Ryan W. Caster Xiaokang Wang Stephanie A. Betzenderfer Claire X. Chen Veasna M. Duong Carolina V. Ryklansky Alp Alpekin Nathan Beaumont Harshul Kapoor Nicole Kim Hosna Mohabbot Boyu Pang Rachel Teel Lillian Whithaus Ilias Tagkopoulos Justin B. Siegel 《PloS one》2016,11(1)
The use of computational modeling algorithms to guide the design of novel enzyme catalysts is a rapidly growing field. Force-field based methods have now been used to engineer both enzyme specificity and activity. However, the proportion of designed mutants with the intended function is often less than ten percent. One potential reason for this is that current force-field based approaches are trained on indirect measures of function rather than direct correlation to experimentally-determined functional effects of mutations. We hypothesize that this is partially due to the lack of data sets for which a large panel of enzyme variants has been produced, purified, and kinetically characterized. Here we report the kcat and KM values of 100 purified mutants of a glycoside hydrolase enzyme. We demonstrate the utility of this data set by using machine learning to train a new algorithm that enables prediction of each kinetic parameter based on readily-modeled structural features. The generated dataset and analyses carried out in this study not only provide insight into how this enzyme functions, they also provide a clear path forward for the improvement of computational enzyme redesign algorithms. 相似文献
86.
Background
Children with antisocial behaviour show deficits in the perception of emotional expressions in others that may contribute to the development and persistence of antisocial and aggressive behaviour. Current treatments for antisocial youngsters are limited in effectiveness. It has been argued that more attention should be devoted to interventions that target neuropsychological correlates of antisocial behaviour. This study examined the effect of emotion recognition training on criminal behaviour.Methods
Emotion recognition and crime levels were studied in 50 juvenile offenders. Whilst all young offenders received their statutory interventions as the study was conducted, a subgroup of twenty-four offenders also took part in a facial affect training aimed at improving emotion recognition. Offenders in the training and control groups were matched for age, SES, IQ and lifetime crime level. All offenders were tested twice for emotion recognition performance, and recent crime data were collected after the testing had been completed.Results
Before the training there were no differences between the groups in emotion recognition, with both groups displaying poor fear, sadness and anger recognition. After the training fear, sadness and anger recognition improved significantly in juvenile offenders in the training group. Although crime rates dropped in all offenders in the 6 months following emotion testing, only the group of offenders who had received the emotion training showed a significant reduction in the severity of the crimes they committed.Conclusions
The study indicates that emotion recognition can be relatively easily improved in youths who engage in serious antisocial and criminal behavior. The results suggest that improved emotion recognition has the potential to reduce the severity of reoffending. 相似文献87.
Abhishek Chatterjee Celia Caballero-Franco Dannika Bakker Stephanie Totten Armando Jardim 《The Journal of biological chemistry》2015,290(42):25579-25594
Enterohemorrhagic Escherichia coli is a causative agent of gastrointestinal and diarrheal diseases. Pathogenesis associated with enterohemorrhagic E. coli involves direct delivery of virulence factors from the bacteria into epithelial cell cytosol via a syringe-like organelle known as the type III secretion system. The type III secretion system protein EspD is a critical factor required for formation of a translocation pore on the host cell membrane. Here, we show that recombinant EspD spontaneously integrates into large unilamellar vesicle (LUV) lipid bilayers; however, pore formation required incorporation of anionic phospholipids such as phosphatidylserine and an acidic pH. Leakage assays performed with fluorescent dextrans confirmed that EspD formed a structure with an inner diameter of ∼2.5 nm. Protease mapping indicated that the two transmembrane helical hairpin of EspD penetrated the lipid layer positioning the N- and C-terminal domains on the extralumenal surface of LUVs. Finally, a combination of glutaraldehyde cross-linking and rate zonal centrifugation suggested that EspD in LUV membranes forms an ∼280–320-kDa oligomeric structure consisting of ∼6–7 subunits. 相似文献
88.
Wilhelm Salzberger Wilfredo F. Garcia-Beltran Haley Dugan Supreetha Gubbala Camille Simoneau Simon B. Gressens Stephanie Jost Marcus Altfeld 《PloS one》2015,10(12)
Viral infections can affect the glycosylation pattern of glycoproteins involved in antiviral immunity. Given the importance of protein glycosylation for immune function, we investigated the effect that modulation of the highly conserved HLA class I N-glycan has on KIR:HLA interactions and NK cell function. We focused on HLA-B*57:01 and its interaction with KIR3DL1, which has been shown to play a critical role in determining the progression of a number of human diseases, including human immunodeficiency virus-1 infection. 721.221 cells stably expressing HLA-B*57:01 were treated with a panel of glycosylation enzyme inhibitors, and HLA class I expression and KIR3DL1 binding was quantified. In addition, the functional outcomes of HLA-B*57:01 N-glycan disruption/modulation on KIR3DL1ζ+ Jurkat reporter cells and primary human KIR3DL1+ NK cells was assessed. Different glycosylation enzyme inhibitors had varying effects on HLA-B*57:01 expression and KIR3DL1-Fc binding. The most remarkable effect was that of tunicamycin, an inhibitor of the first step of N-glycosylation, which resulted in significantly reduced KIR3DL1-Fc binding despite sustained expression of HLA-B*57:01 on 721.221 cells. This effect was paralleled by decreased activation of KIR3DL1ζ+ Jurkat reporter cells, as well as increased degranulation of primary human KIR3DL1+ NK cell clones when encountering HLA-B*57:01-expressing 721.221 cells that were pre-treated with tunicamycin. Overall, these results demonstrate that N-glycosylation of HLA class I is important for KIR:HLA binding and has an impact on NK cell function. 相似文献
89.
Stephanie J. Peacock Andrew W. Bateman Martin Krkošek Mark A. Lewis 《Theoretical Ecology》2016,9(3):365-380
The dynamics of coupled populations have mostly been studied in the context of metapopulation viability with application to, for example, species at risk. However, when considering pests and pathogens, eradication, not persistence, is often the end goal. Humans may intervene to control nuisance populations, resulting in reciprocal interactions between the human and natural systems that can lead to unexpected dynamics. The incidence of these human-natural couplings has been increasing, hastening the need to better understand the emergent properties of such systems in order to predict and manage outbreaks of pests and pathogens. For example, the success of the growing aquaculture industry depends on our ability to manage pathogens and maintain a healthy environment for farmed and wild fish. We developed a model for the dynamics of connected populations subject to control, motivated by sea louse parasites that can disperse among salmon farms. The model includes exponential population growth with a forced decline when populations reach a threshold, representing control interventions. Coupling two populations with equal growth rates resulted in phase locking or synchrony in their dynamics. Populations with different growth rates had different periods of oscillation, leading to quasiperiodic dynamics when coupled. Adding small amounts of stochasticity destabilized quasiperiodic cycles to chaos, while stochasticity was damped for periodic or stable dynamics. Our analysis suggests that strict treatment thresholds, although well intended, can complicate parasite dynamics and hinder control efforts. Synchronizing populations via coordinated management among farms leads to more effective control that is required less frequently. Our model is simple and generally applicable to other systems where dispersal affects the management of pests and pathogens. 相似文献
90.
Pieter Steenhuis Stephanie Herder Suyin Gelis Thomas Braulke Stephan Storch 《Traffic (Copenhagen, Denmark)》2010,11(7):987-1000
CLN7 is a polytopic lysosomal membrane protein deficient in variant late infantile neuronal ceroid lipofuscinosis, a neurodegenerative lysosomal storage disorder. In this study fluorescence protease protection assays and mutational analyses revealed the N‐ and C‐terminal tails of CLN7 in the cytosol and two N‐glycosylation sites at N371 and N376. Both partially and non‐glycosylated CLN7 were correctly transported to lysosomes. To identify lysosomal targeting motifs, we generated CD4‐chimera fused to the N‐ and C‐terminal domains of CLN7. Lysosomal localization of the chimeric proteins requires a consensus acidic dileucine‐based motif in the N‐terminus and two tandem tyrosine‐based signals in the C‐terminus. Mutation of these sorting motifs resulted in cell surface redistribution of CD4 chimeras. However, the dileucine‐based motif is of critical importance for lysosomal localization of the full‐length CLN7 in different cell lines. Cell surface biotinylation revealed that at equilibrium 22% of total CLN7 is localized at the plasma membrane. Mutation of the dileucine motif or the co‐expression of dominant‐negative mutant dynamin K44A led to a further increase of CLN7 at the plasma membrane. Our data demonstrate that CLN7 contains several cytoplasmic lysosomal targeting signals of which the N‐terminal dileucine‐based motif is required for the predominant lysosomal targeting along the indirect pathway and clathrin‐mediated endocytosis of CLN7. 相似文献