全文获取类型
收费全文 | 3295篇 |
免费 | 260篇 |
国内免费 | 2篇 |
专业分类
3557篇 |
出版年
2024年 | 3篇 |
2023年 | 18篇 |
2022年 | 40篇 |
2021年 | 61篇 |
2020年 | 38篇 |
2019年 | 41篇 |
2018年 | 51篇 |
2017年 | 48篇 |
2016年 | 87篇 |
2015年 | 145篇 |
2014年 | 162篇 |
2013年 | 217篇 |
2012年 | 247篇 |
2011年 | 261篇 |
2010年 | 181篇 |
2009年 | 177篇 |
2008年 | 241篇 |
2007年 | 231篇 |
2006年 | 200篇 |
2005年 | 199篇 |
2004年 | 203篇 |
2003年 | 165篇 |
2002年 | 177篇 |
2001年 | 54篇 |
2000年 | 31篇 |
1999年 | 33篇 |
1998年 | 35篇 |
1997年 | 27篇 |
1996年 | 18篇 |
1995年 | 23篇 |
1994年 | 11篇 |
1993年 | 14篇 |
1992年 | 7篇 |
1991年 | 10篇 |
1990年 | 9篇 |
1989年 | 6篇 |
1988年 | 9篇 |
1987年 | 4篇 |
1986年 | 5篇 |
1985年 | 9篇 |
1984年 | 5篇 |
1983年 | 11篇 |
1982年 | 12篇 |
1981年 | 8篇 |
1980年 | 2篇 |
1979年 | 3篇 |
1978年 | 5篇 |
1977年 | 5篇 |
1976年 | 4篇 |
1973年 | 2篇 |
排序方式: 共有3557条查询结果,搜索用时 0 毫秒
41.
Shaimaa Al-Janabi Henk-Jan van Slooten Mike Visser Tjeerd van der Ploeg Paul J. van Diest Mehdi Jiwa 《PloS one》2013,8(12)
Introduction
Mitotic Activity Index (MAI) is an important independent prognostic factor and an integral part of the breast cancer grading system. Thus, correct estimation of this prognostically relevant feature is essential for guiding treatment decision and assessing patient prognosis.The aim of this study was to validate the use of high resolution Whole Slide Images (WSI) in estimating MAI in breast cancer specimens.Methods
MAI was evaluated in 100 consecutive breast cancer specimens by three observers on two occasions, microscopically and on WSI with a wash out period of 4 months. MAI was also translated to mitotic scores as in grading. Inter- and intra-observer agreement between microscopic and digital MAI counts and scores was measured.Results
Almost perfect inter-observer agreements were obtained from counting MAI using a conventional microscope (intra-class correlation coefficient (ICCC) 0.879) as well as on WSI (ICCC 0.924). K coefficients reflected good inter-observer agreements among observers'' microscopic mitotic scores (average kappa 0.642). Comparable results were also observed among digital mitotic scores (average kappa 0.635). There was strong to perfect intra-observer agreements between MAI counts and mitotic scores for the two diagnostic modalities (ICCC 0.716–0.863, kappa 0.506–0.617). There were no significant differences in mitotic scores using both diagnostic modalities.Conclusion
Scoring mitoses using WSI in breast cancer seems to be just as reliable and reproducible as when using a microscope. Further development of software and image quality will definitely encourage the use of WSI in routine pathology practice. 相似文献42.
43.
Accurate multidimensional localization of isolated fluorescent emitters is a time consuming process in single-molecule based super-resolution microscopy. We demonstrate a functional method for real-time reconstruction with automatic feedback control, without compromising the localization accuracy. Compatible with high frame rates of EM-CCD cameras, it relies on a wavelet segmentation algorithm, together with a mix of CPU/GPU implementation. A combination with Gaussian fitting allows direct access to 3D localization. Automatic feedback control ensures optimal molecule density throughout the acquisition process. With this method, we significantly improve the efficiency and feasibility of localization-based super-resolution microscopy. 相似文献
44.
In this study, population genetic and demographic parameters were inferred using sequence data from 151 individuals of Anguilla mossambica originating from continental south and south-east Africa and Madagascar. The analyses were based on a 532 bp segment of the mitochondrial cytochrome b gene. The absence of genetic structuring was observed thereby supporting the hypothesis of panmixia for the endemic A. mossambica. The overall nucleotide diversity π = 0.002 and the haplotype diversity reached h = 0.691. Significant negative values from several tests of neutrality and mismatch analysis pointed to fluctuating historical population sizes. Bayesian averaging resulted in higher support for population growth models vs. a constant population-size model. Population decline and subsequent growth most likely predated the last glacial and were probably related to extended periods of extreme drought followed by wetter and more stable hydroclimate between 150 and 75,000 years before present (kBP). According to this scenario the female effective population size has increased since 110 kBP by c. two orders of magnitude to a recent level of about 650,000 (219,317–2,292,000). 相似文献
45.
John G Hodgson Gabriel Montserrat Marti Bozena er Glynis Jones Amy Bogaard Mike Charles Xavier Font Mohammed Ater Abdelkader Taleb Bianca A Santini Younes Hmimsa Carol Palmer Peter J Wilson Stuart R Band Amy Styring Charlotte Diffey Laura Green Erika Nitsch Elizabeth Stroud Gemma Warham 《Annals of botany》2020,126(7):1109
Background and AimsPlants depend fundamentally on establishment from seed. However, protocols in trait-based ecology currently estimate seed size but not seed number. This can be rectified. For annuals, seed number should simply be a positive function of vegetative biomass and a negative function of seed size.MethodsUsing published values of comparative seed number as the ‘gold standard’ and a large functional database, comparative seed yield and number per plant and per m2 were predicted by multiple regression. Subsequently, ecological variation in each was explored for English and Spanish habitats, newly calculated C-S-R strategies and changed abundance in the British flora.Key ResultsAs predicted, comparative seed mass yield per plant was consistently a positive function of plant size and competitive ability, and largely independent of seed size. Regressions estimating comparative seed number included, additionally, seed size as a negative function. Relationships differed numerically between regions, habitats and C-S-R strategies. Moreover, some species differed in life history over their geographical range. Comparative seed yield per m2 was positively correlated with FAO crop yield, and increasing British annuals produced numerous seeds. Nevertheless, predicted values must be viewed as comparative rather than absolute: they varied according to the ‘gold standard’ predictor used. Moreover, regressions estimating comparative seed yield per m2 achieved low precision.ConclusionsFor the first time, estimates of comparative seed yield and number for >800 annuals and their predictor equations have been produced and the ecological importance of these regenerative traits has been illustrated. ‘Regenerative trait-based ecology’ remains in its infancy, with work needed on determinate vs. indeterminate flowering (‘bet-hedging’), C-S-R methodologies, phylogeny, comparative seed yield per m2 and changing life history. Nevertheless, this has been a positive start and readers are invited to use estimates for >800 annuals, in the Supplementary data, to help advance ‘regenerative trait-based ecology’ to the next level. 相似文献
46.
Laura David Peter Schwan Martin Lobedann Sven-Oliver Borchert Bastian Budde Maike Temming Mike Kuerschner Francisca Maria Alberti Aguilo Kerstin Baumarth Tobias Thüte Benjamin Maiser Andreas Blank Viktorija Kistler Nils Weber Heiko Brandt Martin Poggel Klaus Kaiser Karl Geisen Felix Oehme Gerhard Schembecker 《Biotechnology and bioengineering》2020,117(4):1024-1036
Continuous processing is the future production method for monoclonal antibodies (mAbs). A fully continuous, fully automated downstream process based on disposable equipment was developed and implemented inside the MoBiDiK pilot plant. However, a study evaluating the comparability between batch and continuous processing based on product quality attributes was not conducted before. The work presented fills this gap comparing both process modes experimentally by purifying the same harvest material (side-by-side comparability). Samples were drawn at different time points and positions in the process for batch and continuous mode. Product quality attributes, product-related impurities, as well as process-related impurities were determined. The resulting polished material was processed to drug substance and further evaluated regarding storage stability and degradation behavior. The in-process control data from the continuous process showed the high degree of accuracy in providing relevant process parameters such as pH, conductivity, and protein concentration during the entire process duration. Minor differences between batch and continuous samples are expected as different processing conditions are unavoidable due to the different nature of batch and continuous processing. All tests revealed no significant differences in the intermediates and comparability in the drug substance between the samples of both process modes. The stability study of the final product also showed no differences in the stability profile during storage and forced degradation. Finally, online data analysis is presented as a powerful tool for online-monitoring of chromatography columns during continuous processing. 相似文献
47.
Laura Willen Maria-Gloria Basez Vit Dvorak Francis B. D. Veriegh Frank T. Aboagye Bright Idun Maha Elhadi Osman Mike Y. Osei-Atweneboana Orin Courtenay Petr Volf 《PLoS neglected tropical diseases》2021,15(6)
BackgroundSimulium damnosum sensu lato (s.l.) blackflies transmit Onchocerca volvulus, a filarial nematode that causes human onchocerciasis. Human landing catches (HLCs) is currently the sole method used to estimate blackfly biting rates but is labour-intensive and questionable on ethical grounds. A potential alternative is to measure host antibodies to vector saliva deposited during bloodfeeding. In this study, immunoassays to quantify human antibody responses to S. damnosum s.l. saliva were developed, and the salivary proteome of S. damnosum s.l. was investigated.Methodology/Principal findingsBlood samples from people living in onchocerciasis-endemic areas in Ghana were collected during the wet season; samples from people living in Accra, a blackfly-free area, were considered negative controls and compared to samples from blackfly-free locations in Sudan. Blackflies were collected by HLCs and dissected to extract their salivary glands. An ELISA measuring anti-S. damnosum s.l. salivary IgG and IgM was optimized and used to quantify the humoral immune response of 958 individuals. Both immunoassays differentiated negative controls from endemic participants. Salivary proteins were separated by gel-electrophoresis, and antigenic proteins visualized by immunoblot. Liquid chromatography mass spectrometry (LC–MS/MS) was performed to characterize the proteome of S. damnosum s.l. salivary glands. Several antigenic proteins were recognized, with the major ones located around 15 and 40 kDa. LC–MS/MS identified the presence of antigen 5-related protein, apyrase/nucleotidase, and hyaluronidase.Conclusions/SignificanceThis study validated for the first time human immunoassays that quantify humoral immune responses as potential markers of exposure to blackfly bites. These assays have the potential to facilitate understanding patterns of exposure as well as evaluating the impact of vector control on biting rates. Future studies need to investigate seasonal fluctuations of these antibody responses, potential cross-reactions with other bloodsucking arthropods, and thoroughly identify the most immunogenic proteins. 相似文献
48.
49.
Sean D. Mason Mike J. McLaughlin Caroline Johnston Ann McNeill 《Plant and Soil》2013,373(1-2):711-722
Background and aims
Recent research has demonstrated the high accuracy of a new method for assessment of plant available P in soil called diffusive gradients in thin-films (DGT). The process of P released by additions of bicarbonate to soil samples simulating common soil P tests is yet to be assessed by the new method (DGT). The aim of this study was to identify the pools of soil P extracted by soil test methods (DGT, Colwell and resin) by comparing, in 32P–labelled soils, the specific activity (SA) of phosphorus extracted by common soil test extracts with the SA of wheat plants grown in a range of agricultural soils from southern Australia.Methods
Wheat (cv. Frame) was grown for 4 weeks in 14 soils that were labelled uniformly with carrier-free 32P. The specific activity (SA) of P (MBq 32P kg 31P?1) in each soil test extract was compared to the SA of P in the wheat plants.Results
The SA of P in plants were similar to P extracted by the Colwell extractant in only 4 of the 14 soils; while SA in plants and extractants corresponded in 10 of the soils for the resin method and in 12 of the soils for the DGT method. Phosphorus in the Colwell and resin extract solutions had significantly lower SAs compared to P in the plants for 10 and 4 of the soils, respectively, indicating greater extraction of non-labile P sources (unlabelled 31P). Phosphorus in the DGT extractant had significantly lower SA than the plants for 1 soil and in 1 soil the SA was higher. Overall, across all soils, 25 % of P extracted by the Colwell method was non labile compared to 9 % and 2 % for the resin and DGT methods, respectively.Conclusion
The new DGT method for extraction of soil P has the potential to accurately predict occurrences of P deficiency because it generally extracts the same pool of labile soil P accessed by wheat plants, while methods using bicarbonate solution (e.g. Colwell, Olsen) or water (resin) at wide soil:solution ratios are more likely to measure more non-labile forms of P in soil. 相似文献50.
Capsule King Penguins recognize their mates by voice, but Guillemots do not need acoustic cues even though their calls show individual variation. Aims To determine whether the structure of Guillemot calls could allow individual recognition, as with King Penguin, and whether acoustic cues are used to locate mates among a dense mass of conspecifics at a colony. Methods Observations were made on breeding Guillemots and King Penguins. Calls made by birds returning to their mates were recorded, the signals digitized and the calls analysed. Calls were later played back to the mates of the birds concerned and the effects noted on both them and their neighbours. Results Both Guillemots and King Penguins emitted calls on return to the breeding site which contained individual signatures and were therefore potentially usable for mate recognition. In King Penguins, auditory recognition was essential for finding a mate, whereas in Guillemots most of the arriving birds located their mate in a dense crowd of conspecifics without the help of acoustic signals. Guillemots could differentiate neighbours from strangers without auditory cues. Conclusion Calls are essential for the successful identification of mates by King Penguins but not by Guillemots. 相似文献