全文获取类型
收费全文 | 892篇 |
免费 | 106篇 |
出版年
2022年 | 8篇 |
2021年 | 10篇 |
2020年 | 6篇 |
2019年 | 9篇 |
2018年 | 12篇 |
2017年 | 9篇 |
2016年 | 19篇 |
2015年 | 39篇 |
2014年 | 26篇 |
2013年 | 37篇 |
2012年 | 69篇 |
2011年 | 64篇 |
2010年 | 55篇 |
2009年 | 33篇 |
2008年 | 55篇 |
2007年 | 66篇 |
2006年 | 53篇 |
2005年 | 39篇 |
2004年 | 31篇 |
2003年 | 32篇 |
2002年 | 36篇 |
2001年 | 17篇 |
2000年 | 15篇 |
1999年 | 17篇 |
1998年 | 6篇 |
1996年 | 4篇 |
1995年 | 7篇 |
1994年 | 10篇 |
1993年 | 11篇 |
1992年 | 23篇 |
1991年 | 16篇 |
1990年 | 16篇 |
1989年 | 12篇 |
1988年 | 11篇 |
1987年 | 11篇 |
1986年 | 8篇 |
1985年 | 6篇 |
1984年 | 6篇 |
1983年 | 8篇 |
1982年 | 5篇 |
1981年 | 6篇 |
1979年 | 4篇 |
1978年 | 6篇 |
1976年 | 3篇 |
1974年 | 9篇 |
1973年 | 13篇 |
1972年 | 6篇 |
1971年 | 4篇 |
1968年 | 5篇 |
1913年 | 2篇 |
排序方式: 共有998条查询结果,搜索用时 265 毫秒
41.
Hanna Alastalo Mikaela B. von Bonsdorff Katri R?ikk?nen Anu-Katriina Pesonen Clive Osmond David J. P. Barker Kati Heinonen Eero Kajantie Johan G. Eriksson 《PloS one》2013,8(7)
Background
Severe stress experienced in early life may have long-term effects on adult physiological and psychological health and well-being. We studied physical and psychosocial functioning in late adulthood in subjects separated temporarily from their parents in childhood during World War II.Methods
The 1803 participants belong to the Helsinki Birth Cohort Study, born 1934–44. Of them, 267 (14.8%) had been evacuated abroad in childhood during WWII and the remaining subjects served as controls. Physical and psychosocial functioning was assessed with the Short Form 36 scale (SF-36) between 2001 and 2004. A test for trends was based on linear regression. All analyses were adjusted for age at clinical examination, social class in childhood and adulthood, smoking, alcohol intake, physical activity, body mass index, cardiovascular disease and diabetes.Results
Physical functioning in late adulthood was lower among the separated men compared to non-separated men (b = −0.40, 95% confidence interval [95% CI]: −0.71 to −0.08). Those men separated in school age (>7 years) and who were separated for a duration over 2 years had the highest risk for lower physical functioning (b = −0.89, 95% CI: −1.58 to −0.20) and (b = −0.65, 95% CI: −1.25 to −0.05), respectively). Men separated for a duration over 2 years also had lower psychosocial functioning (b = −0.70, 95% CI: −1.35 to −0.06). These differences in physical and psychosocial functioning were not observed among women.Conclusion
Early life stress may increase the risk for impaired physical functioning in late adulthood among men. Timing and duration of the separation influenced the physical and psychosocial functioning in late adulthood. 相似文献42.
Sophie Garnier Vinh Truong Jessy Brocheton Tanja Zeller Maxime Rovital Philipp S. Wild Andreas Ziegler The Cardiogenics Consortium Thomas Munzel Laurence Tiret Stefan Blankenberg Panos Deloukas Jeannette Erdmann Christian Hengstenberg Nilesh J. Samani Heribert Schunkert Willem H. Ouwehand Alison H. Goodall Fran?ois Cambien David-Alexandre Trégou?t 《PLoS genetics》2013,9(1)
43.
Background
Pyrosequencing can be applied for Single-Nucleotide-Polymorphism (SNP)-based pathogen typing or for providing sequence information of short DNA stretches. However, for some pathogens molecular typing cannot be performed relying on a single SNP or short sequence stretch, necessitating the consideration of several genomic regions. A promising rapid approach is the simultaneous application of multiple sequencing primers, called multiplex pyrosequencing. These primers generate a fingerprint-pyrogram which is constituted by the sum of all individual pyrograms originating from each primer used.Methods
To improve pyrosequencing-based pathogen typing, we have developed the software tool MultiPSQ that expedites the analysis and evaluation of multiplex-pyrograms. As a proof of concept, a multiplex pyrosequencing assay for the typing of orthopoxviruses was developed to analyse clinical samples diagnosed in the German Consultant Laboratory for Poxviruses.Results
The software tool MultiPSQ enabled the analysis of multiplex-pyrograms originating from various pyrosequencing primers. Thus several target regions can be used for pathogen typing based on pyrosequencing. As shown with a proof of concept assay, SNPs present in different orthopoxvirus strains could be identified correctly with two primers by MultiPSQ.Conclusions
Software currently available is restricted to a fixed number of SNPs and sequencing primers, severely limiting the usefulness of this technique. In contrast, our novel software MultiPSQ allows analysis of data from multiplex pyrosequencing assays that contain any number of sequencing primers covering any number of polymorphisms. 相似文献44.
Mu-En Chung Kati Goroncy Alisa Kolesnikova David Schönauer Ulrich Schwaneberg 《Biotechnology and bioengineering》2020,117(12):3699-3711
We report a first of its kind functional cell surface display of nucleic acid polymerase and its directed evolution to efficiently incorporate 2′-O-methyl nucleotide triphosphates (2′-OMe-NTPs). In the development of polymerase cell surface display, two autotransporter proteins (Escherichia coli adhesin involved in diffuse adherence and Pseudomonas aeruginosa esterase A [EstA]) were employed to transport and anchor the 68-kDa Klenow fragment (KF) of E. coli DNA polymerase I on the surface of E. coli. The localization and function of the displayed KF were verified by analysis of cell outer membrane fractions, immunostaining, and fluorometric detection of synthesized DNA products. The EstA cell surface display system was applied to evolve KF for the incorporation of 2′-OMe-NTPs and a KF variant with a 50.7-fold increased ability to successively incorporate 2′-OMe-NTPs was discovered. Expanding the scope of cell-surface displayable proteins to the realm of polymerases provides a novel screening tool for tailoring polymerases to diverse application demands in a polymerase chain reaction and sequencing-based biotechnological and medical applications. Especially, cell surface display enables novel polymerase screening strategies in which the heat-lysis step is bypassed and thus allows the screening of mesophilic polymerases with broad application potentials ranging from diagnostics and DNA sequencing to replication of synthetic genetic polymers. 相似文献
45.
ABSTRACTThe synchrony effect (i.e. superior performance at optimal, inferior performance at suboptimal times of day) has been broadly studied within the context of circadian rhythms. Whether one chronotype copes better with the synchrony effect than the other received only insufficient empirical attention. We report on an applied experimental study investigating the impact of chronotype on the synchrony effect in a semantic analogy task. To detect an analogy, 36 participants (12 males) aged between 18 and 40 had to decide whether the relation between events of a source pair was mirrored by the relation between events of a target pair (e.g. to cook: to eat = to saddle: to ride). Temporal orientation of the relation within each event pair was varied corresponding either to the chronological or reverse order. Response times (RTs), error rates, as well as the psychophysiological parameters pre-experimental pupil baseline and peak pupil dilation replicate findings of a synchrony effect (shorter RTs and allocation of less cognitive resources at optimal times of day) and show an impact of chronotype (morning types generally outperforming evening types). Most importantly, morning types appeared to cope better with the synchrony effect than evening types: At suboptimal times, morning types solved the analogy detection task more efficient; that is faster with the same accuracy and without the investment of more cognitive resources. They also showed greater alertness and wakefulness indexed by greater pre-experimental pupil baselines. At optimal times of day, morning types have more cognitive resources available to allocate these to the more demanding conditions to outperform evening types. We interpret these findings to suggest that morning types are more able to adapt to unfavourable circumstances (for instance, by avoiding wasteful resource allocation when there are less cognitive resources available). Evening types appear less able to adapt to suboptimal times than morning types, because they have to deal with social jetlag and decreased self-control. 相似文献
46.
Andreja Radović Vassiliki Kati Melita Perčec Tadić Damijan Denac Dražen Kotrošan 《Bird Study》2013,60(1):106-114
Capsule Spatial environmental modelling well predicted nesting distribution of the White stork in Southeast Europe and can be used in conservation planning with respect to climate change.Aims To create spatial models for predicting White Stork presence and densities in the Southeast Europe to identify areas of suitable habitat for White Storks.Methods We quantified the habitat used by nesting White storks in Southeast Europe. Using spatial modelling, we defined a set of free and available online environmental variables that predict the breeding localities of the species. We employed pseudo-absences and the kriging of the residuals in order to create predictive models of nest presence and density.Results The presence–absence model was found to be precise in predicting the presence of nests. Both density and presence of breeding pairs were best explained negatively by elevation, slope, minimum temperature during May, and distance to the nearest human settlement and positively by topographic wetness index, total area of human settlement and spring precipitation.Conclusion Our robust and easily repeatable models offer a conservation tool to reveal suitable but unoccupied localities for breeding White Storks pairs which may inform our understanding of how climate change might affect the species' distribution in the future. For example, protecting White Storks on the Dalmatian coast may become even more significant in the future, because the Dalmatian coast is predicted as the only suitable breeding area in Croatia later this century. 相似文献
47.
Functionalized collagen is attractive for the development of synthetic biomaterials. Herein we present the functionalization of azidoproline containing collagen model peptides with various alkynes using click chemistry. The influence on the stability of the collagen triple helix of the stereochemistry of the introduced triazolyl prolines (4R or 4S), the position of their incorporation (Xaa or Yaa) and the substituents attached to them are shown. The results provide a useful guide for the optimal functionalization of collagen using click chemistry. 相似文献
48.
Elhaseen Elamin Ad Masclee Kati Juuti-Uusitalo Sven van IJzendoorn Freddy Troost Harm-Jan Pieters Jan Dekker Daisy Jonkers 《PloS one》2013,8(3)
Background & Aims
Evidence is accumulating that ethanol and its oxidative metabolite, acetaldehyde, can disrupt intestinal epithelial integrity, an important factor contributing to ethanol-induced liver injury. However, ethanol can also be metabolized non-oxidatively generating phosphatidylethanol and fatty acid ethyl esters (FAEEs). This study aims to investigate the effects of FAEEs on barrier function, and to explore the role of oxidative stress as possible mechanism.Methods
Epithelial permeability was assessed by paracellular flux of fluorescein isothiocyanate-conjugated dextran using live cell imaging. Cell integrity was evaluated by lactate dehydrogenase release. Localization and protein levels of ZO-1 and occludin were analyzed by immunofluorescence and cell-based ELISA, respectively. Intracellular oxidative stress and cellular ATP levels were measured by dichlorofluorescein and luciferase driven bioluminescence, respectively.Results
In vitro, ethyl oleate and ethyl palmitate dose dependently increased permeability associated with disruption and decreased ZO-1 and occludin protein levels, respectively, and increased intracellular oxidative stress without compromising cell viability. These effects could partially be attenuated by pretreatment with the antioxidant, resveratrol, pointing to the role of oxidative stress in the FAEEs-induced intestinal barrier dysfunction.Conclusions
These findings show that FAEEs can induce intestinal barrier dysfunction by disrupting the tight junctions, most likely via reactive oxygen species-dependent mechanism. 相似文献49.
Ezrin and HNRNP expression correlate with increased virus release rate and early onset of virus‐induced apoptosis of MDCK suspension cells
下载免费PDF全文
![点击此处可从《Biotechnology journal》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Sabine Kluge Yvonne Genzel Kim Laus Anja Serve Antje Pflugmacher Britta Peschel Erdmann Rapp Udo Reichl 《Biotechnology journal》2016,11(10):1332-1342
With the aim to adapt high‐yield adherent cell lines to suspension growth, Madin Darby canine kidney (MDCK) suspension cells were developed recently that achieved comparable influenza virus yields despite an early induction of apoptosis compared to the parental adherent cell line. For both cell lines, a comprehensive study under comparable infection conditions is performed comprising information on: time course of viral infection, antiviral state of cells, virus‐induced apoptosis, and virus‐induced cellular protein expression for early and late infection with influenza A/PuertoRico/8/34 H1N1. The proteomic analysis is performed with 2D differential gel electrophoreses followed by mass spectrometry. Based on flow cytometric data and on the differential expression of various stress and apoptosis‐related proteins, the earlier onset of virus‐induced apoptosis is confirmed for suspension cells. Surprisingly, the data indicated an increased virus release rate for suspension cells. These observations correlate with an increased expression of the apical marker protein ezrin, known to play a role in influenza‐induced cytoskeletal rearrangement, and the differential expression of heterogeneous nuclear ribonucleoproteins, known to bind actively influenza viral proteins and play a central role in regulating gene expression. Based on these findings, additional studies towards the design of MDCK suspension cells with further increase in influenza virus yields will be performed. 相似文献
50.
Mario Katić 《The journal of the Royal Anthropological Institute》2019,25(2):407-408