全文获取类型
收费全文 | 2981篇 |
免费 | 249篇 |
国内免费 | 2篇 |
专业分类
3232篇 |
出版年
2023年 | 9篇 |
2022年 | 11篇 |
2021年 | 19篇 |
2020年 | 14篇 |
2019年 | 34篇 |
2018年 | 45篇 |
2017年 | 30篇 |
2016年 | 49篇 |
2015年 | 89篇 |
2014年 | 91篇 |
2013年 | 140篇 |
2012年 | 207篇 |
2011年 | 181篇 |
2010年 | 138篇 |
2009年 | 128篇 |
2008年 | 164篇 |
2007年 | 190篇 |
2006年 | 195篇 |
2005年 | 193篇 |
2004年 | 182篇 |
2003年 | 174篇 |
2002年 | 151篇 |
2001年 | 43篇 |
2000年 | 24篇 |
1999年 | 31篇 |
1998年 | 51篇 |
1997年 | 33篇 |
1996年 | 34篇 |
1995年 | 47篇 |
1994年 | 33篇 |
1993年 | 42篇 |
1992年 | 40篇 |
1991年 | 23篇 |
1990年 | 27篇 |
1989年 | 34篇 |
1988年 | 20篇 |
1987年 | 13篇 |
1986年 | 17篇 |
1985年 | 20篇 |
1984年 | 26篇 |
1983年 | 25篇 |
1982年 | 26篇 |
1981年 | 19篇 |
1980年 | 23篇 |
1979年 | 11篇 |
1978年 | 25篇 |
1977年 | 16篇 |
1976年 | 13篇 |
1974年 | 12篇 |
1973年 | 7篇 |
排序方式: 共有3232条查询结果,搜索用时 0 毫秒
101.
Zusammenfassung Eine Methode für die Darstellung von Bakterien-Geißeln im Phasenkontrast wird beschrieben: Fixierung der Ausstriche mit Osmiumsäuredämpfen, Beizung mit Zettnows Antimon-Tannin-Beize und nachfolgende Untersuchung im Phasenkontrastmikroskop in Luft oder Wasser.Dieses Verfahren arbeitet sicher und ist wegen seiner Einfachheit für differentialdiagnostische Untersuchungen besser geeignet als die bisher beschriebenen Methoden. 相似文献
102.
3-Deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthases catalyse the first step of the shikimate pathway. Two unrelated DAHP synthase types have been described in plants and bacteria. Two type II (aroA(A2) and aroA(A5)) and one type I DAHP synthase gene (aroA001) were identified from the myxobacterium Stigmatella aurantiaca Sg a15. Inactivation of aroA(A5) leads to a mutant that is impaired in the biosynthesis of aurachins, which are electron transport inhibitors and contain an anthranilate moiety. Feeding of anthranilic acid to the mutant culture restores production of aurachins. Inactivation of aroA(A2) and aroA001 does not impair production of aurachins or other known secondary metabolites of S. aurantiaca Sg a15. 相似文献
103.
104.
T Rosseel M Scheuch D Höper N De Regge AB Caij F Vandenbussche S Van Borm 《PloS one》2012,7(7):e41967
In 2011, a novel Orthobunyavirus was identified in cattle and sheep in Germany and the Netherlands. This virus was named Schmallenberg virus (SBV). Later, presence of the virus was confirmed using real time RT-PCR in cases of congenital malformations of bovines and ovines in several European countries, including Belgium. In the absence of specific sequencing protocols for this novel virus we confirmed its presence in RT-qPCR positive field samples using DNase SISPA-next generation sequencing (NGS), a virus discovery method based on random amplification and next generation sequencing. An in vitro transcribed RNA was used to construct a standard curve allowing the quantification of viral RNA in the field samples. Two field samples of aborted lambs containing 7.66 and 7.64 log(10) RNA copies per μL total RNA allowed unambiguous identification of SBV. One sample yielded 192 SBV reads covering about 81% of the L segment, 56% of the M segment and 13% of the S segment. The other sample resulted in 8 reads distributed over the L and M segments. Three weak positive field samples (one from an aborted calf, two from aborted lambs) containing virus quantities equivalent to 4.27-4.89 log(10) RNA copies per μL did not allow identification using DNase SISPA-NGS. This partial sequence information was compared to the whole genome sequence of SBV isolated from bovines in Germany, identifying several sequence differences. The applied viral discovery method allowed the confirmation of SBV in RT-qPCR positive brain samples. However, the failure to confirm SBV in weak PCR-positive samples illustrates the importance of the selection of properly targeted and fresh field samples in any virus discovery method. The partial sequences derived from the field samples showed several differences compared to the sequences from bovines in Germany, indicating sequence divergence within the epidemic. 相似文献
105.
Koch B Mitterer V Niederhauser J Stanborough T Murat G Rechberger G Bergler H Kressler D Pertschy B 《The Journal of biological chemistry》2012,287(26):21806-21815
2000 ribosomes have to be synthesized in yeast every minute. Therefore the fast production of ribosomal proteins, their efficient delivery to the nucleus and correct incorporation into ribosomal subunits are prerequisites for optimal growth rates. Here, we report that the ankyrin repeat protein Yar1 directly interacts with the small ribosomal subunit protein Rps3 and accompanies newly synthesized Rps3 from the cytoplasm into the nucleus where Rps3 is assembled into pre-ribosomal subunits. A yar1 deletion strain displays a similar phenotype as an rps3 mutant strain, showing an accumulation of 20S pre-rRNA and a 40S export defect. The combination of an rps3 mutation with a yar1 deletion leads to an enhancement of these phenotypes, while increased expression of RPS3 suppresses the defects of a yar1 deletion strain. We further show that Yar1 protects Rps3 from aggregation in vitro and increases its solubility in vivo. Our data suggest that Yar1 is a specific chaperone for Rps3, which serves to keep Rps3 soluble until its incorporation into the pre-ribosome. 相似文献
106.
Brigitte Lefebvre Simon Lévesque Anne-Marie Bourgault Michael R. Mulvey Laura Mataseje David Boyd Florence Doualla-Bell Cécile Tremblay 《PloS one》2015,10(4)
The emergence and spread of carbapenemase-producing Enterobacteriaceae (CPE) represent a major public health concern because these bacteria are usually extensively resistant to most antibiotics. In order to evaluate their dissemination in Quebec, a surveillance program was introduced in 2010. We report the molecular and epidemiological profiles of CPE isolates collected. Between August 2010 and December 2012, a total of 742 non-duplicate isolates non-susceptible to carbapenems were analysed. AmpC β-lactamase and metallo-β-lactamase production were detected by Etest and carbapenemase production by the modified Hodge test (MHT). Antibiotic susceptibility profiles were determined using broth microdilution or Etest. Clonality of Klebsiella pneumoniae carbapenemase (KPC) strains was analyzed by pulsed-field gel electrophoresis (PFGE). The presence of genes encoding carbapenemases as well as other β-lactamases was detected using PCR. Of the 742 isolates tested, 169 (22.8%) were CPE. Of these 169 isolates, 151 (89.3%) harboured a bla
KPC gene while the remaining isolates carried bla
SME (n = 9), bla
OXA-48 (n = 5), bla
NDM (n = 3), and bla
NMC (n = 1) genes. Among the 93 KPC strains presenting with a unique pattern (unique PFGE pattern and/or unique antibiotics susceptibility profile), 99% were resistant to ertapenem, 95% to imipenem, 87% to meropenem, 97% to aztreonam, 31% to colistin and 2% to tigecycline. In 19 patients, 2 to 5 KPC strains from different species or with a different PFGE pattern were isolated. CPE strains were present in the province of Quebec with the majority of strains harbouring KPC. Alternately, SME, OXA-48 and NMC containing strains were rarely found. 相似文献
107.
Marko MG Ahmed T Bunnell SC Wu D Chung H Huber BT Meydani SN 《Journal of immunology (Baltimore, Md. : 1950)》2007,178(3):1443-1449
Aging is associated with reduced IL-2 production and T cell proliferation. Vitamin E supplementation, in aged animals and humans, increases cell division and IL-2 production by naive T cells. The immune synapse forms at the site of contact between a T cell and an APC and participates in T cell activation. We evaluated whether vitamin E affects the redistribution of signaling proteins to the immune synapse. Purified CD4(+) T cells, from the spleens of young and old mice, were treated with vitamin E before stimulation with a surrogate APC expressing anti-CD3. Using confocal fluorescent microscopy, we observed that CD4(+) T cells from old mice were significantly less likely to recruit signaling proteins to the immune synapse than cells from young mice. Vitamin E increased the percentage of old CD4(+) T cells capable of forming an effective immune synapse. Similar results were found following in vivo supplementation with vitamin E. When compared with memory cells, naive T cells from aged mice were more defective in immune synapse formation and were more responsive to vitamin E supplementation. These data show, for the first time, that vitamin E significantly improves age-related early T cell signaling events in naive CD4(+) T cells. 相似文献
108.
Pivneva T Haas B Reyes-Haro D Laube G Veh RW Nolte C Skibo G Kettenmann H 《Cell calcium》2008,43(6):591-601
Ca(2+) signaling is the astrocyte form of excitability and the endoplasmic reticulum (ER) plays an important role as an intracellular Ca(2+) store. Since the subcellular distribution of the ER influences Ca(2+) signaling, we compared the arrangement of ER in astrocytes of hippocampus tissue and astrocytes in cell culture by electron microscopy. While the ER was usually located in close apposition to the plasma membrane in astrocytes in situ, the ER in cultured astrocytes was close to the nuclear membrane. Activation of metabotropic receptors linked to release of Ca(2+) from ER stores triggered distinct responses in cultured and in situ astrocytes. In culture, Ca(2+) signals were commonly first recorded close to the nucleus and with a delay at peripheral regions of the cells. Store-operated Ca(2+) entry (SOC) as a route to refill the Ca(2+) stores could be easily identified in cultured astrocytes as the Zn(2+)-sensitive component of the Ca(2+) signal. In contrast, such a Zn(2+)-sensitive component was not recorded in astrocytes from hippocampal slices despite of evidence for SOC. Our data indicate that both, astrocytes in situ and in vitro express SOC necessary to refill stores, but that a SOC-related signal is not recorded in the cytoplasm of astrocytes in situ since the stores are close to the plasma membrane and the refill does not affect cytoplasmic Ca(2+) levels. 相似文献
109.
Hoeberichts FA Vaeck E Kiddle G Coppens E van de Cotte B Adamantidis A Ormenese S Foyer CH Zabeau M Inzé D Périlleux C Van Breusegem F Vuylsteke M 《The Journal of biological chemistry》2008,283(9):5708-5718
Eukaryotic phosphomannomutases (PMMs) catalyze the interconversion of mannose 6-phosphate to mannose 1-phosphate and are essential to the biosynthesis of GDP-mannose. As such, plant PMMs are involved in ascorbic acid (AsA) biosynthesis and N-glycosylation. We report on the conditional phenotype of the temperature-sensitive Arabidopsis thaliana pmm-12 mutant. Mutant seedlings were phenotypically similar to wild type seedlings when grown at 16-18 degrees C but died within several days after transfer to 28 degrees C. This phenotype was observed throughout both vegetative and reproductive development. Protein extracts derived from pmm-12 plants had lower PMM protein and enzyme activity levels. In vitro biochemical analysis of recombinant proteins showed that the mutant PMM protein was compromised in its catalytic efficiency (K cat/K m). Despite significantly decreased AsA levels in pmm-12 plants, AsA deficiency could not account for the observed phenotype. Since, at restrictive temperature, total glycoprotein patterns were altered and glycosylation of protein-disulfide isomerase was perturbed, we propose that a deficiency in protein glycosylation is responsible for the observed cell death phenotype. 相似文献
110.
The biosynthesis of the volatiles 2,5‐ and 2,6‐diisopropylpyrazine ( 2 and 3 , resp.) released by the myxobacteria Nannocystis exedens subsp. cinnabarina (Na c29) and Chondromyces crocatus (strains Cm c2 and Cm c5) was studied. Isotopically labeled precursors and proposed pathway intermediates were fed to agar plate cultures of the myxobacteria. Subsequently, the volatiles were collected by use of a closed loop stripping apparatus (CLSA), and incorporation into the pyrazines was followed by GC/MS analysis. [2H8]Valine was smoothly incorporated into both pyrazines clearly establishing their origin from the amino acid pool. The cyclic dipeptide valine anhydride ( 16 ) – a potential intermediate on the biosynthetic pathway to branched dialkylpyrazines – was synthesized containing 2H1 labels in specific positions. Feeding of [2H16]‐ 16 and [2H12]‐ 16 in both valine subunits mainly resulted in the formation of pyrazines derived from only one labeled amino acid, whereas only traces of the expected pyrazines with two labeled subunits were found. To investigate the origin of nitrogen in the pyrazines, a feeding experiment with [15N]valine was performed, resulting in the incorporation of the 15N label. The results contradict a biosynthetic pathway via cyclic dipeptides, but rather point to a pathway on which valine is reduced to valine aldehyde. Its dimerization to 2,5‐diisopropyldihydropyrazine 36 and subsequent oxidation results in 2 . The proposed biosynthetic pathway neatly fits the results of earlier labeling studies and also explains the formation of the regioisomer 2,6‐diisopropylpyrazine 3 by isomerization during the first condensation step of two molecules valine aldehyde. A general biosynthetic pathway to different classes of pyrazines is presented. 相似文献