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991.
Bartos M Hlozek P Svastova P Dvorska L Bull T Matlova L Parmova I Kuhn I Stubbs J Moravkova M Kintr J Beran V Melicharek I Ocepek M Pavlik I 《Journal of microbiological methods》2006,64(3):333-345
From Mycobacterium avium species Mycobacterium avium subsp. paratuberculosis (n=961), Mycobacterium a. avium (n=677), Mycobacterium a. silvaticum (n=5), and Mycobacterium a. hominissuis (n=1566) were examined, and from Mycobacterium tuberculosis complex M. tuberculosis (n=2), Mycobacterium bovis (n=13), M. bovis BCG (n=4), and Mycobacterium caprae (n=10) were examined. From other mycobacterial species Mycobacterium intracellulare (n=60) and atypical mycobacteria (n=256) including Mycobacterium fortuitum, Mycobacterium chelonae, Mycobacterium scrofulaceum, Mycobacterium gastri and other species of conditionally pathogenic mycobacteria were analysed. The internal standard molecules corresponding to insertion sequences IS900, IS901, IS1245, and flanking region (FR300) of IS901 were produced by PCR of alfalfa genome segment and inserted into plasmid vector. The resulting recombinant plasmid molecules were used as internal standards in coamplification with a total of 4729 mycobacterial collection strains and field isolates between 1996 and 2003. The size differences between amplicons obtained from IS900 (258 bp), IS901 (1108 bp), IS1245 (427 bp), and FR300 (300 bp) and from corresponding internal standard molecules ISIS900 (591 bp), ISIS901 (1 336 bp), ISIS1245 (583 bp), and IS901 flanking region of 300 bp ISFR300 (488 bp), respectively, allowed easy discrimination. The internal amplicons were visible by naked aye on agarose gel when 10(1), 10(3), 10(2), and 10(2) molecules for ISIS900, ISIS901, ISIS1245, and ISFR300 were used in the PCR, respectively, when no bacterial DNA was added to the reaction. The system was tested to define the amount of internal standards that could be used in the PCR without affecting the amplification of the specific segment. Non-specific amplifications were observed in M. fortuitum with IS1245 PCR and mixed infections with M. a. avium and M. a. hominissuis from pigs and cattle were found. PCR results of typing were compared with serotyping and Accu-Probes analyses in selected field isolates. 相似文献
992.
Wu CS Greenwood DR Cooney JM Jensen DJ Tatnell MA Cooper GJ Mountjoy KG 《American journal of physiology. Endocrinology and metabolism》2006,291(6):E1372-E1380
Desacetyl alpha-MSH predominates over alpha-MSH during development, but whether it is biologically active and has a physiological role is unclear. We compared the effects of 0.3 microg.g(-1).day(-1) desacetyl alpha-MSH with that of 0.3 microg.g(-1).day(-1) alpha-MSH on postnatal body growth by administering the peptides subcutaneously daily for postnatal days 0-14 and also used a two-dimensional gel electrophoresis gel-based proteomic approach to analyze protein changes in hypothalami, the relay center for body weight and growth regulation, after 14 days of treatment. We found that the growth rate between days 1 and 10 was significantly decreased by desacetyl alpha-MSH but not by alpha-MSH, but by day 14, a time reported for development of a mature pattern of hypothalamic innervation, both peptides had significantly increased neonatal growth compared with PBS-treated control rats. Desacetyl alpha-MSH significantly increased spleen weight, but alpha-MSH had no effect. alpha-MSH significantly decreased kidney weight, but desacetyl alpha-MSH had no effect. Both desacetyl alpha-MSH and alpha-MSH significantly decreased brain weight. By 14 days, both peptides significantly changed expression of a number of hypothalamic proteins, specifically metabolic enzymes, cytoskeleton, signaling, and stress response proteins. We show that peripherally administered desacetyl alpha-MSH is biologically active and induces responses that can differ from those for alpha-MSH. In conclusion, desacetyl alpha-MSH appears to be an important regulator of neonatal rat growth. 相似文献
993.
Common kestrels (Falco tinnunculus) and long-eared owls (Asio otus) in intensively farmed areas in Switzerland decreased markedly as a result of declining vole (Microtus spp.) populations. In order to counteract the loss of biodiversity in intensively farmed areas, the Swiss agri-environment scheme stipulates several types of ecological compensation areas, which together should take up 7% of the farmland. Among them are wild flower and herbaceous strips, which are not mown every year and which in summer support up to 8 times more small mammals than ordinary fields and grassland. This study investigates whether kestrels and long-eared owls preferentially hunt on ecological compensation areas and whether preferred hunting areas are related to the density of small mammals or to the density and height of the vegetation. Both kestrels and long-eared owls mainly hunted on freshly mown low-intensity meadows and artificial grassland, despite low densities of small mammals. Therefore, vegetation structure was more important for the selection of hunting sites than prey abundance. However, both predators preferred to hunt on freshly mown grassland and meadows bordering a wild flower or herbaceous strip. Voles from these strips probably invaded the adjacent freshly mown grassland and became an easy prey for kestrels and owls. In intensively farmed regions, ecological compensation areas, particularly those not mown each year, are an important refuge for small mammals, although in summer the small mammals are not directly accessible to hunting birds. Hence, a mosaic of different habitat types with grassland mown at different times of the year together with undisturbed strips is best suited to provide a year-round supply of accessible food for vole hunters. 相似文献
994.
Tadashi Aogaichi Janine Evans Jerome Gabriel Gerhard W.E. Plaut 《Archives of biochemistry and biophysics》1980,204(1):350-360
(i) The activity of purified NAD-specific isocitrate dehydrogenase from bovine heart was stimulated by free Ca2+ in the presence of ADP and subsaturating levels of magnesium isocitrate, but not in absence of ADP. However, Ca2+ was not absolutely required for ADP activation. This was particularly apparent when free Mg2+ was kept low (0.0024–0.020 mm) and the substrate magnesium dl-isocitrate ranged from 0.07–0.25 mm. When kinetic constants were determined at pH 7.4 under these conditions and in the absence of ethylene glycol bis(β-aminoethyl ether) N,N′-tetraacetate, Ca2+ had little or no effect on Km (app) for ADP; the stimulation of rate by Ca2+ was mainly due to increased V (app). With subsaturating ADP, there was an interdependence in the interaction of the enzyme with substrate and Ca2+. Thus, with ADP constant (0.30 mm) the values of Km (app) for magnesium dl-isocitrate declined from 0.35 mm at zero Ca2+ to 0.19 mm with saturating Ca2+ without affecting V; Km (app) for free Ca2+ declined with increasing magnesium isocitrate to a limiting Km of 0.3 μm. (ii) Ethylene glycol bis(β-aminoethyl ether)-N,N′-tetraacetate, frequently used as a calcium buffer, inhibited enzyme activity with and without ADP. (iii) The enzyme was not inhibited by the calmodulin inhibitors trifluoperazine and chlorpromazine. Inhibition by lanthanide ions of the isocitrate dehydrogenase was competitive with magnesium isocitrate and not with respect to Ca2+. The values of Kis (1.8 to 3.1 μm) for La3+, Yb3+, Gd3+, Eu3+, Tb3+, and Er3+ were about two orders of magnitude smaller than Km for magnesium dl-isocitrate. 相似文献
995.
996.
Andrew J. Griffith Joe Craft Janine Evans Tsuneyo Mimori John A. Hardin 《Molecular biology reports》1992,16(2):91-97
cDNA encoding the p70 polypeptide subunit of the human Ku autoantigen was isolated. In vitro expression analysis of the cDNA demonstrates that it encodes the entire open reading frame. Nucleotide sequence analysis and comparison to other previously described sequences indicate the existence of several single-nucleotide and amino acid polymorphisms. Southern blot analyses demonstrate the presence of multiple copies of homologous DNA sequences in the human genome. These data support the hypothesis that multiple genes encode a family of Ku(p70)-related polypeptides. 相似文献
997.
Janine Bessard Jean-Luc Cracowski Franoise Stanke-Labesque Germain Bessard 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2001,754(2)
F2-Isoprostanes are stable lipid peroxidation products of arachidonic acid, the quantification of which provides an index of oxidative stress in vivo. We describe a method for analysing isoprostaglandin F2α type III (15-F2t-IsoP) in biological fluids. The method involves solid-phase extraction on octadecyl endcapped and aminopropyl cartridges. After conversion to trimethylsilyl ester trimethylsilyl ether derivatives, isoprostaglandin F2α type III is analysed by mass spectrometry, operated in electronic impact selected ion monitoring mode. We have compared enzyme immunoassay (EIA; Cayman, Ann Arbor, MI, USA) to this method with 30 human urine aliquots following the same extraction procedure in order to determine the agreement between both methods. Isoprostaglandin F2α type III concentrations determined with gas chromatography–mass spectrometry (GC–MS) did not agree with those determined with EIA. Our results suggest that GC–MS and EIA do not measure the same compounds. As a consequence, comparison of clinical results using GC–MS and EIA should be avoided. 相似文献
998.
999.
Alexander Rouvinski Sharon Karniely Maria Kounin Sanaa Moussa Miri D. Goldberg Gabriela Warburg Roman Lyakhovetsky Dulce Papy-Garcia Janine Kutzsche Carsten Korth George A. Carlson Susan F. Godsave Peter J. Peters Katarina Luhr Krister Kristensson Albert Taraboulos 《The Journal of cell biology》2014,204(3):423-441
Mammalian prions refold host glycosylphosphatidylinositol-anchored PrPC into β-sheet–rich PrPSc. PrPSc is rapidly truncated into a C-terminal PrP27-30 core that is stable for days in endolysosomes. The nature of cell-associated prions, their attachment to membranes and rafts, and their subcellular locations are poorly understood; live prion visualization has not previously been achieved. A key obstacle has been the inaccessibility of PrP27-30 epitopes. We overcame this hurdle by focusing on nascent full-length PrPSc rather than on its truncated PrP27-30 product. We show that N-terminal PrPSc epitopes are exposed in their physiological context and visualize, for the first time, PrPSc in living cells. PrPSc resides for hours in unexpected cell-surface, slow moving strings and webs, sheltered from endocytosis. Prion strings observed by light and scanning electron microscopy were thin, micrometer-long structures. They were firmly cell associated, resisted phosphatidylinositol-specific phospholipase C, aligned with raft markers, fluoresced with thioflavin, and were rapidly abolished by anti-prion glycans. Prion strings and webs are the first demonstration of membrane-anchored PrPSc amyloids. 相似文献
1000.
Suwa Y Yuichi S Norton JM Bollmann A Klotz MG Stein LY Laanbroek HJ Arp DJ Goodwin LA Chertkov O Held B Bruce D Detter JC Detter JC Tapia R Han CS 《Journal of bacteriology》2011,193(18):5047-5048
Nitrosomonas sp. strain AL212 is an obligate chemolithotrophic ammonia-oxidizing bacterium (AOB) that was originally isolated in 1997 by Yuichi Suwa and colleagues. This organism belongs to Nitrosomonas cluster 6A, which is characterized by sensitivity to high ammonia concentrations, higher substrate affinity (lower K(m)), and lower maximum growth rates than strains in Nitrosomonas cluster 7, which includes Nitrosomonas europaea and Nitrosomonas eutropha. Genome-informed studies of this ammonia-sensitive cohort of AOB are needed, as these bacteria are found in freshwater environments, drinking water supplies, wastewater treatment systems, and soils worldwide. 相似文献