全文获取类型
收费全文 | 9809篇 |
免费 | 801篇 |
国内免费 | 9篇 |
出版年
2023年 | 42篇 |
2022年 | 82篇 |
2021年 | 175篇 |
2020年 | 115篇 |
2019年 | 112篇 |
2018年 | 160篇 |
2017年 | 130篇 |
2016年 | 272篇 |
2015年 | 417篇 |
2014年 | 512篇 |
2013年 | 630篇 |
2012年 | 782篇 |
2011年 | 768篇 |
2010年 | 456篇 |
2009年 | 445篇 |
2008年 | 597篇 |
2007年 | 646篇 |
2006年 | 615篇 |
2005年 | 534篇 |
2004年 | 536篇 |
2003年 | 513篇 |
2002年 | 510篇 |
2001年 | 109篇 |
2000年 | 83篇 |
1999年 | 120篇 |
1998年 | 150篇 |
1997年 | 105篇 |
1996年 | 94篇 |
1995年 | 106篇 |
1994年 | 87篇 |
1993年 | 83篇 |
1992年 | 92篇 |
1991年 | 39篇 |
1990年 | 51篇 |
1989年 | 36篇 |
1988年 | 38篇 |
1987年 | 25篇 |
1986年 | 33篇 |
1985年 | 33篇 |
1984年 | 27篇 |
1983年 | 27篇 |
1982年 | 37篇 |
1981年 | 31篇 |
1980年 | 22篇 |
1979年 | 19篇 |
1978年 | 12篇 |
1977年 | 29篇 |
1976年 | 23篇 |
1974年 | 8篇 |
1973年 | 11篇 |
排序方式: 共有10000条查询结果,搜索用时 31 毫秒
981.
Here we present ORF-selector ESPRIT, a 9-fold enhanced version of our technology for screening incremental truncation libraries to identify soluble high yielding constructs of challenging proteins. Gene fragments are truncated at both termini to access internal domains and the resulting reading frame problem is addressed by an unbiased, intein-based open reading frame selection yielding only in-frame DNA inserts. This enriched library is then subcloned into a standard high-level expression plasmid where tens of thousands of constructs can be assayed in a two-step process using colony- and liquid-handling robots to isolate rare highly expressing clones useful for production of multi milligram quantities of purifiable proteins. The p85α protein was used to benchmark the system resulting in isolation of all known domains, either alone or in tandem. The human kinase IKK1 was then screened resulting in purification of a predicted internal domain. This strategy provides an integrated, facile route to produce soluble proteins from challenging and poorly understood target genes at quantities compatible with structural biology, screening applications and immunisation studies. The high genetic diversity that can be sampled opens the way to study more diverse systems including multisubunit complexes. 相似文献
982.
Tracqui P Broisat A Toczek J Mesnier N Ohayon J Riou L 《Journal of structural biology》2011,174(1):115-123
Several studies have suggested that evolving mechanical stresses and strains drive atherosclerotic plaque development and vulnerability. Especially, stress distribution in the plaque fibrous capsule is an important determinant for the risk of vulnerable plaque rupture. Knowledge of the stiffness of atherosclerotic plaque components is therefore of critical importance. In this work, force mapping experiments using atomic force microscopy (AFM) were conducted in apolipoprotein E-deficient (ApoE(-/-)) mouse, which represents the most widely used experimental model for studying mechanisms underlying the development of atherosclerotic lesions. To obtain the elastic material properties of fibrous caps and lipidic cores of atherosclerotic plaques, serial cross-sections of aortic arch lesions were probed at different sites. Atherosclerotic plaque sub-structures were subdivided into cellular fibrotic, hypocellular fibrotic and lipidic rich areas according to histological staining. Hertz's contact mechanics were used to determine elasticity (Young's) moduli that were related to the underlying histological plaque structure. Cellular fibrotic regions exhibit a mean Young modulus of 10.4±5.7kPa. Hypocellular fibrous caps were almost six-times stiffer, with average modulus value of 59.4±47.4kPa, locally rising up to ~250kPa. Lipid rich areas exhibit a rather large range of Young's moduli, with average value of 5.5±3.5kPa. Such precise quantification of plaque stiffness heterogeneity will allow investigators to have prospectively a better monitoring of atherosclerotic disease evolution, including arterial wall remodeling and plaque rupture, in response to mechanical constraints imposed by vascular shear stress and blood pressure. 相似文献
983.
Philippe Corbeil Evelyn AnakaAuthor vitae 《Journal of electromyography and kinesiology》2011,21(5):734-741
The study of gait initiation (GI) has primarily focused on gait initiated in a forward direction, however, in everyday life, GI is often combined with a directional change. Ten young adults initiated gait with their right foot in four directions (to the left: −15°, straight ahead: 0°, to the right: 15° and 30°) at self-selected and fast gait speeds. The relationship between starting direction of GI and the lateral center of foot pressure displacement for normal (r2 = 0.57) and fast gait speed (r2 = 0.75) indicated that the lateral component plays an important role with regards to controlling the desired direction of gait. At the first step of the swing limb, the progression velocity of the center of mass (CM) remained slower for the 30° condition only, whereas no difference was found between directions for CM velocity perpendicular to the intended direction. These results suggest that postural adjustments are scaled to initiate gait in a predetermined direction. By the first step, the orientation of CM is toward the intended direction of gait, however, when gait is initiated in combination with a large change in direction, additional adjustments may be required to reach the intended progression velocity. 相似文献
984.
Yilmaz P Kottmann R Field D Knight R Cole JR Amaral-Zettler L Gilbert JA Karsch-Mizrachi I Johnston A Cochrane G Vaughan R Hunter C Park J Morrison N Rocca-Serra P Sterk P Arumugam M Bailey M Baumgartner L Birren BW Blaser MJ Bonazzi V Booth T Bork P Bushman FD Buttigieg PL Chain PS Charlson E Costello EK Huot-Creasy H Dawyndt P DeSantis T Fierer N Fuhrman JA Gallery RE Gevers D Gibbs RA San Gil I Gonzalez A Gordon JI Guralnick R Hankeln W Highlander S Hugenholtz P Jansson J Kau AL Kelley ST 《Nature biotechnology》2011,29(5):415-420
Here we present a standard developed by the Genomic Standards Consortium (GSC) for reporting marker gene sequences--the minimum information about a marker gene sequence (MIMARKS). We also introduce a system for describing the environment from which a biological sample originates. The 'environmental packages' apply to any genome sequence of known origin and can be used in combination with MIMARKS and other GSC checklists. Finally, to establish a unified standard for describing sequence data and to provide a single point of entry for the scientific community to access and learn about GSC checklists, we present the minimum information about any (x) sequence (MIxS). Adoption of MIxS will enhance our ability to analyze natural genetic diversity documented by massive DNA sequencing efforts from myriad ecosystems in our ever-changing biosphere. 相似文献
985.
Haen SP Gouttefangeas C Schmidt D Boss A Clasen S von Herbay A Kosan B Aebert H Pereira PL Rammensee HG 《Cell stress & chaperones》2011,16(5):495-504
Due to their adjuvant effect and their ability to chaperone tumor-associated peptides, heat shock proteins constitute a potent
alarm signal for the immune system and can lead to activation of anti-tumor T-cell immunity. Radiofrequency ablation has been
reported to induce heat shock protein expression especially that of heat shock protein 70 in sublethally damaged tumor cells.
In this study, we evaluated the release of heat shock protein 70 into the serum of cancer-bearing patients directly after
radiofrequency ablation. Sera of 22 patients undergoing radiofrequency ablation for the treatment of primary and secondary
malignancies of the liver, kidney, and lung, as well as control sera of 20 patients undergoing diagnostic liver biopsy were
analyzed using a manufactured heat shock protein 70 ELISA. A significant increase in serum levels of heat shock protein 70
was detectable in the patient cohort 1 day after radiofrequency ablation. More than a twofold increase was observed in nine
out of 22 patients, which tended to correlate with favorable clinical outcome. No patient of the control group revealed a
comparable increase. Radiofrequency ablation can lead to a release of heat shock protein 70 into the serum, which is transiently
detectable 1 day after treatment. Elevated heat shock protein 70 serum levels may constitute a biomarker for favorable clinical
outcome. 相似文献
986.
Thébault N Léger LA Passelergue P 《Journal of strength and conditioning research / National Strength & Conditioning Association》2011,25(10):2857-2865
The purpose of this study was to reinvestigate the relationship between aerobic fitness and fatigue indices of repeated-sprint ability (RSA), with special attention to methodological normalization. Soldiers were divided into low (n = 10) and high (n = 9) fitness groups according to a preset maximal aerobic speed (MAS) of 17 km·h(-1) (~60 ml O2·kg(-1)·min) measured with the University of Montreal Track Test (UMTT). Subjects' assessment included the RSA test (3 sets of 5 40-m sprints with 1-minute rest between sprints and 1.5 minutes between sets), a 40-m sprint (criterion test used in the computation of fatigue indices for the RSA test), strength and power measurement of the lower limbs, and the 20-m shuttle run test (20-m SRT) and the UMTT, which are measures of maximal aerobic power. The highest correlation with the RSA fatigue indices was obtained with the 20-m SRT (r = 0.90, p = 0.0001, n = 19), a test with 180° direction changes and accelerations and decelerations. The lower correlation (r = 0.66, p < 0.01, n = 19) with the UMTT (continuous forward running) suggests that some aerobic tests better disclose the importance of aerobic fitness for RSA and that aerobic power is not the sole determinant of RSA. However, neither strength nor vertical jumping power was correlated to the RSA fatigue indices. Subjects with greater MAS were able to maintain almost constant level of speed throughout series of repeated sprints and achieved better recovery between series. A MAS of at least 17 km·h(-1) favors constant and high speed level during repeated sprints. From a practical point of view, a high aerobic fitness is a precious asset in counteracting fatigue in sports with numerous sprint repetitions. 相似文献
987.
Ellingsgaard H Hauselmann I Schuler B Habib AM Baggio LL Meier DT Eppler E Bouzakri K Wueest S Muller YD Hansen AM Reinecke M Konrad D Gassmann M Reimann F Halban PA Gromada J Drucker DJ Gribble FM Ehses JA Donath MY 《Nature medicine》2011,17(11):1481-1489
Exercise, obesity and type 2 diabetes are associated with elevated plasma concentrations of interleukin-6 (IL-6). Glucagon-like peptide-1 (GLP-1) is a hormone that induces insulin secretion. Here we show that administration of IL-6 or elevated IL-6 concentrations in response to exercise stimulate GLP-1 secretion from intestinal L cells and pancreatic alpha cells, improving insulin secretion and glycemia. IL-6 increased GLP-1 production from alpha cells through increased proglucagon (which is encoded by GCG) and prohormone convertase 1/3 expression. In models of type 2 diabetes, the beneficial effects of IL-6 were maintained, and IL-6 neutralization resulted in further elevation of glycemia and reduced pancreatic GLP-1. Hence, IL-6 mediates crosstalk between insulin-sensitive tissues, intestinal L cells and pancreatic islets to adapt to changes in insulin demand. This previously unidentified endocrine loop implicates IL-6 in the regulation of insulin secretion and suggests that drugs modulating this loop may be useful in type 2 diabetes. 相似文献
988.
Hurtado MÁ da Silva PM Le Goïc N Palacios E Soudant P 《Fish & shellfish immunology》2011,31(6):978-984
Most experimental procedures on molluscs are done after acclimatization of wild animals to lab conditions. Similarly, short-term acclimation is often unavoidable in a field survey when biological analysis cannot be done within the day of sample collection. However, acclimatization can affect the general physiological condition and particularly the immune cell responses of molluscs. Our aim was to study the changes in the hemocyte characteristics of the Pacific oyster Crassostrea gigas and the carpet shell clam Ruditapes decussatus acclimated 1 or 2 days under emersed conditions at 14 ± 1 °C and for 1, 2, 7, or 10 days to flowing seawater conditions (submerged) at 9 ± 1 °C, when compared to hemolymph withdrawn from organisms sampled in the field and immediately analyzed in the laboratory (unacclimated). The hemocyte characteristics assessed by flow cytometry were the total (THC) and differential hemocyte count, percentage of dead cells, phagocytosis, and reactive oxygen species (ROS) production. Dead hemocytes were lower in oysters acclimated both in emersed and submerged conditions (1%-5%) compared to those sampled in the field (7%). Compared to oysters, the percentage of dead hemocytes was lower in clams (0.4% vs. 1.1%) and showed a tendency to decrease during acclimatization in both emersed and submerged conditions. In comparison to organisms not acclimated, the phagocytosis of hemocytes decreased in both oysters and clams acclimated under submerged conditions, but was similar in those acclimated in emersed conditions. The ROS production remained stable in both oysters and clams acclimated in emersed conditions, whereas in submerged conditions ROS production did not change in both the hyalinocytes and granulocytes of oysters, but increased in clams. In oysters, the THC decreased when they were acclimated 1 and 2 days in submerged conditions and was mainly caused by a decrease in granulocytes, but the decrease in THC in oysters acclimated 2 days in emersed conditions was caused by a decrease in hyalinocytes and small agranular cells. In clams, the THC was significantly lower in comparison to those not acclimated, regardless of the conditions of the acclimatization. These findings demonstrate that hemocyte characteristics were differentially affected in both species by the tested conditions of acclimatization. The phagocytosis and ROS production in clams and phagocytosis in oysters were not different in those acclimated for 1 day under both conditions, i.e. emersed and submerged, and those sampled in the field (unacclimated). The THC was significantly affected by acclimatization conditions, so the differences between clams and oysters should be considered in studies where important concentrations of hemocytes are required. The difference in the immune response between both species could be related to their habitat (epifaunal vs. infaunal) and their ability of resilience to manipulation and adaptation to captivity. Our results suggest that functional characteristics of hemocytes should be analyzed in both oysters and clams during the first 1 or 2 days, preferably acclimated under emersed rather than submerged conditions. 相似文献
989.
Activation and Function of the MAPKs and Their Substrates, the MAPK-Activated Protein Kinases 总被引:2,自引:0,他引:2
Summary: The mitogen-activated protein kinases (MAPKs) regulate diverse cellular programs by relaying extracellular signals to intracellular responses. In mammals, there are more than a dozen MAPK enzymes that coordinately regulate cell proliferation, differentiation, motility, and survival. The best known are the conventional MAPKs, which include the extracellular signal-regulated kinases 1 and 2 (ERK1/2), c-Jun amino-terminal kinases 1 to 3 (JNK1 to -3), p38 (α, β, γ, and δ), and ERK5 families. There are additional, atypical MAPK enzymes, including ERK3/4, ERK7/8, and Nemo-like kinase (NLK), which have distinct regulation and functions. Together, the MAPKs regulate a large number of substrates, including members of a family of protein Ser/Thr kinases termed MAPK-activated protein kinases (MAPKAPKs). The MAPKAPKs are related enzymes that respond to extracellular stimulation through direct MAPK-dependent activation loop phosphorylation and kinase activation. There are five MAPKAPK subfamilies: the p90 ribosomal S6 kinase (RSK), the mitogen- and stress-activated kinase (MSK), the MAPK-interacting kinase (MNK), the MAPK-activated protein kinase 2/3 (MK2/3), and MK5 (also known as p38-regulated/activated protein kinase [PRAK]). These enzymes have diverse biological functions, including regulation of nucleosome and gene expression, mRNA stability and translation, and cell proliferation and survival. Here we review the mechanisms of MAPKAPK activation by the different MAPKs and discuss their physiological roles based on established substrates and recent discoveries. 相似文献
990.
As a result of an invasion by the native grass Elymus athericus (Link) Kerguélen (Poaceae) in the last 10 years, a major change in vegetation cover has occurred in salt marshes of the Mont Saint-Michel bay, Western France. The impact of such an invasion on carabid assemblages, a dominant group of terrestrial arthropods in these habitats and containing several stenotopic species, is investigated here. In our study site, carabid data are available from 1983 and 1984, allowing a comparison of species distribution ranges in salt marshes before (1983-1984) and after (2002) the Elymus athericus invasion. A total of 16,867 adults belonging to 40 species were caught. By considering the presence-absence of species shared between studies, we show that the invasion by Elymus athericus promoted the progression of non-coastal species (mainly Pterostichus s.l. spp.). This did however not interfere with resident species distributions, finally resulting in higher carabid species richness in the entire area. The species composition and abundances of carabid assemblages were also compared between natural and invaded stations in 2002. The main result is that abundances of some halophilic species decreased in one invaded plot (in case of Pogonus chalceus (Marsham 1802)) whereas the opposite pattern was observed for other species (e.g., Bembidion minimum (Fabricius 1792)). Invaded habitats were characterized by lower percentages of halophilic species and higher total species richness. 相似文献