全文获取类型
收费全文 | 1364篇 |
免费 | 141篇 |
出版年
2022年 | 8篇 |
2021年 | 18篇 |
2020年 | 11篇 |
2019年 | 11篇 |
2018年 | 16篇 |
2017年 | 18篇 |
2016年 | 33篇 |
2015年 | 45篇 |
2014年 | 63篇 |
2013年 | 60篇 |
2012年 | 82篇 |
2011年 | 78篇 |
2010年 | 59篇 |
2009年 | 45篇 |
2008年 | 72篇 |
2007年 | 77篇 |
2006年 | 67篇 |
2005年 | 81篇 |
2004年 | 85篇 |
2003年 | 59篇 |
2002年 | 58篇 |
2001年 | 49篇 |
2000年 | 64篇 |
1999年 | 44篇 |
1998年 | 25篇 |
1997年 | 14篇 |
1996年 | 14篇 |
1995年 | 11篇 |
1994年 | 15篇 |
1993年 | 7篇 |
1992年 | 28篇 |
1991年 | 28篇 |
1990年 | 22篇 |
1989年 | 22篇 |
1988年 | 18篇 |
1987年 | 18篇 |
1986年 | 12篇 |
1985年 | 4篇 |
1984年 | 4篇 |
1983年 | 3篇 |
1981年 | 6篇 |
1980年 | 6篇 |
1979年 | 6篇 |
1978年 | 7篇 |
1977年 | 5篇 |
1976年 | 7篇 |
1975年 | 3篇 |
1968年 | 2篇 |
1965年 | 3篇 |
1959年 | 2篇 |
排序方式: 共有1505条查询结果,搜索用时 15 毫秒
71.
72.
73.
Sequential activation of phosphatidylinositol 3-kinase, beta Pix, Rac1, and Nox1 in growth factor-induced production of H2O2 下载免费PDF全文
Park HS Lee SH Park D Lee JS Ryu SH Lee WJ Rhee SG Bae YS 《Molecular and cellular biology》2004,24(10):4384-4394
The generation of reactive oxygen species (ROS) in cells stimulated with growth factors requires the activation of phosphatidylinositol 3-kinase (PI3K) and the Rac protein. We report here that the COOH-terminal region of Nox1, a protein related to gp91(phox) (Nox2) of phagocytic cells, is constitutively associated with beta Pix, a guanine nucleotide exchange factor for Rac. Both growth factor-induced ROS production and Rac1 activation were completely blocked in cells depleted of beta Pix by RNA interference. Rac1 was also shown to bind to the COOH-terminal region of Nox1 in a growth factor-dependent manner. Moreover, the depletion of Nox1 by RNA interference inhibited growth factor-induced ROS generation. These results suggest that ROS production in growth factor-stimulated cells is mediated by the sequential activation of PI3K, beta Pix, and Rac1, which then binds to Nox1 to stimulate its NADPH oxidase activity. 相似文献
74.
Hwang KC Lim S Kwon HM Bae YS Kang SM Chung KH Graham RM Rhee SG Jang Y 《The Journal of steroid biochemistry and molecular biology》2004,91(3):131-138
Ischemia and simulated ischemic conditions cause intracellular Ca2+ overload in the myocardium. The relationship between ischemia injury and Ca2+ overload has not been fully characterized. The aim of the present study was to investigate the expression and characteristics of PLC isozymes in myocardial infarction-induced cardiac remodeling and heart failure. In normal rat heart tissue, PLC-delta1 (about 44 ng/mg of heart tissue) was most abundant isozymes compared to PLC-gamma1 (6.8 ng/mg) and PLC-beta1 (0.4 ng/mg). In ischemic heart and hypoxic neonatal cardiomyocytes, PLC-delta1, but not PLC-beta1 and PLC-gamma1, was selectively degraded, a response that could be inhibited by the calpain inhibitor, calpastatin, and by the caspase inhibitor, zVAD-fmk. Overexpression of the PLC-delta1 in hypoxic neonatal cardiomyocytes rescued intracellular Ca2+ overload by ischemic conditions. In the border zone and scar region of infarcted myocardium, and in hypoxic neonatal cardiomyocytes, the selective degradation of PLC-delta1 by the calcium sensitive proteases may play important roles in intracellular Ca2+ regulations under the ischemic conditions. It is suggested that PLC isozyme-changes may contribute to the alterations in calcium homeostasis in myocardial ischemia. 相似文献
75.
76.
Dexamethasone treatment causes resistance to insulin-stimulated cellular potassium uptake in the rat 总被引:1,自引:0,他引:1
Rhee MS Perianayagam A Chen P Youn JH McDonough AA 《American journal of physiology. Cell physiology》2004,287(5):C1229-C1237
Patients treated with glucocorticoids have elevated skeletal muscle ouabain binding sites. The major Na+-K+-ATPase (NKA) isoform proteins found in muscle, 2 and 1, are increased by 50% in rats treated for 14 days with the synthetic glucocorticoid dexamethasone (DEX). This study addressed whether the DEX-induced increase in the muscle NKA pool leads to increased insulin-stimulated cellular K+ uptake that could precipitate hypokalemia. Rats were treated with DEX or vehicle via osmotic minipumps at one of two doses: 0.02 mg·kg1·day1 for 14 days (low DEX; n = 5 pairs) or 0.1 mg·kg1·day1 for 7 days (high DEX; n = 6 pairs). Insulin was infused at a rate of 5 mU·kg1·min1 over 2.5 h in conscious rats. Insulin-stimulated cellular K+ and glucose uptake rates were assessed in vivo by measuring the exogenous K+ infusion () and glucose infusion (Ginf) rates needed to maintain constant plasma K+ and glucose concentrations during insulin infusion. DEX at both doses decreased insulin-stimulated glucose uptake as previously reported. Ginf (in mmol·kg1·h1) was 10.2 ± 0.6 in vehicle-treated rats, 5.8 ± 0.8 in low-DEX-treated rats, and 5.2 ± 0.6 in high-DEX-treated rats. High DEX treatment also reduced insulin-stimulated K+ uptake. (in mmol·kg1·h1) was 0.53 ± 0.08 in vehicle-treated rats, 0.49 ± 0.14 in low-DEX-treated rats, and 0.27 ± 0.08 in high-DEX-treated rats. DEX treatment did not alter urinary K+ excretion. NKA 2-isoform levels in the low-DEX-treated group, measured by immunoblotting, were unchanged, but they increased by 38 ± 15% (soleus) and by 67 ± 3% (gastrocnemius) in the high-DEX treatment group. The NKA 1-isoform level was unchanged. These results provide novel evidence for the insulin resistance of K+ clearance during chronic DEX treatment. Insulin-stimulated cellular K+ uptake was significantly depressed despite increased muscle sodium pump pool size. skeletal muscle; sodium pump; Na+-K+-ATPase 相似文献
77.
High-throughput fluorescent tagging of full-length Arabidopsis gene products in planta 总被引:12,自引:0,他引:12
Tian GW Mohanty A Chary SN Li S Paap B Drakakaki G Kopec CD Li J Ehrhardt D Jackson D Rhee SY Raikhel NV Citovsky V 《Plant physiology》2004,135(1):25-38
We developed a high-throughput methodology, termed fluorescent tagging of full-length proteins (FTFLP), to analyze expression patterns and subcellular localization of Arabidopsis gene products in planta. Determination of these parameters is a logical first step in functional characterization of the approximately one-third of all known Arabidopsis genes that encode novel proteins of unknown function. Our FTFLP-based approach offers two significant advantages: first, it produces internally-tagged full-length proteins that are likely to exhibit native intracellular localization, and second, it yields information about the tissue specificity of gene expression by the use of native promoters. To demonstrate how FTFLP may be used for characterization of the Arabidopsis proteome, we tagged a series of known proteins with diverse subcellular targeting patterns as well as several proteins with unknown function and unassigned subcellular localization. 相似文献
78.
Two enzymes in one; two yeast peroxiredoxins display oxidative stress-dependent switching from a peroxidase to a molecular chaperone function 总被引:20,自引:0,他引:20
Jang HH Lee KO Chi YH Jung BG Park SK Park JH Lee JR Lee SS Moon JC Yun JW Choi YO Kim WY Kang JS Cheong GW Yun DJ Rhee SG Cho MJ Lee SY 《Cell》2004,117(5):625-635
Although a great deal is known biochemically about peroxiredoxins (Prxs), little is known about their real physiological function. We show here that two cytosolic yeast Prxs, cPrxI and II, which display diversity in structure and apparent molecular weights (MW), can act alternatively as peroxidases and molecular chaperones. The peroxidase function predominates in the lower MW forms, whereas the chaperone function predominates in the higher MW complexes. Oxidative stress and heat shock exposure of yeasts causes the protein structures of cPrxI and II to shift from low MW species to high MW complexes. This triggers a peroxidase-to-chaperone functional switch. These in vivo changes are primarily guided by the active peroxidase site residue, Cys(47), which serves as an efficient "H(2)O(2)-sensor" in the cells. The chaperone function of these proteins enhances yeast resistance to heat shock. 相似文献
79.
Fennell DE Rhee SK Ahn YB Häggblom MM Kerkhof LJ 《Applied and environmental microbiology》2004,70(2):1169-1175
Terminal restriction fragment length polymorphism analysis of reverse-transcribed 16S rRNA during periods of community flux was used as a tool to delineate the roles of the members of a 2-bromophenol-degrading, sulfate-reducing consortium. Starved, washed cultures were amended with 2-bromophenol plus sulfate, 2-bromophenol plus hydrogen, phenol plus sulfate, or phenol with no electron acceptor and were monitored for substrate use. In the presence of sulfate, 2-bromophenol and phenol were completely degraded. In the absence of sulfate, 2-bromophenol was dehalogenated and phenol accumulated. Direct terminal restriction fragment length polymorphism fingerprinting of the 16S rRNA in the various subcultures indicated that phylotype 2BP-48 (a Desulfovibrio-like sequence) was responsible for the dehalogenation of 2-bromophenol. A stable coculture was established which contained predominantly 2BP-48 and a second Desulfovibrio-like bacterium (designated BP212 based on terminal restriction fragment length polymorphism fingerprinting) that was capable of dehalogenating 2-bromophenol to phenol. Strain 2BP-48 in the coculture could couple reductive dehalogenation to growth with 2-bromophenol, 2,6-dibromophenol, or 2-iodophenol and lactate or formate as the electron donor. In addition to halophenols, strain 2BP-48 appears to use sulfate, sulfite, and thiosulfate as electron acceptors and is capable of simultaneous sulfidogenesis and reductive dehalogenation in the presence of sulfate. 相似文献
80.
Detection of genes involved in biodegradation and biotransformation in microbial communities by using 50-mer oligonucleotide microarrays 总被引:13,自引:0,他引:13
Rhee SK Liu X Wu L Chong SC Wan X Zhou J 《Applied and environmental microbiology》2004,70(7):4303-4317
To effectively monitor biodegrading populations, a comprehensive 50-mer-based oligonucleotide microarray was developed based on most of the 2,402 known genes and pathways involved in biodegradation and metal resistance. This array contained 1,662 unique and group-specific probes with <85% similarity to their nontarget sequences. Based on artificial probes, our results showed that under hybridization conditions of 50 degrees C and 50% formamide, the 50-mer microarray hybridization can differentiate sequences having <88% similarity. Specificity tests with representative pure cultures indicated that the designed probes on the arrays appeared to be specific to their corresponding target genes. The detection limit was approximately 5 to 10 ng of genomic DNA in the absence of background DNA and 50 to 100 ng of pure-culture genomic DNA in the presence of background DNA or 1.3 x 10(7) cells in the presence of background RNA. Strong linear relationships between the signal intensity and the target DNA and RNA were observed (r(2) = 0.95 to 0.99). Application of this type of microarray to analyze naphthalene-amended enrichment and soil microcosms demonstrated that microflora changed differently depending on the incubation conditions. While the naphthalene-degrading genes from Rhodococcus-type microorganisms were dominant in naphthalene-degrading enrichments, the genes involved in naphthalene (and polyaromatic hydrocarbon and nitrotoluene) degradation from gram-negative microorganisms, such as Ralstonia, Comamonas, and Burkholderia, were most abundant in the soil microcosms. In contrast to general conceptions, naphthalene-degrading genes from Pseudomonas were not detected, although Pseudomonas is widely known as a model microorganism for studying naphthalene degradation. The real-time PCR analysis with four representative genes showed that the microarray-based quantification was very consistent with real-time PCR (r(2) = 0.74). In addition, application of the arrays to both polyaromatic-hydrocarbon- and benzene-toluene-ethylbenzene-xylene-contaminated and uncontaminated soils indicated that the developed microarrays appeared to be useful for profiling differences in microbial community structures. Our results indicate that this technology has potential as a specific, sensitive, and quantitative tool in revealing a comprehensive picture of the compositions of biodegradation genes and the microbial community in contaminated environments, although more work is needed to improve detection sensitivity. 相似文献