全文获取类型
收费全文 | 3349篇 |
免费 | 221篇 |
出版年
2022年 | 20篇 |
2021年 | 33篇 |
2020年 | 19篇 |
2019年 | 22篇 |
2018年 | 32篇 |
2017年 | 32篇 |
2016年 | 47篇 |
2015年 | 77篇 |
2014年 | 100篇 |
2013年 | 150篇 |
2012年 | 138篇 |
2011年 | 135篇 |
2010年 | 87篇 |
2009年 | 75篇 |
2008年 | 173篇 |
2007年 | 165篇 |
2006年 | 187篇 |
2005年 | 161篇 |
2004年 | 160篇 |
2003年 | 133篇 |
2002年 | 151篇 |
2001年 | 145篇 |
2000年 | 132篇 |
1999年 | 94篇 |
1998年 | 50篇 |
1997年 | 41篇 |
1996年 | 29篇 |
1995年 | 25篇 |
1994年 | 23篇 |
1993年 | 31篇 |
1992年 | 71篇 |
1991年 | 75篇 |
1990年 | 63篇 |
1989年 | 65篇 |
1988年 | 77篇 |
1987年 | 56篇 |
1986年 | 56篇 |
1985年 | 37篇 |
1984年 | 43篇 |
1983年 | 34篇 |
1982年 | 27篇 |
1981年 | 21篇 |
1980年 | 25篇 |
1979年 | 38篇 |
1978年 | 26篇 |
1977年 | 26篇 |
1976年 | 22篇 |
1975年 | 17篇 |
1974年 | 20篇 |
1973年 | 20篇 |
排序方式: 共有3570条查询结果,搜索用时 15 毫秒
921.
Hirano M Kaneko S Yamashita T Luo H Qin W Shirota Y Nomura T Kobayashi K Murakami S 《The Journal of biological chemistry》2003,278(7):5109-5115
Hepatitis C virus (HCV) NS5B is an RNA-dependent RNA polymerase (RdRP), a central catalytic enzyme in HCV replication. While studying the subcellular localization of a NS5B mutant lacking the C-terminal membrane-anchoring domain, NS5Bt, we found that expression of the green fluorescent protein (GFP)-fused form was exclusively nucleolar. Interestingly, the distribution of endogenous nucleolin changed greatly in the cells expressing GFP-NS5B, with nucleolin colocalized with GFP-NS5B in perinuclear regions in addition to the nucleolus, suggesting that NS5B retains the ability to bind nucleolin. The interaction between nucleolin and NS5B was demonstrated by GST pull-down assay. GST pull-down assay results indicated that C-terminal region of nucleolin was important for its binding to NS5B. Scanning clustered alanine substitution mutants library of NS5B revealed two sites on NS5B that binds nucleolin. NS5B amino acids 208-214 and 500-506 were both found to be indispensable for the nucleolin binding. We reported that the latter sequence is essential for oligomerization of NS5B, which is a prerequisite for the RdRP activity. C-terminal nucleolin inhibited the NS5B RdRP activity in a dose-dependent manner. Taken together, this indicates the binding ability of nucleolin may be involved in NS5B functions. 相似文献
922.
Morishima A Ohkubo N Maeda N Miki T Mitsuda N 《The Journal of biological chemistry》2003,278(40):38188-38193
The levels of plasma HDL cholesterol and apoA-I in NFkappaB p50 subunit-deficient mice were significantly higher than those in wild-type mice under regular and high fat diets, without any significant difference in the level of total cholesterol. To examine the role of NFkappaBin lipid metabolism, we studied its effect on the regulation of apoA-I secretion from human hepatoma HepG2 cells. Lipopolysaccharide-induced activation of NFkappaB reduced the expression of apoA-I mRNA and protein, whereas adenovirus-mediated expression of IkappaBalpha super-repressor ameliorated the reduction. This IkappaBalpha-induced apoA-I increase was blocked by preincubation with MK886, a selective inhibitor of peroxisome proliferator-activated receptor alpha (PPARalpha), suggesting that NFkappaB inactivation induces apoA-I through activation of PPARalpha. To further support this idea, the expression of IkappaBalpha increased apoA-I promoter activity, and this increase was blocked by preincubation with MK886. Mutations in the putative PPARalpha-binding site in the apoA-I promoter or lack of the site abrogated these changes. Taking these results together, inhibition of NFkappaB increases apoA-I and HDL cholesterol through activation of PPARalpha in vivo and in vitro. Our data suggest a new aspect of lipid metabolism and may lead to a new paradigm for prevention and treatment of atherosclerotic disease. 相似文献
923.
The negative-feedback regulation of the IL-13 signal by the IL-13 receptor alpha2 chain in bronchial epithelial cells 总被引:3,自引:0,他引:3
Yasunaga S Yuyama N Arima K Tanaka H Toda S Maeda M Matsui K Goda C Yang Q Sugita Y Nagai H Izuhara K 《Cytokine》2003,24(6):293-303
Bronchial asthma is a complex disease characterized by airway inflammation involving Th2 cytokines. Among Th2 cytokines, the significance of IL-13 in the pathogenesis of bronchial asthma has recently emerged. Particularly, the direct action of IL-13 on bronchial epithelial cells (BECs) is critical for generation of airway hyperresponsiveness. IL-13 has two binding units; the IL-13 receptor alpha1 chain transduces the IL-13 signal comprising a heterodimer with the IL-4R alpha chain, whereas the IL-13 receptor alpha2 chain (IL-13Ralpha2) is thought to act as a decoy receptor. However, it remains obscure how expression of these molecules is regulated in each cell. In this article, we analyzed the expression of these components in BECs. Either IL-4 or IL-13 induced intracellular expression of IL-13Ralpha2 in BECs, which was STAT6-dependent and required de novo protein synthesis. IL-13Ralpha2 expressed on the cell surface as a monomer inhibited the STAT6-dependent IL-13 signal. Furthermore, expression of IL-13Ralpha2 was induced in lung tissues of ovalbumin-induced asthma model mice. Taken together, our results suggested the possibility that IL-13Ralpha2 induced by its ligand is transferred to the cell surface by an unknown mechanism, and it down-regulates the IL-13 signal in BECs, which functions as a unique negative-feedback system for the cytokine signal. 相似文献
924.
PIG-W is critical for inositol acylation but not for flipping of glycosylphosphatidylinositol-anchor 下载免费PDF全文
Murakami Y Siripanyapinyo U Hong Y Kang JY Ishihara S Nakakuma H Maeda Y Kinoshita T 《Molecular biology of the cell》2003,14(10):4285-4295
Many cell surface proteins are anchored to a membrane via a glycosylphosphatidylinositol (GPI), which is attached to the C termini in the endoplasmic reticulum. The inositol ring of phosphatidylinositol is acylated during biosynthesis of GPI. In mammalian cells, the acyl chain is added to glucosaminyl phosphatidylinositol at the third step in the GPI biosynthetic pathway and then is usually removed soon after the attachment of GPIs to proteins. The mechanisms and roles of the inositol acylation and deacylation have not been well clarified. Herein, we report derivation of human and Chinese hamster mutant cells defective in inositol acylation and the gene responsible, PIG-W. The surface expressions of GPI-anchored proteins on these mutant cells were greatly diminished, indicating the critical role of inositol acylation. PIG-W encodes a 504-amino acid protein expressed in the endoplasmic reticulum. PIG-W is most likely inositol acyltransferase itself because the tagged PIG-W affinity purified from transfected human cells had inositol acyltransferase activity and because both mutant cells were complemented with PIG-W homologs of Saccharomyces cerevisiae and Schizosaccharomyces pombe. The inositol acylation is not essential for the subsequent mannosylation, indicating that glucosaminyl phosphatidylinositol can flip from the cytoplasmic side to the luminal side of the endoplasmic reticulum. 相似文献
925.
Maeda M Mizuki E Itami T Ohba M 《In vitro cellular & developmental biology. Animal》2003,39(5-6):208-212
Cell growth in ovarian primary culture of the kuruma shrimp, Marsupenaeus japonicus, was examined under various culture conditions. The best growth of ovarian cells was obtained in a culture system consisting of double strength of Leibovitz-15 medium supplemented with 10% fetal bovine serum, glucose (1 g/L), proline (0.1 g/L), TC-Yeastolate (1 g/L), and lactalbumin hydrolysate (1 g/L). The cells survived in this medium at 25 degrees C for 45 d. The epithelial-like cells predominated in 10-d-old cultures, covering >80% of the surface area on the bottom of flask. Cells in mitosis were often observed. Cell proliferation was monitored by incorporation of 5-bromo-2'-deoxyuridine (BrdU), an analog of thymidine. 5-Bromo-2'-deoxyuridine-associated cells accounted for 11.5 and 35.0% of cell populations at 2 and 24 h, respectively, after BrdU treatment. Our results provide an improved culture technique for ovarian tissue of the kuruma shrimp. 相似文献
926.
Sequence analysis of a cDNA for D-erythrulose reductase from chicken liver showed that the deduced open reading frame encodes the protein with a molecular mass of 26 kDa consisting of 246 amino acids. Although the reductase shares more than 60% identity in the amino acid sequence with the mouse tetrameric carbonyl reductase, these two enzymes have many biochemical differences; their substrate specificity, subcellular localization, organ distribution, etc. A three-dimensional structure of D-erythrulose reductase was predicted by comparative modeling based on the structure of the tetrameric carbonyl reductase (PDB entry = 1CYD). Most of the residues at the active site (within 4 A from the ligand) of the carbonyl reductase were also conserved in the D-erythrulose reductase. Nevertheless, Val190 and Leu146 in the active site of the tetrameric carbonyl reductase were substituted in the D-erythrulose reductase by Asn192 and His148, respectively. The substitutions in the active sites may be related to the difference in substrate specificity of the two enzymes. The phylogenic analysis of D-erythrulose reductase and the other related proteins suggests that the protein described as a carbonyl reductase D-erythrulose reductase. 相似文献
927.
928.
Suzuki T Nakamoto T Ogawa S Seo S Matsumura T Tachibana K Morimoto C Hirai H 《The Journal of biological chemistry》2002,277(17):14933-14941
CasL/HEF1 belongs to the p130(Cas) family. It is tyrosine-phosphorylated following beta(1) integrin and/or T cell receptor stimulation and is thus considered to be important for immunological reactions. CasL has several structural motifs such as an SH3 domain and a substrate domain and interacts with many molecules through these motifs. To obtain more insights on the CasL-mediated signal transduction, we sought proteins that interact with the CasL SH3 domain by far Western screening, and we identified a novel human molecule, MICAL (a Molecule Interacting with CasL). MICAL is a protein of 118 kDa and is expressed in the thymus, lung, spleen, kidney, testis, and hematopoietic cells. MICAL has a calponin homology domain, a LIM domain, a putative leucine zipper motif, and a proline-rich PPKPP sequence. MICAL associates with CasL through this PPKPP sequence. MICAL is a cytoplasmic protein and colocalizes with CasL at the perinuclear area. Through the COOH-terminal region, MICAL also associates with vimentin that is a major component of intermediate filaments. Immunostaining revealed that MICAL localizes along with vimentin intermediate filaments. These results suggest that MICAL may be a cytoskeletal regulator that connects CasL to intermediate filaments. 相似文献
929.
930.
Role of adiponectin in preventing vascular stenosis. The missing link of adipo-vascular axis 总被引:93,自引:0,他引:93
Matsuda M Shimomura I Sata M Arita Y Nishida M Maeda N Kumada M Okamoto Y Nagaretani H Nishizawa H Kishida K Komuro R Ouchi N Kihara S Nagai R Funahashi T Matsuzawa Y 《The Journal of biological chemistry》2002,277(40):37487-37491
Obesity is more linked to vascular disease, including atherosclerosis and restenotic change, after balloon angioplasty. The precise mechanism linking obesity and vascular disease is still unclear. Previously we have demonstrated that the plasma levels of adiponectin, an adipose-derived hormone, decreases in obese subjects, and that hypoadiponectinemia is associated to ischemic heart disease. In current the study, we investigated the in vivo role of adiponectin on the neointimal thickening after artery injury using adiponectin-deficient mice and adiponectin-producing adenovirus. Adiponectin-deficient mice showed severe neointimal thickening and increased proliferation of vascular smooth muscle cells in mechanically injured arteries. Adenovirus-mediated supplement of adiponectin attenuated neointimal proliferation. In cultured smooth muscle cells, adiponectin attenuated DNA synthesis induced by growth factors including platelet-derived growth factor, heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), basic fibroblast growth factor, and EGF and cell proliferation and migration induced by HB-EGF. In cultured endothelial cells, adiponectin attenuated HB-EGF expression stimulated by tumor necrosis factor alpha. The current study suggests an adipo-vascular axis, a direct link between fat and artery. A therapeutic strategy to increase plasma adiponectin should be useful in preventing vascular restenosis after angioplasty. 相似文献