全文获取类型
收费全文 | 2964篇 |
免费 | 310篇 |
国内免费 | 409篇 |
出版年
2024年 | 12篇 |
2023年 | 79篇 |
2022年 | 147篇 |
2021年 | 247篇 |
2020年 | 148篇 |
2019年 | 200篇 |
2018年 | 171篇 |
2017年 | 117篇 |
2016年 | 146篇 |
2015年 | 237篇 |
2014年 | 252篇 |
2013年 | 255篇 |
2012年 | 301篇 |
2011年 | 239篇 |
2010年 | 165篇 |
2009年 | 117篇 |
2008年 | 124篇 |
2007年 | 93篇 |
2006年 | 77篇 |
2005年 | 91篇 |
2004年 | 60篇 |
2003年 | 75篇 |
2002年 | 61篇 |
2001年 | 58篇 |
2000年 | 32篇 |
1999年 | 25篇 |
1998年 | 20篇 |
1997年 | 5篇 |
1996年 | 12篇 |
1995年 | 10篇 |
1994年 | 5篇 |
1993年 | 7篇 |
1992年 | 9篇 |
1991年 | 6篇 |
1990年 | 8篇 |
1989年 | 9篇 |
1988年 | 4篇 |
1987年 | 6篇 |
1986年 | 4篇 |
1985年 | 8篇 |
1984年 | 4篇 |
1983年 | 4篇 |
1979年 | 3篇 |
1978年 | 3篇 |
1976年 | 3篇 |
1974年 | 4篇 |
1973年 | 3篇 |
1972年 | 2篇 |
1970年 | 2篇 |
1960年 | 2篇 |
排序方式: 共有3683条查询结果,搜索用时 31 毫秒
951.
Man Wang Huan Yu Ting Zhang Lihua Cao Yang Du Yuhao Xie Jiafu Ji Jianmin Wu 《Molecular & cellular proteomics : MCP》2022,21(1):100181
Patient-derived organoids recently emerged as promising ex vivo 3D culture models recapitulating histological and molecular characteristics of original tissues, thus proteomic profiling of organoids could be valuable for function investigation and clinical translation. However, organoids are usually cultured in murine Matrigel (served as scaffolds and matrix), which brings an issue to separate organoids from Matrigel. Because of the complex compositions of Matrigel and thousands of identical peptides shared between Matrigel and organoids, insufficiently dissolved Matrigel could influence proteomic analysis of organoids in multiple ways. Thus, how to dissolve Matrigel matrix and recovery organoid cells efficiently is vital for sample preparation. Here, we comprehensively compared three popular Matrigel dissolving methods (cell recovery solution, dispase, and PBS–EDTA buffer) and investigated the effect of undissolved Matrigel proteins on proteomic profiles of organoids. By integrative analysis of label-free proteomes of Matrigel and stable isotope labeling by amino acids in cell culture proteomes of organoids collected by three methods, respectively, we found that dispase showed an optimal efficiency, with the highest peptide yield and the highest incorporation ratio of stable isotope labeling by amino acids in cell culture labels (97.1%), as well as with the least potential Matrigel contaminants. To help analysis of proteomic profiles of organoids collected by the other two methods, we identified 312 high-confidence Matrigel contaminants, which could be filtered out to attenuate Matrigel interference with minimal loss of biological information. Together, our study identifies bioinformatics and experimental approaches to eliminate interference of Matrigel contaminants efficiently, which will be valuable for basic and translational proteomic research using organoid models. 相似文献
952.
953.
Pengchao Guo Yunshi Zhang Li Zhang Haiyang Xu Huan Zhang Zhan Wang Yongliang Jiang David Molloy Ping Zhao Qingyou Xia 《The Journal of biological chemistry》2021,297(5)
Juvenile hormone (JH) acid methyltransferase (JHAMT) is a rate-limiting enzyme that converts JH acids or inactive precursors of JHs to active JHs at the final step of JH biosynthesis in insects and thus presents an excellent target for the development of insect growth regulators or insecticides. However, the three-dimensional properties and catalytic mechanism of this enzyme are not known. Herein, we report the crystal structure of the JHAMT apoenzyme, the three-dimensional holoprotein in binary complex with its cofactor S-adenosyl-l-homocysteine, and the ternary complex with S-adenosyl-l-homocysteine and its substrate methyl farnesoate. These structures reveal the ultrafine definition of the binding patterns for JHAMT with its substrate/cofactor. Comparative structural analyses led to novel findings concerning the structural specificity of the progressive conformational changes required for binding interactions that are induced in the presence of cofactor and substrate. Importantly, structural and biochemical analyses enabled identification of one strictly conserved catalytic Gln/His pair within JHAMTs required for catalysis and further provide a molecular basis for substrate recognition and the catalytic mechanism of JHAMTs. These findings lay the foundation for the mechanistic understanding of JH biosynthesis by JHAMTs and provide a rational framework for the discovery and development of specific JHAMT inhibitors as insect growth regulators or insecticides. 相似文献
954.
955.
956.
957.
Weichao Li Qiuju Liang Ratnesh Chandra Mishra Raul Sanchez-Mu�oz Huan Wang Xin Chen Dominique Van Der Straeten Chunyi Zhang Youli Xiao 《The Plant cell》2021,33(10):3367
Folates are indispensable for plant development, but their molecular mode of action remains elusive. We synthesized a probe, “5-F-THF-Dayne,” comprising 5-formyl-tetrahydrofolate (THF) coupled to a photoaffinity tag. Exploiting this probe in an affinity proteomics study in Arabidopsis thaliana, we retrieved 51 hits. Thirty interactions were independently validated with in vitro expressed proteins to bind 5-F-THF with high or low affinity. Interestingly, the interactors reveal associations beyond one-carbon metabolism, covering also connections to nitrogen (N) metabolism, carbohydrate metabolism/photosynthesis, and proteostasis. Two of the interactions, one with the folate biosynthetic enzyme DIHYDROFOLATE REDUCTASE-THYMIDYLATE SYNTHASE 1 (AtDHFR-TS1) and another with N metabolism-associated glutamine synthetase 1;4 (AtGLN1;4), were further characterized. In silico and experimental analyses revealed G35/K36 and E330 as key residues for the binding of 5-F-THF in AtDHFR-TS1 and AtGLN1;4, respectively. Site-directed mutagenesis of AtGLN1;4 E330, which co-localizes with the ATP-binding pocket, abolished 5-F-THF binding as well as AtGLN1;4 activity. Furthermore, 5-F-THF was noted to competitively inhibit the activities of AtDHFR-TS1 and AtGLN1;4. In summary, we demonstrated a regulatory role for 5-F-THF in N metabolism, revealed 5-F-THF-mediated feedback regulation of folate biosynthesis, and identified a total of 14 previously unknown high-affinity binding cellular targets of 5-F-THF. Together, this sets a landmark toward understanding the role of folates in plant development.Exploration of proteins that interact or bind with folates reveals folate-modulated growth and development in Arabidopsis. 相似文献
958.
Domokos Ger? Petra Szoleczky Katalin Módis John P. Pribis Yousef Al-Abed Huan Yang Sangeeta Chevan Timothy R. Billiar Kevin J. Tracey Csaba Szabo 《PloS one》2013,8(6)
High mobility group box 1 (HMGB1), a highly conserved, ubiquitous protein, is released into the circulation during sterile inflammation (e.g. arthritis, trauma) and circulatory shock. It participates in the pathogenesis of delayed inflammatory responses and organ dysfunction. While several molecules have been identified that modulate the release of HMGB1, less attention has been paid to identify pharmacological inhibitors of the downstream inflammatory processes elicited by HMGB1 (C23-C45 disulfide C106 thiol form). In the current study, a cell-based medium-throughput screening of a 5000+ compound focused library of clinical drugs and drug-like compounds was performed in murine RAW264.7 macrophages, in order to identify modulators of HMGB1-induced tumor-necrosis factor alpha (TNFα) production. Clinically used drugs that suppressed HMGB1-induced TNFα production included glucocorticoids, beta agonists, and the anti-HIV compound indinavir. A re-screen of the NIH clinical compound library identified beta-agonists and various intracellular cAMP enhancers as compounds that potentiate the inhibitory effect of glucocorticoids on HMGB1-induced TNFα production. The molecular pathways involved in this synergistic anti-inflammatory effect are related, at least in part, to inhibition of TNFα mRNA synthesis via a synergistic suppression of ERK/IκB activation. Inhibition of TNFα production by prednisolone+salbutamol pretreatment was also confirmed in vivo in mice subjected to HMGB1 injection; this effect was more pronounced than the effect of either of the agents administered separately. The current study unveils several drug-like modulators of HMGB1-mediated inflammatory responses and offers pharmacological directions for the therapeutic suppression of inflammatory responses in HMGB1-dependent diseases. 相似文献
959.
Does there exist an intrinsic relationship between the flexibility and self-assembly of pepfactants?
Short peptide surfactants (pepfactants) bearing the amphiphilic molecular architecture of hydrophobic and hydrophilic moieties are attractive for a wide range of biological and medical applications. Understanding of structural basis and molecular mechanism underlying the self-assembling behaviour of pepfactants is fundamentally important for rationally designing surfactant-like peptides to function as diverse biomaterials. To date, however, the relationship between the physicochemical properties and self-assembly of pepfactants still remains largely unexplored. In this study, we attempt to elucidate the role of structural flexibility and dynamics in peptide self-assembly. Two fast and reliable quantitative structure–property relationship predictors are carefully developed with the sophisticated genetic algorithm/partial least squares method; the predictors are then applied to estimate molecular flexibility and self-assembling ability for 10,000 randomly generated surfactant-like peptides. As a result, a significant negative correlation between the flexibility and self-assembly is observed, which can be fitted fairly well using an exponential curve. Furthermore, atomistic molecular dynamics simulations also reveal a noticeable difference of flexibility profile between strong and weak self-assembling peptides. All these come together to suggest that the self-assembling behaviour of pepfactants is highly dependent on their structural flexibility. 相似文献
960.
Hu B Wu Z Jin H Hashimoto N Liu T Phan SH 《Journal of immunology (Baltimore, Md. : 1950)》2004,173(7):4661-4668
The role of IL-1beta in inflammation is amply documented, but its ability to inhibit myofibroblast differentiation and, in particular, the suppression of alpha-smooth muscle actin (alpha-SMA) gene expression is less well understood. Because IL-1beta can induce C/EBPbeta expression, the role of C/EBPbeta isoforms in IL-1beta regulation of alpha-SMA gene expression was investigated in rat lung myofibroblasts. The results showed that IL-1beta inhibited alpha-SMA expression in a dose-dependent manner, which was associated with stimulation of the expression of both C/EBPbeta isoforms, liver-enriched activating protein (LAP) and liver-enriched inhibitory protein (LIP). However, a greater increase in LIP relative to LAP expression resulted in a reduced LAP/LIP ratio after IL-1beta treatment. Transfection with an LAP-expressing plasmid stimulated, whereas an LIP-expressing plasmid inhibited, alpha-SMA expression. Cells from C/EBPbeta-deficient mice had reduced levels of alpha-SMA expression and promoter activity, which failed to respond to IL-1beta treatment. Sequence analysis identified the presence of a C/EBPbeta consensus binding sequence in the alpha-SMA promoter, which, when mutated, resulted in diminished promoter activity and abolished its responsiveness to IL-1beta treatment. EMSA revealed binding of C/EBPbeta to this C/EBPbeta consensus binding sequence from the alpha-SMA promoter. Finally, IL-1beta enhanced the expression of eukaryotic initiation factor 4E, a stimulator of LIP expression, which may account for a mechanism by which IL-1beta could alter the LAP/LIP ratio. These data taken together suggest that C/EBPbeta isoforms regulate alpha-SMA gene expression, and that its inhibition by IL-1beta was due to preferential stimulation of LIP expression. 相似文献