全文获取类型
收费全文 | 38657篇 |
免费 | 3001篇 |
国内免费 | 2848篇 |
出版年
2024年 | 64篇 |
2023年 | 440篇 |
2022年 | 885篇 |
2021年 | 2139篇 |
2020年 | 1355篇 |
2019年 | 1712篇 |
2018年 | 1724篇 |
2017年 | 1175篇 |
2016年 | 1642篇 |
2015年 | 2401篇 |
2014年 | 2829篇 |
2013年 | 3076篇 |
2012年 | 3583篇 |
2011年 | 3167篇 |
2010年 | 1995篇 |
2009年 | 1617篇 |
2008年 | 1971篇 |
2007年 | 1724篇 |
2006年 | 1587篇 |
2005年 | 1286篇 |
2004年 | 1053篇 |
2003年 | 911篇 |
2002年 | 758篇 |
2001年 | 664篇 |
2000年 | 588篇 |
1999年 | 630篇 |
1998年 | 353篇 |
1997年 | 369篇 |
1996年 | 346篇 |
1995年 | 316篇 |
1994年 | 336篇 |
1993年 | 264篇 |
1992年 | 312篇 |
1991年 | 242篇 |
1990年 | 213篇 |
1989年 | 189篇 |
1988年 | 128篇 |
1987年 | 101篇 |
1986年 | 92篇 |
1985年 | 86篇 |
1984年 | 59篇 |
1983年 | 53篇 |
1982年 | 34篇 |
1981年 | 10篇 |
1980年 | 9篇 |
1979年 | 11篇 |
1975年 | 1篇 |
1972年 | 1篇 |
1971年 | 1篇 |
1965年 | 1篇 |
排序方式: 共有10000条查询结果,搜索用时 78 毫秒
941.
Absolute Quantification of E1, Ubiquitin-Like Proteins and Nedd8–MLN4924 Adduct by Mass Spectrometry
Xiaofeng Yang James E. Brownell Qing Xu Fengying Zhu Jingya Ma Huay-Keng Loke Neil Rollins Teresa A. Soucy James J. Minissale Michael P. Thomas William D. Mallender Lawrence R. Dick Ping Li Hua Liao 《Cell biochemistry and biophysics》2013,67(1):139-147
Ubiquitin (Ub) and ubiquitin-like (Ubl) proteins regulate a variety of important cellular processes by forming covalent conjugates with target proteins or lipids. Ubl conjugation is catalyzed by a cascade of proteins including activating enzymes (E1), conjugating enzymes (E2), and in many cases ligation enzymes (E3). The discovery of MLN4924 (Brownell et al., Mol Cell 37: 102–111, 1), an investigational small molecule that is a mechanism-based inhibitor of NEDD8-activating enzyme (NAE), reveals a promising strategy of targeting E1/Ubl pathway for therapeutic purposes. In order to better understand, the biochemical dynamics of Ubl conjugation in cells and tissues, we have developed a mass spectrometry-based method to quantify E1 and Ubls using isotope-labeled proteins as internal standards. Furthermore, we have used the described method to quantify levels of the covalent Nedd8-inhibitor adduct formed in MLN4924 treated cells and tissues. The Nedd8–MLN4924 adduct is a tight-binding inhibitor of NAE, and its cellular concentration represents an indirect pharmacodynamic readout of NAE/Nedd8 pathway inhibition. 相似文献
942.
Haiyan Liu Lihong Li Lixian Zhou Yan Xu 《Preparative biochemistry & biotechnology》2013,43(8):725-733
AbstractThe hydrolysates of soy protein and milk protein are nutritional and functional food ingredients. Aspergillus pseudoglaucus aspergillopepsin I (App) is an acidic protease, including signal peptide, propeptide, and catalytic domain. Here, we cloned the catalytic domain App with or without propeptide in Escherichia coli. The results showed that the App without propeptide was not expressed or did not exhibit activity and App with propeptide (proApp) was highly expressed with a specific activity of 903?U/mg. Moreover, the denaturation temperature of proApp was 4.1?°C higher than App’s. The proApp showed 104?U/mg and 252?U/mg hydrolysis activities towards soy protein and milk protein under acidic conditions. By RP-HPLC analysis, the peptides obtained from the hydrolysates of soy protein and milk protein were hydrophilic peptides. This work first demonstrates efficient proteolysis of soy protein and milk protein through the functional expression of full-length proApp, which will likely have valuable industrial applications. 相似文献
943.
944.
Qingye Ou Jian’an He Chunqiao Liu Lei Shi Chunzhong Zhao Yungqing Xu Dayong Gu 《Biotechnology and Bioprocess Engineering》2013,18(5):1031-1037
The aim of this study was to construct a gene chip system based on a surface plasmon resonance technique, where peptide nucleic acid (PNA) oligomers are used as probes. Since the self-assembled monolayer (SAM) technology offers good control at the molecular level, we prepared 2D surface chemistry via SAM for probe attachments. PNA, which was designed according to the bioinformatics, was immobilized on the SAM-modified chip, and subsequently, relevant parameters of the experiment were ensured and optimized. Our results suggest that the ion strength and pH value of the buffer solution do not play significant roles in PNA or its complementary strand hybridization. The PNA probe binds to its complementary nucleic acid strand with a higher sensitivity and specificity compared to those of a traditional DNA probe. The PNA probe combined with surface plasmon resonance (SPR) technology has the benefits of being a label-free and in-real time monitor, as well as having improved hybridization and stability efficiency, which highlight the PNA gene chip detection system as a promising biosensor for clinical applications. 相似文献
945.
Mei Ling Xu Hyoung Jin Kim Don Yong Chang Hong-Jin Kim 《Journal of microbiology (Seoul, Korea)》2013,51(3):389-393
Acidic protein levels in the milk decrease markedly as lactation progresses, suggesting that it is an important part of the colostrum. However, little attention has been paid to their biological function. In this study, we isolated the acidic protein fraction of bovine colostrum (AFC, isoelectric point <5) by anion-exchange chromatography, and investigated the effect of its dietary intake on influenza A (H1N1) virus infection. 100% of mice infected with 1 LD50 of the virus survived when administered AFC for 14 days prior to infection, compared with 33% survival when administered phosphate buffered saline (PBS). Moreover, consumption of AFC reduced the weight loss associated with infection. We propose that dietary intake of AFC has a prophylactic effect on influenza A virus infection. 相似文献
946.
947.
Xiaolei Zhu Lan Ye Huiming Ge Ling Chen Nan Jiang Lai Qian Lingling Li Rong Liu Shen Ji Su Zhang Jiali Jin Dening Guan Wei Fang Renxiang Tan Yun Xu 《Aging cell》2013,12(1):85-92
Increasing evidence demonstrates that amyloid beta (Aβ) elicits mitochondrial dysfunction and oxidative stress, which contributes to the pathogenesis of Alzheimer's disease (AD). Identification of the molecules targeting Aβ is thus of particular significance in the treatment of AD. Hopeahainol A (HopA), a polyphenol with a novel skeleton obtained from Hopea hainanensis, is potentially acetylcholinesterase‐inhibitory and anti‐oxidative in H2O2‐treated PC12 cells. In this study, we reported that HopA might bind to Aβ1–42 directly and inhibit the Aβ1–42 aggregation using a combination of molecular dynamics simulation, binding assay, transmission electron microscopic analysis and staining technique. We also demonstrated that HopA decreased the interaction between Aβ1–42 and Aβ‐binding alcohol dehydrogenase, which in turn reduced mitochondrial dysfunction and oxidative stress in vivo and in vitro. In addition, HopA was able to rescue the long‐term potentiation induction by protecting synaptic function and attenuate memory deficits in APP/PS1 mice. Our data suggest that HopA might be a promising drug for therapeutic intervention in AD. 相似文献
948.
949.
When an γ‐irradiated Dy‐, Tm‐, Sm‐ or Mn‐doped CaSO4 crystal is impulsively deformed, two peaks appear in the ML intensity versus time curve, whereby the first ML peak is found in the deformation region and the second in the post‐deformation region of the crystals. In this study, intensities Im1 and Im2 corresponding to first and second ML peaks, respectively, increased linearly with an impact velocity v0 of the piston used to deform the crystals, and times tm1 and tm2 corresponding to the first and second ML peaks, respectively, decreased with impact velocity. Total ML intensity initially increased with impact velocity and then reached a saturation value for higher values of impact velocity. ML intensity increased with increasing γ‐doses and size of crystals. Results showed that the electric field produced as a result of charging of newly‐created surfaces caused tunneling of electrons to the valence band of the hole‐trapping centres. The free holes generated moved in the valence band and their subsequent recombination with electron trapping centres released energy, thereby resulting in excitation of luminescent centres. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献
950.
Fengzhao Yang Zhancheng Xu Jinjie Wang Feng Zan Chaoqing Dong Jicun Ren 《Luminescence》2013,28(3):392-400
In this study, we report for the first time a one‐pot approach for the synthesis of new CdSeTeS quaternary‐alloyed quantum dots (QDs) in aqueous phase by microwave irradiation. CdCl2 was used as a Cd precursor during synthesis, NaHTe and NaHSe were used as Te and Se precursors and mercaptopropionic acid (MPA) was used as a stabilizer and source of sulfur. A series of quaternary‐alloyed QDs of different sizes were prepared. CdSeTeS QDs exhibited a wide emission range from 549 to 709 nm and high quantum yield (QY) up to 57.7 %. Most importantly, the quaternary‐alloyed QDs possessed significantly long fluorescence lifetimes > 100 ns as well as excellent photostability. Results of high‐resolution transmission electron microscopy (HRTEM), energy dispersive X‐ray spectroscopy (EDX) and powder X‐ray diffraction (XRD) spectroscopy showed that the nanocrystals possessed a quaternary alloy structure with good crystallinity. Fluorescence correlation spectroscopy (FCS) showed that QDs possessed good water solubility and monodispersity in aqueous solution. Furthermore, CdSeTeS QDs were modified with alpha‐thio‐omega‐carboxy poly(ethylene glycol) (HS‐PEG‐COOH) and the modified QDs were linked to anti‐epidermal growth factor receptor (EGFR) antibodies. QDs with the EGFR antibodies as labeling probes were successfully applied to targeted imaging for EGFR on the surface of SiHa cervical cancer cells. We believe that CdSeTeS QDs can become useful probes for in vivo targeted imaging and clinical diagnosis. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献