全文获取类型
收费全文 | 5047篇 |
免费 | 427篇 |
国内免费 | 38篇 |
出版年
2024年 | 5篇 |
2023年 | 27篇 |
2022年 | 73篇 |
2021年 | 108篇 |
2020年 | 85篇 |
2019年 | 126篇 |
2018年 | 150篇 |
2017年 | 134篇 |
2016年 | 198篇 |
2015年 | 309篇 |
2014年 | 343篇 |
2013年 | 381篇 |
2012年 | 454篇 |
2011年 | 430篇 |
2010年 | 331篇 |
2009年 | 249篇 |
2008年 | 322篇 |
2007年 | 318篇 |
2006年 | 252篇 |
2005年 | 236篇 |
2004年 | 226篇 |
2003年 | 192篇 |
2002年 | 202篇 |
2001年 | 30篇 |
2000年 | 46篇 |
1999年 | 55篇 |
1998年 | 33篇 |
1997年 | 31篇 |
1996年 | 20篇 |
1995年 | 17篇 |
1994年 | 16篇 |
1993年 | 11篇 |
1992年 | 10篇 |
1991年 | 15篇 |
1990年 | 5篇 |
1989年 | 6篇 |
1988年 | 4篇 |
1987年 | 7篇 |
1986年 | 3篇 |
1985年 | 4篇 |
1984年 | 4篇 |
1983年 | 3篇 |
1979年 | 5篇 |
1976年 | 3篇 |
1975年 | 4篇 |
1974年 | 3篇 |
1968年 | 3篇 |
1963年 | 2篇 |
1956年 | 2篇 |
1954年 | 2篇 |
排序方式: 共有5512条查询结果,搜索用时 15 毫秒
991.
Kise Y Lee SW Park SG Fukai S Sengoku T Ishii R Yokoyama S Kim S Nureki O 《Nature structural & molecular biology》2004,11(2):149-156
Human tryptophanyl-tRNA synthetase (TrpRS) is secreted into the extracellular region of vascular endothelial cells. The splice variant form (mini TrpRS) functions in vascular endothelial cell apoptosis as an angiostatic cytokine. In contrast, the closely related human tyrosyl-tRNA synthetase (TyrRS) functions as an angiogenic cytokine in its truncated form (mini TyrRS). Here, we determined the crystal structure of human mini TrpRS at a resolution of 2.3 A and compared the structure with those of prokaryotic TrpRS and human mini TyrRS. Deletion of the tRNA anticodon-binding (TAB) domain insertion, consisting of eight residues in the human TrpRS, abolished the enzyme's apoptotic activity for endothelial cells, whereas its translational catalysis and cell-binding activities remained unchanged. Thus, we have identified the inserted peptide motif that activates the angiostatic signaling. 相似文献
992.
基质金属蛋白酶MMP-26能有效促进人绒毛膜上皮癌细胞JEG-3浸润能力 总被引:1,自引:0,他引:1
胚胎植入过程中,滋养层细胞浸润与肿瘤的迁移过程非常相似,但显著的区别在于前者是受严格调控的有节制的浸润,基质金属蛋白酶(MMPs)的许多成员在其中起重要的作用.MMP-26是近年来发现的MMPs家族的新成员,它在滋养层细胞中的作用所知甚少.利用国际常用的人滋养层细胞模型——人绒毛膜上皮癌细胞系(JEG-3)作为体外实验模型,探讨MMP-26在人滋养层细胞浸润调节中的作用.将含有MMP-26全长cDNA的pCR3.1质粒转染到JEG-3细胞中,获得过量表达MMP-26基因的稳定细胞系JEG-3/MMP-26;细胞浸润分析表明JEG/MMP-26细胞的浸润能力较母本细胞明显增强;RT-PCR和明胶酶谱分析显示JEG-3/MMP-26细胞中MMP-9的表达和分泌水平提高;双荧光免疫细胞化学进一步显示MMP-26和MMP-9蛋白在细胞中有共定位现象.上述结果表明MMP-26能有效促进人滋养层细胞浸润,其作用可能是通过与其他MMP分子(如MMP-9)的协调来实现的. 相似文献
993.
An insulin-stimulated ribosomal protein S6 kinase from rabbit liver 总被引:14,自引:0,他引:14
J S Gregory T G Boulton B C Sang M H Cobb 《The Journal of biological chemistry》1989,264(31):18397-18401
In this report we describe an activated form of S6 protein kinase in rabbits treated acutely with insulin. The major insulin-stimulated activity in rabbit liver is increased 2- to 5-fold compared to material from untreated animals based on DEAE-cellulose profiles. The activity observed in DEAE-cellulose fractions can be separated into a major and a minor peak, each having very similar chromatographic behavior. Chromatography on DEAE-cellulose, S-Sepharose, heptyl-Sepharose, heparin-agarose, and Mono Q results in greater than 20,000-fold purification of the insulin-stimulated enzyme with a 12% recovery. The stimulated activity has chromatographic properties similar to an S6 protein kinase studied previously in 3T3-L1 cells (Cobb, M. H. (1986) J. Biol. Chem. 261, 12994-12999) and other systems. The enzyme purified from insulin-treated animals contains a major band that migrates in sodium dodecyl sulfate-polyacrylamide gels with Mr congruent to 70,000; this band also appears in the control preparation. Treatment of the insulin-stimulated S6 kinase with the catalytic subunit of phosphatase 2a reduces its activity by 97%. The activity of the inactivated S6 kinase is stimulated nearly 5-fold by a 15-min preincubation with partially purified insulin-stimulated microtubule-associated protein-2 kinase. 相似文献
994.
Sisi Patricia Lolita Ameria Hye Sook Jung Hee Sook Kim Sang Soo Han Hak Sung Kim Jin Ho Lee 《Biotechnology letters》2015,37(8):1637-1644
Objective
To examine the role of a gene encoding flavin-containing monooxygenase (cFMO) from Corynebacterium glutamicum ATCC13032 when cloned and expressed in Escherichia coli for the production of indigo pigments.Results
The blue pigments produced by recombinant E. coli were identified as indigo and indirubin. The cFMO was purified as a fused form with maltose-binding protein (MBP). The enzyme was optimal at 25 °C and pH 8. From absorption spectrum analysis, the cFMO was classified as a flavoprotein. FMO activity was strongly inhibited by 1 mM Cu2+ and recovered by adding 1–10 mM EDTA. The enzyme catalyzed the oxidation of TMA, thiourea, and cysteamine, but not glutathione or cysteine. MBP-cFMO had an indole oxygenase activity through oxygenation of indole to indoxyl. The recombinant E. coli produced 685 mg indigo l?1 and 103 mg indirubin l?1 from 2.5 g l-tryptophan l?1.Conclusion
The results suggest the cFMO can be used for the microbial production of both indigo and indirubin.995.
996.
P2Y12 antagonist attenuates eosinophilic inflammation and airway hyperresponsiveness in a mouse model of asthma
下载免费PDF全文
![点击此处可从《Journal of cellular and molecular medicine》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Dong‐Hyeon Suh Hoang Kim Tu Trinh Jing‐Nan Liu Le Duy Pham Sang Myun Park Hae‐Sim Park Yoo Seob Shin 《Journal of cellular and molecular medicine》2016,20(2):333-341
Leukotriene E4 (LTE4) that plays a key role in airway inflammation is expressed on platelets and eosinophils. We investigated whether blocking of the P2Y12 receptor can suppress eosinophilic inflammation in a mouse model of asthma because platelets and eosinophils share this receptor to be activated. BALB/c mice were sensitized by intraperitoneal injection of ovalbumin (OVA), followed by OVA nebulization. On each challenge day, clopidogrel, a P2Y12 antagonist was administered 30 min. before each challenge. Forty‐eight hours after the last OVA challenge, mice were assessed for airway hyperresponsiveness (AHR), cell composition and cytokine levels, including chemokine ligand 5 (CCL5), in bronchoalveolar lavage (BAL) fluid. EOL cells were treated with LTE4, with or without clopidogrel treatment, and intracellular and extracellular eosinophil cationic protein (ECP) expressions were measured to find the inhibiting function of P2Y12 antagonist on eosinophilic activation. The levels of P2Y12 expression were increased markedly in the lung homogenates of OVA‐sensitized and ‐challenged mice after platelet depletion. Administration of clopidogrel decreased AHR and the number of airway inflammatory cells, including eosinophils, in BAL fluid following OVA challenge. These results were associated with decreased levels of Th2 cytokines and CCL5. Histological examination showed that inflammatory cells as well as mucus‐containing goblet cells were reduced in clopidogrel‐administered mice compared to vehicle‐treated mice. Clopidogrel inhibited extracellular ECP secretion after LTE4 stimulation in EOL‐1 cells. Clopidogrel could prevent development of AHR and airway inflammation in a mouse model of asthma. P2Y12 can be a novel therapeutic target to the suppression of eosinophils in asthma. 相似文献
997.
本工作将富集小鼠着丝粒DNA克隆于载体EMBL3,得到约2000个克隆,建立了着丝粒基因文库。从中随机挑取20个克隆,鉴定其克隆片段平均分子量大小约为14kb。 相似文献
998.
Designing Active and Stable Silicon Photocathodes for Solar Hydrogen Production Using Molybdenum Sulfide Nanomaterials
下载免费PDF全文
![点击此处可从《Liver Transplantation》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Jesse D. Benck Sang Chul Lee Kara D. Fong Jakob Kibsgaard Robert Sinclair Thomas F. Jaramillo 《Liver Transplantation》2014,4(18)
Silicon is a promising photocathode for tandem photoelectrochemical water splitting devices, but efficient catalysis and long term stability remain key challenges. Here, it is demonstrated that with appropriately engineered interfaces, molybdenum sulfide nanomaterials can provide both corrosion protection and catalytic activity in silicon photocathodes. Using a thin MoS2 surface protecting layer, MoS2‐n+p Si electrodes that show no loss in performance after 100 h of operation are created. Transmission electron microscopy measurements show the atomic structure of the device surface and reveal the characteristics of the MoS2 layer that provide both catalytic activity and excellent stability. In spite of a low concentration of exposed catalytically active sites, these electrodes possess the best performance of any precious metal‐free silicon photocathodes with demonstrated long term stability to date. To further improve efficiency, a second molybdenum sulfide nanomaterial, highly catalytically active [Mo3S13]2? clusters, is incorporated. These photocathodes offer a promising pathway towards sustainable hydrogen production. 相似文献
999.
Proteomics has emerged as an indispensable methodology for large-scale protein analysis in functional genomics. The Escherichia coli proteome has been extensively studied and is well defined in terms of biochemical, biological, and biotechnological data. Even before the entire E. coli proteome was fully elucidated, the largest available data set had been integrated to decipher regulatory circuits and metabolic pathways, providing valuable insights into global cellular physiology and the development of metabolic and cellular engineering strategies. With the recent advent of advanced proteomic technologies, the E. coli proteome has been used for the validation of new technologies and methodologies such as sample prefractionation, protein enrichment, two-dimensional gel electrophoresis, protein detection, mass spectrometry (MS), combinatorial assays with n-dimensional chromatographies and MS, and image analysis software. These important technologies will not only provide a great amount of additional information on the E. coli proteome but also synergistically contribute to other proteomic studies. Here, we review the past development and current status of E. coli proteome research in terms of its biological, biotechnological, and methodological significance and suggest future prospects. 相似文献
1000.
Summary Fed-batch culture was carried out to increase cell mass followed by batch culture for spore production ofbacillus thuringiensis. High cell mass obtained by increasing the feeding glucose concentration in constant fed-batch culture which supported fast cell growth resulted in good sporulation during subsequent batch culture, and the maximum cell mass of 72.6 g/L and spore concentration of 1.25×1010 spores/mL could be obtained. 相似文献