全文获取类型
收费全文 | 13634篇 |
免费 | 888篇 |
国内免费 | 3篇 |
专业分类
14525篇 |
出版年
2023年 | 80篇 |
2022年 | 160篇 |
2021年 | 272篇 |
2020年 | 202篇 |
2019年 | 272篇 |
2018年 | 330篇 |
2017年 | 303篇 |
2016年 | 472篇 |
2015年 | 692篇 |
2014年 | 750篇 |
2013年 | 957篇 |
2012年 | 1148篇 |
2011年 | 1065篇 |
2010年 | 680篇 |
2009年 | 633篇 |
2008年 | 828篇 |
2007年 | 793篇 |
2006年 | 785篇 |
2005年 | 671篇 |
2004年 | 607篇 |
2003年 | 607篇 |
2002年 | 537篇 |
2001年 | 96篇 |
2000年 | 83篇 |
1999年 | 99篇 |
1998年 | 121篇 |
1997年 | 122篇 |
1996年 | 111篇 |
1995年 | 92篇 |
1994年 | 90篇 |
1993年 | 86篇 |
1992年 | 66篇 |
1991年 | 58篇 |
1990年 | 47篇 |
1989年 | 47篇 |
1988年 | 39篇 |
1987年 | 27篇 |
1986年 | 31篇 |
1985年 | 45篇 |
1984年 | 39篇 |
1983年 | 38篇 |
1982年 | 40篇 |
1981年 | 45篇 |
1980年 | 32篇 |
1979年 | 30篇 |
1978年 | 29篇 |
1977年 | 21篇 |
1975年 | 22篇 |
1974年 | 19篇 |
1973年 | 24篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
71.
Molecular basis of the differential interaction with lithium of glycine transporters GLYT1 and GLYT2
Pérez-Siles G Morreale A Leo-Macías A Pita G Ortíz AR Aragón C López-Corcuera B 《Journal of neurochemistry》2011,118(2):195-204
Glycine synaptic levels are controlled by glycine transporters (GLYTs) catalyzing Na(+)/Cl(-)/glycine cotransport. GLYT1 displays a 2:1 :1 stoichiometry and is the main regulator of extracellular glycine concentrations. The neuronal GLYT2, with higher sodium coupling (3:1 :1), supplies glycine to the pre-synaptic terminal to refill synaptic vesicles. In this work, using structural homology modelling and molecular dynamics simulations of GLYTs, we predict the conservation of the two sodium sites present in the template (leucine transporter from Aquifex aeolicus), and confirm its use by mutagenesis and functional analysis. GLYTs Na1 and Na2 sites show differential cation selectivity, as inferred from the action of lithium, a non-transport-supporting ion, on Na(+)-site mutants. GLYTs lithium responses were unchanged in Na1-site mutants, but abolished or inverted in mutants of Na2 site, which binds lithium in the presence of low sodium concentrations and therefore, controls lithium responses. Here, we report, for the first time, that lithium exerts opposite actions on GLYTs isoforms. Glycine transport by GLYT1 is inhibited by lithium whereas GLYT2 transport is stimulated, and this effect is more evident at increased glycine concentrations. In contrast to GLYT1, high and low affinity lithium-binding processes were detected in GLYT2. 相似文献
72.
Moreschi I Bruzzone S Bodrato N Usai C Guida L Nicholas RA Kassack MU Zocchi E De Flora A 《Cell calcium》2008,43(4):344-355
Nicotinic acid adenine dinucleotide phosphate (NAADP+) is an intracellular second messenger releasing Ca2+ from intracellular stores in different cell types. In addition, it is also active in triggering [Ca2+](i) increase when applied extracellularly and various underlying mechanisms have been proposed. Here, we used hP2Y(11)-transfected 1321N1 astrocytoma cells to unequivocally establish whether extracellular NAADP+ is an agonist of the P2Y(11) receptor, as previously reported for beta-NAD+ [I. Moreschi, S. Bruzzone, R.A. Nicholas, et al., Extracellular NAD+ is an agonist of the human P2Y11 purinergic receptor in human granulocytes, J. Biol. Chem. 281 (2006) 31419-31429]. Extracellular NAADP+ triggered a concentration-dependent two-step elevation of [Ca2+](i) in 1321N1-hP2Y(11) cells, but not in wild-type 1321N1 cells, secondary to the intracellular production of IP(3), cAMP and cyclic ADP-ribose (cADPR). Specifically, the transient [Ca2+](i) rise proved to be related to IP(3) overproduction and to consequent Ca2+ mobilization, while the sustained [Ca2+](i) elevation was caused by the cAMP/ADP-ribosyl cyclase (ADPRC)/cADPR signalling cascade and by influx of extracellular Ca2+. In human granulocytes, endogenous P2Y(11) proved to be responsible for the NAADP+-induced cell activation (as demonstrated by the use of NF157, a selective and potent inhibitor of P2Y(11)), unveiling a role of NAADP+ as a pro-inflammatory cytokine. In conclusion, we provide unequivocal evidence for the activation of a member of the P2Y receptor subfamily by NAADP+. 相似文献
73.
74.
Rubio A Rambla JL Santaella M Gómez MD Orzaez D Granell A Gómez-Gómez L 《The Journal of biological chemistry》2008,283(36):24816-24825
Saffron, the processed stigma of Crocus sativus, is characterized by the presence of several apocarotenoids that contribute to the color, flavor, and aroma of the spice. However, little is known about the synthesis of aroma compounds during the development of the C. sativus stigma. The developing stigma is nearly odorless, but before and at anthesis, the aromatic compound beta-ionone becomes the principal norisoprenoid volatile in the stigma. In this study, four carotenoid cleavage dioxygenase (CCD) genes, CsCCD1a, CsCCD1b, CsCCD4a, and CsCCD4b, were isolated from C. sativus. Expression analysis showed that CsCCD1a was constitutively expressed, CsCCD1b was unique to the stigma tissue, but only CsCCD4a and -b had expression patterns consistent with the highest levels of beta-carotene and emission of beta-ionone derived during the stigma development. The CsCCD4 enzymes were localized in plastids and more specifically were present in the plastoglobules. The enzymatic activities of CsCCD1a, CsCCD1b, and CsCCD4 enzymes were determined by Escherichia coli expression, and subsequent analysis of the volatile products was generated by GC/MS. The four CCDs fell in two phylogenetically divergent dioxygenase classes, but all could cleave beta-carotene at the 9,10(9',10') positions to yield beta-ionone. The data obtained suggest that all four C. sativus CCD enzymes may contribute in different ways to the production of beta-ionone. In addition, the location and precise timing of beta-ionone synthesis, together with its known activity as a fragrance and insect attractant, suggest that this volatile may have a role in Crocus pollination. 相似文献
75.
Agroinjection of tomato fruits. A tool for rapid functional analysis of transgenes directly in fruit 总被引:13,自引:0,他引:13 下载免费PDF全文
Transient expression of foreign genes in plant tissues is a valuable tool for plant biotechnology. To shorten the time for gene functional analysis in fruits, we developed a transient methodology that could be applied to tomato (Solanum lycopersicum cv Micro Tom) fruits. It was found that injection of Agrobacterium cultures through the fruit stylar apex resulted in complete fruit infiltration. This infiltration method, named fruit agroinjection, rendered high levels of 35S Cauliflower mosaic virus-driven beta-glucuronidase and yellow fluorescence protein transient expression in the fruit, with higher expression levels around the placenta and moderate levels in the pericarp. Usefulness of fruit agroinjection was assayed in three case studies: (1) the heat shock regulation of an Arabidopsis (Arabidopsis thaliana) promoter, (2) the production of recombinant IgA antibodies as an example of molecular farming, and (3) the virus-induced gene silencing of the carotene biosynthesis pathway. In all three instances, this technology was shown to be efficient as a tool for fast transgene expression in fruits. 相似文献
76.
77.
We have recently reported that a functional alpha-L-fucosidase could be expressed by a single insertional mutation in the region of overlap between the ORFs SSO11867 and SSO3060 of the hyperthermophilic Archaeon Sulfolobus solfataricus [Cobucci-Ponzano et al. J. Biol. Chem. (2003) 278, 14622-14631]. This enzyme, belonging to glycoside hydrolase family 29 (GH29), showed micromolar specificity for p-nitrophenyl-alpha-L-fucoside (pNp-Fuc) and promoted transfucosylation reactions by following a reaction mechanism in which the products retained the anomeric configuration of the substrate. The active site residues in GH29 enzymes are still unknown. We describe here the identification of the catalytic nucleophile of the reaction in the alpha-L-fucosidase from S. solfataricus by reactivation with sodium azide of the mutant Asp242Gly that shows a 10(3)-fold activity reduction on pNp-Fuc. The detailed stereochemical analysis of the fucosyl-azide produced by the mutant reactivated on pNp-Fuc revealed its inverted (beta-fucosyl azide) configuration compared with the substrate. This allows for the first time the unambiguous assignment of Asp242, and its homologous residues, as the nucleophilic catalytic residues of GH29 alpha-L-fucosidases. This is the first time that this approach is used for alpha-L-glycosidases, widening the applicability of this method. 相似文献
78.
Carolina Elejalde-Palmett Ignacio Martinez San Segundo Imène Garroum Laurence Charrier Damien De Bellis Antonio Mucciolo Aurore Guerault Jie Liu Viktoria Zeisler-Diehl Asaph Aharoni Lukas Schreiber Bénédicte Bakan Mads H. Clausen Markus Geisler Christiane Nawrath 《Current biology : CB》2021,31(10):2111-2123.e9
79.
Marcos Amaku Francisco Antonio Bezerra Coutinho Eleazar Chaib Eduardo Massad 《Bulletin of mathematical biology》2013,75(1):82-93
We address the observation that, in some cases, patients infected with the hepatitis C virus (HCV) are cleared of HCV when super-infected with the hepatitis A virus (HAV). We hypothesise that this phenomenon can be explained by the competitive exclusion principle, including the action of the immune system, and show that the inclusion of the immune system explains both the elimination of one virus and the co-existence of both infections for a certain range of parameters. We discuss the potential clinical implications of our findings. 相似文献
80.