全文获取类型
收费全文 | 3129篇 |
免费 | 178篇 |
国内免费 | 1篇 |
出版年
2024年 | 3篇 |
2023年 | 35篇 |
2022年 | 58篇 |
2021年 | 112篇 |
2020年 | 85篇 |
2019年 | 89篇 |
2018年 | 129篇 |
2017年 | 117篇 |
2016年 | 127篇 |
2015年 | 189篇 |
2014年 | 222篇 |
2013年 | 249篇 |
2012年 | 244篇 |
2011年 | 231篇 |
2010年 | 138篇 |
2009年 | 108篇 |
2008年 | 150篇 |
2007年 | 122篇 |
2006年 | 144篇 |
2005年 | 110篇 |
2004年 | 100篇 |
2003年 | 86篇 |
2002年 | 89篇 |
2001年 | 62篇 |
2000年 | 34篇 |
1999年 | 42篇 |
1998年 | 11篇 |
1997年 | 8篇 |
1996年 | 14篇 |
1995年 | 5篇 |
1994年 | 13篇 |
1993年 | 6篇 |
1992年 | 17篇 |
1991年 | 13篇 |
1990年 | 13篇 |
1989年 | 10篇 |
1988年 | 14篇 |
1987年 | 13篇 |
1986年 | 11篇 |
1985年 | 15篇 |
1984年 | 14篇 |
1983年 | 10篇 |
1981年 | 7篇 |
1980年 | 4篇 |
1979年 | 9篇 |
1976年 | 3篇 |
1975年 | 5篇 |
1974年 | 3篇 |
1972年 | 2篇 |
1966年 | 3篇 |
排序方式: 共有3308条查询结果,搜索用时 15 毫秒
101.
Rogerio Faustino Ribeiro Jr Felipe F. Potratz Brunella M. M. Pavan Ludimila Forechi Filipe Lugon Moulin Lima Jonaina Fiorim Aurelia Araujo Fernandes Dalton Valentim Vassallo Ivanita Stefanon 《PloS one》2013,8(1)
Carvedilol has beneficial effects on cardiac function in patients with heart failure but its effect on ovariectomy-induced myocardial contractile dysfunction remains unclear. Estrogen deficiency induces myocardial contractile dysfunction and increases cardiovascular disease risk in postmenopausal women. Our aim was to investigate whether carvedilol, a beta receptor blocker, would prevent ovariectomy-induced myocardial contractile dysfunction. Female rats (8 weeks old) that underwent bilateral ovariectomy were randomly assigned to receive daily treatment with carvedilol (OVX+CAR, 20 mg/kg), placebo (OVX) and SHAM for 58 days. Left ventricle papillary muscle was mounted for isometric tension recordings. The inotropic response to Ca2+ (0.62 to 3.75 mM) and isoproterenol (Iso 10−8 to 10−2 M) were assessed. Expression of calcium handling proteins was measured by western blot analysis. Carvedilol treatment in the OVX animals: prevented weight gain and slight hypertrophy, restored the reduced positive inotropic responses to Ca2+ and isoproterenol, prevented the reduction in SERCA2a expression, abolished the increase in superoxide anion production, normalized the increase in p22phox expression, and decreased serum angiotensin converting enzyme (ACE) activity. This study demonstrated that myocardial contractile dysfunction and SERCA2a down regulation were prevented by carvedilol treatment. Superoxide anion production and NADPH oxidase seem to be involved in this response. 相似文献
102.
Maria Beatriz Calderan Rodrigues Léia Cecília de Lima Fávaro Ana Paula de Souza Pallu Anderson Ferreira Fernanda de Souza Sebastianes Maria Juliana Calderan Rodrigues Marcel Bellato Spósito Welington Luiz de Araújo Aline Aparecida Pizzirani-Kleiner 《Fungal biology》2013,117(7-8):556-568
Guignardia citricarpa is the causal agent of Citrus Black Spot (CBS), an important disease in Citriculture. Due to the expressive value of this activity worldwide, especially in Brazil, understanding more about the functioning of this fungus is of utmost relevance, making possible the elucidation of its infection mechanisms, and providing tools to control CBS. This work describes for the first time an efficient and successful methodology for genetic transformation of G. citricarpa mycelia, which generated transformants expressing the gene encoding for the gfp (green fluorescent protein) and also their interaction with citrus plant. Mycelia of G. citricarpa were transformed via Agrobacterium tumefaciens, which carried the plasmid pFAT-gfp, contains the genes for hygromycin resistance (hph) as well as gfp. The optimization of the agrotransformation protocol was performed testing different conditions (type of membrane; inductor agent concentration [acetosyringone – AS] and cocultivation time). Results demonstrated that the best condition occurred with the utilization of cellulose's ester membrane; 200 μM of AS and 96 h as cocultivation time. High mitotic stability (82 %) was displayed by transformants using Polymerase Chain Reaction (PCR) technique to confirm the hph gene insertion. In addition, the presence of gfp was observed inside mycelia by epifluorescence optical microscopy. This technique easy visualization of the behaviour of the pathogen interacting with the plant for the first time, allowing future studies on the pathogenesis of this fungus. The establishment of a transformation method for G. citricarpa opens a range of possibilities and facilitates the study of insertional mutagenesis and genetic knockouts, in order to identify the most important genes involved in the pathogenesis mechanisms and plant–pathogen interaction. 相似文献
103.
104.
Susana Pascoal Joana Esteves de Lima Jonathan D. Leslie Simon M. Hughes Leonor Saúde 《PloS one》2013,8(6)
Background
Accurate regulation of Notch signalling is central for developmental processes in a variety of tissues, but its function in pectoral fin development in zebrafish is still unknown.Methodology/Principal Findings
Here we show that core elements necessary for a functional Notch pathway are expressed in developing pectoral fins in or near prospective muscle territories. Blocking Notch signalling at different levels of the pathway consistently leads to the formation of thin, wavy, fragmented and mechanically weak muscles fibres and loss of stress fibres in endoskeletal disc cells in pectoral fins. Although the structural muscle genes encoding Desmin and Vinculin are normally transcribed in Notch-disrupted pectoral fins, their proteins levels are severely reduced, suggesting that weak mechanical forces produced by the muscle fibres are unable to stabilize/localize these proteins. Moreover, in Notch signalling disrupted pectoral fins there is a decrease in the number of Pax7-positive cells indicative of a defect in myogenesis.Conclusions/Significance
We propose that by controlling the differentiation of myogenic progenitor cells, Notch signalling might secure the formation of structurally stable muscle fibres in the zebrafish pectoral fin. 相似文献105.
Marcelo Magioli Katia Maria Paschoaletto Micchi de Barros Ferraz Eleonore Zulnara Freire Setz Alexandre Reis Percequillo Michelle Viviane de Sá Santos Rondon Vanessa Villanova Kuhnen Mariana Cristina da Silva Canhoto Karen Evelyn Almeida dos Santos Claudia Zukeran Kanda Gabriela de Lima Fregonezi Helena Alves do Prado Mitra Katherina Ferreira Milton Cezar Ribeiro Priscilla Marqui Schmidt Villela Luiz Lehmann Coutinho Márcia Gonçalves Rodrigues 《European Journal of Wildlife Research》2016,62(4):431-446
106.
Souring of oil fields during secondary oil recovery by water injection occurs mainly due to the action of sulfate-reducing bacteria (SRB) adhered to the rock surface in the vicinity of injection wells. Upflow packed-bed bioreactors have been used in petroleum microbiology because of its similarity to the oil field near the injection wells or production. However, these reactors do not realistically describe the regions near the injection wells, which are characterized by the presence of a saturated zone and a void region close to the well. In this study, the hydrodynamics of the two-compartment packing-free/packed-bed pilot bioreactor that mimics an oil reservoir was studied. The packed-free compartment was modeled using a continuous stirred tank model with mass exchange between active and stagnant zones, whereas the packed-bed compartment was modeled using a piston-dispersion-exchange model. The proposed model adequately represents the hydrodynamic of the packed-free/packed-bed bioreactor while the simulations provide important information about the characteristics of the residence time distribution (RTD) curves for different sets of model parameters. Simulations were performed to represent the control of the sulfate-reducing bacteria activity in the bioreactor with the use of molybdate in different scenarios. The simulations show that increased amounts of molybdate cause an effective inhibition of the souring sulfate-reducing bacteria activity. 相似文献
107.
Rafael Conceição de Souza Gabriela de Medeiros Muniz Andrei Santos Siqueira Adonis de Melo Lima Alessandra Pereira da Silva Evonnildo Costa Gonçalves João Lídio da Silva Gonçalves Vianez Júnior 《Journal of molecular modeling》2016,22(11):269
Human immunodeficiency virus (HIV) infections continue to exert an enormous impact on global human health. This led experts to emphasize the importance of new measures for preventing HIV infections, including the development of vaccines and novel drugs. In this context, a promising approach involves the use of lectins that can bind the surface envelope glycoprotein gp120 of HIV with high affinity, preventing viral entry. The cyanobacterial lectin microvirin (MVN) has been proposed as a candidate for development as a topical microbicide because of its ability to bind to high mannose-type glycans, potently inhibiting HIV-1 entry. Thus, the aim of this computational study was to investigate the effects of four point mutations (D53Q, D53E, D53K, and D53W) on the structure and affinity of MVN with di-mannose (MAN). Molecular dynamics simulations followed by binding free energy calculations using MM-GBSA were employed. The calculated binding free energy of ligand-receptor complexation of MVN with MAN was ?26.02 kcal mol-1. We identified in the wild-type protein that residues I45, T59, and Q81 have a major contribution to the binding free energy of di-mannose. Among the investigated mutants, the most promising one was the D53W mutation, with a theoretical binding free energy value of ?29.16 kcal mol-1. We suggest that this increased stability is due to the introduction of extra rigidity on the hinge region connecting two key structural elements of the MVN binding site. 相似文献
108.
Rachel Carmenta George Alan Blackburn Gemma Davies Claudio de Sassi André Lima Luke Parry Wlodek Tych Jos Barlow 《PloS one》2016,11(2)
Tropical forests are experiencing a growing fire problem driven by climatic change, agricultural expansion and forest degradation. Protected areas are an important feature of forest protection strategies, and sustainable use reserves (SURs) may be reducing fire prevalence since they promote sustainable livelihoods and resource management. However, the use of fire in swidden agriculture, and other forms of land management, may be undermining the effectiveness of SURs in meeting their conservation and sustainable development goals. We analyse MODIS derived hot pixels, TRMM rainfall data, Terra-Class land cover data, socio-ecological data from the Brazilian agro-census and the spatial extent of rivers and roads to evaluate whether the designation of SURs reduces fire occurrence in the Brazilian Amazon. Specifically, we ask (1) a. Is SUR location (i.e., de facto) or (1) b. designation (i.e. de jure) the driving factor affecting performance in terms of the spatial density of fires?, and (2), Does SUR creation affect fire management (i.e., the timing of fires in relation to previous rainfall)? We demonstrate that pre-protection baselines are crucial for understanding reserve performance. We show that reserve creation had no discernible impact on fire density, and that fires were less prevalent in SURs due to their characteristics of sparser human settlement and remoteness, rather than their status de jure. In addition, the timing of fires in relation to rainfall, indicative of local fire management and adherence to environmental law, did not improve following SUR creation. These results challenge the notion that SURs promote environmentally sensitive fire-management, and suggest that SURs in Amazonia will require special attention if they are to curtail future accidental wildfires, particularly as plans to expand the road infrastructure throughout the region are realised. Greater investment to support improved fire management by farmers living in reserves, in addition to other fire users, will be necessary to help ameliorate these threats. 相似文献
109.
Janaína Marques de Almeida Vanderlei Aparecido de Lima Patrícia Carla Giloni de Lima Adriana Knob 《Bioenergy Research》2016,9(3):701-710
Efficient, low-cost enzymatic hydrolysis of lignocellulosic biomass is essential for cost-effective production of bioethanol. The aim of this study was to establish a fungal fermentation-based strategy for the economic enzymatic conversion of pineapple peel into fermentable sugars. Trichoderma viride was grown on passion fruit peel in order to improve its β-glucosidase production, and a crude extract was then used to hydrolyze pineapple peel. The effects of medium pH, cultivation time, and passion fruit peel concentration on β-glucosidase production were evaluated using a central composite rotational design (CCRD) combined with response surface methodology (RSM). Optimal β-glucosidase activity of 2.40 U mL?1 was found after 6.5 days of cultivation in medium at pH 6.0, containing 2.0 % passion fruit peel. Saccharification of pineapple peel was also optimized by RSM and CCRD with respect to pH, temperature, β-glucosidase concentration, and reaction time and proceeded optimally at pH 4.0, 55 °C, with a β-glucosidase loading of 31.25 U g?1 dry feedstock and 75 h of reaction. Under these conditions, T. viride crude extract hydrolyzed pineapple peel with a glucose yield of 65.3 %. This study therefore presents passion fruit peel as an attractive raw material for the production of β-glucosidases. In addition, it describes an improved, effective, and low-cost enzymatic method for the production of fermentable sugars from pineapple peel, an abundant and inexpensive agro-industrial waste. 相似文献
110.
Ricardo Andrade Zampieri Maria Fernanda Laranjeira-Silva Sandra Marcia Muxel Ana Carolina Stocco de Lima Jeffrey Jon Shaw Lucile Maria Floeter-Winter 《PLoS neglected tropical diseases》2016,10(2)