全文获取类型
收费全文 | 635篇 |
免费 | 67篇 |
国内免费 | 196篇 |
出版年
2024年 | 2篇 |
2023年 | 28篇 |
2022年 | 21篇 |
2021年 | 31篇 |
2020年 | 29篇 |
2019年 | 47篇 |
2018年 | 27篇 |
2017年 | 28篇 |
2016年 | 18篇 |
2015年 | 33篇 |
2014年 | 43篇 |
2013年 | 74篇 |
2012年 | 35篇 |
2011年 | 60篇 |
2010年 | 37篇 |
2009年 | 42篇 |
2008年 | 31篇 |
2007年 | 37篇 |
2006年 | 38篇 |
2005年 | 36篇 |
2004年 | 35篇 |
2003年 | 17篇 |
2002年 | 19篇 |
2001年 | 20篇 |
2000年 | 10篇 |
1999年 | 15篇 |
1998年 | 8篇 |
1997年 | 11篇 |
1996年 | 8篇 |
1995年 | 6篇 |
1994年 | 1篇 |
1993年 | 3篇 |
1992年 | 7篇 |
1991年 | 3篇 |
1990年 | 4篇 |
1989年 | 5篇 |
1988年 | 2篇 |
1986年 | 3篇 |
1985年 | 3篇 |
1984年 | 3篇 |
1983年 | 3篇 |
1982年 | 5篇 |
1981年 | 3篇 |
1980年 | 2篇 |
1979年 | 3篇 |
1978年 | 1篇 |
1976年 | 1篇 |
排序方式: 共有898条查询结果,搜索用时 312 毫秒
71.
目的 探讨全程无缝隙护理模式对肿瘤科护理工作质量的影响。方法 2013年1月-12月我院肿瘤科采用无缝隙护理模式的患者120例为观察组,2012年1月-12月采用常规护理的120例肿瘤科患者为对照组,对比两组患者的心理舒适度、患者对护理工作的满意度。结果 观察组患者的安全感、满足感、尊重感所占比例均高于对照组,差异均有统计学意义(P<0.05)。观察组患者对护士的工作能力、关爱与沟通、服务热情主动、病区管理、健康教育满意度均高于对照组,差异均有统计学意义(P<0.05)。结论 全程无缝隙护理模式应用于肿瘤科护理,能够有效改善患者不良情绪,提高护理工作质量及患者满意度。 相似文献
72.
Fabien Sénéchal Mélanie L'Enfant Jean-Marc Domon Emeline Rosiau Marie-Jeanne Crépeau Ogier Surcouf Juan Esquivel-Rodriguez Paulo Marcelo Alain Mareck Fran?ois Guérineau Hyung-Rae Kim Jozef Mravec Estelle Bonnin Elisabeth Jamet Daisuke Kihara Patrice Lerouge Marie-Christine Ralet Jér?me Pelloux Catherine Rayon 《The Journal of biological chemistry》2015,290(38):23320-23335
Pectin methylesterases (PMEs) catalyze the demethylesterification of homogalacturonan domains of pectin in plant cell walls and are regulated by endogenous pectin methylesterase inhibitors (PMEIs). In Arabidopsis dark-grown hypocotyls, one PME (AtPME3) and one PMEI (AtPMEI7) were identified as potential interacting proteins. Using RT-quantitative PCR analysis and gene promoter::GUS fusions, we first showed that AtPME3 and AtPMEI7 genes had overlapping patterns of expression in etiolated hypocotyls. The two proteins were identified in hypocotyl cell wall extracts by proteomics. To investigate the potential interaction between AtPME3 and AtPMEI7, both proteins were expressed in a heterologous system and purified by affinity chromatography. The activity of recombinant AtPME3 was characterized on homogalacturonans (HGs) with distinct degrees/patterns of methylesterification. AtPME3 showed the highest activity at pH 7.5 on HG substrates with a degree of methylesterification between 60 and 80% and a random distribution of methyl esters. On the best HG substrate, AtPME3 generates long non-methylesterified stretches and leaves short highly methylesterified zones, indicating that it acts as a processive enzyme. The recombinant AtPMEI7 and AtPME3 interaction reduces the level of demethylesterification of the HG substrate but does not inhibit the processivity of the enzyme. These data suggest that the AtPME3·AtPMEI7 complex is not covalently linked and could, depending on the pH, be alternately formed and dissociated. Docking analysis indicated that the inhibition of AtPME3 could occur via the interaction of AtPMEI7 with a PME ligand-binding cleft structure. All of these data indicate that AtPME3 and AtPMEI7 could be partners involved in the fine tuning of HG methylesterification during plant development. 相似文献
73.
Expression of a bacterial 3‐dehydroshikimate dehydratase reduces lignin content and improves biomass saccharification efficiency 下载免费PDF全文
Aymerick Eudes Noppadon Sathitsuksanoh Edward E. K. Baidoo Anthe George Yan Liang Fan Yang Seema Singh Jay D. Keasling Blake A. Simmons Dominique Loqué 《Plant biotechnology journal》2015,13(9):1241-1250
Lignin confers recalcitrance to plant biomass used as feedstocks in agro‐processing industries or as source of renewable sugars for the production of bioproducts. The metabolic steps for the synthesis of lignin building blocks belong to the shikimate and phenylpropanoid pathways. Genetic engineering efforts to reduce lignin content typically employ gene knockout or gene silencing techniques to constitutively repress one of these metabolic pathways. Recently, new strategies have emerged offering better spatiotemporal control of lignin deposition, including the expression of enzymes that interfere with the normal process for cell wall lignification. In this study, we report that expression of a 3‐dehydroshikimate dehydratase (QsuB from Corynebacterium glutamicum) reduces lignin deposition in Arabidopsis cell walls. QsuB was targeted to the plastids to convert 3‐dehydroshikimate – an intermediate of the shikimate pathway – into protocatechuate. Compared to wild‐type plants, lines expressing QsuB contain higher amounts of protocatechuate, p‐coumarate, p‐coumaraldehyde and p‐coumaryl alcohol, and lower amounts of coniferaldehyde, coniferyl alcohol, sinapaldehyde and sinapyl alcohol. 2D‐NMR spectroscopy and pyrolysis‐gas chromatography/mass spectrometry (pyro‐GC/MS) reveal an increase of p‐hydroxyphenyl units and a reduction of guaiacyl units in the lignin of QsuB lines. Size‐exclusion chromatography indicates a lower degree of lignin polymerization in the transgenic lines. Therefore, our data show that the expression of QsuB primarily affects the lignin biosynthetic pathway. Finally, biomass from these lines exhibits more than a twofold improvement in saccharification efficiency. We conclude that the expression of QsuB in plants, in combination with specific promoters, is a promising gain‐of‐function strategy for spatiotemporal reduction of lignin in plant biomass. 相似文献
74.
对低聚壳聚糖进行N-酰化改性,制得取代度相同的N-马来酰低聚壳聚糖(NMCOS),N-琥珀酰低聚壳聚糖(NSCOS),N-邻苯二甲酰低聚壳聚糖(NPCOS),其中NMCOS1、NSCOS1、NPCOS1的取代度均为0.25;NMCOS2、NSCOS2、NPCOS2的取代度均为0.49。考察了6种N-酰化低聚壳聚糖衍生物的还原能力。结果表明:当取代度相同时,N-邻苯二甲酰低聚壳聚糖的还原能力最强,其次是N-马来酰低聚壳聚糖,N-琥珀酰低聚壳聚糖的还原能力最差。这可能是由取代基的性质不同所致。 相似文献
75.
苏南地区典型城镇建设用地扩展的时空分异 总被引:3,自引:0,他引:3
选择江苏省常熟市南部的辛庄镇为研究区,采用该区1980、1991、2001和2009年4期高分辨率遥感影像,利用GIS技术,结合现有的建设用地扩展指标构建了综合扩展程度指数模型,定量分析了研究区1980-2009年间3个时段建设用地扩展的总体特征和时空分异特征.结果表明:随着农村城镇化和工业化进程的加快,辛庄镇建设用地显著扩展,1980-2009年间共增加19.24 km2,其中,2001-2009年该区建设用地进入高速扩展期,扩展面积、扩展贡献率和扩展强度均最大.研究区建设用地扩展具有明显的空间分异特征:1980-1991年,新增建设用地主要集中于镇区所在村域,1991年后,辛庄镇建设重心逐渐向工业发达的村庄转移.研究期间,辛庄镇的新增建设用地主要源于水田和旱地,二者共占建设用地总增加面积的88.1%,其余土地类型对建设用地的转入贡献率相对较小. 相似文献
76.
1949-2008年黄土高原沟壑区农业生态经济系统耦合分析——以陕西长武县为例 总被引:3,自引:0,他引:3
在分析农业经济系统与农业生态系统相互胁迫、动态演进关系的基础上,采用改进的熵值法确定了位于黄土高原沟壑区的长武县农业生态经济系统评价指标权重,利用耦合度模型分析了1949-2008年长武县农业生态经济系统耦合规律.结果表明:1949-2008年,长武县农业生态经济系统的发展经历了"粗放式传统农业阶段-农业机械化进程阶段-前现代化农业协调发展阶段"3个历史时期,与国家宏观经济政策调控效应基本吻合,系统耦合在"协调发展"和"极限发展"间经历了两起两落的急剧变化过程.1955-1961年、1984-1992年,长武县农业生态经济系统处于极限化耦合态势;1962-1965年、1981-1983年,为纠正前阶段宏观政策后的快速过渡期;1949-1954年、1966-1980年为生态系统与经济系统的低水平协调发展阶段;1993年以后,农业生态经济系统经过螺旋式发展,进入高水平协调发展阶段.目前,长武县农业生态经济系统有突破协调发展水平,并向极限化发展的潜在危险.良好的农业政策和外部环境能够促进农业生态经济系统在协调发展的水平上稳定发展,反之,容易出现系统相悖发展的局面. 相似文献
77.
Climate change effects on walnut pests in California 总被引:1,自引:0,他引:1
EIKE LUEDELING KIMBERLY P. STEINMANN MINGHUA ZHANG PATRICK H. BROWN JOSEPH GRANT EVAN H. GIRVETZ 《Global Change Biology》2011,17(1):228-238
Increasing temperatures are likely to impact ectothermic pests of fruits and nuts. This paper aims to assess changes to pest pressure in California's US$0.7 billion walnut industry due to recent historic and projected future temperature changes. For two past (1950 and 2000) and 18 future climate scenarios (2041–2060 and 2080–2099; each for three General Circulation Models and three greenhouse gas emissions scenarios), 100 years of hourly temperature were generated for 205 locations. Degree‐day models were used to project mean generation numbers for codling moth (Cydia pomonella L.), navel orangeworm (Amyelois transitella Walker), two‐spotted spider mite (Tetranychus urticae Koch), and European red mite (Panonychus ulmi Koch). In the Central Valley, the number of codling moth generations predicted for degree days accumulated between April 1 and October 1 rose from 2–4 in 1950 to 3–5 among all future scenarios. Generation numbers increased from 10–18 to 14–24 for two‐spotted spider mite, from 9–14 to 14–20 for European red mite, and from 2–4 to up to 5 for navel orangeworm. Overall pest pressure can thus be expected to increase substantially. Our study did not include the possibility of higher winter survival rates, leading to higher initial pest counts in spring, or of extended pest development times in the summer, factors that are likely to exacerbate future pest pressure. On the other hand, initiation of diapause may prevent an extension of the season length for arthropods, and higher incidence of heat death in summer may constrain pest population sizes. More information on the impact of climate change on complex agroecological food webs and on the response of pests to high temperatures is needed for improving the reliability of projections. 相似文献
78.
Pizzut-Serin S Potocki-Véronèse G van der Veen BA Albenne C Monsan P Remaud-Simeon M 《FEBS letters》2005,579(6):1405-1410
The BLAST search for amylosucrases has yielded several gene sequences of putative amylosucrases, however, with various questionable annotations. The putative encoded proteins share 32-48% identity with Neisseria polysaccharea amylosucrase (AS) and contain several amino acid residues proposed to be involved in AS specificity. First, the B-domains of the putative proteins and AS are highly similar. In addition, they also reveal additional residues between putative beta-strand 7 and alpha-helix 7 which could correspond to the AS B'-domain, which turns the active site into a deep pocket. Finally, conserved Asp and Arg residues could form a salt bridge similar to that found in AS, which is responsible for the glucosyl unit transfer specificity. Among these found genes, locus NP_294657.1 (dras) identified in the Deinococcus radiodurans genome was initially annotated as an alpha-amylase encoding gene. The putative encoded protein (DRAS) shares 42% identity with N. polysaccharea AS. To investigate the activity of this protein, gene NP_294657.1 was cloned and expressed in Escherichia coli. When acting on sucrose, the pure recombinant enzyme was shown to catalyse insoluble amylose polymer synthesis accompanied by side-reactions (sucrose hydrolysis, sucrose isomer and soluble maltooligosaccharide formation). Kinetic analyses further showed that DRAS follows a non-Michaelian behaviour toward sucrose substrate and is activated by glycogen, as is AS. This demonstrates that gene NP_294657.1 encodes an amylosucrase. 相似文献
79.
Impact and efficiency of GH10 and GH11 thermostable endoxylanases on wheat bran and alkali-extractable arabinoxylans 总被引:1,自引:0,他引:1
Beaugrand J Chambat G Wong VW Goubet F Rémond C Paës G Benamrouche S Debeire P O'Donohue M Chabbert B 《Carbohydrate research》2004,339(15):2529-2540
The results of a comparative study of two thermostable (1-->4)-beta-xylan endoxylanases using a multi-technical approach indicate that a GH11 xylanase is more useful than a GH10 xylanase for the upgrading of wheat bran into soluble oligosaccharides. Both enzymes liberated complex mixtures of xylooligosaccharides. 13C NMR analysis provided evidence that xylanases cause the co-solubilisation of beta-glucan, which is a result of cell-wall disassembly. The simultaneous use of both xylanases did not result in a synergistic action on wheat bran arabinoxylans, but instead led to the production of a product mixture whose profile resembled that produced by the action of the GH10 xylanase alone. Upon treatment with either xylanase, the diferulic acid levels in residual bran were unaltered, whereas content in ferulic and p-coumaric acids were unequally decreased. With regard to the major differences between the enzymes, the products resulting from the action of the GH10 xylanase were smaller in size than those produced by the GH11 xylanase, indicating a higher proportion of cleavage sites for the GH10 xylanase. The comparison of the kinetic parameters of each xylanase using various alkali-extractable arabinoxylans indicated that the GH10 xylanase was most active on soluble arabinoxylans. In contrast, probably because GH11 xylanase can better penetrate the cell-wall network, this enzyme was more efficient than the GH10 xylanase in the hydrolysis of wheat bran. Indeed the former enzyme displayed a nearly 2-fold higher affinity and a 6.8-fold higher turnover rate in the presence of this important by-product of the milling industry. 相似文献
80.
M. Abou Hachem S. Bozonnet M. Willemoë s B. Kramh ft K. Fukuda B. C. B nsager M. T. Jensen J. N hr S. Tranier N. Juge X. Robert R. Haser N. Aghajari B. Svensson 《Biocatalysis and Biotransformation》2006,24(1):83-93
-Amylases are endo-acting retaining enzymes of glycoside hydrolase family 13 with a catalytic (β/)8-domain containing an inserted loop referred to as domain B and a C-terminal anti-parallel β-sheet termed domain C. New insights integrate the roles of Ca2 + , different substrates, and proteinaceous inhibitors for -amylases. Isozyme specific effects of Ca2 + on the 80% sequence identical barley -amylases AMY1 and AMY2 are not obvious from the two crystal structures, containing three superimposable Ca2 + with identical ligands. A fully hydrated fourth Ca2 + at the interface of the AMY2/barley -amylase/subtilisin inhibitor (BASI) complex interacts with catalytic groups in AMY2, and Ca2 + occupies an identical position in AMY1 with thiomaltotetraose bound at two surface sites. EDTA-treatment, DSC, and activity assays indicate that AMY1 has the highest affinity for Ca2 + . Subsite mapping has revealed that AMY1 has ten functional subsites which can be modified by means protein engineering to modulate the substrate specificity. Other mutational analyses show that surface carbohydrate binding sites are critical for interaction with polysaccharides. The conserved Tyr380 in the newly discovered 'sugar tongs' site in domain C of AMY1 is thus critical for binding to starch granules. Furthermore, mutations of binding sites mostly reduced the degree of multiple attack in amylose hydrolysis. AMY1 has higher substrate affinity than AMY2, but isozyme chimeras with AMY2 domain C and other regions from AMY1 have higher substrate affinity than both parent isozymes. The latest revelations addressing various structural and functional aspects that govern the mode of action of barley -amylases are reported in this review. 相似文献