首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   13649篇
  免费   1041篇
  国内免费   859篇
  15549篇
  2024年   32篇
  2023年   141篇
  2022年   344篇
  2021年   621篇
  2020年   380篇
  2019年   500篇
  2018年   441篇
  2017年   338篇
  2016年   500篇
  2015年   833篇
  2014年   889篇
  2013年   1070篇
  2012年   1234篇
  2011年   1052篇
  2010年   647篇
  2009年   579篇
  2008年   782篇
  2007年   611篇
  2006年   619篇
  2005年   493篇
  2004年   415篇
  2003年   350篇
  2002年   293篇
  2001年   270篇
  2000年   223篇
  1999年   221篇
  1998年   155篇
  1997年   127篇
  1996年   140篇
  1995年   129篇
  1994年   117篇
  1993年   94篇
  1992年   135篇
  1991年   117篇
  1990年   81篇
  1989年   102篇
  1988年   60篇
  1987年   48篇
  1986年   58篇
  1985年   64篇
  1984年   18篇
  1983年   24篇
  1982年   28篇
  1981年   12篇
  1980年   25篇
  1979年   20篇
  1978年   16篇
  1977年   19篇
  1974年   9篇
  1972年   13篇
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
91.
摘要:【目的】结合纳米技术建立检测大肠杆菌(Escherichia coli)O157︰H7高灵敏检测技术。【方法】采用化学共沉淀法制备出核心粒径约为10 nm的免疫纳米磁颗粒,柠檬酸钠还原法制备粒径约为20 nm的免疫胶体金。压电免疫传感器通过金黄色葡萄球菌蛋白A(Protein A from Staphylococcus aureus SPA)法将抗体固定于石英晶振上,两种免疫纳米颗粒借助不同的抗体连接于传感器上对检测频率信号进行放大。【结果】SPA在石英晶振上的最佳固定浓度和时间为1.2 mg/mL和40 min,抗体的最佳固定浓度和时间为1.0 mg/mL和60 min。压电免疫传感器通过两种免疫纳米颗粒的放大作用,使其对大肠杆菌O157︰H7的检测限从104 cfu/mL提高到101 cfu/mL。【结论】免疫纳米颗粒强化对压电免疫传感器的检测频率信号具有很好的放大效应,可以明显提高其检测灵敏度。  相似文献   
92.
The abundance–adaptation hypothesis argues that taxa with more individuals and faster generation times will have more evolutionary ‘experiments’ allowing expansion into, and diversification within, novel habitats. Thus, as older taxa have produced more individuals over time, and smaller taxa have higher population sizes and faster generation times, the Latitudinal Diversity Gradients (LDGs) of these clades should show shallower slopes. We describe the LDGs for archaea, bacteria, fungi, invertebrates and trees from six North American forests. For three focal groups – bacteria, ants, and trees – older taxa had shallower LDG slopes than the more recent, terminal taxa. Across 12 orders of magnitude of body mass, LDG slopes were steeper in larger taxa. The slopes of LDGs vary systematically with body size and clade age, underscoring the non‐canonical nature of LDGs. The steepest LDG slopes were found for the largest organisms while the smallest, from bacteria to small litter‐soil invertebrates, have shallower‐ to zero‐slope LDGs. If tropical niche conservatism is the failure of clades to adapt to, and diversify in temperate habitats, then the steep LDGs of chordates and plants likely arise from the decreased ability of clades with large individuals to adapt to the multiple challenges of extra‐tropical life.  相似文献   
93.
Quorum sensing (QS) is a ubiquitous cell–cell communication mechanism in microbes that coordinates population‐level cell behaviors, such as biofilm production, virulence, swarming motility, and bacterial persistence. Efforts to engineer QS systems to take part in metabolic network regulation represent a promising strategy for synthetic biology and pathway engineering. Recently, design, construction, and implementation of QS circuits for programmed control of bacterial phenotypes and metabolic pathways have gained much attention, but have not been reviewed recently. In this article, the architectural organizations and genetic contributions of the naturally occurring QS components to understand the mechanisms are summarized. Then, the most recent progress in application of QS toolkits to develop synthetic networks for novel cell behaviors creation and metabolic pathway engineering is highlighted. The current challenges in large‐scale application of these QS circuits in synthetic biology and metabolic engineering fields are discussed and future perspectives for further engineering efforts are provided.  相似文献   
94.
Lipoxygenases (LOXs) are enzymes involved in lipid peroxidation. Here we reported the identification, molecular and functional characterization of the gene encoding rice (Oryza sativa L.) seed LOX3 (sLOX3). Via a map-based cloning strategy we identified Os03g0700400 as the candidate gene encoding sLOX3. Further functional complementary test and biochemical characterization of the recombinant Os03g0700400 protein verified the identification. The sLOX3 gene was highly expressed in roots, moderately in embryos and very weakly in leaves, leaf sheaths and stems. Transient expression experiment (in rice protoplasts) and subsequent laser confocal microscopic analysis demonstrated that the sLOX3 protein was localized into the cytosol. We next showed that overexpression of sLOX3 in a japonica sLOX3-normal rice cultivar, Wuyunjing 7 accelerated the decrease of seed germination ability when the seeds were routinely stored, which demonstrated that sLOX3 had a negative effect on seed longevity (storability). Meanwhile, an increased occurrence of embryo decay was observed in the same transgenic seeds, suggesting that sLOX3 might negatively affect seed longevity by facilitating colonization of particular seed pathogens. Our result forwarded the understanding of the effects of 9-LOX on rice seed longevity.  相似文献   
95.
滞育和非滞育棉铃虫前胸腺的形态解剖学比较研究   总被引:4,自引:0,他引:4  
该文用解剖镜和电子显微镜对滞育和非滞育棉铃虫Helicoverpa armigera前胸腺的形态解剖结构进行了比较研究。结果发现滞育棉铃虫的前胸腺细胞及其细胞间隙相对较小,不易着色,细胞核规则,较小,细胞质中几乎见不到光滑内质网和粗面内质网,线粒体极少,这些观察到的现象充分说明滞育棉铃虫的前胸腺活性较低。  相似文献   
96.
Microbes are key components of the soil environment, playing an important role in maintaining soil health, sustainability, and productivity. The composition and structure of soil bacterial communities were examined in winter wheat–rice (WR) and winter wheat–maize (WM) cropping systems derived from five locations in the Low-Middle Yangtze River plain and the Huang-Huai-Hai plain by pyrosequencing of the 16S ribosomal RNA gene amplicons. A total of 102,367 high quality sequences were used for multivariate statistical analysis and to test for correlation between community structure and environmental variables such as crop rotations, soil properties, and locations. The most abundant phyla across all soil samples were Proteobacteria, Acidobacteria, and Bacteroidetes. Similar patterns of bacterial diversity and community structure were observed within the same cropping systems, and a higher relative abundance of anaerobic bacteria was found in WR compared to WM cropping systems. Variance partitioning analysis revealed complex relationships between bacterial community and environmental variables. The effect of crop rotations was low but significant, and interactions among soil properties, locations, and crop rotations accounted for most of the explained variation in the structure of bacterial communities. Soil properties such as pH, available P, and available K showed higher correlations (positive or negative) with the majority of the abundant taxa. Bacterial diversity (the Shannon index) and richness (Chao1 and ACE) were higher under WR than WM cropping systems.  相似文献   
97.
Filamin-A cross-links actin filaments into dynamic orthogonal networks, and interacts with an array of proteins of diverse cellular functions. Because several filamin-A interaction partners are implicated in signaling of cell mobility regulation, we tested the hypothesis that filamin-A plays a role in cancer metastasis. Using four pairs of filamin-A proficient and deficient isogenic cell lines, we found that filamin-A deficiency in cancer cells significantly reduces their migration and invasion. Using a xenograft tumor model with subcutaneous and intracardiac injections of tumor cells, we found that the filamin-A deficiency causes significant reduction of lung, splenic and systemic metastasis in nude mice. We evaluated the expression of filamin-A in breast cancer tissues by immunohistochemical staining, and found that low levels of filamin-A expression in cancer cells of the tumor tissues are associated with a better distant metastasis-free survival than those with normal levels of filamin-A. These data not only validate filamin-A as a prognostic marker for cancer metastasis, but also suggest that inhibition of filamin-A in cancer cells may reduce metastasis and that filamin-A can be used as a therapeutic target for filamin-A positive cancer.  相似文献   
98.
陕西食源性沙门氏菌耐药及相关基因   总被引:10,自引:0,他引:10  
【目的】研究食源性沙门氏菌对常用抗生素的药敏性及相关耐药基因,更好的了解耐药性的产生和传播途径,确保食品安全。【方法】使用the Clinical and Laboratory Standards Institute推荐的琼脂稀释法测定沙门氏菌的药敏性,PCR和基因序列测定方法确定耐药沙门氏菌中整合子及其携带的耐药基因、与头孢菌素抗性相关的基因、沙门氏菌基因岛及与氟喹诺酮类抗生素耐药相关的基因突变。【结果】359株沙门氏菌中,67%的菌株对磺胺甲恶唑产生抗性,对甲氧苄啶/磺胺甲恶唑、四环素、卡那霉素、萘啶酮酸、氨苄西林、阿莫西林/克拉维酸、链霉素、氯霉素和庆大霉素、环丙沙星、头孢曲松、头孢西丁和头孢哌酮的耐药率分别为58%、56%、37%、35%、33%、32%、29%、26%、21%、16%、9%和8%。284株耐药菌中,79%的菌株可抗至少1种抗生素,25.9%可抗10种以上抗生素,2.5%可抗14种抗生素。耐药的Ⅰ类整合子以1.4kb最为常见,携带的耐药基因有aadA1、aadA2、aadA5、tetR、blaPSE-1、blaDHA-1、blaVEB-1、dhfrⅠ、dhfrⅤ、dhfrⅦ和dhfr17等。62株耐头孢曲松和/或头孢哌酮的沙门氏菌中,blaTEM和blaCMY-2基因的检出率分别为51.6%和56.5%。13.6%的沙门氏菌中检出了沙门氏菌基因岛。35株耐氟喹诺酮类抗生素的沙门氏菌的gyrA、parC和parE基因中共检出68个点突变,gyrA基因中常见突变为Ser83Phe、Ser83Tyr、Asp87Gly和Asp87Asn,parC基因中为Ser80Arg。parE基因中检出了Lys441Ile、Lys428Gln、Asp494Asn、Lys428Gln和Gly442Ser突变,这些点突变均为首次在食源性沙门氏菌中检出。【结论】陕西食源性沙门氏菌耐药状况严重,整合子、沙门氏菌基因岛和β-内酰胺酶编码基因的存在及解旋酶和拓扑异构酶基因突变是导致沙门氏菌耐药的重要机制。  相似文献   
99.
Riboprinting was used to determine the relationships among strains belonging to 15 species of the genusKluyveromyces. The small subunit ribosomal RNA gene (SSU rDNA) was amplified using the Polymerase Chain Reaction (PCR) and subjected to a battery of nine restriction enzymes. Similarity coefficients between strains were calculated based on shared and unique restriction fragments. Cluster analysis revealed three major groups that generally correlated with previously reported relationships based on other molecular data. Variations in SSU rDNA restriction fragments may be used for differentiation of theKluyveromyces strains included in this study.The U.S. Government right to retain a non-exclusive, royalty free licence in and to any copyright is acknowledged.  相似文献   
100.
木本植物木质部空穴和栓塞化研究(综述)   总被引:10,自引:0,他引:10  
木本植物木质部空穴和栓塞化现象的发现至今已近80年历史,国外学者对此做了较多的研究,国内此方面的研究极少。木质部空穴和栓塞化研究是木本植物体内水分传输研究的前沿所在,不同学者在不同地区对不同材料的研究结果各异,争议颇多。本文对这一研究方向近年来的资料作了概括和总结,包括栓塞化现象的发现、检测方法、诱因及形成机理、木质部栓塞化同输水结构间的关系等方面。  相似文献   
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号