全文获取类型
收费全文 | 2226篇 |
免费 | 117篇 |
出版年
2023年 | 6篇 |
2022年 | 23篇 |
2021年 | 43篇 |
2020年 | 15篇 |
2019年 | 26篇 |
2018年 | 35篇 |
2017年 | 30篇 |
2016年 | 44篇 |
2015年 | 74篇 |
2014年 | 71篇 |
2013年 | 155篇 |
2012年 | 154篇 |
2011年 | 152篇 |
2010年 | 94篇 |
2009年 | 92篇 |
2008年 | 154篇 |
2007年 | 174篇 |
2006年 | 144篇 |
2005年 | 120篇 |
2004年 | 177篇 |
2003年 | 130篇 |
2002年 | 121篇 |
2001年 | 26篇 |
2000年 | 25篇 |
1999年 | 21篇 |
1998年 | 29篇 |
1997年 | 21篇 |
1996年 | 13篇 |
1995年 | 16篇 |
1994年 | 10篇 |
1993年 | 13篇 |
1992年 | 13篇 |
1991年 | 20篇 |
1990年 | 8篇 |
1989年 | 9篇 |
1988年 | 18篇 |
1987年 | 4篇 |
1986年 | 10篇 |
1985年 | 3篇 |
1984年 | 4篇 |
1983年 | 3篇 |
1982年 | 4篇 |
1980年 | 3篇 |
1979年 | 4篇 |
1977年 | 2篇 |
1976年 | 6篇 |
1975年 | 5篇 |
1972年 | 2篇 |
1969年 | 3篇 |
1967年 | 5篇 |
排序方式: 共有2343条查询结果,搜索用时 437 毫秒
71.
Polyglutamine (polyQ) expansion mutation causes conformational, neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases. These diseases are characterized by the aggregation of misfolded proteins, such as amyloid fibrils, which are toxic to cells. Amyloid fibrils are formed by a nucleated growth polymerization reaction. Unexpectedly, the critical nucleus of polyQ aggregation was found to be a monomer, suggesting that the rate-limiting nucleation process of polyQ aggregation involves the folding of mutated protein monomers. The monoclonal antibody 1C2 selectively recognizes expanded pathogenic and aggregate-prone glutamine repeats in polyQ diseases, including Huntington's disease (HD), as well as binding to polyleucine. We have therefore assayed the in vitro and in vivo aggregation kinetics of these monomeric proteins. We found that the repeat-length-dependent differences in aggregation lag times of variable lengths of polyQ and polyleucine tracts were consistently related to the integration of the length-dependent intensity of anti-1C2 signal on soluble monomers of these proteins. Surprisingly, the correlation between the aggregation lag times of polyQ tracts and the intensity of anti-1C2 signal on soluble monomers of huntingtin precisely reflected the repeat-length dependent age-of-onset of HD patients. These data suggest that the alterations in protein surface structure due to polyQ expansion mutation in soluble monomers of the mutated proteins act as an amyloid-precursor epitope. This, in turn, leads to nucleation, a key process in protein aggregation, thereby determining HD onset. These findings provide new insight into the gain-of-function mechanisms of polyQ diseases, in which polyQ expansion leads to nucleation rather than having toxic effects on the cells. 相似文献
72.
Isawa H Orito Y Jingushi N Iwanaga S Morita A Chinzei Y Yuda M 《The FEBS journal》2007,274(16):4271-4286
Two plasma kallikrein-kinin system inhibitors in the salivary glands of the kissing bug Triatoma infestans, designated triafestin-1 and triafestin-2, have been identified and characterized. Reconstitution experiments showed that triafestin-1 and triafestin-2 inhibit the activation of the kallikrein-kinin system by inhibiting the reciprocal activation of factor XII and prekallikrein, and subsequent release of bradykinin. Binding analyses showed that triafestin-1 and triafestin-2 specifically interact with factor XII and high molecular weight kininogen in a Zn2+-dependent manner, suggesting that they specifically recognize Zn2+-induced conformational changes in factor XII and high molecular weight kininogen. Triafestin-1 and triafestin-2 also inhibit factor XII and high molecular weight kininogen binding to negatively charged surfaces. Furthermore, they interact with both the N-terminus of factor XII and domain D5 of high molecular weight kininogen, which are the binding domains for biological activating surfaces. These results suggest that triafestin-1 and triafestin-2 inhibit activation of the kallikrein-kinin system by interfering with the association of factor XII and high molecular weight kininogen with biological activating surfaces, resulting in the inhibition of bradykinin release in an animal host during insect blood-feeding. 相似文献
73.
BACKGROUND: Regulatory factors and detailed physiology of in vivo microcirculation have remained not fully clarified after many different modalities of imaging had invented. While many macroscopic parameters of blood flow reflect flow velocity, changes in blood flow velocity and red blood cell (RBC) flux does not hold linear relationship in the microscopic observations. There are reports of discrepancy between RBC velocity and RBC flux, RBC flux and plasma flow volume, and of spatial and temporal heterogeneity of flow regulation in the peripheral tissues in microscopic observations, a scientific basis for the requirement of more detailed studies in microcirculatory regulation using intravital microscopy. METHODS: We modified Jeff Lichtman''s method of in vivo microscopic observation of mouse sternomastoid muscles. Mice are anesthetized,
ventilated, and injected with PKH26L-fluorescently labeled RBCs for microscopic observation.RESULT & CONCLUSIONS: Fluorescently labeled RBCs are detected and distinguished well by a wide-field microscope. Muscle contraction evoked by electrical stimulation induced increase in RBC flux. Quantification of other parameters including RBC velocity and capillary density were feasible. Mice tolerated well the surgery, injection of stained RBCs, microscopic observation, and electrical stimulation. No muscle or blood vessel damage was observed, suggesting that our method is relatively less invasive and suited for long-term observations.Download video file.(92M, mpg) 相似文献
74.
Sugiyama N Masuda T Shinoda K Nakamura A Tomita M Ishihama Y 《Molecular & cellular proteomics : MCP》2007,6(6):1103-1109
We developed novel methods for phosphopeptide enrichment using aliphatic hydroxy acid-modified metal oxide chromatography (MOC). Titania and zirconia were successfully applied to enrich phosphopeptides with the aid of aliphatic hydroxy acids, such as lactic acid and beta-hydroxypropanoic acid, to reduce the interaction between acidic non-phosphopeptides and the metal oxides. These methods removed the vast majority of non-phosphopeptides from phosphoprotein standard digests, and large numbers of phosphopeptides could be readily identified. The methods were coupled with nano-LC-MS/MS systems without difficulty. Recovery of phosphopeptides in MOC varied greatly from peptide to peptide, ranging from a few percent to 100%, and the average was almost 50%. Repeatability and linearity were satisfactory. In an examination of the cytoplasmic fraction of HeLa cells, more than 1000 phosphopeptides were identified using lactic acid-modified titania MOC and beta-hydroxypropanoic acid-modified zirconia MOC, respectively. The overlap between phosphopeptides enriched by these two methods was 40%, and the combined results provided 1646 unique phosphopeptides. To our knowledge, this is the first successful application of a single MOC-based approach to phosphopeptide enrichment from complex biological samples such as cell lysates. 相似文献
75.
王明强 罗阿蓉 周青松 陈婧婷 谢婷婷 李逸 Douglas Chesters 石晓宇 肖晖 刘桓吉 丁强 周璇 罗一平 路园园 佟一杰 赵政宇 白明 郭鹏飞 陈思翀 中村彰宏 彭艳琼 赵延会 魏淑花 林晓龙 陈华燕 罗世孝 陆宴辉 鲁亮 余建平 周欣 邹怡 路浩 朱朝东 《生物多样性》2022,30(10):22454-236
当前, 全球昆虫数量和多样性均处于下降趋势, 而导致这一趋势的原因主要包括人为干扰及气候变化。本文基于森林、草地、农业、水生和土壤生态系统, 以植食性、访花、捕食性、寄生性、食果以及食腐昆虫为重点功能昆虫群, 综述了近三十年来国内外昆虫多样性研究领域的主要进展, 并分析了发展趋势。近年来, 昆虫多样性的研究维度不断拓展, 形态多样性研究不断深入, 系统发生多样性、功能多样性和遗传多样性等研究也显著加强。此外, 昆虫多样性研究的空间尺度也逐步扩大, 大尺度区域性研究甚至全球范围的调查持续增长。昆虫进化历史也被引入多样性格局研究中, 并随着系统发生信息学方法的普及而被整合到生态系统建成和生物多样性形成机制研究中。未来需要加强关键昆虫类群整合分类学研究、功能性状多样性、林冠昆虫多样性、互作网络结构等方向的研究。 相似文献
76.
77.
78.
Akihiro Hirata Yoko Miyamoto Akihisa Kaneko Hiroki Sakai Kyoko Yoshizaki Tokuma Yanai Takako Miyabe‐Nishiwaki Juri Suzuki 《Journal of medical primatology》2019,48(2):137-140
Primary neuroendocrine neoplasm of the liver is extremely rare in both humans and non‐human primates. The present report describes the clinical and pathological findings of an aged Japanese macaque (Macaca fuscata) with hepatic neuroendocrine carcinoma. To our knowledge, this is the first report of hepatic neuroendocrine neoplasm in macaques. 相似文献
79.
Sakamoto Kazunori Ogiwara Natsuko Kaji Tomomitsu Sugimoto Yurie Ueno Mitsuru Sonoda Masatoshi Matsui Akihiro Ishida Junko Tanaka Maho Totoki Yasushi Shinozaki Kazuo Seki Motoaki 《Journal of plant research》2019,132(4):541-568
Journal of Plant Research - Soybean (Glycine max) roots establish associations with nodule-inducing rhizobia and arbuscular mycorrhizal (AM) fungi. Both rhizobia and AM fungi have been shown to... 相似文献
80.
Akihiro Kuno Yoshihisa Ikeda Shinya Ayabe Kanako Kato Kotaro Sakamoto Sayaka R. Suzuki Kento Morimoto Arata Wakimoto Natsuki Mikami Miyuki Ishida Natsumi Iki Yuko Hamada Megumi Takemura Yoko Daitoku Yoko Tanimoto Tra Thi Huong Dinh Kazuya Murata Michito Hamada Masafumi Muratani Atsushi Yoshiki Fumihiro Sugiyama Satoru Takahashi Seiya Mizuno 《PLoS biology》2022,20(1)
Genome editing can introduce designed mutations into a target genomic site. Recent research has revealed that it can also induce various unintended events such as structural variations, small indels, and substitutions at, and in some cases, away from the target site. These rearrangements may result in confounding phenotypes in biomedical research samples and cause a concern in clinical or agricultural applications. However, current genotyping methods do not allow a comprehensive analysis of diverse mutations for phasing and mosaic variant detection. Here, we developed a genotyping method with an on-target site analysis software named Determine Allele mutations and Judge Intended genotype by Nanopore sequencer (DAJIN) that can automatically identify and classify both intended and unintended diverse mutations, including point mutations, deletions, inversions, and cis double knock-in at single-nucleotide resolution. Our approach with DAJIN can handle approximately 100 samples under different editing conditions in a single run. With its high versatility, scalability, and convenience, DAJIN-assisted multiplex genotyping may become a new standard for validating genome editing outcomes.Genome editing can introduce designed mutations into a target genomic site, but also into unintended off-target sites. DAJIN, a novel nanopore sequencing data analysis tool, identifies and quantifies allele numbers and their mutation patterns, reporting consensus sequences and visualizing mutations in alleles at single-nucleotide resolution. 相似文献