全文获取类型
收费全文 | 2906篇 |
免费 | 158篇 |
出版年
2023年 | 10篇 |
2022年 | 42篇 |
2021年 | 48篇 |
2020年 | 31篇 |
2019年 | 61篇 |
2018年 | 82篇 |
2017年 | 62篇 |
2016年 | 84篇 |
2015年 | 129篇 |
2014年 | 151篇 |
2013年 | 250篇 |
2012年 | 226篇 |
2011年 | 243篇 |
2010年 | 144篇 |
2009年 | 130篇 |
2008年 | 204篇 |
2007年 | 157篇 |
2006年 | 166篇 |
2005年 | 137篇 |
2004年 | 125篇 |
2003年 | 110篇 |
2002年 | 93篇 |
2001年 | 26篇 |
2000年 | 22篇 |
1999年 | 23篇 |
1998年 | 21篇 |
1997年 | 25篇 |
1996年 | 20篇 |
1995年 | 16篇 |
1994年 | 16篇 |
1993年 | 13篇 |
1992年 | 22篇 |
1991年 | 20篇 |
1990年 | 19篇 |
1989年 | 17篇 |
1988年 | 16篇 |
1987年 | 12篇 |
1986年 | 9篇 |
1985年 | 12篇 |
1984年 | 12篇 |
1983年 | 8篇 |
1982年 | 4篇 |
1980年 | 3篇 |
1979年 | 6篇 |
1977年 | 4篇 |
1974年 | 3篇 |
1973年 | 4篇 |
1970年 | 4篇 |
1969年 | 4篇 |
1968年 | 4篇 |
排序方式: 共有3064条查询结果,搜索用时 369 毫秒
961.
Pufferfishes of the Family Tetraodontidae are the most speciose group in the Order Tetraodontiformes and mainly inhabit coastal waters along continents. Although no members of other tetraodontiform families have fully discarded their marine lives, approximately 30 tetraodontid species spend their entire lives in freshwaters in disjunct tropical regions of South America, Central Africa, and Southeast Asia. To investigate the interrelationships of tetraodontid pufferfishes and thereby elucidate the evolutionary origins of their freshwater habitats, we performed phylogenetic analysis based on whole mitochondrial genome sequences from 50 tetraodontid species and closely related species (including 31 newly determined sequences). The resulting phylogenies reveal that the family is composed of four major lineages and that freshwater species from the different continents are independently nested in two of the four lineages. A monophyletic origin of the use of freshwater habitats was statistically rejected, and ancestral habitat reconstruction on the resulting tree demonstrates that tetraodontids independently entered freshwater habitats in different continents at least three times. Relaxed molecular-clock Bayesian divergence time estimation suggests that the timing of these invasions differs between continents, occurring at 0-10 million years ago (MA) in South America, 17-38 MA in Central Africa, and 48-78 MA in Southeast Asia. These timings are congruent with geological events that could facilitate adaptation to freshwater habitats in each continent. 相似文献
962.
Background
Influenza virus infection causes highly contagious, severe respiratory disorders and gives rise to thousands of deaths every year; however, the efficacy of currently approved defense strategies, including vaccines and neuraminidase inhibitors, is limited because the virus frequently acquires resistance via antigen drift and reassortment. It is therefore important to establish a novel, effective therapeutic strategy that is effective irrespective of viral subtype.Methodology/Principal Findings
Here, we identify the Ras–phosphoinositide 3-kinase (PI3K) signaling pathway as a host-cell regulatory mechanism for influenza virus entry. The binding of Ras to PI3K is specifically involved in clathrin-independent endocytosis, endosomal maturation, and intracellular transport of viruses, which result in decreased infectious efficacy of different subtypes of influenza viruses in cells lacking the Ras–PI3K interaction. Moreover, influenza virus infection indeed triggered Ras activation and subsequent PI3K activation in early endosomes.Conclusions/Significance
Taken together, these results demonstrate that the Ras–PI3K signaling axis acts as a host-oriented mechanism for viral internalization. Given that virus incorporation is a process conserved among virus subtypes and species, this signaling pathway may provide a target for potent, well-tolerated prophylactics and therapeutics against a broad range of viruses. 相似文献963.
Maasa Tsuge Yusuke MasudaHidenori Kaneoka Shunsuke KidaniKatsuhide Miyake Shinji Iijima 《Biochemical and biophysical research communications》2013
Damaged DNA-binding protein (DDB) is a heterodimer composed of two subunits, p127 and p48, which have been designated DDB1 and DDB2, respectively. DDB2 recognizes and binds to UV-damaged DNA during nucleotide excision repair. Here, we demonstrated that DDB2 was SUMOylated in a UV-dependent manner, and its major SUMO E3 ligase was PIASy as determined by RNA interference-mediated knockdown. The UV-induced physical interaction between DDB2 and PIASy supported this notion. PIASy knockdown reduced the removal of cyclobutane pyrimidine dimers (CPDs) from total genomic DNA, but did not affect that of 6-4 pyrimidine pyrimidone photoproducts (6-4PPs). Thus, DDB2 plays an indispensable role in CPD repair, but not in 6-4PP repair, which is consistent with the observation that DDB2 was SUMOylated by PIASy. These results suggest that the SUMOylation of DDB2 facilitates CPD repair. 相似文献
964.
PingGen Cai Yusuke Mizutani Masahiro Tsuchiya John M. Maloney Ben Fabry Krystyn J. Van Vliet Takaharu Okajima 《Biophysical journal》2013
Among individual cells of the same source and type, the complex shear modulus G∗ exhibits a large log-normal distribution that is the result of spatial, temporal, and intrinsic variations. Such large distributions complicate the statistical evaluation of pharmacological treatments and the comparison of different cell states. However, little is known about the characteristic features of cell-to-cell variation. In this study, we investigated how this variation depends on the spatial location within the cell and on the actin filament cytoskeleton, the organization of which strongly influences cell mechanics. By mechanically probing fibroblasts arranged on a microarray, via atomic force microscopy, we observed that the standard deviation σ of G∗ was significantly reduced among cells in which actin filaments were depolymerized. The parameter σ also exhibited a subcellular spatial dependence. Based on our findings regarding the frequency dependence of σ of the storage modulus G′, we proposed two types of cell-to-cell variation in G′ that arise from the purely elastic and the frequency-dependent components in terms of the soft glassy rheology model of cell deformability. We concluded that the latter inherent cell-to-cell variation can be reduced greatly by disrupting actin networks, by probing at locations within the cell nucleus boundaries distant from the cell center, and by measuring at high loading frequencies. 相似文献
965.
Yoko Aoki Tetsuya Niihori Toshihiro Banjo Nobuhiko Okamoto Seiji Mizuno Kenji Kurosawa Tsutomu Ogata Fumio Takada Michihiro Yano Toru Ando Tadataka Hoshika Christopher Barnett Hirofumi Ohashi Hiroshi Kawame Tomonobu Hasegawa Takahiro Okutani Tatsuo Nagashima Satoshi Hasegawa Ryo Funayama Takeshi Nagashima Keiko Nakayama Shin-ichi Inoue Yusuke Watanabe Toshihiko Ogura Yoichi Matsubara 《American journal of human genetics》2013,93(1):173-180
RAS GTPases mediate a wide variety of cellular functions, including cell proliferation, survival, and differentiation. Recent studies have revealed that germline mutations and mosaicism for classical RAS mutations, including those in HRAS, KRAS, and NRAS, cause a wide spectrum of genetic disorders. These include Noonan syndrome and related disorders (RAS/mitogen-activated protein kinase [RAS/MAPK] pathway syndromes, or RASopathies), nevus sebaceous, and Schimmelpenning syndrome. In the present study, we identified a total of nine missense, nonsynonymous mutations in RIT1, encoding a member of the RAS subfamily, in 17 of 180 individuals (9%) with Noonan syndrome or a related condition but with no detectable mutations in known Noonan-related genes. Clinical manifestations in the RIT1-mutation-positive individuals are consistent with those of Noonan syndrome, which is characterized by distinctive facial features, short stature, and congenital heart defects. Seventy percent of mutation-positive individuals presented with hypertrophic cardiomyopathy; this frequency is high relative to the overall 20% incidence in individuals with Noonan syndrome. Luciferase assays in NIH 3T3 cells showed that five RIT1 alterations identified in children with Noonan syndrome enhanced ELK1 transactivation. The introduction of mRNAs of mutant RIT1 into 1-cell-stage zebrafish embryos was found to result in a significant increase of embryos with craniofacial abnormalities, incomplete looping, a hypoplastic chamber in the heart, and an elongated yolk sac. These results demonstrate that gain-of-function mutations in RIT1 cause Noonan syndrome and show a similar biological effect to mutations in other RASopathy-related genes. 相似文献
966.
Hideaki Nakamura Daisuke HattoriDaisuke Tokunaga Yusuke Suzuki 《Analytical biochemistry》2013,441(2):140-146
A simple and rapid isothermal absorptiometric assay for detection of viable microbes using the redox color indicator 2,6-dichlorophenolindophenol (DCPIP) was studied. The absorbance of DCPIP decreased at 600 nm because of a redox reaction occurring between DCPIP and the surface membrane of viable microbes and was inversely proportional to the viable cell density. The redox reaction was found not only with bacteria, but also with yeast and a mixture of bacteria and yeast. In this assay, the influence of light scattering and absorption caused by microbial cells and coexisting substances in the sample was excluded by a time difference method. The assay required only 10 min for one incubation mixture, and highly repeatable results from three consecutive measurements were obtained by isothermal incubation for specific times at 30 °C using a thermostable three-cuvette-stir system. Thus, the cell density of microbial cell suspensions or growth medium was successfully determined, and a practical lower detection limit for food inspection was obtained at 104–106 cfu/ml. Single-cell effects on DCPIP reduction were evaluated and compared between species. Consequently, this assay is expected to be a useful tool for the rapid measurement of viable microbes as a preliminary assay for the Hazard Analysis Critical Control Point program. 相似文献
967.
Yusuke Kato Hisashi Yagi Yuichi Kaji Tetsuro Oshika Yuji Goto 《The Journal of biological chemistry》2013,288(35):25109-25118
Corneal dystrophies are genetic disorders resulting in progressive corneal clouding due to the deposition of amyloid fibrils derived from keratoepithelin, also called transforming growth factor β-induced protein (TGFBI). The formation of amyloid fibrils is often accelerated by surfactants such as sodium dodecyl sulfate (SDS). Most eye drops contain benzalkonium chloride (BAC), a cationic surfactant, as a preservative substance. In the present study, we aimed to reveal the role of BAC in the amyloid fibrillation of keratoepithelin-derived peptides in vitro. We used three types of 22-residue synthetic peptides covering Leu110-Glu131 of the keratoepithelin sequence: an R-type peptide with wild-type R124, a C-type peptide with C124 associated with lattice corneal dystrophy type I, and a H-type peptide with H124 associated with granular corneal dystrophy type II. The time courses of spontaneous amyloid fibrillation and seed-dependent fibril elongation were monitored in the presence of various concentrations of BAC or SDS using thioflavin T fluorescence. BAC and SDS accelerated the fibrillation of all synthetic peptides in the absence and presence of seeds. Optimal acceleration occurred near the CMC, which suggests that the unstable and dynamic interactions of keratoepithelin peptides with amphipathic surfactants led to the formation of fibrils. These results suggest that eye drops containing BAC may deteriorate corneal dystrophies and that those without BAC are preferred especially for patients with corneal dystrophies. 相似文献
968.
Mai Nakashima Yuta Shirogane Takao Hashiguchi Yusuke Yanagi 《The Journal of biological chemistry》2013,288(12):8085-8091
Measles virus (MV), an enveloped RNA virus belonging to the Paramyxoviridae family, enters the cell through membrane fusion mediated by two viral envelope proteins, an attachment protein hemagglutinin (H) and a fusion (F) protein. The crystal structure of the receptor-binding head domain of MV-H bound to its cellular receptor revealed that the MV-H head domain forms a tetrameric assembly (dimer of dimers), which occurs in two forms (forms I and II). In this study, we show that mutations in the putative dimer-dimer interface of the head domain in either form inhibit the ability of MV-H to support membrane fusion, without greatly affecting its cell surface expression, receptor binding, and interaction with the F protein. Notably, some anti-MV-H neutralizing monoclonal antibodies are directed to the region around the dimer-dimer interface in form I rather than receptor-binding sites. These observations suggest that the dimer-dimer interactions of the MV-H head domain, especially that in form I, contribute to triggering membrane fusion, and that conformational shift of head domain tetramers plays a role in the process. Furthermore, our results indicate that although the stalk and transmembrane regions may be mainly responsible for the tetramer formation of MV-H, the head domain alone can form tetramers, albeit at a low efficiency. 相似文献
969.
Jun Zhang Yusuke Nakatsu Takanori Shinjo Ying Guo Hideyuki Sakoda Takeshi Yamamotoya Yuichiro Otani Hirofumi Okubo Akifumi Kushiyama Midori Fujishiro Toshiaki Fukushima Yoshihiro Tsuchiya Hideaki Kamata Misaki Iwashita Fusanori Nishimura Hideki Katagiri Shin-ichiro Takahashi Hiroki Kurihara Takafumi Uchida Tomoichiro Asano 《The Journal of biological chemistry》2013,288(28):20692-20701
Pin1 and Par14 are parvulin-type peptidyl-prolyl cis/trans isomerases. Although numerous proteins have been identified as Pin1 substrates, the target proteins of Par14 remain largely unknown. Par14 expression levels are increased in the livers and embryonic fibroblasts of Pin1 KO mice, suggesting a compensatory relationship between the functions of Pin1 and Par14. In this study, the association of Par14 with insulin receptor substrate 1 (IRS-1) was demonstrated in HepG2 cells overexpressing both as well as endogenously in the mouse liver. The analysis using deletion-mutated Par14 and IRS-1 constructs revealed the N-terminal portion containing the basic domain of Par14 and the two relatively C-terminal portions of IRS-1 to be involved in these associations, in contrast to the WW domain of Pin1 and the SAIN domain of IRS-1. Par14 overexpression in HepG2 markedly enhanced insulin-induced IRS-1 phosphorylation and its downstream events, PI3K binding with IRS-1 and Akt phosphorylation. In contrast, treating HepG2 cells with Par14 siRNA suppressed these events. In addition, overexpression of Par14 in the insulin-resistant ob/ob mouse liver by adenoviral transfer significantly improved hyperglycemia with normalization of hepatic PEPCK and G6Pase mRNA levels, and gene suppression of Par14 using shRNA adenovirus significantly exacerbated the glucose intolerance in Pin1 KO mice. Therefore, although Pin1 and Par14 associate with different portions of IRS-1, the prolyl cis/trans isomerization in multiple sites of IRS-1 by these isomerases appears to be critical for efficient insulin receptor-induced IRS-1 phosphorylation. This process is likely to be one of the major mechanisms regulating insulin sensitivity and also constitutes a potential therapeutic target for novel insulin-sensitizing agents. 相似文献
970.
Atsushi Kuno Yusuke S. Hori Ryusuke Hosoda Masaya Tanno Tetsuji Miura Kazuaki Shimamoto Yoshiyuki Horio 《The Journal of biological chemistry》2013,288(8):5963-5972
Cardiomyopathy is the main cause of death in Duchenne muscular dystrophy. Here, we show that oral administration of resveratrol, which leads to activation of an NAD+-dependent protein deacetylase SIRT1, suppresses cardiac hypertrophy and fibrosis and restores cardiac diastolic function in dystrophin-deficient mdx mice. The pro-hypertrophic co-activator p300 protein but not p300 mRNA was up-regulated in the mdx heart, and resveratrol administration down-regulated the p300 protein level. In cultured cardiomyocytes, cardiomyocyte hypertrophy induced by the α1-agonist phenylephrine was inhibited by the overexpression of SIRT1 as well as resveratrol, both of which down-regulated p300 protein levels but not p300 mRNA levels. In addition, activation of atrial natriuretic peptide promoter by p300 was inhibited by SIRT1. We found that SIRT1 induced p300 down-regulation via the ubiquitin-proteasome pathway by deacetylation of lysine residues for ubiquitination. These findings indicate the pathological significance of p300 up-regulation in the dystrophic heart and indicate that SIRT1 activation has therapeutic potential for dystrophic cardiomyopathy. 相似文献