全文获取类型
收费全文 | 2328篇 |
免费 | 179篇 |
国内免费 | 2篇 |
专业分类
2509篇 |
出版年
2023年 | 13篇 |
2022年 | 28篇 |
2021年 | 52篇 |
2020年 | 22篇 |
2019年 | 26篇 |
2018年 | 56篇 |
2017年 | 42篇 |
2016年 | 58篇 |
2015年 | 104篇 |
2014年 | 96篇 |
2013年 | 159篇 |
2012年 | 150篇 |
2011年 | 191篇 |
2010年 | 114篇 |
2009年 | 114篇 |
2008年 | 145篇 |
2007年 | 120篇 |
2006年 | 116篇 |
2005年 | 122篇 |
2004年 | 122篇 |
2003年 | 118篇 |
2002年 | 93篇 |
2001年 | 15篇 |
2000年 | 23篇 |
1999年 | 24篇 |
1998年 | 28篇 |
1997年 | 20篇 |
1996年 | 24篇 |
1995年 | 22篇 |
1994年 | 14篇 |
1993年 | 23篇 |
1992年 | 14篇 |
1991年 | 11篇 |
1990年 | 13篇 |
1989年 | 9篇 |
1988年 | 14篇 |
1987年 | 13篇 |
1986年 | 11篇 |
1985年 | 13篇 |
1984年 | 15篇 |
1983年 | 15篇 |
1982年 | 15篇 |
1981年 | 11篇 |
1980年 | 15篇 |
1979年 | 14篇 |
1978年 | 9篇 |
1977年 | 6篇 |
1976年 | 12篇 |
1975年 | 9篇 |
1973年 | 5篇 |
排序方式: 共有2509条查询结果,搜索用时 15 毫秒
21.
Modulation of DNA end joining by nuclear proteins 总被引:6,自引:0,他引:6
Liang L Deng L Chen Y Li GC Shao C Tischfield JA 《The Journal of biological chemistry》2005,280(36):31442-31449
DNA double strand breaks in mammalian cells are primarily repaired by homologous recombination and non-homologous end joining (NHEJ). NHEJ may either be error-free or mutagenic with deletions or insertions at the joint. Recent studies showed that DNA ends can also be joined via microhomologous sequences flanking the break point especially when proteins responsible for NHEJ, such as Ku, are absent. Microhomology-mediated end joining (MHEJ) is always accompanied by a deletion that spans one of the two homologous sequences and the intervening sequence, if any. In this study we evaluated several factors affecting the relative contribution of MHEJ to DNA end joining using nuclear extracts and DNA substrates containing 10-bp repeats at the ends. We found that the occurrence of MHEJ is determined by the relative abundance of nuclear proteins. At low DNA/protein ratios, an error-free end-joining mechanism predominated over MHEJ. As the DNA/protein ratio increased, MHEJ became predominant. We show that the nuclear proteins that contribute to the inhibition of the error-prone MHEJ include Ku and histone H1. Treatment of extracts with flap endonuclease 1 antiserum significantly reduced MHEJ. Addition of a 17-bp intervening sequence between the microhomologous sequences significantly reduced the efficiency of MHEJ. Thus, this cell-free assay provides a platform for evaluating factors modulating end joining. 相似文献
22.
Here, we report the systematic exploration and modeling of interactions between light and sugar signaling. The data set analyzed explores the interactions of sugar (sucrose) with distinct light qualities (white, blue, red, and far-red) used at different fluence rates (low or high) in etiolated seedlings and mature green plants. Boolean logic was used to model the effect of these carbon/light interactions on three target genes involved in nitrogen assimilation: asparagine synthetase (ASN1 and ASN2) and glutamine synthetase (GLN2). This analysis enabled us to assess the effects of carbon on light-induced genes (GLN2/ASN2) versus light-repressed genes (ASN1) in this pathway. New interactions between carbon and blue-light signaling were discovered, and further connections between red/far-red light and carbon were modeled. Overall, light was able to override carbon as a major regulator of ASN1 and GLN2 in etiolated seedlings. By contrast, carbon overrides light as the major regulator of GLN2 and ASN2 in light-grown plants. Specific examples include the following: Carbon attenuated the blue-light induction of GLN2 in etiolated seedlings and also attenuated the white-, blue-, and red-light induction of GLN2 and ASN2 in light-grown plants. By contrast, carbon potentiated far-red-light induction of GLN2 and ASN2 in light-grown plants. Depending on the fluence rate of far-red light, carbon either attenuated or potentiated light repression of ASN1 in light-grown plants. These studies indicate the interaction of carbon with blue, red, and far-red-light signaling and set the stage for further investigation into modeling this complex web of interacting pathways using systems biology approaches. 相似文献
23.
Carla Malaquias Almeida Jos A. Manso Ana C. Figueiredo Liliana Antunes Rui Cruz Bruno Manadas Daniel Bur Pedro Jos Barbosa Pereira Carlos Faro Isaura Simes 《Applied microbiology and biotechnology》2017,101(18):6951-6968
The potential of using a synthetic cardosin-based rennet in cheese manufacturing was recently demonstrated with the development and optimization of production of a recombinant form of cardosin B in Kluyveromyces lactis. With the goal of providing a more detailed characterization of this rennet, we herein evaluate the impact of the plant-specific insert (PSI) on cardosin B secretion in this yeast, and provide a thorough analysis of the specificity requirements as well as the biochemical and structural properties of the isolated recombinant protease. We demonstrate that the PSI domain can be substituted by different linker sequences without substantially affecting protein secretion and milk clotting activity. However, the presence of small portions of the PSI results in dramatic reductions of secretion yields in this heterologous system. Kinetic characterization and specificity profiling results clearly suggest that synthetic cardosin B displays lower catalytic efficiency and is more sequence selective than native cardosin B. Elucidation of the structure of synthetic cardosin B confirms the canonical fold of an aspartic protease with the presence of two high mannose-type, N-linked glycan structures; however, there are some differences in the conformation of the flap region when compared to cardosin A. These subtle variations in catalytic properties and the more stringent substrate specificity of synthetic cardosin B help to explain the observed suitability of this rennet for cheese production. 相似文献
24.
L Pereira R Zamudio G Soares-Souza P Herrera L Cabrera CC Hooper J Cok JM Combe G Vargas WA Prado S Schneider F Kehdy MR Rodrigues SJ Chanock DE Berg RH Gilman E Tarazona-Santos 《PloS one》2012,7(8):e41200
Gastric cancer is one of the most lethal types of cancer and its incidence varies worldwide, with the Andean region of South America showing high incidence rates. We evaluated the genetic structure of the population from Lima (Peru) and performed a case-control genetic association study to test the contribution of African, European, or Native American ancestry to risk for gastric cancer, controlling for the effect of non-genetic factors. A wide set of socioeconomic, dietary, and clinic information was collected for each participant in the study and ancestry was estimated based on 103 ancestry informative markers. Although the urban population from Lima is usually considered as mestizo (i.e., admixed from Africans, Europeans, and Native Americans), we observed a high fraction of Native American ancestry (78.4% for the cases and 74.6% for the controls) and a very low African ancestry (<5%). We determined that higher Native American individual ancestry is associated with gastric cancer, but socioeconomic factors associated both with gastric cancer and Native American ethnicity account for this association. Therefore, the high incidence of gastric cancer in Peru does not seem to be related to susceptibility alleles common in this population. Instead, our result suggests a predominant role for ethnic-associated socioeconomic factors and disparities in access to health services. Since Native Americans are a neglected group in genomic studies, we suggest that the population from Lima and other large cities from Western South America with high Native American ancestry background may be convenient targets for epidemiological studies focused on this ethnic group. 相似文献
25.
Abstract: In an article on the role of temporal information in life-cycle assessment in this journal, Field and colleagues argued that frequently it is not the single product but the "fleet" (or cohort) of products that "is the appropriate unit of analysis," and that in focusing on the fleet one "explicitly introduces the notion of time as a critical element of comparative life-cycle assessments. …" Major transitions, such as replacement of one fleet of products by an alternative fleet, correspond to a system in a transient rather than steady state, and explicit consideration of time is central to transient analysis.
One tool increasingly used as part of life-cycle assessment, economic input-output (EIO) analysis, at best deals with time in an implicit fashion. This article illustrates how the sequential interindustry model (SIM), a formulation of the EIOmodel that explicitly represents time, might be utilized in life-cycle assessment. SIM introduces this temporal component by explicitly accounting for the time required by production activities and the resulting sequencing of the inputs. This can be thought of as engineering rather than accounting information. The data demands of such a model are not likely to be met at present or at any time in the near future. Even so, simulation methods and the use of so-called synthetic data have a history of productive use in a number of fields, including the social sciences.
SIM also utilizes the contribution of Joshi on the application of the EIO model to environmental impact and the inclusion of the use as well as the production phases of a product in EIO analysis. The possibility of accounting for discounting of future events, with its impact on decision making, is also briefly discussed. 相似文献
One tool increasingly used as part of life-cycle assessment, economic input-output (EIO) analysis, at best deals with time in an implicit fashion. This article illustrates how the sequential interindustry model (SIM), a formulation of the EIOmodel that explicitly represents time, might be utilized in life-cycle assessment. SIM introduces this temporal component by explicitly accounting for the time required by production activities and the resulting sequencing of the inputs. This can be thought of as engineering rather than accounting information. The data demands of such a model are not likely to be met at present or at any time in the near future. Even so, simulation methods and the use of so-called synthetic data have a history of productive use in a number of fields, including the social sciences.
SIM also utilizes the contribution of Joshi on the application of the EIO model to environmental impact and the inclusion of the use as well as the production phases of a product in EIO analysis. The possibility of accounting for discounting of future events, with its impact on decision making, is also briefly discussed. 相似文献
26.
Koenitzer JR Isbell TS Patel HD Benavides GA Dickinson DA Patel RP Darley-Usmar VM Lancaster JR Doeller JE Kraus DW 《American journal of physiology. Heart and circulatory physiology》2007,292(4):H1953-H1960
Hydrogen sulfide (H(2)S) has recently been shown to have a signaling role in vascular cells. Similar to nitric oxide (NO), H(2)S is enzymatically produced by amino acid metabolism and can cause posttranslational modification of proteins, particularly at thiol residues. Molecular targets for H(2)S include ATP-sensitive K(+) channels, and H(2)S may interact with NO and heme proteins such as cyclooxygenase. It is well known that the reactions of NO in the vasculature are O(2) dependent, but this has not been addressed in most studies designed to elucidate the role of H(2)S in vascular function. This is important, since H(2)S reactions can be dramatically altered by the high concentrations of O(2) used in cell culture and organ bath experiments. To test the hypothesis that the effects of H(2)S on the vasculature are O(2) dependent, we have measured real-time levels of H(2)S and O(2) in respirometry and vessel tension experiments, as well as the associated vascular responses. A novel polarographic H(2)S sensor developed in our laboratory was used to measure H(2)S levels. Here we report that, in rat aorta, H(2)S concentrations that mediate rapid contraction at high O(2) levels cause rapid relaxation at lower physiological O(2) levels. At high O(2), the vasoconstrictive effect of H(2)S suggests that it may not be H(2)S per se but, rather, a putative vasoactive oxidation product that mediates constriction. These data are interpreted in terms of the potential for H(2)S to modulate vascular tone in vivo. 相似文献
27.
28.
Lupi A Messana I Denotti G Schininà ME Gambarini G Fadda MB Vitali A Cabras T Piras V Patamia M Cordaro M Giardina B Castagnola M 《Proteomics》2003,3(4):461-467
Human salivary cystatins, five major (S, S1, S2, SA, SN) and two minor (C and D), are multifunctional proteins playing a different role in the oral environment. Salivary cystatin SN is able to effectively inhibit lysosomal cathepsins B, C, H and L and cystatin SA inhibits cathepsins C and L in vitro. These activities suggest, particularly for cystatin SN, an important role in the control of proteolytic events in vivo. Differently, cystatins S are involved, together with statherin, in the mineral balance of the tooth. Due to their distinct role, a reliable method for identification and quantification of the different cystatins, as well as of possible truncated and derived forms, could be helpful for the assessment of the status of the oral cavity. To this purpose high-performance liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI MS) was applied to the analysis of human saliva obtained from healthy subjects. All known salivary cystatins, with the exception of cystatin C, were detected. Strong evidence was also obtained for the presence in saliva of post-translational modified isoforms of cystatins, which may be related to donor habits. Cystatin SN and cystatins S, S1 and S2 were well separated by HPLC-ESI MS coupling from other components and thus this approach can be successfully applied to their quantification. 相似文献
29.
CpG-DNA and its related synthetic CpG oligodeoxynucleotides (CpG-ODNs) play an important role in immune cell survival. It has been suggested that Akt is one of the CpG-DNA-responsive serine/threonine kinases; however, the target protein of CpG-DNA that leads to Akt activation has not been elucidated. Here, we report that ex vivo stimulation of bone marrow-derived macrophages (BMDMs) from mice lacking the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) results in defective phosphorylation and activation of Akt by CpG-DNA. Unexpectedly, loss of the Toll-like receptor 9 has a minimal effect on Akt activation in response to CpG-DNA. Further in vitro analysis using purified DNA-PK and recombinant Akt proteins reveals that DNA-PK directly induces phosphorylation and activation of Akt. In addition, in BMDMs, DNA-PKcs associates with Akt upon CpG-DNA stimulation and triggers transient nuclear translocation of Akt. Thus, our findings establish a novel role for DNA-PKcs in CpG-DNA signaling and define a CpG-DNA/DNA-PKcs/Akt pathway. 相似文献
30.