全文获取类型
收费全文 | 6266篇 |
免费 | 404篇 |
出版年
2023年 | 16篇 |
2022年 | 58篇 |
2021年 | 105篇 |
2020年 | 63篇 |
2019年 | 88篇 |
2018年 | 130篇 |
2017年 | 108篇 |
2016年 | 199篇 |
2015年 | 312篇 |
2014年 | 359篇 |
2013年 | 483篇 |
2012年 | 600篇 |
2011年 | 534篇 |
2010年 | 334篇 |
2009年 | 292篇 |
2008年 | 359篇 |
2007年 | 355篇 |
2006年 | 344篇 |
2005年 | 306篇 |
2004年 | 286篇 |
2003年 | 286篇 |
2002年 | 266篇 |
2001年 | 46篇 |
2000年 | 48篇 |
1999年 | 54篇 |
1998年 | 84篇 |
1997年 | 52篇 |
1996年 | 50篇 |
1995年 | 54篇 |
1994年 | 43篇 |
1993年 | 38篇 |
1992年 | 45篇 |
1991年 | 12篇 |
1990年 | 24篇 |
1989年 | 15篇 |
1988年 | 19篇 |
1987年 | 19篇 |
1986年 | 12篇 |
1985年 | 18篇 |
1984年 | 25篇 |
1983年 | 10篇 |
1982年 | 12篇 |
1981年 | 11篇 |
1980年 | 7篇 |
1978年 | 7篇 |
1977年 | 13篇 |
1976年 | 13篇 |
1975年 | 9篇 |
1974年 | 9篇 |
1966年 | 5篇 |
排序方式: 共有6670条查询结果,搜索用时 953 毫秒
171.
Fran?ois Téoulé Cynthia Brisac Isabelle Pelletier Pierre-Olivier Vidalain Sophie Jégouic Carmen Mirabelli Ma?l Bessaud Nicolas Combelas Arnaud Autret Frédéric Tangy Francis Delpeyroux Bruno Blondel 《Journal of virology》2013,87(20):11031-11046
We have shown that the circulating vaccine-derived polioviruses responsible for poliomyelitis outbreaks in Madagascar have recombinant genomes composed of sequences encoding capsid proteins derived from poliovaccine Sabin, mostly type 2 (PVS2), and sequences encoding nonstructural proteins derived from other human enteroviruses. Interestingly, almost all of these recombinant genomes encode a nonstructural 3A protein related to that of field coxsackievirus A17 (CV-A17) strains. Here, we investigated the repercussions of this exchange, by assessing the role of the 3A proteins of PVS2 and CV-A17 and their putative cellular partners in viral replication. We found that the Golgi protein acyl-coenzyme A binding domain-containing 3 (ACBD3), recently identified as an interactor for the 3A proteins of several picornaviruses, interacts with the 3A proteins of PVS2 and CV-A17 at viral RNA replication sites, in human neuroblastoma cells infected with either PVS2 or a PVS2 recombinant encoding a 3A protein from CV-A17 [PVS2-3A(CV-A17)]. The small interfering RNA-mediated downregulation of ACBD3 significantly increased the growth of both viruses, suggesting that ACBD3 slowed viral replication. This was confirmed with replicons. Furthermore, PVS2-3A(CV-A17) was more resistant to the replication-inhibiting effect of ACBD3 than the PVS2 strain, and the amino acid in position 12 of 3A was involved in modulating the sensitivity of viral replication to ACBD3. Overall, our results indicate that exchanges of nonstructural proteins can modify the relationships between enterovirus recombinants and cellular interactors and may thus be one of the factors favoring their emergence. 相似文献
172.
Lucas Sánchez-Sampedro Carmen Elena Gómez Ernesto Mejías-Pérez Eva Pérez-Jiménez Juan Carlos Oliveros Mariano Esteban 《Journal of virology》2013,87(12):6955-6974
Replication-competent poxvirus vectors with an attenuation phenotype and with a high immunogenic capacity of the foreign expressed antigen are being pursued as novel vaccine vectors against different pathogens. In this investigation, we have examined the replication and immunogenic characteristics of two vaccinia virus (VACV) mutants, M65 and M101. These mutants were generated after 65 and 101 serial passages of persistently infected Friend erythroleukemia (FEL) cells. In cultured cells of different origins, the mutants are replication competent and have growth kinetics similar to or slightly reduced in comparison with those of the parental Western Reserve (WR) virus strain. In normal and immune-suppressed infected mice, the mutants showed different levels of attenuation and pathogenicity in comparison with WR and modified vaccinia Ankara (MVA) strains. Wide genome analysis after deep sequencing revealed selected genomic deletions and mutations in a number of viral open reading frames (ORFs). Mice immunized in a DNA prime/mutant boost regimen with viral vectors expressing the LACK (Leishmania homologue for receptors of activated C kinase) antigen of Leishmania infantum showed protection or a delay in the onset of cutaneous leishmaniasis. Protection was similar to that triggered by MVA-LACK. In immunized mice, both polyfunctional CD4+ and CD8+ T cells with an effector memory phenotype were activated by the two mutants, but the DNA-LACK/M65-LACK protocol preferentially induced CD4+ whereas DNA-LACK/M101-LACK preferentially induced CD8+ T cell responses. Altogether, our findings showed the adaptive changes of the WR genome during long-term virus-host cell interaction and how the replication competency of M65 and M101 mutants confers distinct biological properties and immunogenicity in mice compared to those of the MVA strain. These mutants could have applicability for understanding VACV biology and as potential vaccine vectors against pathogens and tumors. 相似文献
173.
Isabel Marques Antonio José Díaz‐Pérez José Ángel López‐Rodríguez Victoria Mirones Ana Sus Félix Llamas Alicia Alonso Ernesto Pérez‐Collazos Juan Viruel Elvira Sahuquillo Maria Del Carmen Sancho Benjamin Komac José Antonio Manso José Gabriel Segarra‐Moragues David Draper Luis Villar Pilar Catalán 《Botanical journal of the Linnean Society. Linnean Society of London》2013,173(4):676-706
The Iberian mountain spiny fescues are a reticulate group of five diploid grass taxa consisting of three parental species and two putative hybrids: F. × souliei (F. eskia × F. quadriflora) and F. × picoeuropeana (F. eskia × F. gautieri). Phenotypic and molecular studies were conducted with the aim of determining the taxonomic boundaries and genetic relationships of the five taxa and disentangling the origins of the two hybrids. Statistical analyses of 31 selected phenotypic traits were conducted on individuals from 159 populations and on nine type specimens. Molecular analyses of random amplified polymorphic DNA (RAPD) markers were performed on 29 populations. The phenotypic analyses detected significant differences between the five taxa and demonstrated the overall intermediacy of the F. × picoeuropeana and F. × souliei between their respective parents. The RAPD analysis corroborated the genetic differentiation of F. eskia, F. gautieri and F. quadriflora and the intermediate nature of the two hybrids; however, they also detected genetic variation within F. × picoeuropeana. These results suggest distinct origins for F. × picoeuropeana in the Cantabrian and Pyrenean mountains, with the sporadic Pyrenean populations having potentially resulted from recent hybridizations and the stabilized Cantabrian ones from older events followed by potential displacements of the parents. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 676–706. 相似文献
174.
Carla Pérez-Cruz Ornella Carrión Lidia Delgado Gemma Martinez Carmen López-Iglesias Elena Mercade 《Applied and environmental microbiology》2013,79(6):1874-1881
Outer membrane vesicles (OMVs) from Gram-negative bacteria are known to be involved in lateral DNA transfer, but the presence of DNA in these vesicles has remained difficult to explain. An ultrastructural study of the Antarctic psychrotolerant bacterium Shewanella vesiculosa M7T has revealed that this Gram-negative bacterium naturally releases conventional one-bilayer OMVs through a process in which the outer membrane is exfoliated and only the periplasm is entrapped, together with a more complex type of OMV, previously undescribed, which on formation drag along inner membrane and cytoplasmic content and can therefore also entrap DNA. These vesicles, with a double-bilayer structure and containing electron-dense material, were visualized by transmission electron microscopy (TEM) after high-pressure freezing and freeze-substitution (HPF-FS), and their DNA content was fluorometrically quantified as 1.8 ± 0.24 ng DNA/μg OMV protein. The new double-bilayer OMVs were estimated by cryo-TEM to represent 0.1% of total vesicles. The presence of DNA inside the vesicles was confirmed by gold DNA immunolabeling with a specific monoclonal IgM against double-stranded DNA. In addition, a proteomic study of purified membrane vesicles confirmed the presence of plasma membrane and cytoplasmic proteins in OMVs from this strain. Our data demonstrate the existence of a previously unobserved type of double-bilayer OMV in the Gram-negative bacterium Shewanella vesiculosa M7T that can incorporate DNA, for which we propose the name outer-inner membrane vesicle (O-IMV). 相似文献
175.
Carmen R. Lugo-Lugo 《Ethnic and racial studies》2013,36(3):611-628
This paper intends to open a dialogue about the use of two specific labels in the US (i.e. ‘Chicanas’ and ‘Latinas’), with a special emphasis on how they are used in the US academy as both markers and identities. More specifically, this paper explores recent trends in the US academy by which women of Latin American descent are lumped together under the rubric Latinas, and in many cases assumed to be equivalent to Chicanas. Arguing that academia must learn to be more thoughtful when creating, defining and adopting categories, the article warns against recreating the very power dynamics we find in mainstream US society by way of these specific labels. Finally, ‘So you are a mestiza’ reminds its readers that, as a practice, social justice asks for a fundamental recognition of a culturally pluralist, democratic society. In this kind of society, histories must endure and multiple realities must be acknowledged. 相似文献
176.
177.
This investigation was designed to explore the relationships between lichen symbionts (phycobiont and mycobiont) and the substrate on which they grow by examining the chemical and ultrastructural features of the lichen-soil interface. These lichens form an integral part of microbiotic soil crusts. Fragments of three different lichen biotypes growing over gypsum crystals and marls were fixed and embedded in resin. The lichen-substratum interface was then examined by scanning electron microscopy with backscattered electron imaging. In situ observation, microanalytical (EDS), and FT-Raman plus infrared spectroscopy of the lichen-substratum interface indicated that different ultrastructural features of the mycobiont were related to biogeochemical processes and Ca 2+ distribution in the soil crust. Phycobionts were observed to make direct contact with the substratum and to be surrounded by a nondifferentiated thallus structure. These observations suggest that they can grow outside the thallus in the early stages of lichen development in the semi-arid conditions of their habitat. The particular ultrastructural features of the lichen thallus and of the lichen-substratum interface appear to have marked effects on runoff phenomena and ponding generation of the surface. 相似文献
178.
José M. Pastor Vicente Bernal Manuel Salvador Montserrat Argando?a Carmen Vargas Laszlo Csonka ángel Sevilla José L. Iborra Joaquín J. Nieto Manuel Cánovas 《The Journal of biological chemistry》2013,288(24):17769-17781
Bacterial osmoadaptation involves the cytoplasmic accumulation of compatible solutes to counteract extracellular osmolarity. The halophilic and highly halotolerant bacterium Chromohalobacter salexigens is able to grow up to 3 m NaCl in a minimal medium due to the de novo synthesis of ectoines. This is an osmoregulated pathway that burdens central metabolic routes by quantitatively drawing off TCA cycle intermediaries. Consequently, metabolism in C. salexigens has adapted to support this biosynthetic route. Metabolism of C. salexigens is more efficient at high salinity than at low salinity, as reflected by lower glucose consumption, lower metabolite overflow, and higher biomass yield. At low salinity, by-products (mainly gluconate, pyruvate, and acetate) accumulate extracellularly. Using [1-13C]-, [2-13C]-, [6-13C]-, and [U-13C6]glucose as carbon sources, we were able to determine the main central metabolic pathways involved in ectoines biosynthesis from glucose. C. salexigens uses the Entner-Doudoroff pathway rather than the standard glycolytic pathway for glucose catabolism, and anaplerotic activity is high to replenish the TCA cycle with the intermediaries withdrawn for ectoines biosynthesis. Metabolic flux ratios at low and high salinity were similar, revealing a certain metabolic rigidity, probably due to its specialization to support high biosynthetic fluxes and partially explaining why metabolic yields are so highly affected by salinity. This work represents an important contribution to the elucidation of specific metabolic adaptations in compatible solute-accumulating halophilic bacteria. 相似文献
179.
Carmen F. Ludwig Florian Ullrich Lilia Leisle Tobias Stauber Thomas J. Jentsch 《The Journal of biological chemistry》2013,288(40):28611-28619
CLC anion transporters form dimers that function either as Cl− channels or as electrogenic Cl−/H+ exchangers. CLC channels display two different types of “gates,” “protopore” gates that open and close the two pores of a CLC dimer independently of each other and common gates that act on both pores simultaneously. ClC-7/Ostm1 is a lysosomal 2Cl−/1H+ exchanger that is slowly activated by depolarization. This gating process is drastically accelerated by many CLCN7 mutations underlying human osteopetrosis. Making use of some of these mutants, we now investigate whether slow voltage activation of plasma membrane-targeted ClC-7/Ostm1 involves protopore or common gates. Voltage activation of wild-type ClC-7 subunits was accelerated by co-expressing an excess of ClC-7 subunits carrying an accelerating mutation together with a point mutation rendering these subunits transport-deficient. Conversely, voltage activation of a fast ClC-7 mutant could be slowed by co-expressing an excess of a transport-deficient mutant. These effects did not depend on whether the accelerating mutation localized to the transmembrane part or to cytoplasmic cystathionine-β-synthase (CBS) domains of ClC-7. Combining accelerating mutations in the same subunit did not speed up gating further. No currents were observed when ClC-7 was truncated after the last intramembrane helix. Currents and slow gating were restored when the C terminus was co-expressed by itself or fused to the C terminus of the β-subunit Ostm1. We conclude that common gating underlies the slow voltage activation of ClC-7. It depends on the CBS domain-containing C terminus that does not require covalent binding to the membrane domain of ClC-7. 相似文献
180.
Michela Campagna Laura Marcos-Villar Francesca Arnoldi Carlos F. de la Cruz-Herrera Pedro Gallego José González-Santamaría Dolores González Fernando Lopitz-Otsoa Manuel S. Rodriguez Oscar R. Burrone Carmen Rivas 《Journal of virology》2013,87(2):807-817
Posttranslational modification by SUMO provides functional flexibility to target proteins. Viruses interact extensively with the cellular SUMO modification system in order to improve their replication, and there are numerous examples of viral proteins that are SUMOylated. However, thus far the relevance of SUMOylation for rotavirus replication remains unexplored. In this study, we report that SUMOylation positively regulates rotavirus replication and viral protein production. We show that SUMO can be covalently conjugated to the viroplasm proteins VP1, VP2, NSP2, VP6, and NSP5. In addition, VP1, VP2, and NSP2 can also interact with SUMO in a noncovalent manner. We observed that an NSP5 SUMOylation mutant protein retains most of its activities, such as its interaction with VP1 and NSP2, the formation of viroplasm-like structures after the coexpression with NSP2, and the ability to complement in trans the lack of NSP5 in infected cells. However, this mutant is characterized by a high degree of phosphorylation and is impaired in the formation of viroplasm-like structures when coexpressed with VP2. These results reveal for the first time a positive role for SUMO modification in rotavirus replication, describe the SUMOylation of several viroplasm resident rotavirus proteins, and demonstrate a requirement for NSP5 SUMOylation in the production of viroplasm-like structures. 相似文献