全文获取类型
收费全文 | 1519篇 |
免费 | 89篇 |
专业分类
1608篇 |
出版年
2024年 | 3篇 |
2023年 | 6篇 |
2022年 | 30篇 |
2021年 | 51篇 |
2020年 | 32篇 |
2019年 | 34篇 |
2018年 | 48篇 |
2017年 | 34篇 |
2016年 | 66篇 |
2015年 | 84篇 |
2014年 | 105篇 |
2013年 | 107篇 |
2012年 | 121篇 |
2011年 | 131篇 |
2010年 | 84篇 |
2009年 | 66篇 |
2008年 | 102篇 |
2007年 | 88篇 |
2006年 | 93篇 |
2005年 | 55篇 |
2004年 | 61篇 |
2003年 | 56篇 |
2002年 | 52篇 |
2001年 | 17篇 |
2000年 | 6篇 |
1999年 | 12篇 |
1998年 | 12篇 |
1997年 | 1篇 |
1996年 | 11篇 |
1995年 | 9篇 |
1994年 | 4篇 |
1993年 | 3篇 |
1992年 | 3篇 |
1990年 | 3篇 |
1989年 | 3篇 |
1987年 | 1篇 |
1985年 | 2篇 |
1984年 | 2篇 |
1980年 | 1篇 |
1978年 | 1篇 |
1977年 | 1篇 |
1976年 | 1篇 |
1975年 | 2篇 |
1974年 | 1篇 |
1972年 | 1篇 |
1971年 | 1篇 |
1963年 | 1篇 |
排序方式: 共有1608条查询结果,搜索用时 15 毫秒
71.
The Impact of Genetic Removal of GFAP and/or Vimentin on Glutamine Levels and Transport of Glucose and Ascorbate in Astrocytes 总被引:12,自引:0,他引:12
Pekny Milos Eliasson Camilla Siushansian Ramin Ding Mei Dixon S. Jeffrey Pekna Marcela Wilson John X. Hamberger Anders 《Neurochemical research》1999,24(11):1357-1362
The importance of the intermediate filament (IF) proteins glial fibrillary acidic protein (GFAP) and vimentin for astrocyte function was studied by investigating astrocytes prepared from GFAP-/-and/or vimentin-/- mice. The rate of glucose uptake through facilitative hexose transporters was not affected by depletion of GFAP or vimentin. Similarly, the absence of these IF proteins did not affect ascorbate uptake, under control or cyclic AMP-stimulated conditions, or ascorbate efflux through volume-sensitive organic anion channels. However, compared with wild-type astrocytes, glutamine concentrations were increased up to 200% in GFAP-/- astrocytes and up to 150% in GFAP+/-astrocytes and this increase was not dependent on the presence of vimentin. GFAP-/- astrocytes in culture still contain IFs (made of vimentin and nestin), whereas GFAP-/-vim-/- cultured astrocytes lack IFs. Thus, glutamine levels appear to correlate inversely with GFAP, rather than depend on the presence of IFs per se. Furthermore, the effect of GFAP is dose-dependent since the glutamine concentration in GFAP+/- astrocytes falls between those in wild-type and GFAP-/-astrocytes. 相似文献
72.
Arabidopsis thaliana defective in induced resistance 1 (At-DIR1) has been characterized as a protein responsible for the generation or transmission of the still unknown signal involved in systemic acquired resistance. This acidic apoplastic protein is a member of the family of lipid transfer proteins and was detected in vascular fluids. To our knowledge, no DIR1-like protein has been described in other plant species. Hence, we have performed data mining to identify a putative ortholog of DIR1 in tomato. This strategy allowed the detection of a few gene products displaying sequence similarity to At-DIR1 whose structural features were further analysed in silico. The best match (unigene SGN-327306) encoded a protein with an acidic pI, a peculiar characteristic of DIR1 among lipid transfer proteins, and was hence selected as a putative tomato ortholog of At-DIR1. This sequence, named Le-DIR1, served for the design of a specific antigenic peptide and the generation of polyclonal antibodies. The antiserum anti-Le-DIR1 recognized a peptide of the expected size (7kDa) in phloem sap of tomato plants, hence confirming the existence of the predicted protein in vascular fluids. This result supports the notion of the existence of common systemic acquired resistance (SAR) signaling molecules in different species. 相似文献
73.
Claudine Dherin Emeric Gueneau Mathilde Francin Marcela Nunez Simona Miron Sascha Emilie Liberti Lene Juel Rasmussen Sophie Zinn-Justin Bernard Gilquin Jean-Baptiste Charbonnier Serge Boiteux 《Molecular and cellular biology》2009,29(3):907-918
Mlh1 is an essential factor of mismatch repair (MMR) and meiotic recombination. It interacts through its C-terminal region with MutL homologs and proteins involved in DNA repair and replication. In this study, we identified the site of yeast Mlh1 critical for the interaction with Exo1, Ntg2, and Sgs1 proteins, designated as site S2 by reference to the Mlh1/Pms1 heterodimerization site S1. We show that site S2 is also involved in the interaction between human MLH1 and EXO1 or BLM. Binding at this site involves a common motif on Mlh1 partners that we called the MIP-box for the Mlh1 interacting protein box. Direct and specific interactions between yeast Mlh1 and peptides derived from Exo1, Ntg2, and Sgs1 and between human MLH1 and peptide derived from EXO1 and BLM were measured with Kd values ranging from 8.1 to 17.4 μM. In Saccharomyces cerevisiae, a mutant of Mlh1 targeted at site S2 (Mlh1-E682A) behaves as a hypomorphic form of Exo1. The site S2 in Mlh1 mediates Exo1 recruitment in order to optimize MMR-dependent mutation avoidance. Given the conservation of Mlh1 and Exo1 interaction, it may readily impact Mlh1-dependent functions such as cancer prevention in higher eukaryotes. 相似文献
74.
We compared the in vitro growth of promastigotes from two Leishmania species in TC-100 and Schneider media. Leishmania (Leishmania) amazonensis replication rates were similar in both tissue culture media and reached maximum rates by 48 h. In contrast Leishmania (Viannia) braziliensis growth was significantly greater in TC-100 but maximum rates were achieved by 96 h. Folic acid appears to be the limiting factor and supplementation of Schneider media with this nutrient improved L. (V.) braziliensis replication rates and decreased the time of maximum replication to 48 h. 相似文献
75.
Katarina J?rlestedt Catherine I. Rousset Maryam Faiz Ulrika Wilhelmsson Anders St?hlberg Hana Sourkova Marcela Pekna Carina Mallard Henrik Hagberg Milos Pekny 《PloS one》2010,5(4)
Background
Astroglial cells are activated following injury and up-regulate the expression of the intermediate filament proteins glial fibrillary acidic protein (GFAP) and vimentin. Adult mice lacking the intermediate filament proteins GFAP and vimentin (GFAP−/−Vim−/−) show attenuated reactive gliosis, reduced glial scar formation and improved regeneration of neuronal synapses after neurotrauma. GFAP−/−Vim−/− mice exhibit larger brain infarcts after middle cerebral artery occlusion suggesting protective role of reactive gliosis after adult focal brain ischemia. However, the role of astrocyte activation and reactive gliosis in the injured developing brain is unknown.Methodology/Principal Findings
We subjected GFAP−/−Vim−/− and wild-type mice to unilateral hypoxia-ischemia (HI) at postnatal day 9 (P9). Bromodeoxyuridine (BrdU; 25 mg/kg) was injected intraperitoneally twice daily from P9 to P12. On P12 and P31, the animals were perfused intracardially. Immunohistochemistry with MAP-2, BrdU, NeuN, and S100 antibodies was performed on coronal sections. We found no difference in the hemisphere or infarct volume between GFAP−/−Vim−/− and wild-type mice at P12 and P31, i.e. 3 and 22 days after HI. At P31, the number of NeuN+ neurons in the ischemic and contralateral hemisphere was comparable between GFAP−/−Vim−/− and wild-type mice. In wild-type mice, the number of S100+ astrocytes was lower in the ipsilateral compared to contralateral hemisphere (65.0±50.1 vs. 85.6±34.0, p<0.05). In the GFAP−/−Vim−/− mice, the number of S100+ astrocytes did not differ between the ischemic and contralateral hemisphere at P31. At P31, GFAP−/−Vim−/− mice showed an increase in NeuN+BrdU+ (surviving newly born) neurons in the ischemic cortex compared to wild-type mice (6.7±7.7; n = 29 versus 2.9±3.6; n = 28, respectively, p<0.05), but a comparable number of S100+BrdU+ (surviving newly born) astrocytes.Conclusions/Significance
Our results suggest that attenuation of reactive gliosis in the developing brain does not affect the hemisphere or infarct volume after HI, but increases the number of surviving newborn neurons. 相似文献76.
The amount of iron within the cell is carefully regulated in order to provide an adequate level of micronutrient while preventing its accumulation and toxicity. Iron excess is believed to generate oxidative stress, understood as an increase in the steady-state concentration of oxygen radical intermediates. Nitric oxide (NO) is an inorganic free-radical gaseous molecule which has been shown over the last decade to play an unprecedented variety of roles in biological systems. The effect of nitrogen reactive species may explain the iron sequestration pattern that characterizes macrophages under inflammatory conditions. From a patho-physiological viewpoint, further studies are required to assess the usefulness of this mechanism to minimize formation and release of free radicals in diseased tissues. However, contrary to the deleterious effects of the reactive nitrogen oxide species formed from either NO/O(2) and NO/O(2)(-), it has been pointed out that NO shows antioxidant properties. A number of studies have described the complex relationships between iron and NO, but controversy remains as to the influence and significance of iron on inflammatory NO production. To explore the initial steps of the effects triggered by LPS administration in the presence of excess iron, male Wistar rats were treated with: lipopolysaccharide from Escherichia coli (serotype 0127:B8) (LPS); iron-dextran; or iron-dextran plus LPS and liver samples were taken after 6 h. EPR spectra of NO-Hb in the venous blood were determined at 77 K. Iron-dextran administered to rats intraperitoneally resulted predominantly in iron uptake by the liver Kupffer cells and led to an increased NO level in blood in the presence of LPS. Further studies will be required to assess the complex role of the Kupffer cells on iNOS induction and NO production. 相似文献
77.
Norma Binsztein Marcela C. Costagliola Mariana Pichel Vernica Jurquiza Fernando C. Ramírez Rut Akselman Marta Vacchino Anwarul Huq Rita Colwell 《Applied microbiology》2004,70(12):7481-7486
In Argentina, as in other countries of Latin America, cholera has occurred in an epidemic pattern. Vibrio cholerae O1 is native to the aquatic environment, and it occurs in both culturable and viable but nonculturable (VNC) forms, the latter during interepidemic periods. This is the first report of the presence of VNC V. cholerae O1 in the estuarine and marine waters of the Río de la Plata and the Argentine shelf of the Atlantic Ocean, respectively. Employing immunofluorescence and PCR methods, we were able to detect reservoirs of V. cholerae O1 carrying the virulence-associated genes ctxA and tcpA. The VNC forms of V. cholerae O1 were identified in samples of water, phytoplankton, and zooplankton; the latter organisms were mainly the copepods Acartia tonsa, Diaptomus sp., Paracalanus crassirostris, and Paracalanus parvus. We found that under favorable conditions, the VNC form of V. cholerae can revert to the pathogenic, transmissible state. We concluded that V. cholerae O1 is a resident of Argentinean waters, as has been shown to be the case in other geographic regions of the world. 相似文献
78.
José Ronnie Vasconcelos Oscar Bru?a–Romero Adriano F. Araújo Mariana R. Dominguez Jonatan Ersching Bruna C. G. de Alencar Alexandre V. Machado Ricardo T. Gazzinelli Karina R. Bortoluci Gustavo P. Amarante-Mendes Marcela F. Lopes Mauricio M. Rodrigues 《PLoS pathogens》2012,8(5)
MHC class Ia-restricted CD8+ T cells are important mediators of the adaptive immune response against infections caused by intracellular microorganisms. Whereas antigen-specific effector CD8+ T cells can clear infection caused by intracellular pathogens, in some circumstances, the immune response is suboptimal and the microorganisms survive, causing host death or chronic infection. Here, we explored the cellular and molecular mechanisms that could explain why CD8+ T cell-mediated immunity during infection with the human protozoan parasite Trypanosoma cruzi is not optimal. For that purpose, we compared the CD8+ T-cell mediated immune responses in mice infected with T. cruzi or vaccinated with a recombinant adenovirus expressing an immunodominant parasite antigen. Several functional and phenotypic characteristics of specific CD8+ T cells overlapped. Among few exceptions was an accelerated expansion of the immune response in adenoviral vaccinated mice when compared to infected ones. Also, there was an upregulated expression of the apoptotic-signaling receptor CD95 on the surface of specific T cells from infected mice, which was not observed in the case of adenoviral-vaccinated mice. Most importantly, adenoviral vaccine provided at the time of infection significantly reduced the upregulation of CD95 expression and the proapoptotic phenotype of pathogen-specific CD8+ cells expanded during infection. In parallel, infected adenovirus-vaccinated mice had a stronger CD8 T-cell mediated immune response and survived an otherwise lethal infection. We concluded that a suboptimal CD8+ T-cell response is associated with an upregulation of CD95 expression and a proapoptotic phenotype. Both can be blocked by adenoviral vaccination. 相似文献
79.
80.
Aerobic bacteria, such as Burkholderia xenovorans LB400, are able to degrade a wide range of polychlorobiphenyls (PCBs). Generally, these bacteria are not able to transform
chlorobenzoates (CBAs), which accumulate during PCB degradation. In this study, the effects of CBAs on the growth, the morphology
and the proteome of Burkholderia
xenovorans LB400 were analysed. 4-CBA and 2-CBA were observed to inhibit the growth of strain LB400 on glucose. Strain LB400 exposed
to 4-CBA exhibited increased number and size of electron-dense granules in the cytoplasm, which could be polyphosphates. Two-dimensional
(2-D) polyacrylamide gel electrophoresis was used to characterise the molecular response of strain LB400 to 4-CBA. This compound
induced the enzymes BenD and CatA of benzoate and catechol catabolic pathways. The induction of molecular chaperones DnaK
and HtpG by 4-CBA indicated that the exposure to this compound constitutes a stressful condition for this bacterium. Additionally,
the induction of some Krebs cycle enzymes was observed, probably as response to cellular energy requirements. This study contributes
to the knowledge on the effects of CBA on the PCB-degrader Burkholderia xenovorans LB400. 相似文献