全文获取类型
收费全文 | 1873篇 |
免费 | 117篇 |
国内免费 | 1篇 |
出版年
2023年 | 8篇 |
2022年 | 35篇 |
2021年 | 62篇 |
2020年 | 37篇 |
2019年 | 66篇 |
2018年 | 50篇 |
2017年 | 55篇 |
2016年 | 73篇 |
2015年 | 114篇 |
2014年 | 132篇 |
2013年 | 135篇 |
2012年 | 153篇 |
2011年 | 121篇 |
2010年 | 98篇 |
2009年 | 70篇 |
2008年 | 101篇 |
2007年 | 85篇 |
2006年 | 96篇 |
2005年 | 71篇 |
2004年 | 60篇 |
2003年 | 50篇 |
2002年 | 45篇 |
2001年 | 45篇 |
2000年 | 45篇 |
1999年 | 34篇 |
1998年 | 27篇 |
1997年 | 20篇 |
1996年 | 15篇 |
1995年 | 9篇 |
1994年 | 9篇 |
1993年 | 4篇 |
1992年 | 5篇 |
1991年 | 13篇 |
1990年 | 5篇 |
1989年 | 3篇 |
1986年 | 1篇 |
1985年 | 3篇 |
1984年 | 5篇 |
1983年 | 4篇 |
1982年 | 1篇 |
1981年 | 4篇 |
1980年 | 3篇 |
1979年 | 2篇 |
1978年 | 2篇 |
1977年 | 2篇 |
1975年 | 2篇 |
1974年 | 4篇 |
1972年 | 2篇 |
1969年 | 1篇 |
1968年 | 2篇 |
排序方式: 共有1991条查询结果,搜索用时 15 毫秒
121.
Marked seasonality and high spatial variability of protist communities in shallow freshwater systems
Marianne Simon Purificación López-García Philippe Deschamps David Moreira Gwendal Restoux Paola Bertolino Ludwig Jardillier 《The ISME journal》2015,9(9):1941-1953
Small eukaryotes have key roles in aquatic ecosystems, influencing their local environment, global biogeochemical cycles and climate. Their impact depends on community structure, which varies along time. However, very few studies take into account temporal variation. This is especially true for small, shallow freshwater systems, which remain largely understudied despite their wide variety, global surface and intense microbial activity. We have monthly followed changes in the community structure of small microbial eukaryotes (0.2–5 μm cell diameter) for 2 years in four ponds and one brook located in North-Western France based on massive 18S rDNA amplicon 454 pyrosequencing. We detected a total of 3742 stringently defined operational taxonomic units (OTUs) encompassing all recognized eukaryotic supergroups and lineages of uncertain affiliation. Although geographically close, protist communities in the five ecosystems were contrasting, with very few shared OTUs, suggesting that environmental selection mainly drives community structure. The temporal dynamics of different high-rank taxa appeared complex and rapid at monthly scales. Despite this, a clear and reproducible seasonality was observed. As expected, low-abundance OTUs dominated the community. Although some of them appeared sporadically or remained at low frequencies during the survey, others occasionally reached relatively high abundances, sometimes recurrently. This shows that at least a fraction of low-abundance eukaryotes constitutes a seed bank. The annual proportion of primary producers, free-living heterotrophs and parasites appeared remarkably constant among the different ecosystems, suggesting underlying trends of ecosystem carrying capacity for these functional groups. 相似文献
122.
Isabela Resende Pereira Glaucia Vilar-Pereira Virgínia Marques Andrea Alice da Silva Bráulia Caetano Otacilio Cruz Moreira Alexandre Vieira Machado Oscar Bruna-Romero Maurício Martins Rodrigues Ricardo Tostes Gazzinelli Joseli Lannes-Vieira 《PLoS pathogens》2015,11(1)
Chagas disease (CD), caused by the protozoan Trypanosoma cruzi, is a prototypical neglected tropical disease. Specific immunity promotes acute phase survival. Nevertheless, one-third of CD patients develop chronic chagasic cardiomyopathy (CCC) associated with parasite persistence and immunological unbalance. Currently, the therapeutic management of patients only mitigates CCC symptoms. Therefore, a vaccine arises as an alternative to stimulate protective immunity and thereby prevent, delay progression and even reverse CCC. We examined this hypothesis by vaccinating mice with replication-defective human Type 5 recombinant adenoviruses (rAd) carrying sequences of amastigote surface protein-2 (rAdASP2) and trans-sialidase (rAdTS) T. cruzi antigens. For prophylactic vaccination, naïve C57BL/6 mice were immunized with rAdASP2+rAdTS (rAdVax) using a homologous prime/boost protocol before challenge with the Colombian strain. For therapeutic vaccination, rAdVax administration was initiated at 120 days post-infection (dpi), when mice were afflicted by CCC. Mice were analyzed for electrical abnormalities, immune response and cardiac parasitism and tissue damage. Prophylactic immunization with rAdVax induced antibodies and H-2Kb-restricted cytotoxic and interferon (IFN)γ-producing CD8+ T-cells, reduced acute heart parasitism and electrical abnormalities in the chronic phase. Therapeutic vaccination increased survival and reduced electrical abnormalities after the prime (analysis at 160 dpi) and the boost (analysis at 180 and 230 dpi). Post-therapy mice exhibited less heart injury and electrical abnormalities compared with pre-therapy mice. rAdVax therapeutic vaccination preserved specific IFNγ-mediated immunity but reduced the response to polyclonal stimuli (anti-CD3 plus anti-CD28), CD107a+ CD8+ T-cell frequency and plasma nitric oxide (NO) levels. Moreover, therapeutic rAdVax reshaped immunity in the heart tissue as reduced the number of perforin+ cells, preserved the number of IFNγ+ cells, increased the expression of IFNγ mRNA but reduced inducible NO synthase mRNA. Vaccine-based immunostimulation with rAd might offer a rational alternative for re-programming the immune response to preserve and, moreover, recover tissue injury in Chagas’ heart disease. 相似文献
123.
Diana Moreira Vasco Rodrigues Maria Abengozar Luis Rivas Eduardo Rial Mireille Laforge Xiaoling Li Marc Foretz Benoit Viollet Jér?me Estaquier Anabela Cordeiro da Silva Ricardo Silvestre 《PLoS pathogens》2015,11(3)
Metabolic manipulation of host cells by intracellular pathogens is currently recognized to play an important role in the pathology of infection. Nevertheless, little information is available regarding mitochondrial energy metabolism in Leishmania infected macrophages. Here, we demonstrate that during L. infantum infection, macrophages switch from an early glycolytic metabolism to an oxidative phosphorylation, and this metabolic deviation requires SIRT1 and LKB1/AMPK. SIRT1 or LBK1 deficient macrophages infected with L. infantum failed to activate AMPK and up-regulate its targets such as Slc2a4 and Ppargc1a, which are essential for parasite growth. As a result, impairment of metabolic switch caused by SIRT1 or AMPK deficiency reduces parasite load in vitro and in vivo. Overall, our work demonstrates the importance of SIRT1 and AMPK energetic sensors for parasite intracellular survival and proliferation, highlighting the modulation of these proteins as potential therapeutic targets for the treatment of leishmaniasis. 相似文献
124.
?ngela Moreira Sofia S. Pereira Madalena Costa Tiago Morais Ana Pinto Rúben Fernandes Mariana P. Monteiro 《PloS one》2015,10(4)
Obesity has been associated with increased incidence and risk of mortality of prostate cancer. One of the proposed mechanisms underlying this risk association is the change in adipokines expression that could promote the development and progression of the prostate tumor cells. The main goal of this study was to evaluate the effect of preadipocyte and adipocyte secretome in the proliferation, migration and invasion of androgen independent prostate carcinoma cells (RM1) and to assess cell proliferation in the presence of the adiposity signals leptin and insulin. RM1 cells were co-cultured in with preadipocytes, adipocytes or cultured in their respective conditioned medium. Cell proliferation was assessed by flow cytometry and XTT viability test. Cell migration was evaluated using a wound healing injury assay of RM1 cells cultured with conditioned media. Cellular invasion of RM1 cells co-cultured with adipocytes and preadipocytes was assessed using matrigel membranes. Preadipocyte conditioned medium was associated with a small increase in RM1 proliferation, while adipocytes conditioned media significantly increased RM1 cell proliferation (p<0.01). Adipocytes also significantly increased the RM1 cells proliferation in co-culture (p <0.01). Cell migration was higher in RM1 cells cultured with preadipocyte and adipocyte conditioned medium. RM1 cell invasion was significantly increased after co-culture with preadipocytes and adipocytes (p <0.05). Insulin also increased significantly the cell proliferation in contrast to leptin, which showed no effect. In conclusion, prostate carcinoma cells seem to be influenced by factors secreted by adipocytes that are able to increase their ability to proliferate, migrate and invade. 相似文献
125.
126.
Eduardo M. Moreira Henning Gall Maarten J. G. Leening Lies Lahousse Daan W. Loth Bouwe P. Krijthe Jessica C. Kiefte-de Jong Guy G. Brusselle Albert Hofman Bruno H. Stricker Hossein A. Ghofrani Oscar H. Franco Janine F. Felix 《PloS one》2015,10(6)
Background
Pulmonary hypertension is characterized by increased pulmonary artery pressure and carries an increased mortality. Population-based studies into pulmonary hypertension are scarce and little is known about its prevalence in the general population. We aimed to describe the distribution of echocardiographically-assessed pulmonary artery systolic pressure (ePASP) in the general population, to estimate the prevalence of pulmonary hypertension, and to identify associated factors.Methods
Participants (n = 3381, mean age 76.4 years, 59% women) from the Rotterdam Study, a population-based cohort, underwent echocardiography. Echocardiographic pulmonary hypertension was defined as ePASP>40 mmHg.Results
Mean ePASP was 26.3 mmHg (SD 7.0). Prevalence of echocardiographic pulmonary hypertension was 2.6% (95%CI: 2.0; 3.2). Prevalence was higher in older participants compared to younger ones (8.3% in those over 85 years versus 0.8% in those between 65 and 70), and in those with underlying disorders versus those without (5.9% in subjects with COPD versus 2.3%; 9.2% in those with left ventricular systolic dysfunction versus 2.3%; 23.1% in stages 3 or 4 left ventricular diastolic dysfunction versus 1.9% in normal or stage 1). Factors independently associated with higher ePASP were older age, higher BMI, left ventricular diastolic dysfunction, COPD and systemic hypertension.Conclusion
In this large population-based study, we show that pulmonary hypertension as measured by echocardiography has a low prevalence in the overall general population in the Netherlands, but estimates may be higher in specific subgroups, especially in those with underlying diseases. Increased pulmonary arterial pressure is likely to gain importance in the near future due to population aging and the accompanying prevalences of underlying disorders. 相似文献127.
128.
Continuous removal of endocrine disruptors by versatile peroxidase using a two‐stage system 下载免费PDF全文
Roberto Taboada‐Puig Thelmo A. Lu‐Chau Gemma Eibes Gumersindo Feijoo Maria T. Moreira Juan M. Lema 《Biotechnology progress》2015,31(4):908-916
The oxidant Mn3+‐malonate, generated by the ligninolytic enzyme versatile peroxidase in a two‐stage system, was used for the continuous removal of endocrine disrupting compounds (EDCs) from synthetic and real wastewaters. One plasticizer (bisphenol‐A), one bactericide (triclosan) and three estrogenic compounds (estrone, 17β‐estradiol, and 17α‐ethinylestradiol) were removed from wastewater at degradation rates in the range of 28–58 µg/L·min, with low enzyme inactivation. First, the optimization of three main parameters affecting the generation of Mn3+‐malonate (hydraulic retention time as well as Na‐malonate and H2O2 feeding rates) was conducted following a response surface methodology (RSM). Under optimal conditions, the degradation of the EDCs was proven at high (1.3–8.8 mg/L) and environmental (1.2–6.1 µg/L) concentrations. Finally, when the two‐stage system was compared with a conventional enzymatic membrane reactor (EMR) using the same enzyme, a 14‐fold increase of the removal efficiency was observed. At the same time, operational problems found during EDCs removal in the EMR system (e.g., clogging of the membrane and enzyme inactivation) were avoided by physically separating the stages of complex formation and pollutant oxidation, allowing the system to be operated for a longer period (~8 h). This study demonstrates the feasibility of the two‐stage enzymatic system for removing EDCs both at high and environmental concentrations. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:908–916, 2015 相似文献
129.
Júlia D. Moreira Luisa Knorr Marcelo Ganzella Ana Paula Thomazi Carolina G. de Souza Débora G. de Souza Carolina F. Pitta Tadeu Mello e Souza Susana Wofchuk Elaine Elisabetsky Lúcia Vinadé Marcos L.S. Perry Diogo O. Souza 《Neurochemistry international》2010,56(6-7):753-759
Essential omega-3 polyunsaturated fatty acids (ω3) are crucial to brain development and function, being relevant for behavioral performance. In the present study we examined the influence of dietary ω3 in the development of the glutamatergic system and on behavior parameters in rats. Female rats received isocaloric diets, either with ω3 (ω3 group) or a ω3 deficient diet (D group). In ontogeny experiments of their litters, hippocampal immunocontent of ionotropic NMDA and AMPA glutamatergic receptors subunits (NR2 A\B and GluR1, respectively) and the alpha isoform of the calcium-calmodulin protein kinase type II (αCaMKII) were evaluated. Additionally, hippocampal [3H]glutamate binding and uptake were assessed. Behavioral performance was evaluated when the litters were adult (60 days old), through the open-field, plus-maze, inhibitory avoidance and flinch-jump tasks. The D group showed decreased immunocontent of all proteins analyzed at 02 days of life (P2) in comparison with the ω3 group, although the difference disappeared at 21 days of life (except for αCaMKII, which content normalized at 60 days old). The same pattern was found for [3H]glutamate binding, whereas [3H]glutamate uptake was not affected. The D group also showed memory deficits in the inhibitory avoidance, increased in the exploratory pattern in open-field, and anxiety-like behavior in plus-maze. Taken together, our results suggest that dietary ω3 content is relevant for glutamatergic system development and for behavioral performance in adulthood. The putative correlation among the neurochemical and behavioral alterations caused by dietary ω3 deficiency is discussed. 相似文献
130.
Tiago Collares Vinicius Farias Campos Fabiana Kömmling Seixas Paulo V. Cavalcanti Odir A. Dellagostin Heden Luiz M. Moreira João Carlos Deschamps 《Journal of biosciences》2010,35(1):39-47
The silver catfish (Rhamdia quelen) is an endemic American fish species. The sperm of each species has its own peculiarities and biological characteristics,
which influence the success of mass DNA transfer methods. Our objective in this study was to evaluate different sperm-mediated
gene transfer (SMGT) methods to obtain transgenic silver catfish. Different treatments for the incorporation of a foreign
pEGFP plasmid group were used: (1) dehydrated/rehydrated (DR), (2) dehydrated/rehydrated/electroporated (DRE), (3) electroporated
(E), (4) incubated with seminal plasma (INC); and (5) incubated in the absence of seminal plasma (INCSP). Sperm motility,
time of activity duration (TAD), fertilization rate (FR), hatching rate (HR) and sperm morphology were also evaluated. The
polymerase chain reaction (PCR) positivity rates for the presence of the transgene were: DRE 60%; DR 40%; E 25%; INC 5% and
INCSP 25%. The rates of embryo EGFP expression were: DRE 63%; DR 44%; E 34%; INC 8% and INCSP 38%. The fertilization rate
in the control and DRE treatments groups were higher than in the DR group, but the E, INC and INCSP treatment groups had the
lowest rate. The hatching rates of the DRE, DR and control groups were higher than in the INCSP, INC and E treatment groups
(P>0.05). There were no differences among the DRE and DR, E and DR, E and INCSP groups in expression and PCR positivity rates
of enhanced green fluorescent protein (EGFP) in embryos. Scanning electron microscopy also did not show any change in sperm
morphology among treatment groups. To the best of our knowledge, this is the first report on transgene transmission of exogenous
DNA into silver catfish larvae through SMGT technology. 相似文献