全文获取类型
收费全文 | 1166篇 |
免费 | 75篇 |
出版年
2023年 | 7篇 |
2022年 | 25篇 |
2021年 | 37篇 |
2020年 | 20篇 |
2019年 | 32篇 |
2018年 | 31篇 |
2017年 | 30篇 |
2016年 | 45篇 |
2015年 | 70篇 |
2014年 | 71篇 |
2013年 | 76篇 |
2012年 | 85篇 |
2011年 | 93篇 |
2010年 | 44篇 |
2009年 | 57篇 |
2008年 | 55篇 |
2007年 | 53篇 |
2006年 | 56篇 |
2005年 | 57篇 |
2004年 | 38篇 |
2003年 | 38篇 |
2002年 | 30篇 |
2001年 | 15篇 |
2000年 | 23篇 |
1999年 | 20篇 |
1998年 | 9篇 |
1997年 | 12篇 |
1996年 | 7篇 |
1995年 | 8篇 |
1994年 | 3篇 |
1993年 | 7篇 |
1992年 | 9篇 |
1991年 | 6篇 |
1990年 | 9篇 |
1989年 | 4篇 |
1988年 | 2篇 |
1987年 | 9篇 |
1986年 | 5篇 |
1985年 | 4篇 |
1984年 | 3篇 |
1982年 | 4篇 |
1980年 | 2篇 |
1979年 | 2篇 |
1977年 | 2篇 |
1976年 | 2篇 |
1971年 | 2篇 |
1969年 | 4篇 |
1967年 | 5篇 |
1966年 | 2篇 |
1928年 | 1篇 |
排序方式: 共有1241条查询结果,搜索用时 109 毫秒
71.
Tselios T Daliani I Probert L Deraos S Matsoukas E Roy S Pires J Moore G Matsoukas J 《Bioorganic & medicinal chemistry》2000,8(8):1903-1909
Experimental autoimmune encephalomyelitis (EAE) is an inflammatory and demyelinating disease of the central nervous system and is an animal model of multiple sclerosis (MS). In the present report, a linear analogue and a series of cyclic semi-mimetic peptides were designed and synthesized based on the human myelin basic protein (MBP(87-99)) epitope (Val87-His-Phe-Phe-Lys-Asn-Ile-Val-Thr-Pro-Arg-Thr-Pro90) and on Copolymer I (a mixture of random polymers of Ala, Gln, Lys and Tyr used to treat MS). These analogues were designed looking for suppressors of EAE induced by guinea pig MBP(72-85) epitope (Gln-Lys-Ser-Gln-Arg-Ser-Gln-Asp-Glu-Asn-Pro-Val) in Lewis rats. The linear analogue [Arg91,Ala96]MBP(87-99), in which Arg substitutes Lys91 and Ala substitutes Pro96, was found to be a strong inhibitor which when administered to Lewis rats together with the encephalitogenic agonist MBP(72-85) completely prevented the induction of EAE. In contrast, three N- and C-termini amide-linked cyclic semi-mimetic peptides, [cyclo-Phe-Arg-Asn-Ile-Val-Thr-Ala-Acp (1), cyclo-Phe-Ala-Arg-Gln-Acp (2), cyclo-Tyr-Ala-Lys-Gln-Acp (3)] as well as a Lys side chain and C-terminous cyclic semi mimetic peptide cyclo(Lys, Acp)-Phe-Lys-Asn-Ile-Val-Thr-Ala-Acp (4) which contain segments of MBP(87-99) or are constituted from immunophoric residues of copolymer 1, were ineffective in inducing or inhibiting EAE in Lewis rats. However co-injection of cyclic analogues with MBP(72-85) delayed the onset of EAE indicating a modulatory effect on the EAE activity of MBP(72-85). These findings suggest that molecule length, size of cyclic moiety and backbone conformation are important elements for immunogenic activity. Moreover blockade of MBP(72-85) induced EAE by the unrelated peptide [Arg91,Ala56]MBP(87-99) could indicate that the mechanism of inhibition is not due to binding competition but rather due to the delivery of a negative signal by the antagonist which overcomes the agonist response possibly through the activation of antigen specific regulatory T cells. 相似文献
72.
M. C. V. Egas M. S. da Costa Don A. Cowan Euclides M. V. Pires 《Extremophiles : life under extreme conditions》1998,2(1):23-32
An extracellular α-amylase produced by the thermophilic bacterium Thermus filiformis Ork A2 was purified from cell-free culture supernatant by ion exchange chromatography. The molecular mass was estimated to
be 60 000 Da by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme was rich in both basic and hydrophobic
amino acids, presenting the following NH2-terminal amino acid sequence: Thr-Ala-Asp-Leu-Ile-Val-Lys-Ile-Asn-Phe. Amylolytic activity on soluble starch was optimal
at pH 5.5–6.0 and 95°C, and the enzyme was stable in the pH range of 4.0–8.0. Calcium enhanced thermostability at temperatures
above 80°C, increasing the half-life of activity to more than 8 h at 85°C, 80 min at 90°C, and 19 min at 95°C. Ethylenediaminetetraacetic
acid (EDTA) inhibited amylase activity, the inhibition being reversed by the addition of calcium or strontium ions. The α-amylase
was also inhibited by copper and mercuric ions, and p-chloromercuribenzoic acid, the latter being reversed in the presence of dithiothreitol. Dithiothreitol and β-mercaptoethanol
activated the enzyme. The α-amylase exhibited Michaelis-Menten kinetics for starch, with a K
m of 5.0 mg·ml−1 and k
cat/K
m of 5.2 × 105 ml·mg−1 s−1. Similar values were obtained for amylose, amylopectin, and glycogen. The hydrolysis pattern was similar for maltooligosaccharides
and polysaccharides, with maltose being the major hydrolysis product. Glucose and maltotriose were generated as secondary
products, although glucose was produced in high levels after a 6-h digestion. To our knowledge this is the first report of
the characterization of an α-amylase from a strain of the genus Thermus.
Received: June 2, 1997 / Accepted: September 16, 1997 相似文献
73.
Reactions of the adduct UCl3·(THF)x (THF = tetrahydrofuran) with compounds of the type K[HnBL4?n], where L = pyrazole or 3,5-dimethylpyrazole, are presented. Based on the results obtained the two compounds UCl2H2BPz2·THF and UCl2HBPz3·THF were isolated. 相似文献
74.
Ana Sílvia Franco Pinheiro Moreira José Pires de Lemos Filho Gerhard Zotz Rosy Mary dos Santos Isaias 《Flora》2009,204(8):604-611
This study compares photosynthetic and structural features of Dichaea cogniauxiana and Epidendrum secundum leaves and roots. The diurnal titratable acidity fluctuations indicated crassulacean acid metabolism (CAM) in E. secundum leaves, associated with anatomical features like thick cuticle, large and vacuolated cells, and reduced stomata size and frequency. Roots of both species had chloroplasts in their cortical parenchyma. However, neither the roots nor D. cogniauxiana leaves did show tissue sap acidity fluctuations. This indicates C3 metabolism in these organs. This lack of oscillation of organic acids in Epidendrum roots was at odds with a CAM-like 13C ratio, suggesting that in spite of active CO2 fixation in roots during the day, the bulk of carbon is imported from the leaves. Roots of both species showed Fv/Fm, ΔF/Fm′, ETR values similar to reports from other non-foliar photosynthetic organs. Besides reducing root carbon cost, root photosynthesis may also be important by alleviating potential hypoxia, since water-saturated velamen severely impedes the gas exchange between radicular cortex. 相似文献
75.
Suely Roizenblatt Altay L. Souza Luciana Palombini Luciana M. Godoy Sergio Tufik Lia Rita A. Bittencourt 《PloS one》2015,10(11)
BackgroundWe are witnessing the growth of urban populations, particularly in the developing world. São Paulo, the largest city in South America, continues to grow, and this growth is dramatically effecting the environment and human health. The aim of this study was to estimate the point prevalence of chronic pain in São Paulo city dwellers and to explore the influence of aspects related to urbanicity.MethodsA two-stage cluster randomized sample included 1100 individuals of the city of Sao Paulo, representing the population proportionally in terms of gender, age and social classes in 2007. For this observational cross-sectional study, the household sample was interviewed using validated questionnaires for sociodemographic aspects, the Beck inventories for anxiety and depression, the WHOQoL-REF for quality of life, the Chalder Fatigue Scale. Musculoskeletal pain was defined as diffuse pain or pain located in the back, joints or limbs. Data regarding sleep complaints and polysomnography were obtained from the Epidemiologic Sleep Study conducted in São Paulo city in 2007.ResultsThe prevalence estimate of chronic musculoskeletal pain was approximately 27%, with a female/male ratio of approximately 2.6/1. The predictors were being in the age-range of 30–39 years, low socioeconomic and schooling levels, obesity, sedentarism, fatigue, non-restorative sleep, daytime sleepiness, poor sleep quality, poor life quality, anxiety and depression symptoms. Psychological wellbeing was the main discriminator between responders with chronic musculoskeletal pain and the controls, followed by depression for the participants with poor psychological wellbeing, and fatigue, for the remaining ones. Insomnia syndrome was the third-level discriminator for those with fatigue, whereas sleep quality for those without fatigue.ConclusionsMusculoskeletal pain was frequently reported by São Paulo city dwellers and its correlates with psychological and sleep aspects are suggestive of a response to urbanicity.
Trial Registration
ClinicalTrials.gov NCT00596713 相似文献76.
Débora P. Paula Benjamin Linard David A. Andow Edison R. Sujii Carmen S. S. Pires Alfried P. Vogler 《Molecular ecology resources》2015,15(4):880-892
DNA methods are useful to identify ingested prey items from the gut of predators, but reliable detection is hampered by low amounts of degraded DNA. PCR‐based methods can retrieve minute amounts of starting material but suffer from amplification biases and cross‐reactions with the predator and related species genomes. Here, we use PCR‐free direct shotgun sequencing of total DNA isolated from the gut of the harlequin ladybird Harmonia axyridis at five time points after feeding on a single pea aphid Acyrthosiphon pisum. Sequence reads were matched to three reference databases: Insecta mitogenomes of 587 species, including H. axyridis sequenced here; A. pisum nuclear genome scaffolds; and scaffolds and complete genomes of 13 potential bacterial symbionts. Immediately after feeding, multicopy mtDNA of A. pisum was detected in tens of reads, while hundreds of matches to nuclear scaffolds were detected. Aphid nuclear DNA and mtDNA decayed at similar rates (0.281 and 0.11 h?1 respectively), and the detectability periods were 32.7 and 23.1 h. Metagenomic sequencing also revealed thousands of reads of the obligate Buchnera aphidicola and facultative Regiella insecticola aphid symbionts, which showed exponential decay rates significantly faster than aphid DNA (0.694 and 0.80 h?1, respectively). However, the facultative aphid symbionts Hamiltonella defensa, Arsenophonus spp. and Serratia symbiotica showed an unexpected temporary increase in population size by 1–2 orders of magnitude in the predator guts before declining. Metagenomics is a powerful tool that can reveal complex relationships and the dynamics of interactions among predators, prey and their symbionts. 相似文献
77.
Brecht Devleesschauwer Juanita A. Haagsma Frederick J. Angulo David C. Bellinger Dana Cole D?rte D?pfer Aamir Fazil Eric M. Fèvre Herman J. Gibb Tine Hald Martyn D. Kirk Robin J. Lake Charline Maertens de Noordhout Colin D. Mathers Scott A. McDonald Sara M. Pires Niko Speybroeck M. Kate Thomas Paul R. Torgerson Felicia Wu Arie H. Havelaar Nicolas Praet 《PloS one》2015,10(12)
Background
The Foodborne Disease Burden Epidemiology Reference Group (FERG) was established in 2007 by the World Health Organization to estimate the global burden of foodborne diseases (FBDs). This paper describes the methodological framework developed by FERG''s Computational Task Force to transform epidemiological information into FBD burden estimates.Methods and Findings
The global and regional burden of 31 FBDs was quantified, along with limited estimates for 5 other FBDs, using Disability-Adjusted Life Years in a hazard- and incidence-based approach. To accomplish this task, the following workflow was defined: outline of disease models and collection of epidemiological data; design and completion of a database template; development of an imputation model; identification of disability weights; probabilistic burden assessment; and estimating the proportion of the disease burden by each hazard that is attributable to exposure by food (i.e., source attribution). All computations were performed in R and the different functions were compiled in the R package ''FERG''. Traceability and transparency were ensured by sharing results and methods in an interactive way with all FERG members throughout the process.Conclusions
We developed a comprehensive framework for estimating the global burden of FBDs, in which methodological simplicity and transparency were key elements. All the tools developed have been made available and can be translated into a user-friendly national toolkit for studying and monitoring food safety at the local level. 相似文献78.
Selective Cytotoxicity of 1,3,4-Thiadiazolium Mesoionic Derivatives on Hepatocarcinoma Cells (HepG2)
Gustavo Jabor Gozzi Amanda do Rocio Andrade Pires Glaucio Valdameri Maria Eliane Merlin Rocha Glaucia Regina Martinez Guilhermina Rodrigues Noleto Alexandra Acco Carlos Eduardo Alves de Souza Aurea Echevarria Camilla Moretto dos Reis Attilio Di Pietro Sílvia Maria Suter Correia Cadena 《PloS one》2015,10(6)
In this work, we evaluated the cytotoxicity of mesoionic 4-phenyl-5-(2-Y, 4-X or 4-X-cinnamoyl)-1,3,4-thiadiazolium-2-phenylamine chloride derivatives (MI-J: X=OH, Y=H; MI-D: X=NO2, Y=H; MI-4F: X=F, Y=H; MI-2,4diF: X=Y=F) on human hepatocellular carcinoma (HepG2), and non-tumor cells (rat hepatocytes) for comparison. MI-J, M-4F and MI-2,4diF reduced HepG2 viability by ~ 50% at 25 μM after 24-h treatment, whereas MI-D required a 50 μM concentration, as shown by 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. The cytotoxicity was confirmed with lactate dehydrogenase assay, of which activity was increased by 55, 24 and 16% for MI-J, MI-4F and MI-2,4diF respectively (at 25 μM after 24 h). To identify the death pathway related to cytotoxicity, the HepG2 cells treated by mesoionic compounds were labeled with both annexin V and PI, and analyzed by flow cytometry. All compounds increased the number of doubly-stained cells at 25 μM after 24 h: by 76% for MI-J, 25% for MI-4F and MI-2,4diF, and 11% for MI-D. It was also verified that increased DNA fragmentation occurred upon MI-J, MI-4F and MI-2,4diF treatments (by 12%, 9% and 8%, respectively, at 25 μM after 24 h). These compounds were only weakly, or not at all, transported by the main multidrug transporters, P-glycoprotein, ABCG2 and MRP1, and were able to slightly inhibit their drug-transport activity. It may be concluded that 1,3,4-thiadiazolium compounds, especially the hydroxy derivative MI-J, constitute promising candidates for future investigations on in-vivo treatment of hepatocellular carcinoma. 相似文献
79.
Frederico Figueiredo Amancio Tiago Pires Heringer Cristina da Cunha Hueb Barata de Oliveira Liliane Boaventura Fassy Frederico Bruzzi de Carvalho Daniela Pagliari Oliveira Claudio Dornas de Oliveira Fernando Otoni Botoni Fernanda do Carmo Magalh?es José Roberto Lambertucci Mariangela Carneiro 《PloS one》2015,10(6)
The purpose of our study was to describe the clinical profile of dengue-infected patients admitted to Brazilian intensive care units (ICU) and evaluate factors associated with death. A longitudinal, multicenter case series study was conducted with laboratory-confirmed dengue patients admitted to nine Brazilian ICUs situated in Minas Gerais state, southeastern Brazil from January 1, 2008, to December 31, 2013. Demographic, clinical and laboratory data; disease severity scores; and mortality were evaluated. A total of 97 patients were studied. The in-ICU and in-hospital mortality rates were 18.6% and 19.6%, respectively. Patients classified as having severe dengue according to current World Health Organization classifications showed an increased risk of death in a univariate analysis. Nonsurvivors were older, exhibited lower serum albumin concentrations and higher total leukocyte counts and serum creatinine levels. Other risk factors (vomiting, lethargy/restlessness, dyspnea/respiratory distress) were also associated with death in a univariate analysis. Multivariate analysis indicated that in-hospital mortality was significantly associated with Acute Physiology and Chronic Health Evaluation II and the Sequential Organ Failure Assessment score. The ICU and in-hospital mortality observed in this study were higher than values reported in similar studies. An increased frequency of ICU admission due to severe organ dysfunction, higher severity indices and scarcity of ICU beds may partially explain the higher mortality. 相似文献
80.
Fabricio Fernandes Fontana Celso Tadeu Barbosa dos Santos Flavia Maria Esteves Ademir Rocha Geisa Ferreira Fernandes Cristiane Candida do Amaral Marcos Abel Domingues Zoilo Pires De Camargo Mario León Silva-Vergara 《Mycopathologia》2010,169(3):159-165
There is some evidence that dogs can be naturally infected by Paracoccidioides brasiliensis in endemic areas of paracoccidioidomycosis. In order to evaluate canine infection with this fungus, a survey with 149 urban
and 126 rural dogs was carried out using ELISA and intradermal tests with the gp43 antigen of P. brasiliensis in Uberaba, Minas Gerais state of Brazil. Forty-one out of 149 urban dogs were euthanatized and had their lungs, liver and
spleen removed. One slice from each viscera was processed for histopathological examination and the remaining was homogenized and then cultivated on mycobiotic agar
at room temperature and Fava-Netto medium at 35°C and observed for 12 weeks. Of urban dogs, 75 (50.3%) were small adult females,
56 (36%) were strays, while 93 (64%) had been donated to the municipal zoonosis control center. Nine (6.2%) had a positive
intradermal test without statistical differences regarding gender, race, nutritional status or origin. No colonies with microscopic
or morphology appearances resembling P. brasiliensis were isolated, nor granulomatous process or fungal structures were observed from histopathological examination. Eighty (53.6%)
of the urban dogs presented seroreactivity, without statistical differences regarding gender, race, nutritional state, origin,
or positive intradermal test. Of 126 rural dogs, 102 (80.5%) presented antibodies against gp43 antigen, and this was statistically
significant in relation to the reactivity detected in urban dogs (P = 0.0001). Thus, dogs are commonly infected with P. brasiliensis, but they probably present natural resistance to develop paracoccidioidomycosis. 相似文献