全文获取类型
收费全文 | 2515篇 |
免费 | 171篇 |
专业分类
2686篇 |
出版年
2023年 | 14篇 |
2022年 | 25篇 |
2021年 | 77篇 |
2020年 | 45篇 |
2019年 | 52篇 |
2018年 | 65篇 |
2017年 | 60篇 |
2016年 | 86篇 |
2015年 | 127篇 |
2014年 | 132篇 |
2013年 | 176篇 |
2012年 | 225篇 |
2011年 | 183篇 |
2010年 | 127篇 |
2009年 | 81篇 |
2008年 | 120篇 |
2007年 | 113篇 |
2006年 | 112篇 |
2005年 | 98篇 |
2004年 | 98篇 |
2003年 | 77篇 |
2002年 | 68篇 |
2001年 | 48篇 |
2000年 | 63篇 |
1999年 | 42篇 |
1998年 | 14篇 |
1997年 | 11篇 |
1996年 | 16篇 |
1995年 | 22篇 |
1993年 | 14篇 |
1992年 | 21篇 |
1991年 | 33篇 |
1990年 | 12篇 |
1989年 | 28篇 |
1988年 | 14篇 |
1987年 | 16篇 |
1986年 | 11篇 |
1985年 | 23篇 |
1984年 | 16篇 |
1983年 | 11篇 |
1981年 | 5篇 |
1980年 | 5篇 |
1979年 | 8篇 |
1975年 | 9篇 |
1974年 | 8篇 |
1973年 | 9篇 |
1972年 | 9篇 |
1971年 | 7篇 |
1970年 | 5篇 |
1968年 | 5篇 |
排序方式: 共有2686条查询结果,搜索用时 15 毫秒
71.
M. L. Gonçalves F. Pinto J. A. Ribeiro 《Nucleosides, nucleotides & nucleic acids》2013,32(5):1161-1163
Abstract The effect of adenosine on 45Ca uptake by rat brain synaptosomes electrically stimulated was studied as function of time of stimulation (10, 30, 120 s). Inhibition of 45Ca uptake was more evident for 120 s. 相似文献
72.
Ricardo Ewbank Steffen Rosangela Caetano Márcia Pinto Diogo Chaves Rossini Ferrari Mayara Bastos Sandra Teixeira de Abreu Dick Menzies Anete Trajman 《PloS one》2013,8(4)
Background
Latent tuberculosis infection (LTBI) is a reservoir for new TB cases. Isoniazid preventive therapy (IPT) reduces the risk of active TB by as much as 90%, but LTBI screening has limitations. Unlike tuberculin skin testing (TST), interferon-gamma release assays are not affected by BCG vaccination, and have been reported to be cost-effective in low-burden countries. The goal of this study was to perform a cost-effectiveness analysis from the health system perspective, comparing three strategies for LTBI diagnosis in TB contacts: tuberculin skin testing (TST), QuantiFERON®-TB Gold-in-Tube (QFT-GIT) and TST confirmed by QFT-GIT if positive (TST/QFT-GIT) in Brazil, a middle-income, high-burden country with universal BCG coverage.Methodology/Principal Findings
Costs for LTBI diagnosis and treatment of a hypothetical cohort of 1,000 adult immunocompetent close contacts were considered. The effectiveness measure employed was the number of averted TB cases in two years. Health system costs were US$ 105,096 for TST, US$ 121,054 for QFT-GIT and US$ 101,948 for TST/QFT-GIT; these strategies averted 6.56, 6.63 and 4.59 TB cases, respectively. The most cost-effective strategy was TST (US$ 16,021/averted case). The incremental cost-effectiveness ratio was US$ 227,977/averted TB case for QFT-GIT. TST/QFT-GIT was dominated.Conclusions
Unlike previous studies, TST was the most cost-effective strategy for averting new TB cases in the short term. QFT-GIT would be more cost-effective if its costs could be reduced to US$ 26.95, considering a TST specificity of 59% and US$ 18 considering a more realistic TST specificity of 80%. Nevertheless, with TST, 207.4 additional people per 1,000 will be prescribed IPT compared with QFT. 相似文献73.
Olga Martinho Filipe Pinto Sara Granja Vera Miranda-Gon?alves Marise A. R. Moreira Luis F. J. Ribeiro Celso di Loreto Marsha R. Rosner Adhemar Longatto-Filho Rui Manuel Reis 《PloS one》2013,8(3)
Cervical cancer is one of the most common cancers in women worldwide, being high-risk group the HPV infected, the leading etiological factor. The raf kinase inhibitory protein (RKIP) has been associated with tumor progression and metastasis in several human neoplasms, however its role on cervical cancer is unclear. In the present study, 259 uterine cervix tissues, including cervicitis, cervical intraepithelial lesions and carcinomas, were analyzed for RKIP expression by immunohistochemistry. We found that RKIP expression was significantly decreased during malignant progression, being highly expressed in non-neoplastic tissues (54% of the samples; 73/135), and expressed at low levels in the cervix invasive carcinomas (∼15% (19/124). Following in vitro downregulation of RKIP, we observed a viability and proliferative advantage of RKIP-inhibited cells over time, which was associated with an altered cell cycle distribution and higher colony number in a colony formation assay. An in vitro wound healing assay showed that RKIP abrogation is associated with increased migratory capability. RKIP downregulation was also associated with an increased vascularization of the tumors in vivo using a CAM assay. Furthermore, RKIP inhibition induced cervical cancer cells apoptotic resistance to cisplatin treatment. In conclusion, we described that RKIP protein is significantly depleted during the malignant progression of cervical tumors. Despite the lack of association with patient clinical outcome, we demonstrate, in vitro and in vivo, that loss of RKIP expression can be one of the factors that are behind the aggressiveness, malignant progression and chemotherapy resistance of cervical cancer. 相似文献
74.
Camila Fernandes Heitor Sá Gon?alves Paula Brito Cabral Helena Camara Pinto Maria Isabel Moraes Pinto Lilia Maria Carneiro Camara 《PloS one》2013,8(11)
Background
Leprosy is a chronic disease, caused by Mycobacterium leprae, which poses a serious public health problem worldwide. Its high incidence in people under 15 years old in Ceará state, Brazil, reflects the difficulty of its control. The spectrum of clinical manifestations is associated with the immune response developed, with the Th1 and Th2 responses being related to the paucibacillary and multibacillary forms, respectively. Regulatory T cells (Treg), which can suppress Th1 and Th2 response, have received special attention in the literature and have been associated with development of chronic infections. However, their role in leprosy in individuals under 15 years old has not yet been elucidated. We evaluated the frequency of CD4+/CD8+CD25highFOXP3+ and CD4+/CD8+CD25highFOXP3high cells in leprosy patients and household contacts, in both cases under 15 years old.Methodology/Principal Findings
PBMC from 12 patients and 17 contacts were cultured for 72 hours with anti-CD3 and anti-CD28 (activators) or with activators associated with total sonicated fraction of M. leprae. After culture, the frequency of CD4+/CD8+ Treg was identified by flow cytometry. Cells stimulated by activators and antigen from multibacillary patients showed Treg frequencies almost two times that of the contacts: CD4+FOXP3+ (21.93±8.43 vs. 13.79±8.19%, p = 0.0500), CD4+FOXP3high (10.33±5.69 vs. 5.57±4.03%, p = 0.0362), CD8+FOXP3+ (13.88±9.19 vs. 6.18±5.56%, p = 0.0230) and CD8+FOXP3high (5.36±4.17 vs. 2.23±2.68%, p = 0.0461). Furthermore, the mean fluorescence intensity of FOXP3 in Treg was higher in multibacillary patients than in the contacts. Interestingly, there was a positive correlation of the bacillary index and number of lesions with the frequency of all Treg evaluated in patients.Conclusions/Significance
We have demonstrated for the first time that multibacillary leprosy patients under 15 years old have greater CD4+ and CD8+ Treg frequencies and these correlate with clinical and laboratorial aspects of disease. These findings suggest the involvement of these cells in the perpetuation of M. leprae infection. 相似文献75.
Mónica Costa Eugénia Cruz James C. Barton Ketil Thorstensen Sandra Morais Berta M. da Silva Jorge P. Pinto Cristina P. Vieira Jorge Vieira Ronald T. Acton Gra?a Porto 《PloS one》2013,8(11)
Hereditary Hemochromatosis (HH) is a recessively inherited disorder of iron overload occurring commonly in subjects homozygous for the C282Y mutation in HFE gene localized on chromosome 6p21.3 in linkage disequilibrium with the human leukocyte antigen (HLA)-A locus. Although its genetic homogeneity, the phenotypic expression is variable suggesting the presence of modifying factors. One such genetic factor, a SNP microhaplotype named A-A-T, was recently found to be associated with a more severe phenotype and also with low CD8+T-lymphocyte numbers. The present study aimed to test whether the predictive value of the A-A-T microhaplotype remained in other population settings. In this study of 304 HH patients from 3 geographically distant populations (Porto, Portugal 65; Alabama, USA 57; Nord-Trøndelag, Norway 182), the extended haplotypes involving A-A-T were studied in 608 chromosomes and the CD8+ T-lymphocyte numbers were determined in all subjects. Patients from Porto had a more severe phenotype than those from other settings. Patients with A-A-T seemed on average to have greater iron stores (p = 0.021), but significant differences were not confirmed in the 3 separate populations. Low CD8+ T-lymphocytes were associated with HLA-A*03-A-A-T in Porto and Alabama patients but not in the greater series from Nord-Trøndelag. Although A-A-T may signal a more severe iron phenotype, this study was unable to prove such an association in all population settings, precluding its use as a universal predictive marker of iron overload in HH. Interestingly, the association between A-A-T and CD8+ T-lymphocytes, which was confirmed in Porto and Alabama patients, was not observed in Nord-Trøndelag patients, showing that common HLA haplotypes like A*01–B*08 or A*03–B*07 segregating with HFE/C282Y in the three populations may carry different messages. These findings further strengthen the relevance of HH as a good disease model to search for novel candidate loci associated with the genetic transmission of CD8+ T-lymphocyte numbers. 相似文献
76.
77.
Jorge Almarza Luis Rincón Alí Bahsas María Angela Pinto Francisco Brito 《The protein journal》2013,32(2):118-125
Understanding of protein–urea interactions is one of the greatest challenges to modern structural protein chemistry. Based in enzyme kinetics experiments and 1H NMR spectroscopic analysis we proposed that urea, at low concentrations, directly interacts with the protonated histidines of the active center of RNase A, following a simple model of competitive inhibition. These results were supported by theoretical analysis based on the frontier molecular orbital theory and suggest that urea might establish a favorable interaction with the cationic amino acids. Our experimental evidence and theoretical analysis indicate that the initials steps of the molecular mechanism of Urea–RNase A interaction passes through the establishment of a three center four electron adduct. Also, our results would explain the observed disruption of the 1H NMR signals corresponding to H12 and H119 (involved in catalysis) of the RNase A studied in the presence of urea. Our interaction model of urea–amino acids (cationic) can be extended to explain the inactivation of other enzymes with cationic amino acids at the active site. 相似文献
78.
Brain fatty acid-binding protein (B-FABP) interacts with biological membranes and delivers polyunsaturated fatty acids (FAs) via a collisional mechanism. The binding of FAs in the protein and the interaction with membranes involve a motif called “portal region”, formed by two small α-helices, A1 and A2, connected by a loop. We used a combination of site-directed mutagenesis and electron spin resonance to probe the changes in the protein and in the membrane model induced by their interaction. Spin labeled B-FABP mutants and lipidic spin probes incorporated into a membrane model confirmed that B-FABP interacts with micelles through the portal region and led to structural changes in the protein as well in the micelles. These changes were greater in the presence of LPG when compared to the LPC models. ESR spectra of B-FABP labeled mutants showed the presence of two groups of residues that responded to the presence of micelles in opposite ways. In the presence of lysophospholipids, group I of residues, whose side chains point outwards from the contact region between the helices, had their mobility decreased in an environment of lower polarity when compared to the same residues in solution. The second group, composed by residues with side chains situated at the interface between the α-helices, experienced an increase in mobility in the presence of the model membranes. These modifications in the ESR spectra of B-FABP mutants are compatible with a less ordered structure of the portal region inner residues (group II) that is likely to facilitate the delivery of FAs to target membranes. On the other hand, residues in group I and micelle components have their mobilities decreased probably as a result of the formation of a collisional complex. Our results bring new insights for the understanding of the gating and delivery mechanisms of FABPs. 相似文献
79.
Objectives
It is unclear whether sedentary behaviour, and the domain in which it occurs, is related to body mass index (BMI) change. We aim to elucidate whether sedentary behaviour is prospectively related to BMI change using markers from three domains (leisure, work and commuting).Methods
Among employed 1958 British birth cohort members (n = 6,562), we analysed whether TV-viewing, work sitting (six categories: 0 h/d to >4 h/d) and motorised commuting (at 45 y) were related to BMI (at 45 y and 50 y) and BMI change 45–50 y, after adjusting for lifestyle and socioeconomic factors.Results
Per category higher TV-viewing, 45 y and 50 y BMI were higher by 0.69 kg/m2 (95% CI: 0.59,0.80) and 0.75 kg/m2 (0.64,0.86) respectively. A category higher TV-viewing was associated with 0.11 kg/m2 (0.06,0.17) increased BMI 45–50 y, attenuating to 0.06 kg/m2 (0.01,0.12) after adjustment. There was no trend for work sitting with 45 y or 50 y BMI, nor, after adjustment, for BMI change. However, those sitting 2–3 h/d had greater BMI gain by 0.33 kg/m2 (0.10,0.56) compared to those sitting 0–1 h/d. Associations between TV-viewing and BMI change were independent of work sitting. Motorised commuting was associated with 45 y, but not 50 y BMI or change.Conclusions
TV-viewing is associated with BMI gain in mid-adulthood; evidence is weaker for other sedentary behaviours. 相似文献80.
Lorenzo Maria Donini Eleonora Poggiogalle Veronica Mosca Alessandro Pinto Amelia Brunani Paolo Capodaglio 《PloS one》2013,8(10)