全文获取类型
收费全文 | 1832篇 |
免费 | 108篇 |
国内免费 | 2篇 |
出版年
2022年 | 26篇 |
2021年 | 28篇 |
2020年 | 16篇 |
2019年 | 28篇 |
2018年 | 29篇 |
2017年 | 27篇 |
2016年 | 47篇 |
2015年 | 110篇 |
2014年 | 80篇 |
2013年 | 107篇 |
2012年 | 171篇 |
2011年 | 140篇 |
2010年 | 90篇 |
2009年 | 80篇 |
2008年 | 109篇 |
2007年 | 108篇 |
2006年 | 115篇 |
2005年 | 118篇 |
2004年 | 98篇 |
2003年 | 73篇 |
2002年 | 77篇 |
2001年 | 20篇 |
2000年 | 23篇 |
1999年 | 9篇 |
1998年 | 22篇 |
1997年 | 9篇 |
1996年 | 7篇 |
1995年 | 7篇 |
1994年 | 7篇 |
1993年 | 7篇 |
1992年 | 21篇 |
1991年 | 5篇 |
1990年 | 7篇 |
1989年 | 8篇 |
1988年 | 5篇 |
1985年 | 4篇 |
1984年 | 3篇 |
1983年 | 7篇 |
1982年 | 5篇 |
1980年 | 5篇 |
1979年 | 3篇 |
1978年 | 6篇 |
1977年 | 4篇 |
1976年 | 3篇 |
1974年 | 5篇 |
1968年 | 3篇 |
1965年 | 4篇 |
1956年 | 3篇 |
1943年 | 3篇 |
1929年 | 3篇 |
排序方式: 共有1942条查询结果,搜索用时 312 毫秒
101.
Clara Braian Mattias Svensson Susanna Brighenti Maria Lerm Venkata R. Parasa 《Journal of visualized experiments : JoVE》2015,(104)
Tuberculosis (TB) still holds a major threat to the health of people worldwide, and there is a need for cost-efficient but reliable models to help us understand the disease mechanisms and advance the discoveries of new treatment options. In vitro cell cultures of monolayers or co-cultures lack the three-dimensional (3D) environment and tissue responses. Herein, we describe an innovative in vitro model of a human lung tissue, which holds promise to be an effective tool for studying the complex events that occur during infection with Mycobacterium tuberculosis (M. tuberculosis). The 3D tissue model consists of tissue-specific epithelial cells and fibroblasts, which are cultured in a matrix of collagen on top of a porous membrane. Upon air exposure, the epithelial cells stratify and secrete mucus at the apical side. By introducing human primary macrophages infected with M. tuberculosis to the tissue model, we have shown that immune cells migrate into the infected-tissue and form early stages of TB granuloma. These structures recapitulate the distinct feature of human TB, the granuloma, which is fundamentally different or not commonly observed in widely used experimental animal models. This organotypic culture method enables the 3D visualization and robust quantitative analysis that provides pivotal information on spatial and temporal features of host cell-pathogen interactions. Taken together, the lung tissue model provides a physiologically relevant tissue micro-environment for studies on TB. Thus, the lung tissue model has potential implications for both basic mechanistic and applied studies. Importantly, the model allows addition or manipulation of individual cell types, which thereby widens its use for modelling a variety of infectious diseases that affect the lungs. 相似文献
102.
103.
Raluca Buzdugan Sandra I. McCoy Constancia Watadzaushe Mi-Suk Kang Dufour Maya Petersen Jeffrey Dirawo Angela Mushavi Hilda Angela Mujuru Agnes Mahomva Reuben Musarandega Anna Hakobyan Owen Mugurungi Frances M. Cowan Nancy S. Padian 《PloS one》2015,10(8)
Objective
We estimated HIV-free infant survival and mother-to-child HIV transmission (MTCT) rates in Zimbabwe, some of the first community-based estimates from a UNAIDS priority country.Methods
In 2012 we surveyed mother-infant pairs residing in the catchment areas of 157 health facilities randomly selected from 5 of 10 provinces in Zimbabwe. Enrolled infants were born 9–18 months before the survey. We collected questionnaires, blood samples for HIV testing, and verbal autopsies for deceased mothers/infants. Estimates were assessed among i) all HIV-exposed infants, as part of an impact evaluation of Option A of the 2010 WHO guidelines (rolled out in Zimbabwe in 2011), and ii) the subgroup of infants unexposed to Option A. We compared province-level MTCT rates measured among women in the community with MTCT rates measured using program monitoring data from facilities serving those communities.Findings
Among 8568 women with known HIV serostatus, 1107 (12.9%) were HIV-infected. Among all HIV-exposed infants, HIV-free infant survival was 90.9% (95% confidence interval (CI): 88.7–92.7) and MTCT was 8.8% (95% CI: 6.9–11.1). Sixty-six percent of HIV-exposed infants were still breastfeeding. Among the 762 infants born before Option A was implemented, 90.5% (95% CI: 88.1–92.5) were alive and HIV-uninfected at 9–18 months of age, and 9.1% (95%CI: 7.1–11.7) were HIV-infected. In four provinces, the community-based MTCT rate was higher than the facility-based MTCT rate. In Harare, the community and facility-based rates were 6.0% and 9.1%, respectively.Conclusion
By 2012 Zimbabwe had made substantial progress towards the elimination of MTCT. Our HIV-free infant survival and MTCT estimates capture HIV transmissions during pregnancy, delivery and breastfeeding regardless of whether or not mothers accessed health services. These estimates also provide a baseline against which to measure the impact of Option A guidelines (and subsequently Option B+). 相似文献104.
Seema Singh Susanna A. Braus-Stromeyer Christian Timpner Van Tuan Tran Gertrud Lohaus Michael Reusche Jessica Knüfer Thomas Teichmann Andreas von Tiedemann Gerhard H. Braus 《Applied microbiology and biotechnology》2010,85(6):1961-1976
The first leaky auxotrophic mutant for aromatic amino acids of the near-diploid fungal plant pathogen Verticillium longisporum (VL) has been generated. VL enters its host Brassica napus through the roots and colonizes the xylem vessels. The xylem contains little nutrients including low concentrations of amino
acids. We isolated the gene Vlaro2 encoding chorismate synthase by complementation of the corresponding yeast mutant strain. Chorismate synthase produces the
first branch point intermediate of aromatic amino acid biosynthesis. A novel RNA-mediated gene silencing method reduced gene
expression of both isogenes by 80% and resulted in a bradytrophic mutant, which is a leaky auxotroph due to impaired expression
of chorismate synthase. In contrast to the wild type, silencing resulted in increased expression of the cross-pathway regulatory
gene VlcpcA (similar to cpcA/GCN4) during saprotrophic life. The mutant fungus is still able to infect the host plant B. napus and the model Arabidopsis thaliana with reduced efficiency. VlcpcA expression is increased in planta in the mutant and the wild-type fungus. We assume that xylem colonization requires induction
of the cross-pathway control, presumably because the fungus has to overcome imbalanced amino acid supply in the xylem. 相似文献
105.
106.
Magda Gioia Giovanni Francesco Fasciglione Susanna Monaco Riccardo Iundusi Diego Sbardella Stefano Marini Umberto Tarantino Massimo Coletta 《Journal of biological inorganic chemistry》2010,15(8):1219-1232
The proteolytic processing of collagen I by three matrix metalloproteinases (MMPs), a collagenase (MMP-1), a gelatinase (MMP-2),
and the ectodomain of a membrane-type metalloproteinase (MMP-14), has been investigated at 37 °C between pH 6.0 and 9.2, a
pH range reflecting conditions found in different body compartments under various physiopathological processes. In the proteolytic
degradation the native collagen triple helix must be partially unwound to allow the binding of α chains to the protease’s
active-site cleft. We have found that MMP-1 interacts with the two types of collagen I α chains in a similar fashion, whereas
both MMP-2 and MMP-14 bind the two α chains in a different way. The overall enzymatic activity is higher on the α-2 chain
for both MMP-1 and MMP-2, whereas the MMP-14 ectodomain preferentially cleaves the α-1 chain. In MMP-2 a marked difference
for substrate affinity (higher for the α-1 chain) is overwhelmed by an even more marked propensity to cleave the α-2 chain.
As a whole, the three classes of MMPs investigated appear to process collagen I in a significantly different fashion, so various
MMPs play different roles in the collagen homeostasis in various compartments (such as bloodstream, synovial fluid, normal
and tumoral tissues), where different pH values are observed. 相似文献
107.
Sonia Lewycka Charles Mwansambo Peter Kazembe Tambosi Phiri Andrew Mganga Mikey Rosato Hilda Chapota Florida Malamba Stefania Vergnano Marie-Louise Newell David Osrin Anthony Costello 《Trials》2010,11(1):1-15
Background
Paclitaxel-eluting stents decrease angiographic and clinical restenosis following percutaneous coronary intervention compared to bare metal stents. TAXUS Element is a third-generation paclitaxel-eluting stent which incorporates a novel, thinner-strut, platinum-enriched metal alloy platform. The stent is intended to have enhanced radiopacity and improved deliverability compared to other paclitaxel-eluting stents. The safety and efficacy of the TAXUS Element stent are being evaluated in the pivotal PERSEUS clinical trials.Methods/Design
The PERSEUS trials include two parallel studies of the TAXUS Element stent in single, de novo coronary atherosclerotic lesions. The PERSEUS Workhorse study is a prospective, randomized (3:1), single-blind, non-inferiority trial in subjects with lesion length ≤28 mm and vessel diameter ≥2.75 mm to ≤4.0 mm which compares TAXUS Element to the TAXUS Express2 paclitaxel-eluting stent system. The Workhorse study employs a novel Bayesian statistical approach that uses prior information to limit the number of study subjects exposed to the investigational device and thus provide a safer and more efficient analysis of the TAXUS Element stent. PERSEUS Small Vessel is a prospective, single-arm, superiority trial in subjects with lesion length ≤20 mm and vessel diameter ≥2.25 mm to <2.75 mm that compares TAXUS Element with a matched historical bare metal Express stent control.Discussion
The TAXUS PERSEUS clinical trial program uses a novel statistical approach to evaluate whether design and metal alloy iterations in the TAXUS Element stent platform provide comparable safety and improved procedural performance compared to the previous generation Express stent. PERSEUS trial enrollment is complete and primary endpoint data are expected in 2010. PERSEUS Workhorse and Small Vessel are registered at http://www.clinicaltrials.gov, identification numbers NCT00484315 and NCT00489541. 相似文献108.
Cilurzo F Cupone IE Minghetti P Buratti S Selmin F Gennari CG Montanari L 《AAPS PharmSciTech》2010,11(4):1511-1517
This work aimed to develop a fast-dissolving film made of low dextrose equivalent maltodextrins (MDX) containing nicotine
hydrogen tartrate salt (NHT). Particular attention was given to the selection of the suitable taste-masking agent (TMA) and
the characterisation of the ductility and flexibility under different mechanical stresses. MDX with two different dextrose
equivalents (DEs), namely DE 6 and DE 12, were selected in order to evaluate the effect of polymer molecular weight on film
tensile properties. The bitterness and astringency intensity of NHT and the suppression effect of several TMA were evaluated
by a Taste-Sensing System. The films were characterised in term of NHT content, tensile properties, disintegration time and
drug dissolution test. As expected, placebo films made of MDX DE 6 appeared stiffer and less ductile than film prepared using
MDX DE 12. The films disintegrated within 10 s. Among the tested TMA, the milk and mint flavours resulted particularly suitable
to mask the taste of NHT. The addition of NHT and taste-masking agents affected film tensile properties; however, the effect
of the addition of these components can be counterweighted by modulating the glycerine content and/or the MDX molecular weight.
The feasibility of NHT loaded fast-dissolving films was demonstrated. 相似文献
109.
110.
Among 21 human strains of Laribacter hongkongensis, small plasmids were observed in four strains, and large ones in six strains. The smallest, 3264-bp plasmid, pHLHK19, has only one ORF that encodes a putative replication initiator protein and a predicted origin of replication (ori) with a DnaA box, three 18-bp direct repeats and five pairs of inverted repeats. An Escherichia coli-L. hongkongensis shuttle vector was constructed by ligating the HindIII-digested pHLHK19, containing the replication initiator protein and ori of pHLHK19, to HindIII-digested pBK-CMV. This shuttle vector can propagate in E. coli and L. hongkongensis with good transformation efficiencies. 相似文献