全文获取类型
收费全文 | 4555篇 |
免费 | 448篇 |
国内免费 | 1篇 |
出版年
2023年 | 16篇 |
2022年 | 48篇 |
2021年 | 114篇 |
2020年 | 67篇 |
2019年 | 73篇 |
2018年 | 91篇 |
2017年 | 88篇 |
2016年 | 200篇 |
2015年 | 319篇 |
2014年 | 299篇 |
2013年 | 318篇 |
2012年 | 439篇 |
2011年 | 331篇 |
2010年 | 226篇 |
2009年 | 159篇 |
2008年 | 215篇 |
2007年 | 218篇 |
2006年 | 208篇 |
2005年 | 165篇 |
2004年 | 159篇 |
2003年 | 129篇 |
2002年 | 112篇 |
2001年 | 66篇 |
2000年 | 85篇 |
1999年 | 74篇 |
1998年 | 36篇 |
1997年 | 27篇 |
1996年 | 30篇 |
1995年 | 25篇 |
1994年 | 19篇 |
1993年 | 33篇 |
1992年 | 40篇 |
1991年 | 37篇 |
1990年 | 37篇 |
1989年 | 26篇 |
1988年 | 25篇 |
1987年 | 21篇 |
1986年 | 21篇 |
1985年 | 23篇 |
1984年 | 26篇 |
1983年 | 29篇 |
1982年 | 22篇 |
1981年 | 22篇 |
1979年 | 20篇 |
1977年 | 20篇 |
1974年 | 16篇 |
1973年 | 21篇 |
1972年 | 16篇 |
1969年 | 21篇 |
1968年 | 15篇 |
排序方式: 共有5004条查询结果,搜索用时 15 毫秒
111.
Megan K. Mueller 《应用发育科学》2013,17(1):5-16
Human-animal relationships are ubiquitous and diverse across the life span and may be especially salient among children and adolescents. However, there is little information regarding whether human-animal interaction (HAI) is actually linked to young people's positive development in normative, nontherapeutic settings. Therefore, the purpose of this study was to explore if and how HAI may be linked to positive youth development (PYD) using data from the 4-H Study of PYD. Results suggested that emotions and cognitions about animals were related to indices of positive development. The implications of these findings, and suggestions for new areas of inquiry regarding the role of HAI as an important developmental context, are discussed. 相似文献
112.
Global Analysis of Apicomplexan Protein S‐Acyl Transferases Reveals an Enzyme Essential for Invasion
Karine Frénal Chwen L. Tay Christina Mueller Ellen S. Bushell Yonggen Jia Arnault Graindorge Oliver Billker Julian C. Rayner Dominique Soldati‐Favre 《Traffic (Copenhagen, Denmark)》2013,14(8):895-911
The advent of techniques to study palmitoylation on a whole proteome scale has revealed that it is an important reversible modification that plays a role in regulating multiple biological processes. Palmitoylation can control the affinity of a protein for lipid membranes, which allows it to impact protein trafficking, stability, folding, signalling and interactions. The publication of the palmitome of the schizont stage of Plasmodium falciparum implicated a role for palmitoylation in host cell invasion, protein export and organelle biogenesis. However, nothing is known so far about the repertoire of protein S‐acyl transferases (PATs) that catalyse this modification in Apicomplexa. We undertook a comprehensive analysis of the repertoire of Asp‐His‐His‐Cys cysteine‐rich domain (DHHC‐CRD) PAT family in Toxoplasma gondii and Plasmodium berghei by assessing their localization and essentiality. Unlike functional redundancies reported in other eukaryotes, some apicomplexan‐specific DHHCs are essential for parasite growth, and several are targeted to organelles unique to this phylum. Of particular interest is DHHC7, which localizes to rhoptry organelles in all parasites tested, including the major human pathogen P. falciparum. TgDHHC7 interferes with the localization of the rhoptry palmitoylated protein TgARO and affects the apical positioning of the rhoptry organelles. This PAT has a major impact on T. gondii host cell invasion, but not on the parasite's ability to egress. 相似文献
113.
114.
Galliano Zanello Ashleigh Goethel Katharina Forster Kaoru Geddes Dana J. Philpott Kenneth Croitoru 《PloS one》2013,8(12)
Although the etiology of Crohn''s disease (CD) remains elusive this disease is characterized by T cell activation that leads to chronic inflammation and mucosal damage. A potential role for maladaptation between the intestinal microbiota and the mucosal immune response is suggested by the fact that mutations in the pattern recognition receptor Nod2 are associated with higher risks for developing CD. Although Nod2 deletion in CD4+ T cells has been shown to impair the induction of colitis in the murine T cell transfer model, the analysis of T cell intrinsic Nod2 function in T cell differentiation and T cell-mediated immunity is inconsistent between several studies. In addition, the role of T cell intrinsic Nod2 in regulatory T cell (Treg) development and function during colitis remain to be analyzed. In this study, we show that Nod2 expression is higher in activated/memory CD4+ T cells and its expression was inducible after T cell receptor (TCR) ligation. Nod2 stimulation with muramyl dipeptide (MDP) led to a nuclear accumulation of c-Rel NF-kB subunit. Although functionally active in CD4+ T cells, the deletion of Nod2 did not impair the induction and the prevention of colitis in the T cell transfer model. Moreover, Nod2 deletion did not affect the development of Foxp3+ Treg cells in the spleen of recipient mice and Nod2 deficient CD4 T cells expressing the OVA specific transgenic TCR were able to differentiate in Foxp3+ Treg cells after OVA feeding. In vitro, CD25+ Nod2 deficient T cells suppressed T cell proliferation as well as wild type counter parts and T cell stimulation with MDP did not affect the proliferation and the cytokine secretion of T cells. In conclusion, our data indicate that Nod2 is functional in murine CD4+ T cells but its expression is dispensable for the T cell regulation of colitis. 相似文献
115.
116.
Darshini Govindasamy Katharina Kranzer Nienke van Schaik Farzad Noubary Robin Wood Rochelle P. Walensky Kenneth A. Freedberg Ingrid V. Bassett Linda-Gail Bekker 《PloS one》2013,8(11)
Background
HIV counseling and testing may serve as an entry point for non-communicable disease screening.Objectives
To determine the yield of newly-diagnosed HIV, tuberculosis (TB) symptoms, diabetes and hypertension, and to assess CD4 count testing, linkage to care as well as correlates of linkage and barriers to care from a mobile testing unit.Methods
A mobile unit provided screening for HIV, TB symptoms, diabetes and hypertension in Cape Town, South Africa between March 2010 and September 2011. The yield of newly-diagnosed cases of these conditions was measured and clients were followed-up between January and November 2011 to assess linkage. Linkage to care was defined as accessing care within one, three or six months post-HIV diagnosis (dependent on CD4 count) and one month post-diagnosis for other conditions. Clinical and socio-demographic correlates of linkage to care were evaluated using Poisson regression and barriers to care were determined.Results
Of 9,806 clients screened, the yield of new diagnoses was: HIV (5.5%), TB suspects (10.1%), diabetes (0.8%) and hypertension (58.1%). Linkage to care for HIV-infected clients, TB suspects, diabetics and hypertensives was: 51.3%, 56.7%, 74.1% and 50.0%. Only disclosure of HIV-positive status to family members or partners (RR=2.6, 95% CI: 1.04-6.3, p=0.04) was independently associated with linkage to HIV care. The main barrier to care reported by all groups was lack of time to access a clinic.Conclusion
Screening for HIV, TB symptoms and hypertension at mobile units in South Africa has a high yield but inadequate linkage. After-hours and weekend clinics may overcome a major barrier to accessing care. 相似文献117.
Jessica L. Barrington-Trimis Susan Searles Nielsen Susan Preston-Martin W. James Gauderman Elizabeth A. Holly Federico M. Farin Beth A. Mueller Roberta McKean-Cowdin 《PloS one》2013,8(11)
Background
A recent meta-analysis suggested an association between exposure to paternal smoking during pregnancy and childhood brain tumor risk, but no studies have evaluated whether this association differs by polymorphisms in genes that metabolize tobacco-smoke chemicals.Methods
We assessed 9 functional polymorphisms in 6 genes that affect the metabolism of polycyclic aromatic hydrocarbons (PAH) to evaluate potential interactions with parental smoking during pregnancy in a population-based case-control study of childhood brain tumors. Cases (N = 202) were ≤10 years old, diagnosed from 1984–1991 and identified in three Surveillance, Epidemiology, and End Results (SEER) registries in the western U.S. Controls in the same regions (N = 286) were frequency matched by age, sex, and study center. DNA for genotyping was obtained from archived newborn dried blood spots.Results
We found positive interaction odds ratios (ORs) for both maternal and paternal smoking during pregnancy, EPHX1 H139R, and childhood brain tumors (P interaction = 0.02; 0.10), such that children with the high-risk (greater PAH activation) genotype were at a higher risk of brain tumors relative to children with the low-risk genotype when exposed to tobacco smoke during pregnancy. A dose-response pattern for paternal smoking was observed among children with the EPHX1 H139R high-risk genotype only (ORno exposure = 1.0; OR≤3 hours/day = 1.32, 95% CI: 0.52–3.34; OR>3hours/day = 3.18, 95% CI: 0.92–11.0; P trend = 0.07).Conclusion
Parental smoking during pregnancy may be a risk factor for childhood brain tumors among genetically susceptible children who more rapidly activate PAH in tobacco smoke. 相似文献118.
The design and generation of DNA constructs is among the necessary but generally tedious tasks for molecular biologists and, typically, the cloning strategy is restricted by available restriction sites. However, increasingly sophisticated experiments require increasingly complex DNA constructs, with an intricacy that exceeds what is achievable using standard cloning procedures. Many transgenes such as inducible gene cassettes or recombination elements consist of multiple components that often require precise in-frame fusions. Here, we present an efficient protocol that facilitates the generation of these complex constructs. The golden GATEway cloning approach presented here combines two established cloning methods, namely golden Gate cloning and Multisite GatewayTM cloning. This allows efficient and seamless assembly as well as reuse of predefined DNA elements. The golden Gate cloning procedure follows clear and simple design rules and allows the assembly of multiple fragments with different sizes into one open reading frame. The final product can be directly integrated into the widely used Multisite GatewayTM cloning system, granting more flexibility when using a transgene in the context of multiple species. This adaptable and streamlined cloning procedure overcomes restrictions of “classical construct generation” and allows focusing on construct design. 相似文献
119.
Katrin Bomans Antje Lang Veronika Roedl Lisa Adolf Kyrillos Kyriosoglou Katharina Diepold Gabriele Eberl Michael M?lh?j Ulrike Strauss Christian Schmalz Rudolf Vogel Dietmar Reusch Harald Wegele Michael Wiedmann Patrick Bulau 《PloS one》2013,8(11)
Biotherapeutics are often produced in non-human host cells like Escherichia coli, yeast, and various mammalian cell lines. A major focus of any therapeutic protein purification process is to reduce host cell proteins to an acceptable low level. In this study, various E. coli host cell proteins were identified at different purifications steps by HPLC fractionation, SDS-PAGE analysis, and tryptic peptide mapping combined with online liquid chromatography mass spectrometry (LC-MS). However, no host cell proteins could be verified by direct LC-MS analysis of final drug substance material. In contrast, the application of affinity enrichment chromatography prior to comprehensive LC-MS was adequate to identify several low abundant host cell proteins at the final drug substance level. Bacterial alkaline phosphatase (BAP) was identified as being the most abundant host cell protein at several purification steps. Thus, we firstly established two different assays for enzymatic and immunological BAP monitoring using the cobas® technology. By using this strategy we were able to demonstrate an almost complete removal of BAP enzymatic activity by the established therapeutic protein purification process. In summary, the impact of fermentation, purification, and formulation conditions on host cell protein removal and biological activity can be conducted by monitoring process-specific host cell proteins in a GMP-compatible and high-throughput (> 1000 samples/day) manner. 相似文献
120.
Cristian Koepfli Kathryn L. Colborn Benson Kiniboro Enmoore Lin Terence P. Speed Peter M. Siba Ingrid Felger Ivo Mueller 《PLoS neglected tropical diseases》2013,7(9)