全文获取类型
收费全文 | 2963篇 |
免费 | 232篇 |
专业分类
3195篇 |
出版年
2023年 | 8篇 |
2022年 | 27篇 |
2021年 | 53篇 |
2020年 | 24篇 |
2019年 | 40篇 |
2018年 | 71篇 |
2017年 | 57篇 |
2016年 | 90篇 |
2015年 | 117篇 |
2014年 | 153篇 |
2013年 | 204篇 |
2012年 | 273篇 |
2011年 | 256篇 |
2010年 | 153篇 |
2009年 | 141篇 |
2008年 | 211篇 |
2007年 | 182篇 |
2006年 | 179篇 |
2005年 | 169篇 |
2004年 | 140篇 |
2003年 | 157篇 |
2002年 | 128篇 |
2001年 | 26篇 |
2000年 | 27篇 |
1999年 | 13篇 |
1998年 | 19篇 |
1997年 | 21篇 |
1996年 | 14篇 |
1995年 | 8篇 |
1994年 | 10篇 |
1993年 | 14篇 |
1992年 | 11篇 |
1991年 | 8篇 |
1988年 | 9篇 |
1987年 | 6篇 |
1986年 | 8篇 |
1985年 | 7篇 |
1984年 | 6篇 |
1982年 | 13篇 |
1980年 | 7篇 |
1979年 | 11篇 |
1978年 | 9篇 |
1977年 | 9篇 |
1976年 | 5篇 |
1974年 | 5篇 |
1973年 | 11篇 |
1971年 | 9篇 |
1970年 | 6篇 |
1969年 | 6篇 |
1967年 | 5篇 |
排序方式: 共有3195条查询结果,搜索用时 0 毫秒
991.
Becker J Semler O Gilissen C Li Y Bolz HJ Giunta C Bergmann C Rohrbach M Koerber F Zimmermann K de Vries P Wirth B Schoenau E Wollnik B Veltman JA Hoischen A Netzer C 《American journal of human genetics》2011,(3):11-371
Osteogenesis imperfecta (OI) is a heterogeneous genetic disorder characterized by bone fragility and susceptibility to fractures after minimal trauma. After mutations in all known OI genes had been excluded by Sanger sequencing, we applied next-generation sequencing to analyze the exome of a single individual who has a severe form of the disease and whose parents are second cousins. A total of 26,922 variations from the human reference genome sequence were subjected to several filtering steps. In addition, we extracted the genotypes of all dbSNP130-annotated SNPs from the exome sequencing data and used these 299,494 genotypes as markers for the genome-wide identification of homozygous regions. A single homozygous truncating mutation, affecting SERPINF1 on chromosome 17p13.3, that was embedded into a homozygous stretch of 2.99 Mb remained. The mutation was also homozygous in the affected brother of the index patient. Subsequently, we identified homozygosity for two different truncating SERPINF1 mutations in two unrelated patients with OI and parental consanguinity. All four individuals with SERPINF1 mutations have severe OI. Fractures of long bones and severe vertebral compression fractures with resulting deformities were observed as early as the first year of life in these individuals. Collagen analyses with cultured dermal fibroblasts displayed no evidence for impaired collagen folding, posttranslational modification, or secretion. SERPINF1 encodes pigment epithelium-derived factor (PEDF), a secreted glycoprotein of the serpin superfamily. PEDF is a multifunctional protein and one of the strongest inhibitors of angiogenesis currently known in humans. Our data provide genetic evidence for PEDF involvement in human bone homeostasis. 相似文献
992.
Scavenius C Sanggaard KW Nikolajsen CL Bak S Valnickova Z Thøgersen IB Jensen ON Højrup P Enghild JJ 《Biochimica et biophysica acta》2011,1814(12):1624-1630
In this study, we show that inter-α-inhibitor is a substrate for both factor XIIIa and tissue transglutaminase. These enzymes catalyze the incorporation of dansylcadaverine and biotin-pentylamine, revealing that inter-α-inhibitor contains reactive Gln residues within all three subunits. These findings suggest that transglutaminases catalyze the covalent conjugation of inter-α-inhibitor to other proteins. This was demonstrated by the cross-linking between inter-α-inhibitor and fibrinogen by either factor XIIIa or tissue transglutaminase. Finally, using quantitative mass spectrometry, we show that inter-α-inhibitor is cross-linked to the fibrin clot in a 1:20 ratio relative to the known factor XIIIa substrate α2-antiplasmin. This interaction may protect fibrin or other Lys-donating proteins from adventitious proteolysis by increasing the local concentration of bikunin. In addition, the reaction may influence the TSG-6/heavy Chain 2-mediated transfer of heavy chains observed during inflammation. 相似文献
993.
Rbfox-regulated alternative splicing is critical for zebrafish cardiac and skeletal muscle functions
Gallagher TL Arribere JA Geurts PA Exner CR McDonald KL Dill KK Marr HL Adkar SS Garnett AT Amacher SL Conboy JG 《Developmental biology》2011,(2):251-261
Rbfox RNA binding proteins are implicated as regulators of phylogenetically-conserved alternative splicing events important for muscle function. To investigate the function of rbfox genes, we used morpholino-mediated knockdown of muscle-expressed rbfox1l and rbfox2 in zebrafish embryos. Single and double morphant embryos exhibited changes in splicing of overlapping sets of bioinformatically-predicted rbfox target exons, many of which exhibit a muscle-enriched splicing pattern that is conserved in vertebrates. Thus, conservation of intronic Rbfox binding motifs is a good predictor of Rbfox-regulated alternative splicing. Morphology and development of single morphant embryos were strikingly normal; however, muscle development in double morphants was severely disrupted. Defects in cardiac muscle were marked by reduced heart rate and in skeletal muscle by complete paralysis. The predominance of wavy myofibers and abnormal thick and thin filaments in skeletal muscle revealed that myofibril assembly is defective and disorganized in double morphants. Ultra-structural analysis revealed that although sarcomeres with electron dense M- and Z-bands are present in muscle fibers of rbfox1l/rbox2 morphants, they are substantially reduced in number and alignment. Importantly, splicing changes and morphological defects were rescued by expression of morpholino-resistant rbfox cDNA. Additionally, a target-blocking MO complementary to a single UGCAUG motif adjacent to an rbfox target exon of fxr1 inhibited inclusion in a similar manner to rbfox knockdown, providing evidence that Rbfox regulates the splicing of target exons via direct binding to intronic regulatory motifs. We conclude that Rbfox proteins regulate an alternative splicing program essential for vertebrate heart and skeletal muscle functions. 相似文献
994.
Günther?Joseph?JirikowskiEmail author Stefan?Richter Carsten?Wolff 《Frontiers in zoology》2013,10(1):76
Background
Malacostracan evolutionary history has seen multiple transformations of ontogenetic mode. For example direct development in connection with extensive brood care and development involving planktotrophic nauplius larvae, as well as intermediate forms are found throughout this taxon. This makes the Malacostraca a promising group for study of evolutionary morphological diversification and the role of heterochrony therein. One candidate heterochronic phenomenon is represented by the concept of the ‘egg-nauplius’, in which the nauplius larva, considered plesiomorphic to all Crustacea, is recapitulated as an embryonic stage.Results
Here we present a comparative investigation of embryonic muscle differentiation in four representatives of Malacostraca: Gonodactylaceus falcatus (Stomatopoda), Neocaridina heteropoda (Decapoda), Neomysis integer (Mysida) and Parhyale hawaiensis (Amphipoda). We describe the patterns of muscle precursors in different embryonic stages to reconstruct the sequence of muscle development, until hatching of the larva or juvenile. Comparison of the developmental sequences between species reveals extensive heterochronic and heteromorphic variation. Clear anticipation of muscle differentiation in the nauplius segments, but also early formation of longitudinal trunk musculature independently of the teloblastic proliferation zone, are found to be characteristic to stomatopods and decapods, all of which share an egg-nauplius stage.Conclusions
Our study provides a strong indication that the concept of nauplius recapitulation in Malacostraca is incomplete, because sequences of muscle tissue differentiation deviate from the chronological patterns observed in the ectoderm, on which the egg-nauplius is based. However, comparison of myogenic sequences between taxa supports the hypothesis of a zoea-like larva that was present in the last common ancestor of Eumalacostraca (Malacostraca without Leptostraca). We argue that much of the developmental sequences of larva muscle patterning were retained in the eumalacostracan lineage despite the reduction of free swimming nauplius larvae, but was severely reduced in the peracaridean clade.995.
David J. Pedersen Barbara Diakanastasis Jacqueline St?ckli Carsten Schmitz-Peiffer 《PloS one》2013,8(3)
We have previously shown that deletion of protein kinase C epsilon (PKCε) in mice results in protection against glucose intolerance caused by a high fat diet. This was in part due to reduced insulin uptake by hepatocytes and insulin clearance, which enhanced insulin availability. Here we employed mouse embryonic fibroblasts (MEFs) derived from wildtype (WT) and PKCε-deficient (PKCε−/−) mice to examine this mechanistically. PKCε−/− MEFs exhibited reduced insulin uptake which was associated with decreased insulin receptor phosphorylation, while downstream signalling through IRS-1 and Akt was unaffected. Cellular fractionation demonstrated that PKCε deletion changed the localization of the insulin receptor, a greater proportion of which co-fractionated with flotillin-1, a marker of membrane microdomains. Insulin stimulation resulted in redistribution of the receptor in WT cells, while this was markedly reduced in PKCε−/− cells. These alterations in insulin receptor trafficking were associated with reduced expression of CEACAM1, a receptor substrate previously shown to modulate insulin clearance. Virally-mediated reconstitution of PKCε in MEFs increased CEACAM1 expression and partly restored the sensitivity of the receptor to insulin-stimulated redistribution. These data indicate that PKCε can affect insulin uptake in MEFs through promotion of receptor-mediated endocytosis, and that this may be mediated by regulation of CEACAM1 expression. 相似文献
996.
Isabelle Beck Susanne Jochner Stefanie Gilles Mareike McIntyre Jeroen T. M. Buters Carsten Schmidt-Weber Heidrun Behrendt Johannes Ring Annette Menzel Claudia Traidl-Hoffmann 《PloS one》2013,8(11)
Background
Evidence is compelling for a positive correlation between climate change, urbanisation and prevalence of allergic sensitisation and diseases. The reason for this association is not clear to date. Some data point to a pro-allergenic effect of anthropogenic factors on susceptible individuals.Objectives
To evaluate the impact of urbanisation and climate change on pollen allergenicity.Methods
Catkins were sampled from birch trees from different sites across the greater area of Munich, pollen were isolated and an urbanisation index, NO2 and ozone exposure were determined. To estimate pollen allergenicity, allergen content and pollen-associated lipid mediators were measured in aqueous pollen extracts. Immune stimulatory and modulatory capacity of pollen was assessed by neutrophil migration assays and the potential of pollen to inhibit dendritic cell interleukin-12 response. In vivo allergenicity was assessed by skin prick tests.Results
The study revealed ozone as a prominent environmental factor influencing the allergenicity of birch pollen. Enhanced allergenicity, as assessed in skin prick tests, was mirrored by enhanced allergen content. Beyond that, ozone induced changes in lipid composition and chemotactic and immune modulatory potential of the pollen. Higher ozone-exposed pollen was characterised by less immune modulatory but higher immune stimulatory potential.Conclusion
It is likely that future climate change along with increasing urbanisation will lead to rising ozone concentrations in the next decades. Our study indicates that ozone is a crucial factor leading to clinically relevant enhanced allergenicity of birch pollen. Thus, with increasing temperatures and increasing ozone levels, also symptoms of pollen allergic patients may increase further. 相似文献997.
C. Alexander Valencia Arunkanth Ankala Devin Rhodenizer Shruti Bhide Martin Robert Littlejohn Lisa Mari Keong Anne Rutkowski Susan Sparks Carsten Bonnemann Madhuri Hegde 《PloS one》2013,8(1)
The congenital muscular dystrophies (CMDs) comprise a heterogeneous group of heritable muscle disorders with often difficult to interpret muscle pathology, making them challenging to diagnose. Serial Sanger sequencing of suspected CMD genes, while the current molecular diagnostic method of choice, can be slow and expensive. A comprehensive panel test for simultaneous screening of mutations in all known CMD-associated genes would be a more effective diagnostic strategy. Thus, the CMDs are a model disorder group for development and validation of next-generation sequencing (NGS) strategies for diagnostic and clinical care applications. Using a highly multiplexed PCR-based target enrichment method (RainDance) in conjunction with NGS, we performed mutation detection in all CMD genes of 26 samples and compared the results with Sanger sequencing. The RainDance NGS panel showed great consistency in coverage depth, on-target efficiency, versatility of mutation detection, and genotype concordance with Sanger sequencing, demonstrating the test''s appropriateness for clinical use. Compared to single tests, a higher diagnostic yield was observed by panel implementation. The panel''s limitation is the amplification failure of select gene-specific exons which require Sanger sequencing for test completion. Successful validation and application of the CMD NGS panel to improve the diagnostic yield in a clinical laboratory was shown. 相似文献
998.
Silke Dornieden Andreas Müller-Schiffmann Heinrich Sticht Nan Jiang Yeliz Cinar Michael W?rdehoff Carsten Korth Susanne Aileen Funke Dieter Willbold 《PloS one》2013,8(3)
Alzheimer’s disease (AD) is a progressive neurodegenerative disorder with devastating effects. Currently, therapeutic options are limited to symptomatic treatment. For more than a decade, research focused on immunotherapy for the causal treatment of AD. However, clinical trials with active immunization using Aβ encountered severe complications, for example meningoencephalitis. Consequently, attention focused on passive immunization using antibodies. As an alternative to large immunoglobulins (IgGs), Aβ binding single-chain variable fragments (scFvs) were used for diagnostic and therapeutic research approaches. scFvs can be expressed in E. coli and may provide improved pharmacokinetic properties like increased blood-brain barrier permeability or reduced side-effects in vivo. In this study, we constructed an scFv from an Aβ binding IgG, designated IC16, which binds the N-terminal region of Aβ (Aβ(1-8)). scFv-IC16 was expressed in E. coli, purified and characterized with respect to its interaction with different Aβ species and its influence on Aβ fibril formation. We were able to show that scFv-IC16 strongly influenced the aggregation behavior of Aβ and could be applied as an Aβ detection probe for plaque staining in the brains of transgenic AD model mice. The results indicate potential for therapy and diagnosis of AD. 相似文献
999.
1000.
Carsten Gründemann Kathrin Thell Karin Lengen Manuel Garcia-K?ufer Yen-Hua Huang Roman Huber David J. Craik Gernot Schabbauer Christian W. Gruber 《PloS one》2013,8(6)
Cyclotides are a diverse and abundant group of ribosomally synthesized plant peptides containing a unique cyclic cystine-knotted topology that confers them with remarkable stability. Kalata B1, a representative member of this family of mini-proteins, has been found to inhibit the proliferation of human peripheral blood mononuclear cells. Analysis of T-cell proliferation upon treatment with chemically synthesized kalata B1 mutants revealed a region comprising inter-cysteine loops 1 and 2 of the cyclotide framework to be important for biological activity. Cytokine signaling analysis using an ‘active’ kalata B1 mutant [T20K], and the reference drug cyclosporin A (CsA) demonstrated that treatment of activated T-lymphocytes with these compounds decreased the expression of the interleukin-2 (IL-2) surface receptor as well as IL-2 cytokine secretion and IL-2 gene expression, whereas the ‘inactive’ kalata B1 mutant [V10K] did not cause any effects. The anti-proliferative activity of [T20K] kalata B1 was antagonized by addition of exogenous IL-2. Furthermore, treatment with [T20K] kalata B1 led to an initial reduction of the effector function, as indicated by the reduced IFN-γ and TNF-α production, but the levels of both cytokines stabilized over time and returned to their normal levels. On the other hand, the degranulation activity remained reduced. This indicated that cyclotides interfere with T-cell polyfunctionality and arrest the proliferation of immune-competent cells through inhibiting IL-2 biology at more than one site. The results open new avenues to utilize native and synthetically-optimized cyclotides for applications in immune-related disorders and as immunosuppressant peptides. 相似文献