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971.
Katharina Reglinski Marina Keil Sabrina Altendorf Dominic Waithe Christian Eggeling Wolfgang Schliebs Ralf Erdmann 《PloS one》2015,10(10)
The human deubiquitinating enzyme ubiquitin-specific protease 2 (USP2) regulates multiple cellular pathways, including cell proliferation and apoptosis. As a result of alternative splicing four USP2 isoenzymes are expressed in human cells of which all contain a weak peroxisome targeting signal of type 1 (PTS1) at their C-termini. Here, we systematically analyzed apoptotic effects induced by overexpression and intracellular localization for each isoform. All isoforms exhibit proapoptotic activity and are post-translationally imported into the matrix of peroxisomes in a PEX5-dependent manner. However, a significant fraction of the USP2 pool resides in the cytosol due to a weaker PTS1 and thus low affinity to the PTS receptor PEX5. Blocking of peroxisomal import did not interfere with the proapoptotic activity of USP2, suggesting that the enzyme performs its critical function outside of this compartment. Instead, increase of the efficiency of USP2 import into peroxisomes either by optimization of its peroxisomal targeting signal or by overexpression of the PTS1 receptor did result in a reduction of the apoptotic rate of transfected cells. Our studies suggest that peroxisomal import of USP2 provides additional control over the proapoptotic activity of cytosolic USP2 by spatial separation of the deubiquitinating enzymes from their interaction partners in the cytosol and nucleus. 相似文献
972.
While it is generally agreed that perception can occur without awareness, there continues to be debate about the type of representational content that is accessible when awareness is minimized or eliminated. Most investigations that have addressed this issue evaluate access to well-learned representations. Far fewer studies have evaluated whether or not associations encountered just once prior to testing might also be accessed and influence behavior. Here, eye movements were used to examine whether or not memory for studied relationships is evident following the presentation of subliminal cues. Participants assigned to experimental or control groups studied scene-face pairs and test trials evaluated implicit and explicit memory for these pairs. Each test trial began with a subliminal scene cue, followed by three visible studied faces. For experimental group participants, one face was the studied associate of the scene (implicit test); for controls none were a match. Subsequently, the display containing a match was presented to both groups, but now it was preceded by a visible scene cue (explicit test). Eye movements were recorded and recognition memory responses were made. Participants in the experimental group looked disproportionately at matching faces on implicit test trials and participants from both groups looked disproportionately at matching faces on explicit test trials, even when that face had not been successfully identified as the associate. Critically, implicit memory-based viewing effects seemed not to depend on residual awareness of subliminal scene cues, as subjective and objective measures indicated that scenes were successfully masked from view. The reported outcomes indicate that memory for studied relationships can be expressed in eye movement behavior without awareness. 相似文献
973.
Darren M. Crayn Klaus Winter Katharina Schulte J. Andrew C. Smith 《Botanical journal of the Linnean Society. Linnean Society of London》2015,178(2):169-221
A comprehensive analysis of photosynthetic pathways in relation to phylogeny and elevational distribution was conducted in Bromeliaceae, an ecologically diverse Neotropical family containing large numbers of both terrestrial and epiphytic species. Tissue carbon isotope ratio (δ13C) was used to determine the occurrence of crassulacean acid metabolism (CAM) and C3 photosynthesis in 1893 species, representing 57% of species and all 56 genera in the family. The frequency of δ13C values showed a strongly bimodal distribution: 1074 species (57%) had values more negative than −20‰ (mode = −26.7‰), typical of predominantly daytime carbon fixation via the C3 pathway, whereas 819 species (43%) possessed values less negative than −20‰ (mode = −13.3‰), indicative of predominantly nocturnal fixation of carbon via the CAM pathway. Amongst the six almost exclusively terrestrial subfamilies in Bromeliaceae, Brocchinioideae, Lindmanioideae and Navioideae consisted entirely of C3 species, with CAM species being restricted to Hechtioideae (all species of Hechtia tested), Pitcairnioideae (all species belonging to a xeric clade comprising Deuterocohnia, Dyckia and Encholirium) and Puyoideae (21% of Puya spp.). Of the other two subfamilies, in the overwhelmingly epiphytic (plus lithophytic) Tillandsioideae, 28% of species possessed CAM photosynthesis, all restricted to the derived genus Tillandsia and tending towards the more extreme epiphytic ‘atmospheric’ life‐form. In Bromelioideae, with comparable numbers of terrestrial and epiphytic species, 90% of taxa showed CAM; included in these are the first records of CAM photosynthesis in Androlepis, Canistropsis, Deinacanthon, Disteganthus, Edmundoa, Eduandrea, Hohenbergiopsis, Lymania, Pseudananas, Ronnbergia and Ursulaea. With respect to elevational gradients, the greatest number of C3 bromeliad species were found at mid‐elevations between 500 and 1500 m, whereas the frequency of CAM species declined monotonically with increasing elevation. However, in Puya, at least ten CAM species have been recorded at elevations > 3000 m, showing that CAM photosynthesis is not necessarily incompatible with low temperatures. This survey identifies five major origins of CAM photosynthesis at a higher taxonomic level in Bromeliaceae, but future phylogenetic work is likely to reveal a more fine‐scale pattern of gains and losses of this trait, especially in ecologically diverse and widely distributed genera such as Tillandsia and Puya. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 178, 169–221. 相似文献
974.
G-Andre Banat Aleksandra Tretyn Soni Savai Pullamsetti Jochen Wilhelm Andreas Weigert Catherine Olesch Katharina Ebel Thorsten Stiewe Friedrich Grimminger Werner Seeger Ludger Fink Rajkumar Savai 《PloS one》2015,10(9)
Recent studies indicate that the abnormal microenvironment of tumors may play a critical role in carcinogenesis, including lung cancer. We comprehensively assessed the number of stromal cells, especially immune/inflammatory cells, in lung cancer and evaluated their infiltration in cancers of different stages, types and metastatic characteristics potential. Immunohistochemical analysis of lung cancer tissue arrays containing normal and lung cancer sections was performed. This analysis was combined with cyto-/histomorphological assessment and quantification of cells to classify/subclassify tumors accurately and to perform a high throughput analysis of stromal cell composition in different types of lung cancer. In human lung cancer sections we observed a significant elevation/infiltration of total-T lymphocytes (CD3+), cytotoxic-T cells (CD8+), T-helper cells (CD4+), B cells (CD20+), macrophages (CD68+), mast cells (CD117+), mononuclear cells (CD11c+), plasma cells, activated-T cells (MUM1+), B cells, myeloid cells (PD1+) and neutrophilic granulocytes (myeloperoxidase+) compared with healthy donor specimens. We observed all of these immune cell markers in different types of lung cancers including squamous cell carcinoma, adenocarcinoma, adenosquamous cell carcinoma, small cell carcinoma, papillary adenocarcinoma, metastatic adenocarcinoma, and bronchioloalveolar carcinoma. The numbers of all tumor-associated immune cells (except MUM1+ cells) in stage III cancer specimens was significantly greater than those in stage I samples. We observed substantial stage-dependent immune cell infiltration in human lung tumors suggesting that the tumor microenvironment plays a critical role during lung carcinogenesis. Strategies for therapeutic interference with lung cancer microenvironment should consider the complexity of its immune cell composition. 相似文献
975.
Activation loop phosphorylation regulates B‐Raf in vivo and transformation by B‐Raf mutants 下载免费PDF全文
Björn Schorch Katharina Heilmann Natalie Stickel Gina J Fiala Lisa C Schmitt Sandra Braun Sophia Ehrenfeld Franziska M Uhl Thorsten Kaltenbacher Florian Weinberg Sebastian Herzog Robert Zeiser Wolfgang W Schamel Hassan Jumaa Tilman Brummer 《The EMBO journal》2016,35(2):143-161
Despite being mutated in cancer and RASopathies, the role of the activation segment (AS) has not been addressed for B‐Raf signaling in vivo. Here, we generated a conditional knock‐in mouse allowing the expression of the B‐RafAVKA mutant in which the AS phosphoacceptor sites T599 and S602 are replaced by alanine residues. Surprisingly, despite producing a kinase‐impaired protein, the BrafAVKA allele does not phenocopy the lethality of Braf‐knockout or paradoxically acting knock‐in alleles. However, BrafAVKA mice display abnormalities in the hematopoietic system, a distinct facial morphology, reduced ERK pathway activity in the brain, and an abnormal gait. This phenotype suggests that maximum B‐Raf activity is required for the proper development, function, and maintenance of certain cell populations. By establishing conditional murine embryonic fibroblast cultures, we further show that MEK/ERK phosphorylation and the immediate early gene response toward growth factors are impaired in the presence of B‐RafAVKA. Importantly, alanine substitution of T599/S602 impairs the transformation potential of oncogenic non‐V600E B‐Raf mutants and a fusion protein, suggesting that blocking their phosphorylation could represent an alternative strategy to ATP‐competitive inhibitors. 相似文献
976.
Dianne F. Newbury Laura Winchester Laura Addis Silvia Paracchini Ann Clark Wendy Cohen Hilary Cowie Katharina Dworzynski Andrea Everitt Ian M. Goodyer Elizabeth Hennessy A. David Kindley Laura L. Miller Anne O'Hare Duncan Shaw Zoe Simkin Emily Simonoff Vicky Slonims Jocelynne Watson Jiannis Ragoussis Jonathon R. Seckl Peter J. Helms Patrick F. Bolton Andrew Pickles Gina Conti-Ramsden Gillian Baird Dorothy V.M. Bishop Anthony P. Monaco 《American journal of human genetics》2009,85(2):264-245
Specific language impairment (SLI) is a common developmental disorder characterized by difficulties in language acquisition despite otherwise normal development and in the absence of any obvious explanatory factors. We performed a high-density screen of SLI1, a region of chromosome 16q that shows highly significant and consistent linkage to nonword repetition, a measure of phonological short-term memory that is commonly impaired in SLI. Using two independent language-impaired samples, one family-based (211 families) and another selected from a population cohort on the basis of extreme language measures (490 cases), we detected association to two genes in the SLI1 region: that encoding c-maf-inducing protein (CMIP, minP = 5.5 × 10−7 at rs6564903) and that encoding calcium-transporting ATPase, type2C, member2 (ATP2C2, minP = 2.0 × 10−5 at rs11860694). Regression modeling indicated that each of these loci exerts an independent effect upon nonword repetition ability. Despite the consistent findings in language-impaired samples, investigation in a large unselected cohort (n = 3612) did not detect association. We therefore propose that variants in CMIP and ATP2C2 act to modulate phonological short-term memory primarily in the context of language impairment. As such, this investigation supports the hypothesis that some causes of language impairment are distinct from factors that influence normal language variation. This work therefore implicates CMIP and ATP2C2 in the etiology of SLI and provides molecular evidence for the importance of phonological short-term memory in language acquisition. 相似文献
977.
Willecke F Zeschky K Ortiz Rodriguez A Colberg C Auwärter V Kneisel S Hutter M Lozhkin A Hoppe N Wolf D von zur Mühlen C Moser M Hilgendorf I Bode C Zirlik A 《PloS one》2011,6(4):e19405
Background
Strong evidence supports a protective role of the cannabinoid receptor 2 (CB2) in inflammation and atherosclerosis. However, direct proof of its involvement in lesion formation is lacking. Therefore, the present study aimed to characterize the role of the CB2 receptor in Murine atherogenesis.Methods and Findings
Low density lipoprotein receptor-deficient (LDLR−/−) mice subjected to intraperitoneal injections of the selective CB2 receptor agonist JWH-133 or vehicle three times per week consumed high cholesterol diet (HCD) for 16 weeks. Surprisingly, intimal lesion size did not differ between both groups in sections of the aortic roots and arches, suggesting that CB2 activation does not modulate atherogenesis in vivo. Plaque content of lipids, macrophages, smooth muscle cells, T cells, and collagen were also similar between both groups. Moreover, CB2 −/−/LDLR−/− mice developed lesions of similar size containing more macrophages and lipids but similar amounts of smooth muscle cells and collagen fibers compared with CB2 +/+/LDLR−/− controls. While JWH-133 treatment reduced intraperitoneal macrophage accumulation in thioglycollate-illicited peritonitis, neither genetic deficiency nor pharmacologic activation of the CB2 receptor altered inflammatory cytokine expression in vivo or inflammatory cell adhesion in the flow chamber in vitro.Conclusion
Our study demonstrates that both activation and deletion of the CB2 receptor do not relevantly modulate atherogenesis in mice. Our data do not challenge the multiple reports involving CB2 in other inflammatory processes. However, in the context of atherosclerosis, CB2 does not appear to be a suitable therapeutic target for reduction of the atherosclerotic plaque. 相似文献978.
Hüttner KG Breuer SK Paul P Majdic O Heitger A Felzmann T 《Cancer immunology, immunotherapy : CII》2005,54(1):67-77
To induce cytolytic immunity, dendritic cells (DCs) need to release bioactive interleukin-12 (IL-12) p70 heterodimeric molecules. To study the role of IL-12 for the generation of an anti-tumor immune response, we generated two classes of DCs. (1) DCs were initiated to secrete IL-12 by exposure to LPS/IFN- for 2 h resulting, as demonstrated in vitro, in continued IL-12 release for another 24 h (termed active DCs). (2) DCs were exposed to LPS/IFN- for 24 h and injected into mice at a time point when IL-12 production had ceased (termed exhausted DCs). These two classes of DCs were probed for their capacity to induce a cytolytic anti-tumor immune response in vivo in a syngeneic mouse tumor model. The mouse tumor cell line K-Balb was engineered to express neomycin phosphotransferase (NPT) as a model tumor antigen. DCs were charged with various NPT-derived antigens, including recombinant NPT protein, whole tumor cell lysate and NPT-derived synthetic peptides, and the induction of in vivo anti-tumor immunity was determined by measuring tumor growth. Only the injection of active DCs, i.e., cells that maintained the capacity to secrete IL-12, but not exhausted DCs that had lost the ability to produce IL-12, resulted in a measurable deceleration of growth of K-Balb-NPT tumors. This anti-tumor immune response was most pronounced when using recombinant protein as an antigen source, which was evident in a prophylactic as well as in a therapeutic setting. The absence of a response to parental K-Balb tumors confirmed the antigen specificity of the anti-tumor immune response. Together these data provide evidence for the unique capacity of actively IL-12 secreting DCs to trigger effective anti-tumor immunity using exogenous tumor antigens. 相似文献
979.
Brigitte Mack Carola Eggert Katharina Eder Sannia Imrich Philipp Baumeister Ulrich Harréus Olivier Gires 《PloS one》2013,8(1)
The study of tumourigenesis commonly involves the use of established cell lines or single cell suspensions of primary tumours. Standard methods for the generation of short-term tumour cell cultures include the disintegration of tissue based on enzymatic and mechanical stress. Here, we describe a simple and rapid method for the preparation of single cells from primary carcinomas, which is independent of enzymatic treatment and feeder cells. Tumour biopsies are processed to 1 mm3 cubes termed explants, which are cultured 1–3 days on agarose-coated well plates in specified medium. Through incisions generated in the explants, single cells are retrieved and collected from the culture supernatant and can be used for further analysis including in vitro and in vivo studies. Collected cells retain tumour-forming capacity in xenotransplantation assays, mimic the phenotype of the primary tumour, and facilitate the generation of cell lines. 相似文献
980.