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61.
Background
Continuing efforts in development of non-invasive prenatal genetic tests have focused on the isolation of fetal nucleated red blood cells (NRBCs) from maternal blood for decades. Because no fetal cell-specific antibody has been described so far, the present study focused on the development of monoclonal antibodies (mAbs) to antigens that are expressed exclusively on fetal NRBCs.Methods: Mice were immunized with fetal erythroid cell membranes and hybridomas screened for Abs using a multi-parameter fluorescence-activated cell sorting (FACS). Selected mAbs were evaluated by comparative FACS analysis involving Abs known to bind erythroid cell surface markers (CD71, CD36, CD34), antigen-i, galactose, or glycophorin-A (GPA). Specificity was further confirmed by extensive immunohistological and immunocytological analyses of NRBCs from umbilical cord blood and fetal and adult cells from liver, bone marrow, peripheral blood, and lymphoid tissues.Results: Screening of 690 hybridomas yielded three clones of which Abs from 4B8 and 4B9 clones demonstrated the desired specificity for a novel antigenic structure expressed on fetal erythroblast cell membranes. The antigenic structure identified is different from known surface markers (CD36, CD71, GPA, antigen-i, and galactose), and is not present on circulating adult erythroid cells, except for occasional detectability in adult bone marrow cells.Conclusions:The new mAbs specifically bind the same or highly overlapping epitopes of a surface antigen that is almost exclusively expressed on fetal erythroid cells. The high specificity of the mAbs should facilitate development of simple methods for reliable isolation of fetal NRBCs and their use in non-invasive prenatal diagnosis of fetal genetic status. 相似文献62.
63.
Masato Ohtani Toshiaki Kondo Naoki Tani Saneyoshi Ueno Leong S. Lee Kevin K. S. Ng Norwati Muhammad Reiner Finkeldey Mohamad Na'iem Sapto Indrioko Koichi Kamiya Ko Harada Bibian Diway Eyen Khoo Kensuke Kawamura Yoshihiko Tsumura 《Molecular ecology》2013,22(8):2264-2279
Tropical rainforests in South‐East Asia have been affected by climatic fluctuations during past glacial eras. To examine how the accompanying changes in land areas and temperature have affected the genetic properties of rainforest trees in the region, we investigated the phylogeographic patterns of a widespread dipterocarp species, Shorea leprosula. Two types of DNA markers were used: expressed sequence tag‐based simple sequence repeats and chloroplast DNA (cpDNA) sequence variations. Both sets of markers revealed clear genetic differentiation between populations in Borneo and those in the Malay Peninsula and Sumatra (Malay/Sumatra). However, in the south‐western part of Borneo, genetic admixture of the lineages was observed in the two marker types. Coalescent simulation based on cpDNA sequence variation suggested that the two lineages arose 0.28–0.09 million years before present and that following their divergence migration from Malay/Sumatra to Borneo strongly exceeded migration in the opposite direction. We conclude that the genetic structure of S. leprosula was largely formed during the middle Pleistocene and was subsequently modified by eastward migration across the subaerially exposed Sunda Shelf. 相似文献
64.
65.
Carboxypeptidase D is the only enzyme responsible for antibody C‐terminal lysine cleavage in Chinese hamster ovary (CHO) cells 下载免费PDF全文
66.
The small GTPase Rac1 is implicated in various cellular processes that are essential for normal cell function. Deregulation of Rac1 signaling has also been linked to a number of diseases, including cancer. The diversity of Rac1 functioning in cells is mainly attributed to its ability to bind to a multitude of downstream effectors following activation by Guanine nucleotide Exchange Factors (GEFs). Despite the identification of a large number of Rac1 binding partners, factors influencing downstream specificity are poorly defined, thus hindering the detailed understanding of both Rac1's normal and pathological functions. In a recent study, we demonstrated a role for 2 Rac-specific GEFs, Tiam1 and P-Rex1, in mediating Rac1 anti- versus pro-migratory effects, respectively. Importantly, via conducting a quantitative proteomic screen, we identified distinct changes in the Rac1 interactome following activation by either GEF, indicating that these opposing effects are mediated through GEF modulation of the Rac1 interactome. Here, we present the full list of identified Rac1 interactors together with functional annotation of the differentially regulated Rac1 binding partners. In light of this data, we also provide additional insights into known and novel signaling cascades that might account for the GEF-mediated Rac1-driven cellular effects. 相似文献
67.
《Molecular & cellular proteomics : MCP》2019,18(10):2108-2120
Highlights
- •Bayesian Beta-Binomial model integrates ion statistics with peptide ratio agreement.
- •Model appropriately interprets information from low signal peptides.
- •Confidence can be assigned even without replicates.
- •Model adds sensitivity to detection of small changes.
68.
《Molecular & cellular proteomics : MCP》2019,18(1):51-64
Highlights
- •microRNA-222 attenuates TGEV-induced mitochondrial dysfunction.
- •microRNA-222 downregulates THBS1 and CD47.
- •THBS1 is the target of microRNA-222 during TGEV infection.
- •THBS1 and CD47 increase mitochondrial Ca2+ level and reduced mitochondrial membrane potential (MMP).
69.
《Molecular & cellular proteomics : MCP》2019,18(2):383-390
Highlights
- •Fast and simple capillary column packing protocol.
- •Low-pressure packing at <100 bars from ultrahigh sorbent suspension concentration.
- •Sorbent particle aggregation leading to blocking of the column entrance is avoided.
- •Effective for long capillary UHPLC column packing with a wide range of sorbents.
70.
Qingping Xu Matthew Biancalana Joanna C. Grant Hsiu‐Ju Chiu Lukasz Jaroszewski Mark W. Knuth Scott A. Lesley Adam Godzik Marc‐Andr Elsliger Ashley M. Deacon Ian A. Wilson 《Protein science : a publication of the Protein Society》2019,28(9):1676-1689
Free‐standing single‐layer β‐sheets are extremely rare in naturally occurring proteins, even though β‐sheet motifs are ubiquitous. Here we report the crystal structures of three homologous, single‐layer, anti‐parallel β‐sheet proteins, comprised of three or four twisted β‐hairpin repeats. The structures reveal that, in addition to the hydrogen bond network characteristic of β‐sheets, additional hydrophobic interactions mediated by small clusters of residues adjacent to the turns likely play a significant role in the structural stability and compensate for the lack of a compact hydrophobic core. These structures enabled identification of a family of secreted proteins that are broadly distributed in bacteria from the human gut microbiome and are putatively involved in the metabolism of complex carbohydrates. A conserved surface patch, rich in solvent‐exposed tyrosine residues, was identified on the concave surface of the β‐sheet. These new modular single‐layer β‐sheet proteins may serve as a new model system for studying folding and design of β‐rich proteins. 相似文献