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Fabian Benz Christoph Roderburg David Vargas Cardenas Mihael Vucur Jérémie Gautheron Alexander Koch Henning Zimmermann J?rn Janssen Lukas Nieuwenhuijsen Mark Luedde Norbert Frey Frank Tacke Christian Trautwein Tom Luedde 《Experimental & molecular medicine》2013,45(9):e42
MicroRNA (miRNA) levels in serum have recently emerged as potential novel biomarkers for various diseases. miRNAs are routinely measured by standard quantitative real-time PCR (qPCR); however, the high sensitivity of qPCR demands appropriate normalization to correct for nonbiological variation. Presently, RNU6B (U6) is used for data normalization of circulating miRNAs in many studies. However, it was suggested that serum levels of U6 themselves might differ between individuals. Therefore, no consensus has been reached on the best normalization strategy in ‘circulating miRNA''. We analyzed U6 levels as well as levels of spiked-in SV40-RNA in sera of 44 healthy volunteers, 203 intensive care unit patients and 64 patients with liver fibrosis. Levels of U6 demonstrated a high variability in sera of healthy donors, patients with critical illness and liver fibrosis. This high variability could also be confirmed in sera of mice after the cecal ligation and puncture procedure. Most importantly, levels of circulating U6 were significantly upregulated in sera of patients with critical illness and sepsis compared with controls and correlated with established markers of inflammation. In patients with liver fibrosis, U6 levels were significantly downregulated. In contrast, levels of spiked-in SV40 displayed a significantly higher stability both in human cohorts (healthy, critical illness, liver fibrosis) and in mice. Thus, we conclude that U6 levels in the serum are dysregulated in a disease-specific manner. Therefore, U6 should not be used for data normalization of circulating miRNAs in inflammatory diseases and previous studies using this approach should be interpreted with caution. Further studies are warranted to identify specific regulatory processes of U6 levels in sepsis and liver fibrosis. 相似文献
67.
A novel approach for the syntheses of carbene adduct 4 is reported. A ring-enlargement strategy was employed for the synthesis of precursors 5 and 6 and established the mechanism of the formation of azulene derivative 6. Synthesis of target precursor 13, a novel precursor for the synthesis of new mesogenic materials, and its various halogenated derivatives (14–16) was carried out. 相似文献
68.
Schulte RF Henning A Tsao J Boesiger P Pruessmann KP 《Journal of magnetic resonance (San Diego, Calif. : 1997)》2007,186(2):167-175
The achievable bandwidth of common linear-phase RF pulses is limited by the maximum feasible B1 amplitude of the MR system. It has been shown previously, that this limitation can be circumvented by overlaying a quadratic phase in the frequency domain, which spreads the power across the pulse duration. Quadratic-phase RF pulses are near optimal in terms of achieving minimal B1max. In this work, it is demonstrated that further B1max reduction can be achieved by combining quadratic with higher-order polynomial-phase functions. RF pulses with a phase response up to tenth order were designed using the Shinnar-Le Roux transformation, yielding considerable increases in bandwidth and selectivity as compared to pure quadratic-phase pulses. These benefits are studied for a range of pulse specifications and demonstrated experimentally. For B1max = 20 microT and a pulse duration of 2.1 ms, it was possible to increase the bandwidth from 3.1 kHz for linear and 3.8 kHz for a quadratic to 9.9 kHz for a polynomial-phase pulse. 相似文献
69.
Click‐Tag and Amine‐Tag: Chemical Tag Approaches for Efficient Protein Labeling In Vitro and on Live Cells using the Naturally Split Npu DnaE Intein 下载免费PDF全文
Dr. Vivien Schütz Prof. Dr. Henning D. Mootz 《Angewandte Chemie (International ed. in English)》2014,53(16):4113-4117
Protein labeling with synthetic moieties remains in many cases a technically challenging or unresolved task. Two new and simple concepts are presented. In both approaches, a very short tag of only a few amino acids is prepared with the desired chemical modification and, in a second step, it is transferred to the protein of interest by protein trans‐splicing. For the amine‐tag, a recombinant intein fragment free of lysine residues was generated such that the amine group of the N terminus could be selectively modified with regular amine‐reactive reagents. Thus, standard bioconjugation procedures without any chemical synthesis could be applied without modification of lysines in the protein of interest. For the click‐tag, protein trans‐splicing was combined with unnatural amino acid mutagenesis and subsequent bioorthogonal side chain modification, as demonstrated for click chemistry using p‐azidophenylalanine. By the two‐step strategy, exposure of the protein of interest to the copper catalyst was avoided. 相似文献
70.
Benedikt Jedlitzke Zahide Yilmaz Wolfgang Drner Henning D. Mootz 《Angewandte Chemie (International ed. in English)》2020,59(4):1506-1510
Photocaged antibody fragments, termed photobodies, have been developed that are impaired in their antigen‐binding capacity and can be activated by irradiation with UV light (365 nm). This rational design concept builds on the selective photocaging of a single tyrosine in a nanobody (a single‐domain antibody fragment). Tyrosine is a frequently occurring residue in central positions of the paratope region. o‐Nitrobenzyl‐protected tyrosine variants were incorporated into four nanobodies, including examples directed against EGFR and HER2, and photodeprotection restores the native sequence. An anti‐GFP photobody exhibited an at least 10 000‐fold impaired binding affinity before photodeprotection compared with the parent nanobody. A bispecific nanobody–photobody fusion protein was generated to trigger protein heterodimerization by light. Photoactivatable antibodies are expected to become versatile protein reagents and to enable novel approaches in diagnostic and therapeutic applications. 相似文献