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431.
Kimon K Frousios Vassiliki A Iconomidou Carolina-Maria Karletidi Stavros J Hamodrakas 《BMC structural biology》2009,9(1):44
Background
Amyloidoses are a group of usually fatal diseases, probably caused by protein misfolding and subsequent aggregation into amyloid fibrillar deposits. The mechanisms involved in amyloid fibril formation are largely unknown and are the subject of current, intensive research. In an attempt to identify possible amyloidogenic regions in proteins for further experimental investigation, we have developed and present here a publicly available online tool that utilizes five different and independently published methods, to form a consensus prediction of amyloidogenic regions in proteins, using only protein primary structure data. 相似文献432.
Vassiliki Kotoula Elpida Charalambous Bart Biesmans Andigoni Malousi Eleni Vrettou George Fountzilas George Karkavelas 《PloS one》2009,4(11)
Background
Testing for tumor specific mutations on routine formalin-fixed paraffin-embedded (FFPE) tissues may predict response to treatment in Medical Oncology and has already entered diagnostics, with KRAS mutation assessment as a paradigm. The highly sensitive real time PCR (Q-PCR) methods developed for this purpose are usually standardized under optimal template conditions. In routine diagnostics, however, suboptimal templates pose the challenge. Herein, we addressed the applicability of sequencing and two Q-PCR methods on prospectively assessed diagnostic cases for KRAS mutations.Methodology/Principal Findings
Tumor FFPE-DNA from 135 diagnostic and 75 low-quality control samples was obtained upon macrodissection, tested for fragmentation and assessed for KRAS mutations with dideoxy-sequencing and with two Q-PCR methods (Taqman-minor-groove-binder [TMGB] probes and DxS-KRAS-IVD). Samples with relatively well preserved DNA could be accurately analyzed with sequencing, while Q-PCR methods yielded informative results even in cases with very fragmented DNA (p<0.0001) with 100% sensitivity and specificity vs each other. However, Q-PCR efficiency (Ct values) also depended on DNA-fragmentation (p<0.0001). Q-PCR methods were sensitive to detect ≤1% mutant cells, provided that samples yielded cycle thresholds (Ct) <29, but this condition was met in only 38.5% of diagnostic samples. In comparison, FFPE samples (>99%) could accurately be analyzed at a sensitivity level of 10% (external validation of TMGB results). DNA quality and tumor cell content were the main reasons for discrepant sequencing/Q-PCR results (1.5%).Conclusions/Significance
Diagnostic targeted mutation assessment on FFPE-DNA is very efficient with Q-PCR methods in comparison to dideoxy-sequencing. However, DNA fragmentation/amplification capacity and tumor DNA content must be considered for the interpretation of Q-PCR results in order to provide accurate information for clinical decision making. 相似文献433.
434.
Teris A. van Beek Kishore K. R. Tetala Irina I. Koleva Airidas Dapkevicius Vassiliki Exarchou Suzanne M. F. Jeurissen Frank W. Claassen Elbert J. C. van der Klift 《Phytochemistry Reviews》2009,8(2):387-399
Natural products chemistry has witnessed many new developments in the last 5 years like extractions with subcritical water
and ionic liquids, LC/HRMS and LC/SPE/cryo-NMR, UHPLC, TLC/MS, MS-based preparative HPLC, comprehensive chromatography (GC × GC,
LC × LC), high-throughput screening, introduction of monolithic columns, miniaturisation, and automated structure identification.
Nevertheless identifying bioactive constituents in complex plant extracts remains a tedious process. The classical approach
of bioassay guided fractionation is time-consuming while off-line screening of extracts does not provide information on individual
compounds and sometimes suffers from false positives or negatives. One way out of this is by coupling chromatography with
chemical or biochemical assays, so called high resolution screening. An example is the development of HPLC on-line assays
for antioxidants. By the post-column addition of a relatively stable coloured radical like DPPH• or ABTS•+, radical scavengers are detected as negative peaks because in a reaction coil they reduce the model radical to its reduced,
non-coloured form. When combined with LC/DAD/MS and LC/SPE/NMR, reliable identification of active constituents becomes possible
without the necessity of ever isolating them in a classical sense. Also for finding leads for new drugs, combining HPLC with
biochemical assays is interesting but technically more difficult. Most enzymes do not work at the organic modifier concentrations
commonly encountered in RP-HPLC and the reaction time is often longer requiring dilution and lengthy coils respectively. Therefore,
new techniques have to be implemented to gain the required sensitivity for on-line enzyme assays. For stable analytes, high
temperature LC offers a solution to the organic modifier problem. When enzymes are highly expensive, like those used in the
screening for Cytochrome P450 inhibitors, miniaturisation to chip format may offer a way out. Microreactors (chips) are not
only useful for miniaturising larger assays but also offer completely new prospects in phytochemical analysis. One such application
is in the sample clean-up of acids and bases like alkaloids. In a lay-out of three parallel channels of 100 μm width with
the middle one containing organic phase and the two outer ones water of high pH (feed phase) and low pH (trapping phase) such
a chip replaces two classical LLE steps but is much faster and requires less solvents and less manpower input. 相似文献
435.
Konstantina Zografou Spyros Sfenthourakis Andrew Pullin Vassiliki Kati 《Journal of Insect Conservation》2009,13(5):505-514
We tested the surrogate value of butterflies, red-listed butterflies and grasshoppers for each other in terms of diversity
patterns congruence and complementarity at a site in the NATURA 2000 network. Grammos Mountain is proposed as a new Prime
Butterfly Area for Greece: it supports a total of 56 grasshopper species and 112 butterfly species, 24 of which are of European
conservation concern (SPEC) and two of Prime Butterflies Area Project. We found a strong congruence in the species richness
patterns of SPEC butterflies, butterflies and grasshoppers, because three common ecological factors influenced them: number
of flower heads, altitude, and cover of low trees or bushes (Redundancy Analysis, CANOCO). Each complementarity network maintained
quite well the species richness of the other two target groups (<18% average species loss). SPEC butterflies were the best
surrogate group overall and therefore we propose that they should be monitored on a permanent basis. 相似文献
436.
Smocovitis VB 《American journal of botany》2006,93(7):942-952
This paper offers highlights from the 100 (plus) years of the Botanical Society of America (BSA) and draws extensively on the archives of the BSA. In addition to examining the founding of the society and the attempt to "professionalize" botany in late 19th century America, the paper also explores the complex relations between the BSA and a number of related societies in the United States, the Society's struggle to create a coherent identity for itself, the place of botany as a whole in the context of the burgeoning biological sciences in the 20th century, and the changing role of the BSA in an international context. The paper assesses both the achievements and the challenges facing the BSA. It closes by offering some historical reflections on the status of "botany" as a science and the historical significance of terms like "plant biology" and "plant science." 相似文献
437.
The function of mutations rdn1A, rdn1T, and rdn2 in 18S rRNA of Saccharomyces cerevisiae is investigated. The mutations correspond to substitutions C1054A, C1054U in helix 34, and G517A in helix 18 of 16S rRNA in Escherichia coli, respectively, in which the first and third mutations caused nonsense suppression, while C1054U caused no suppression. In yeast, rdn1A caused phenotypic suppression at nonsense codons, whereas rdn1T and rdn2 caused antisuppression. We provide in vitro evidence that, in addition, rdn1A decreases translational accuracy at sense codons as well, by a factor of 8, accompanied by extreme sensitivity to paromomycin, compatible with its error-prone character. Mutations rdn1T andrdn2 exhibit hyperaccuracy and paromomycin resistance. Thus, mutations in conserved rRNA regions may affect the same functions in the various species but in opposite directions. Mutation rdn1A, but not rdn1T or rdn2, affected also the catalytic activity of the ribosome, a 60S subunit activity. The rate of peptide bond formation was reduced to half its normal value, indicating a communication between the two subunits. Moreover, error-prone mutation rdn1A was less susceptible to oxidative modifications than wild type, indicated by decreased lipid peroxidation and nonprotein/protein disulfides, as well as by increased protein thiols. In contrast, hyperaccurate mutations rdn1T and rdn2 displayed increased oxidative stress. Our results suggest that the cells may consume more energy to achieve hyperaccuracy leading to increased oxidative modifications. 相似文献
438.
439.
Safina Gadeock Jimmy N.S.N. Tran Jennifer G. Georgiou Iman Jalilian James S. Wiley Ronald Sluyter 《生物化学与生物物理学报:生物膜》2010,1798(11):2058-2066
The P2X7 receptor is an extracellular ATP-gated cation channel critical in inflammation and immunity, and can be up-regulated by IFN-γ and LPS. This study aimed to examine the effect of TGF-β1 on the up-regulation of P2X7 function and expression in leukemic THP-1 monocytes differentiated with IFN-γ and LPS. Cell-surface molecules including P2X7 were examined by immunofluorescence staining. Total P2X7 protein and mRNA was assessed by immunoblotting and RT-PCR respectively. P2X7 function was evaluated by ATP-induced cation dye uptake measurements. Cell-surface P2X7 was present on THP-1 cells differentiated for 3 days with IFN-γ and LPS but not on undifferentiated THP-1 cells. ATP induced ethidium+ uptake into differentiated but not undifferentiated THP-1 cells, and the P2X7 antagonist, KN-62, impaired ATP-induced ethidium+ uptake. Co-incubation of cells with TGF-β1 plus IFN-γ and LPS prevented the up-regulation of P2X7 expression and ATP-induced ethidium+ uptake in a concentration-dependent fashion with a maximum effect at 5 ng/ml and with an IC50 of ~ 0.4 ng/ml. Moreover, ATP-induced YO-PRO-12+ uptake and IL-1β release were abrogated in cells co-incubated with TGF-β1. TGF-β1 also abrogated the amount of total P2X7 protein and mRNA induced by IFN-γ and LPS. Finally, TGF-β1 prevented the up-regulation of cell-surface CD86, but not CD14 and MHC class II, by IFN-γ and LPS. These results indicate that TGF-β1 prevents the up-regulation of P2X7 function and expression by IFN-γ and LPS in THP-1 monocytes. This suggests that TGF-β1 may limit P2X7-mediated processes in inflammation and immunity. 相似文献
440.