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11.
Xuehui Yang Yan Gong Yuefeng Tang Hongfang Li Qing He Lindsey Gower Lucy Liaw Robert E. Friesel 《PloS one》2013,8(3)
Background
Changes in the vascular smooth muscle cell (VSMC) contractile phenotype occur in pathological states such as restenosis and atherosclerosis. Multiple cytokines, signaling through receptor tyrosine kinases (RTK) and PI3K/Akt and MAPK/ERK pathways, regulate these phenotypic transitions. The Spry proteins are feedback modulators of RTK signaling, but their specific roles in VSMC have not been established.Methodology/Principal Findings
Here, we report for the first time that Spry1, but not Spry4, is required for maintaining the differentiated state of human VSMC in vitro. While Spry1 is a known MAPK/ERK inhibitor in many cell types, we found that Spry1 has little effect on MAPK/ERK signaling but increases and maintains Akt activation in VSMC. Sustained Akt signaling is required for VSMC marker expression in vitro, while ERK signaling negatively modulates Akt activation and VSMC marker gene expression. Spry4, which antagonizes both MAPK/ERK and Akt signaling, suppresses VSMC differentiation marker gene expression. We show using siRNA knockdown and ChIP assays that FoxO3a, a downstream target of PI3K/Akt signaling, represses myocardin promoter activity, and that Spry1 increases, while Spry4 decreases myocardin mRNA levels.Conclusions
Together, these data indicate that Spry1 and Spry4 have opposing roles in VSMC phenotypic modulation, and Spry1 maintains the VSMC differentiation phenotype in vitro in part through an Akt/FoxO/myocardin pathway. 相似文献12.
Periphytic biomass in a downriver riffle of the Grand River in Southern Ontario, Canada was measured with concrete-block glass slide samplers from May to December 1971. Average rate of accumulation of periphyton on glass slides was 266.2 mg/m2/day ash-free weight, 120.9 mg/m2/day carbon, 20.4 mg/m2/day nitrogen, or 1.11 mg/m2/day chlorophyll a and on concrete blocks was 590.0 mg/m2/day ash-free weight, 202.9 mg/m2/day carbon, 22.8 mg/m2/day nitrogen, or 2.95 mg/m2/day chlorophyll a. The standing biomass of periphyton on glass slides varied from a maximum of 21.6 g/m2 between May 28 and July 15 to a minimum of 0.7 g/m2 in December with an average of 8.6 g/m2. The biomass on concrete blocks ranged from a high of 132.5 g/m2 in August to a low of 29.9 g/m2 in October with a mean of 66.1 g/m2. The well established periphyton on concrete blocks towards the end of the study period was similar to that from native rocks with respect to biomass, carbon and nitrogen contents, and ratios of biomass to chlorophyll a. This suggests that concrete blocks are a better substrate than glass slides for measuring riverbed periphyton. By comparing biomass of periphyton on slides and on concrete blocks, the average rate of loss of periphyton from concrete blocks was estimated to be 2.9 g/m2/day, representing 63% of the mean total accumulation rate of the periphyton. The magnitude of the estimated total loss and the high standing biomass of the periphyton on concrete blocks manifest the importance of the periphyton as a source of organic matter in the river. 相似文献
13.
The present study analyzed the drosophilid assemblages in different levels of urbanization in the city of Porto Alegre, Rio Grande do Sul, Brazil. Collections were carried out in 2008 in three different environments: a highly urbanized area????Jardim Botanico,?? a forested area with intermediary urbanization????Parque Gabriel Knijnik,?? and in a relatively well-preserved forested area, although threatened by the urban growth????Morro Santana.?? In Jardim Botanico, 36 species belonging to four genera were found, with high abundance of exotic species as Drosophila simulans Sturtevant and Zaprionus indianus (Gupta). In Parque Gabriel Knijnik, 33 species that belonged to four genera were found, with higher abundances of native species belonging to the Drosophila tripunctata species group and Drosophila willistoni species subgroup, and lower abundance of exotic species. As for Morro Santana, 32 species and three genera were found, with higher abundances of native groups, low representativeness of exotic species, and absence of Zaprionus indianus. The analysis of the Jaccard index showed higher similarity in the species composition between samples collected in summer and autumn, and between samples collected in winter and spring. On the other hand, the Morisita index differentiated Jardim Botanico from the other two studied sites. Our results show that Morro Santana is an important area of native biodiversity, reinforcing, therefore, the inclusion of this area in the project for the creation of an ecological corridor as proposed by the Ministry of the Environment of Brazil. 相似文献
14.
Directed Evolution of Streptomyces clavuligerus Deacetoxycephalosporin C Synthase for Enhancement of Penicillin G Expansion 下载免费PDF全文
Chia-Li Wei Yunn-Bor Yang Chan-Hui Deng Wen-Chi Liu Jyh-Shing Hsu Yu-Ching Lin Shwu-Huey Liaw Ying-Chieh Tsai 《Applied microbiology》2005,71(12):8873-8880
The deacetoxycephalosporin C synthase from Streptomyces clavuligerus was directly modified for enhancement of penicillin G expansion into phenylacetyl-7-aminodeacetoxycephalosporanic acid, an important intermediate in the industrial manufacture of cephalosporin antibiotics. Nine new mutants, mutants M73T, T91A, A106T, C155Y, Y184H, M188V, M188I, H244Q, and L277Q with 1.4- to 5.7-fold increases in the kcat/Km ratio, were obtained by screening 6,364 clones after error-prone PCR-based random mutagenesis. Subsequently, DNA shuffling was carried out to screen possible combinations of substitutions, including previous point mutations. One quaternary mutant, the C155Y/Y184H/V275I/C281Y mutant, which had a kcat/Km ratio that was 41-fold higher was found after 10,572 clones were assayed. The distinct mutants obtained using different mutagenesis methods demonstrated the complementarity of the techniques. Interestingly, most of the mutated residues that result in enhanced activities are located within or near the unique small barrel subdomain, suggesting that manipulation of this subdomain may be a constructive strategy for improvement of penicillin expansion. Several mutations had very distinct effects on expansion of penicillins N and G, perhaps due to different penicillin-interacting modes within the enzyme. Thus, the present study provided not only promising enzymes for cephalosporin biosynthesis but also a large number of mutants, which provided new insights into the structure-function relationship of the protein that should lead to further rational engineering. 相似文献
15.
Wen-Feng Liaw Chung-Hung HsiehShie-Ming Peng Gene-Hsiang Lee 《Inorganica chimica acta》2002,332(1):153-159
The SS bond-activation of diorganyl disulfide by the anionic metal carbonyl fragment [Mn(CO)5]− gives rise to an extensive chemistry. Oxidative decarbonylation addition of 2,2′-dithiobis(pyridine-N-oxide) to [Mn(CO)5]−, followed by chelation and metal-center oxidation, led to the formation of [MnII(SC5H4NO)3]− (1). The effective magnetic moment in solid state by SQUID magnetometer was 5.88 μB for complex 1, which is consistent with the MnII having a high-spin d5 electronic configuration in an octahedral ligand field. The average Mn(II)S, SC and NO bond lengths of 2.581(1), 1.692(4) and 1.326(4) Å, respectively, indicate that the negative charge of the bidentate 1-oxo-2-thiopyridinato [SC5H4NO]− ligand in complex 1 is mainly localized on the oxygen atom. The results are consistent with thiolate-donor [SC5H4NO]− stabilization of the lower oxidation state of manganese (Mn(I)), while the O,S-chelating [SC5H4NO]− ligand enhances the stability of manganese in the higher oxidation state (Mn(II)). Activation of SS bond as well as OH bond of 2,2′-dithiosalicylic acid by [Mn(CO)5]− yielded [(CO)3Mn(μ-SC6H4C(O)O)2Mn(CO)3]2− (4). Oxidative addition of bis(o-benzamidophenyl) disulfide to [Mn(CO)5]− resulted in the formation of cis-[Mn(CO)4(SR)2]− (R=C6H4NHCOPh) which was employed as a chelating metallo ligand to synthesize heterotrinuclear [(CO)3Mn(μ-SR)3Co(μ-SR)3Mn(CO)3]− (8) possessing a homoleptic hexathiolatocobalt(III) core. 相似文献
16.
Trace elements and lipid peroxidation in 26 patients with chronic renal failure treated with hemodialysis and 25 healthy subjects
were observed. Both plasma and erythrocyte trace elements and plasma malon dialdehyde (MDA) were examined immediately before
and after hemodialysis. Increased levels of plasma Cu, MDA, and erythrocyte Pb, Mn, Zn, and a significantly decreased plasma
Se, Zn and erythrocyte Se were found in patients before hemodialysis. After a single hemodialysis, erythrocyte Mn, Cu, Zn,
and plasma Cu, Al, and MDA were significantly increased whereas both plasma and erythrocyte Se were lower in patients than
in healthy subjects. The level of MDA was not significantly changed during the single hemodialysis. Both plasma and erythrocyte
Zn levels and plasma Cu and Al were significantly higher after hemodialysis than before hemodialysis. In conclusion, levels
of trace elements are altered by hemodialysis, which may increase patient susceptibility to lipid peroxidation in uremia. 相似文献
17.
Background
In the analysis of complex traits, genetic effects can be confounded with non-genetic effects, especially when using full-sib families. Dominance and epistatic effects are typically confounded with additive genetic and non-genetic effects. This confounding may cause the estimated genetic variance components to be inaccurate and biased.Methods
In this study, we constructed genetic covariance structures from whole-genome marker data, and thus used realized relationship matrices to estimate variance components in a heterogenous population of ~ 2200 mice for which four complex traits were investigated. These mice were genotyped for more than 10,000 single nucleotide polymorphisms (SNP) and the variances due to family, cage and genetic effects were estimated by models based on pedigree information only, aggregate SNP information, and model selection for specific SNP effects.Results and conclusions
We show that the use of genome-wide SNP information can disentangle confounding factors to estimate genetic variances by separating genetic and non-genetic effects. The estimated variance components using realized relationship were more accurate and less biased, compared to those based on pedigree information only. Models that allow the selection of individual SNP in addition to fitting a relationship matrix are more efficient for traits with a significant dominance variance. 相似文献18.
19.
Structural characterization of the 60-kDa bermuda grass pollen isoallergens, a covalent flavoprotein
Liaw S Lee DY Chow LP Lau GX Su SN Chow L 《Biochemical and biophysical research communications》2001,280(3):738-743
Our studies suggest a tripartite structure for the 60-kDa allergen of Bermuda grass pollen (BG60) including a short N-terminal segment, a FAD-binding domain, and a C-terminal domain. The lower molecular weight isoallergens lack the N-terminal segment. The higher protease susceptibility and the lower melting temperature of approximately 20 degrees C of the lower molecular weight isoforms suggest that the N-terminal segment is essential for a compact structure. Database screening reveals that the protease-digested peptide sequences (approximately 180 residues in total) share 40% identity with the plant berberine bridge enzymes. In particular, a 24-residue peptide sequence displays high similarity to a conserved FAD-binding motif. The spectroscopic and SDS-PAGE analyses suggest that the cofactor FAD is covalently linked to the central domain. Therefore, we conclude that BG60 is identified as the first flavinylated allergen. 相似文献
20.
Liaw PC Mather T Oganesyan N Ferrell GL Esmon CT 《The Journal of biological chemistry》2001,276(11):8364-8370
The endothelial cell protein C receptor (EPCR) is an endothelial cell-specific transmembrane protein that binds both protein C and activated protein C (APC). EPCR regulates the protein C anticoagulant pathway by binding protein C and augmenting protein C activation by the thrombin-thrombomodulin complex. EPCR is homologous to the MHC class 1/CD1 family, members of which contain two alpha-helices that sit upon an 8-stranded beta-sheet platform. In this study, we identified 10 residues that, when mutated to alanine, result in the loss of protein C/APC binding (Arg-81, Leu-82, Val-83, Glu-86, Arg-87, Phe-146, Tyr-154, Thr-157, Arg-158, and Glu-160). Glutamine substitutions at the four N-linked carbohydrate attachment sites of EPCR have little affect on APC binding, suggesting that the carbohydrate moieties of EPCR are not critical for ligand recognition. We then mapped the epitopes for four anti-human EPCR monoclonal antibodies (mAbs), two of which block EPCR/Fl-APC (APC labeled at the active site with fluorescein) interactions, whereas two do not. These epitopes were localized by generating human-mouse EPCR chimeric proteins, since the mAbs under investigation do not recognize mouse EPCR. We found that 5 of the 10 candidate residues for protein C/APC binding (Arg-81, Leu-82, Val-83, Glu-86, Arg-87) colocalize with the epitope for one of the blocking mAbs. Three-dimensional molecular modeling of EPCR indicates that the 10 protein C/APC binding candidate residues are clustered at the distal end of the two alpha-helical segments. Protein C activation studies on 293 cells that coexpress EPCR variants and thrombomodulin demonstrate that protein C binding to EPCR is necessary for the EPCR-dependent enhancement in protein activation by the thrombin-thrombomodulin complex. These studies indicate that EPCR has exploited the MHC class 1 fold for an alternative and possibly novel mode of ligand recognition. These studies are also the first to identify the protein C/APC binding region of EPCR and may provide useful information about molecular defects in EPCR that could contribute to cardiovascular disease susceptibility. 相似文献