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31.
Inferences for a semiparametric model with panel data   总被引:1,自引:0,他引:1  
Cheng  SC; Wei  LJ 《Biometrika》2000,87(1):89-97
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Background

The recognition of microbial molecular patterns via Toll-like receptors (TLRs) is critical for mucosal defenses.

Methods

Using well-differentiated primary cultures of human airway epithelia, we investigated the effects of exposure of the cells to cytokines (TNF-α and IFN-γ) and dexamethasone (dex) on responsiveness to the TLR2/TLR1 ligand Pam3CSK4. Production of IL-8, CCL20, and airway surface liquid antimicrobial activity were used as endpoints.

Results

Microarray expression profiling in human airway epithelia revealed that first response cytokines markedly induced TLR2 expression. Real-time PCR confirmed that cytokines (TNF-α and IFN-γ), dexamethasone (dex), or cytokines + dex increased TLR2 mRNA abundance. A synergistic increase was seen with cytokines + dex. To assess TLR2 function, epithelia pre-treated with cytokines ± dex were exposed to the TLR2/TLR1 ligand Pam3CSK4 for 24 hours. While cells pre-treated with cytokines alone exhibited significantly enhanced IL-8 and CCL20 secretion following Pam3CSK4, mean IL-8 and CCL20 release decreased in Pam3CSK4 stimulated cells following cytokines + dex pre-treatment. This marked increase in inflammatory gene expression seen after treatment with cytokines followed by the TLR2 ligand did not correlate well with NF-κB, Stat1, or p38 MAP kinase pathway activation. Cytokines also enhanced TLR2 agonist-induced beta-defensin 2 mRNA expression and increased the antimicrobial activity of airway surface liquid. Dex blocked these effects.

Conclusion

While dex treatment enhanced TLR2 expression, co-administration of dex with cytokines inhibited airway epithelial cell responsiveness to TLR2/TLR1 ligand over cytokines alone. Enhanced functional TLR2 expression following exposure to TNF-α and IFN-γ may serve as a dynamic means to amplify epithelial innate immune responses during infectious or inflammatory pulmonary diseases.  相似文献   
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There has been considerable interest in identifying the neural correlates of action naming, but the bulk of previous work on this topic has utilized static stimuli. Recent research comparing the visual processing of dynamic versus static actions suggests that these two types of stimuli engage largely overlapping neural systems, raising the possibility that the higher-order processing requirements for naming dynamic and static actions might not be very different. To explore this issue in greater depth, we developed the Dynamic Action Naming Test (DANT), which consists of 158 video clips 3-5s in length, for each of which the participant is asked to produce the most appropriate verb. We administered the DANT to 78 brain-damaged patients drawn from our Patient Registry, and to a demographically matched group of 50 normal participants. Out of the 16 patients who performed defectively on the DANT, nearly all (15/16) had damage in the left hemisphere. Lesion analysis indicated that the frontal operculum was the most frequent area of damage in the 15 patients; also, damage to the posterolateral temporal-occipital sector (in and near MT) was specifically related to defective dynamic action naming. Most of the brain-damaged participants (n=71) also received our Static Action Naming Test (SANT), and we found that performances on verb items that were common across the DANT and SANT were highly correlated (R=.91). Moreover, patients who failed the DANT almost invariably also failed the SANT. These findings lend further support to the hypothesis that there is considerable commonality in the neural systems underlying the use of verbs to orally name dynamic and static actions, a conclusion that is in turn compatible with the concept of "representational momentum". Our results also contribute more generally to the rapidly growing field of research on embodied cognition.  相似文献   
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为建立鸭乙型肝炎病毒LJ-76的转染细胞系,将LJ-76病毒DNA插入到pUC19的EcoRⅠ位点上,分离得到含有双拷贝LJ-76DNA的重组质粒.通过磷酸钙沉淀方法,将经CsCl等密度离心纯化的LJ-76DNA双体导入到人肝癌细胞BEL7402中.收集转染细胞的培养液进行蔗糖密度梯度离心,所得沉淀经检测发现含有LJ-76DNA并具有特异性DHBV内源性DNA多聚酶活性;对上述样品通过DotEIA检测DHBV核心抗原及表面抗原结果为阳性.Southernblot分析表明转染细胞内存在病毒DNA复制中间体cccDNA、ssDNA和rcDNA,而cccDNA被认为是复制活动较为活跃的标志.电镜观察转染细胞的上清发现有病毒颗粒的存在.  相似文献   
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Culturable bacteria from the deep subsurface (179 m) at Cerro Negro, New Mexico were isolated and characterized. The average number of viable aerobic bacteria was estimated to be 5×105g–1 of sediment, but only about 0.1% of these could be recovered on agar medium when incubated under aerobic conditions. Of 158 strains isolated from this depth, 92 were characterized by cellular fatty acid profiles (FAME), 36 by analysis of partial 16S rDNA sequences, and 44 by rep-PCR genome fingerprint analysis using three different sets of oligonucleotide primers (REP, BOX, or ERIC). These analyses showed the majority of isolates (67%) were Gram-positive bacteria and primarily members of genera with a high %G+C DNA. The remaining isolates were -subdivisionProteobacteria (19%) and members of the flavobacteria group (14%). The diversity indices based on these different methods of characterization were very high suggesting this subsurface habitat harbors a highly diverse microbial community.  相似文献   
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The effect of abscisic acid on growth, ultrastructure and nucleic acid biosynthesis was studied in tissue culture of spinach (Spinacia oleracea L.). Low concentration (0.01 mg l?1) of abscisic acid increased fresh and dry weight of calluses, whereas 1.0 mg l?1 was inhibitory. The stimulating effect was observed only in the presence of a relatively high concentration of kinetin (1 mg l?1). The inhibitory effect was partly overcome by the same kinetin concentration. The low concentration of abscisic acid probably accelerated the induction of callus growth after subculture and stimulated cell division in the exponential phase of growth. Electron microscopy showed the presence of numerous polysomes and rough endoplasmic reticulum in callus cells grown at the stimulating abscisic acid concentration. Control cells and cells at the inhibitory concentration had slightly hyaline cytoplasm and were more vacuolated. Incubation of callus tissue with 32P in the presence of stimulating concentration of abscisic acid showed a significant increase in the rate of biosynthesis of all nucleic acid classes after 8 h, whereas inhibitory concentration produced a decrease in 32P incorporation. However, when the tissue was grown in the presence of abscisic acid for 20 days, both concentrations decreased the rate of nucleic acid biosynthesis, as compared to the controls.  相似文献   
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Intracellular calcium buffering capacity in isolated squid axons   总被引:12,自引:7,他引:5       下载免费PDF全文
Changes in ionized calcium were studied in axons isolated from living squid by measuring absorbance of the Ca binding dye Arsenazo III using multiwavelength differential absorption spectroscopy. Absorption changes measured in situ were calibrated in vitro with media of ionic composition similar to axoplasm containing CaEGTA buffers. Calcium loads of 50-2,500 μmol/kg axoplasm were induced by microinjection, by stimulation in 112 mM Ca seawater, or by soaking in choline saline with 1-10 mM Ca. Over this range of calcium loading of intact axoplasm, the ionized calcium in the axoplasm rose about 0.6 nM/μM load. Similar loading in axons preteated with carbonyl cyanide 4- trifluoromethoxyphenylhydrazone (FCCP) to inhibit the mitochondrial proton gradient increased ionized calcium by 5-7 percent of the imposed load, i.e. 93-95 percent of the calcium load was buffered by a process insensitive to FCCP. This FCCP- insensitive buffer system was not saturated by the largest calcium loads imposed, indicating a capacity of at least several millimolar. Treatment of previously loaded axons with FCCP or apyrase plus cyanide produced rises in ionized calcium which could be correlated with the extent of the load. Analysis of results indicated that, whereas only 6 percent of the endogenous calcium in fresh axons is stored in the FCCP-sensitive (presumably mitochondrial) buffer system, about 30 percent of an imposed exogenous load in the range of 50-2,500 μM is taken up by this system.  相似文献   
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