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61.
Melo AC Valle D Machado EA Salerno AP Paiva-Silva GO Cunha E Silva NL de Souza W Masuda H 《Insect biochemistry and molecular biology》2000,30(7):549-557
The synthesis and secretion of vitellogenin by the ovary of Rhodnius prolixus was investigated. Using whole ovary or epithelial cells isolated from follicles of different sizes, it is shown that the follicle cells are a site of synthesis for this protein in the ovary. The ovaries or follicle cells were incubated in vitro with [(35)S]-methionine or (32)Pi and the secretion of newly synthesized ovarian vitellogenin (O-Vg) was estimated by the radioactivity associated with the immunoprecipitate or acid-precipitate proteins in the culture medium. The radioactive O-Vg was analyzed by SDS-PAGE followed by autoradiography or after elution from a DEAE-Toyopearl column. The presence of O-Vg inside the follicle cells was detected by immunofluorescence and immunogold labels. Both methods revealed strong labeling inside the follicle cells. While the capacity for total protein synthesis by the follicle cells was maximal during the early phase of vitellogenesis (in small follicles), the synthesis of O-Vg reached its peak during the late phase of oocyte growth, just before formation of the chorion. A possible role for ovarian vitellogenin in Rhodnius and its relationship with Vg synthesis by the fat body is discussed. 相似文献
62.
Angular dependences of perpendicular and parallel mode electron paramagnetic resonance of oxidized beef heart cytochrome c oxidase
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Hunter DJ Oganesyan VS Salerno JC Butler CS Ingledew WJ Thomson AJ 《Biophysical journal》2000,78(1):439-450
Cytochrome c oxidase catalyzes the reduction of oxygen to water with a concomitant conservation of energy in the form of a transmembrane proton gradient. The enzyme has a catalytic site consisting of a binuclear center of a copper ion and a heme group. The spectroscopic parameters of this center are unusual. The origin of broad electron paramagnetic resonance (EPR) signals in the oxidized state at rather low resonant field, the so-called g' = 12 signal, has been a matter of debate for over 30 years. We have studied the angular dependence of this resonance in both parallel and perpendicular mode X-band EPR in oriented multilayers containing cytochrome c oxidase to resolve the assignment. The "slow" form and compounds formed by the addition of formate and fluoride to the oxidized enzyme display these resonances, which result from transitions between states of an integer-spin multiplet arising from magnetic exchange coupling between the five unpaired electrons of high spin Fe(III) heme a(3) and the single unpaired electron of Cu(B). The first successful simulation of similar signals observed in both perpendicular and parallel mode X-band EPR spectra in frozen aqueous solution of the fluoride compound of the closely related enzyme, quinol oxidase or cytochrome bo(3), has been reported recently (Oganesyan et al., 1998, J. Am. Chem. Soc. 120:4232-4233). This suggested that the exchange interaction between the two metal ions of the binuclear center is very weak (|J| approximately 1 cm(-1)), with the axial zero-field splitting (D approximately 5 cm(-1)) of the high-spin heme dominating the form of the ground state. We show that this model accounts well for the angular dependences of the X-band EPR spectra in both perpendicular and parallel modes of oriented multilayers of cytochrome c oxidase derivatives and that the experimental results are inconsistent with earlier schemes that use exchange coupling parameters of several hundred wavenumbers. 相似文献
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65.
Blanda Di Luccia Raffaella Crescenzo Arianna Mazzoli Luisa Cigliano Paola Venditti Jean-Claude Walser Alex Widmer Loredana Baccigalupi Ezio Ricca Susanna Iossa 《PloS one》2015,10(8)
A fructose-rich diet can induce metabolic syndrome, a combination of health disorders that increases the risk of diabetes and cardiovascular diseases. Diet is also known to alter the microbial composition of the gut, although it is not clear whether such alteration contributes to the development of metabolic syndrome. The aim of this work was to assess the possible link between the gut microbiota and the development of diet-induced metabolic syndrome in a rat model of obesity. Rats were fed either a standard or high-fructose diet. Groups of fructose-fed rats were treated with either antibiotics or faecal samples from control rats by oral gavage. Body composition, plasma metabolic parameters and markers of tissue oxidative stress were measured in all groups. A 16S DNA-sequencing approach was used to evaluate the bacterial composition of the gut of animals under different diets. The fructose-rich diet induced markers of metabolic syndrome, inflammation and oxidative stress, that were all significantly reduced when the animals were treated with antibiotic or faecal samples. The number of members of two bacterial genera, Coprococcus and Ruminococcus, was increased by the fructose-rich diet and reduced by both antibiotic and faecal treatments, pointing to a correlation between their abundance and the development of the metabolic syndrome. Our data indicate that in rats fed a fructose-rich diet the development of metabolic syndrome is directly correlated with variations of the gut content of specific bacterial taxa. 相似文献
66.
M. Giovanna Parisi Matteo Cammarata Gigliola Benenati Giuseppina Salerno Valentina Mangano Aiti Vizzini Nicolò Parrinello 《Cell and tissue research》2010,341(2):279-288
The purification, cloning, sequencing, molecular properties and expression of a fucose-binding lectin from the serum of Dicentrarchus labrax (DlFBL) have been previously reported. We now describe the distribution and expression of DlFBL during fish ontogeny. Immunohistochemistry
and in situ hybridization assays were carried out at various developmental stages (from 10 days post-hatching larvae to juveniles).
Another fucose-binding lectin, similar to DlFBL in biochemical, immunochemical and agglutinating properties, was extracted
and purified from eggs and appeared to be localized in the embryo yolk sack residual. DlFBL was found in columnar and goblet
cells of the intestinal epithelium of larvae (from 20 days post-hatching) and juveniles and in parenchymal tissue of juveniles.
DlFBL mRNA and protein were detected in the intestinal epithelium and in hepatocytes. An amplification product from degenerate
primers indicates that lectin isotypes with DlFBL epitopes are expressed in eggs and embryos. Whether the lectin fraction
isolated from eggs and embryos includes DlFBL of maternal origin remains unclear. 相似文献
67.
Prospero Di Pierro Loredana Mariniello Angela Sorrentino Reynaldo Villalonga Belkis Chico Raffaele Porta 《Amino acids》2010,38(2):669-675
Putrescine (1,4-diaminobutane) was covalently linked to alginate and low-methoxyl pectin to synthesize new aminated polysaccharides.
Both putrescine–pectin and –alginate conjugates, although the latter at higher concentrations, were found to be able to act
as effective acyl acceptor transglutaminase substrates in vitro using both dimethylated casein and soy flour proteins as acyl
donors. Monodansylcadaverine, a well known acyl acceptor transglutaminase substrate, dose-dependently counteracted the covalent
binding of the aminated polysaccharides to the proteins. Putrescine–pectin conjugate was also tested to prepare, in combination
with soy flour proteins, edible films in the presence of purified microbial transglutaminase. Characterization of the enzymatically
crosslinked films showed a significant decreased water vapor permeability, with respect to the ones obtained with non-aminated
pectin in the presence of transglutaminase, as well as improved mechanical properties, such as high extensibility. Possible
biotechnological applications of hydrocolloid films containing putrescine–polysaccharide derivatives enzymatically crosslinked
to proteins were suggested. 相似文献
68.
Nacira Darghal Nadia Bouchemal Carlos F.G.C. Geraldes Milena Salerno 《Journal of inorganic biochemistry》2010,104(1):47-54
Alzheimer’s disease (AD) is the most commonly form of dementia in the elderly. The development of molecules able to detect biomarkers characteristic of AD is critical to its understanding and treatment. However, such molecules must be able to pass blood-brain barrier (BBB) which is a major impediment to the entry of many therapeutic drugs into the brain. Such a limitation applies to the development of magnetic resonance imaging molecular neuroimaging agents using biomarkers of AD-like β-amyloid deposits, as the common extracellular contrast agents (CAs) are not able to cross an intact BBB. In this work, we have studied the ability of a series of simple Eu3+ complexes to enter cells overexpressing or not the ABCB1 (P-gp or P-glycoprotein) protein, which is expressed at the BBB and in human embryonic astrocytes. The intracellular uptake of the Eu3+ complexes of linear and macrocyclic polyaminocarboxylate ligands with different charges and lipophilicities was followed by atomic absorption spectrometry. Based on biochemical argument, we propose that lipophilic contrast agents can be efficiently taken up by cells and accumulate inside mitochondria when they are positively charged. The important point is that they are not P-gp substrates, which is one of the major obstacles for them to cross the BBB. 相似文献
69.
Lucia Peluso Cristiana de Luca Silvia Bozza Antonio Leonardi Gloria Giovannini Alfonso Lavorgna Gaetano De Rosa Massimo Mascolo Loredana Ortega De Luna Maria Rosaria Catania Luigina Romani Fabio Rossano 《BMC microbiology》2010,10(1):1-11
Background
Porcine reproductive and respiratory syndrome (PRRS) has now been widely recognized as an economically important disease. The objective of this study was to compare the molecular and biological characteristics of porcine reproductive and respiratory syndrome virus (PRRSV) field isolates in China to those of the modified live virus (MLV) PRRS vaccine and its parent strain (ATCC VR2332).Results
Five genes (GP2, GP3, GP4, GP5 and NSP2) of seven isolates of PRRSV from China, designated LS-4, HM-1, HQ-5, HQ-6, GC-2, GCH-3 and ST-7/2008, were sequenced and analyzed. Phylogenetic analyses based on the nucleotide sequence of the ORF2-5 and NSP2 showed that the seven Chinese isolates belonged to the same genetic subgroup and were related to the North American PRRSV genotype. Comparative analysis with the relevant sequences of another Chinese isolate (BJ-4) and North American (VR2332 and MLV) viruses revealed that these isolates have 80.8-92.9% homology with VR-2332, and 81.3-98.8% identity with MLV and 80.7-92.9% with BJ-4. All Nsp2 nonstructural protein of these seven isolates exhibited variations (a 29 amino acids deletion) in comparison with other North American PRRSV isolates. Therefore, these isolates were novel strain with unique amino acid composition. However, they all share more than 97% identity with other highly pathogenic Chinese PRRSV strains. Additionally, there are extensive amino acid (aa) mutations in the GP5 protein and the Nsp2 protein when compared with the previous isolates.Conclusions
These results might be useful to study the genetic diversity of PRRSV in China and to track the infection sources as well as for vaccines development. 相似文献70.
The present study aimed to evaluate different dosage forms, emulsions, emulgels, lipogels, and thickened microemulsion-based
hydrogel, as fluconazole topical delivery systems with the purpose of determining a formulation with the capacity to deliver
the whole active compound and maintain it within the skin so as to be considered a useful formulation either for topical mycosis
treatment or as adjuvant in a combined therapy for Cutaneous Leishmaniasis. Propylene glycol and diethyleneglycol monoethyl
ether were used for each dosage form as solvent for the drug and also as penetration enhancers. In vitro drug release after application of a clinically relevant dose of each formulation was evaluated and then microemulsions and
lipogels were selected for the in vitro penetration and permeation study. Membranes of mixed cellulose esters and full-thickness pig ear skin were used for the in vitro studies. Candida albicans was used to test antifungal activity. A microemulsion containing diethyleneglycol monoethyl ether was found to be the optimum
formulation as it was able to deliver the whole contained dose and enhance its skin penetration. Also this microemulsion showed
the best performance in the antifungal activity test compared with the one containing propylene glycol. These results are
according to previous reports of the advantages of microemulsions for topical administration and they are very promising for
further clinical evaluation. 相似文献