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341.
342.
Santolaya-Forgas J Edwin S Zeiter K Pitt A Pineles B Tarca AL Espinoza J Kusanovic JP Erez O Wolf R Romero R 《Journal of medical primatology》2007,36(6):370-374
BACKGROUND: During pregnancy, mechanisms that allow for regulation of continuous fetal and placental vasculogenesis with prevention of maternal neo-vascularization remain elusive. The vascular endothelial growth factor (VEGF) biological system has a key role during vasculogenesis. The aims of this study were to validate a bioassay for soluble vascular endothelial growth factor receptors 1 and 2 (sVEGFR-1 and sVEGFR-2) in baboon plasma and to determine the maternal and fetal plasma concentration of these receptors at the end of the baboon pregnancy. METHODS: Maternal peripheral blood samples were obtained from eight baboons (Papio anubis) prior to elective cesarean section and from the umbilical cord after the fetuses were delivered. Spike and recovery experiments at various concentrations in pooled baboon maternal plasma were used to validate a human quantitative sandwich immunoassay for sVEGFR-1 and -2. Concentrations of sVEGFR-1 and -2 were then determined in maternal and fetal plasma samples. RESULTS: No significant correlations were observed between sVEGFR-1 or -2 concentrations in maternal and fetal circulations. The concentration of sVEGFR-1 was at least 30 times greater and that of sVEGFR-2 approximately two times greater, in maternal than in cord plasma (both P < 0.01). CONCLUSION: These findings suggest that baboons can be used to study the regulation of vasculogenesis during pregnancy. 相似文献
343.
Modupeola A. Sowole John A. Alexopoulos Yi-Qiang Cheng Joaquin Ortega Lars Konermann 《Journal of molecular biology》2013
The bacterial protease ClpP consists of 14 subunits that assemble into two stacked heptameric rings. The central degradation chamber can be accessed via axial pores. In free ClpP, these pores are obstructed by the N-terminal regions of the seven subunits at either end of the barrel. Acyldepsipeptides (ADEPs) are antibacterial compounds that bind in hydrophobic clefts surrounding the pore region, causing the pores to open up. The ensuing uncontrolled degradation of intracellular proteins is responsible for the antibiotic activity of ADEPs. Recently published X-ray structures yielded conflicting models regarding the conformation adopted by the N-terminal regions in the open state. Here, we use hydrogen/deuterium exchange (HDX) mass spectrometry to obtain complementary insights into the ClpP behavior with and without ADEP1. Ligand binding causes rigidification of the equatorial belt, accompanied by destabilization in the vicinity of the binding clefts. The N-terminal regions undergo rapid deuteration with only minor changes after ADEP1 binding, revealing a lack of stable H-bonding. Our data point to a mechanism where the pore opening mechanism is mediated primarily by changes in the packing of N-terminal nonpolar side chains. We propose that a “hydrophobic plug” causes pore blockage in ligand-free ClpP. ADEP1 binding provides new hydrophobic anchor points that nonpolar N-terminal residues can interact with. In this way, ADEP1 triggers the transition to an open conformation, where nonpolar moieties are clustered around the rim of the pore. This proposed mechanism helps reconcile the conflicting models that had been put forward earlier. 相似文献
344.
Alberto Mariscal Rosa M. Lopez-Gigosos Manuel Carnero-Varo Joaquin Fernandez-Crehuet 《Applied microbiology and biotechnology》2009,82(4):773-783
A new, quick method, using the resazurin dye test as a bacterial respiration indicator, has been developed to assay the antibacterial
activity of various substances used as disinfectants against bacterial biofilm growth on clinical devices. Resazurin was used
to measure the presence of active biofilm bacteria, after adding disinfectant, in relation to a standard curve generated from
inocula in suspension of the same organism used to grow the biofilm. The biofilm was quantified indirectly by measuring the
fluorescent, water-soluble resorufin product produced when resazurin is reduced by reactions associated with respiration. Four products used as disinfectants and the
biofilm growth of five bacterial species on carriers made of materials commonly found in clinical devices were studied. Under
test conditions, chlorhexidine, NaOCl, ethanol, and Perasafe at concentrations of 0.2, 0.01, 350, and 0.16 mg/ml, respectively,
all produced 5-log reductions in biofilm cell numbers on the three different carriers. The redox-driven test depends on bacterial
catabolism, for which reason resazurin reduction produces an analytic signal of the bacterial activity in whole cells, and
therefore could be used for determining disinfectant efficacy in an assay based on the metabolic activity of microorganisms
grown as biofilm or in suspension. 相似文献
345.
Anita Mitico Tanaka Aparecida Sadae Tanaka Alexandre Paulo Yague Lopes Mickie Takagi Joaquin Cabrera-Crespo Isaias Raw 《Biotechnology letters》2000,22(4):257-260
Factor VIII was purified from porcine plasma using adsorption on aluminium hydroxide with CM-cellulose as a filtration aid, cold ethanol precipitation, and two anion-exchange (Q-Sepharose fast flow) chromatographies. The final product was purified 264-fold and had a specific activity of 10 U mg–1. The method is suitable to produce purified porcine FVIII by an easy process where all steps can be scaled up. The final product is free of von Willebrand factor that is responsible for the main side effects in patients. Finally, this method can be used to obtain purified porcine plasma FVIII for use in haemophilic patients with inhibitors. 相似文献
346.
Tongyu Cao Wikramanayake Elizabeth Alvarez-Connelly Jessica Simon Lucia M. Mauro Javier Guzman George Elgart Lawrence A. Schachner Juan Chen Lisa R. Plano Joaquin J. Jimenez 《Cell stress & chaperones》2010,15(6):985-991
Alopecia areata (AA) is a common autoimmune disease characterized by non-scarring hair loss. Previous studies have demonstrated
an association between AA and physiological/psychological stress. In this study, we investigated the effects of heat treatment,
a physiological stress, on AA development in C3H/HeJ mice. Whereas this strain of mice are predisposed to AA at low incidence
by 18 months of age, we observed a significant increase in the incidence of hair loss in heat-treated 8-month-old C3H/HeJ
mice compared with sham-treated mice. Histological analysis detected mononuclear cell infiltration in anagen hair follicles,
a characteristic of AA, in heat-treated mouse skin. As expected, increased expression of induced HSPA1A/B (formerly called
HSP70i) was detected in skin samples from heat-treated mice. Importantly, increased HSPA1A/B expression was also detected
in skin samples from C3H/HeJ mice that developed AA spontaneously. Our results suggest that induction of HSPA1A/B may precipitate
the development of AA in C3H/HeJ mice. For future studies, the C3H/HeJ mice with heat treatment may prove a useful model to
investigate stress response in AA. 相似文献
347.
Gisela Nogales-Gadea Emma Mormeneo Inés García-Consuegra Juan C. Rubio Anna Orozco Joaquin Arenas Miguel A. Martín Alejandro Lucia Anna M. Gómez-Foix Ramon Martí Antoni L. Andreu 《PloS one》2010,5(10)
Background
Mutations in the PYGM gene encoding skeletal muscle glycogen phosphorylase (GP) cause a metabolic disorder known as McArdle''s disease. Previous studies in muscle biopsies and cultured muscle cells from McArdle patients have shown that PYGM mutations abolish GP activity in skeletal muscle, but that the enzyme activity reappears when muscle cells are in culture. The identification of the GP isoenzyme that accounts for this activity remains controversial.Methodology/Principal Findings
In this study we present two related patients harbouring a novel PYGM mutation, p.R771PfsX33. In the patients'' skeletal muscle biopsies, PYGM mRNA levels were ∼60% lower than those observed in two matched healthy controls; biochemical analysis of a patient muscle biopsy resulted in undetectable GP protein and GP activity. A strong reduction of the PYGM mRNA was observed in cultured muscle cells from patients and controls, as compared to the levels observed in muscle tissue. In cultured cells, PYGM mRNA levels were negligible regardless of the differentiation stage. After a 12 day period of differentiation similar expression of the brain and liver isoforms were observed at the mRNA level in cells from patients and controls. Total GP activity (measured with AMP) was not different either; however, the active GP activity and immunoreactive GP protein levels were lower in patients'' cell cultures. GP immunoreactivity was mainly due to brain and liver GP but muscle GP seemed to be responsible for the differences.Conclusions/Significance
These results indicate that in both patients'' and controls'' cell cultures, unlike in skeletal muscle tissue, most of the protein and GP activities result from the expression of brain GP and liver GP genes, although there is still some activity resulting from the expression of the muscle GP gene. More research is necessary to clarify the differential mechanisms of metabolic adaptations that McArdle cultures undergo in vitro. 相似文献348.
Complex coherent dynamics is present in a wide variety of neural systems. A typical example is the voltage transitions between up and down states observed in cortical areas in the brain. In this work, we study this phenomenon via a biologically motivated stochastic model of up and down transitions. The model is constituted by a simple bistable rate dynamics, where the synaptic current is modulated by short-term synaptic processes which introduce stochasticity and temporal correlations. A complete analysis of our model, both with mean-field approaches and numerical simulations, shows the appearance of complex transitions between high (up) and low (down) neural activity states, driven by the synaptic noise, with permanence times in the up state distributed according to a power-law. We show that the experimentally observed large fluctuation in up and down permanence times can be explained as the result of sufficiently noisy dynamical synapses with sufficiently large recovery times. Static synapses cannot account for this behavior, nor can dynamical synapses in the absence of noise. 相似文献
349.
Mari Carmen Gil Borlado David Moreno Lastres Maritza Gonzalez Hoyuela Maria Moran Alberto Blazquez Rosa Pello Lorena Marin Buera Toni Gabaldon Juan Jose Garcia Pe?as Miguel A. Martín Joaquin Arenas Cristina Ugalde 《PloS one》2010,5(9)
Background
In recent years clinical evidence has emphasized the importance of the mtDNA genetic background that hosts a primary pathogenic mutation in the clinical expression of mitochondrial disorders, but little experimental confirmation has been provided. We have analyzed the pathogenic role of a novel homoplasmic mutation (m.15533 A>G) in the cytochrome b (MT-CYB) gene in a patient presenting with lactic acidosis, seizures, mild mental delay, and behaviour abnormalities.Methodology
Spectrophotometric analyses of the respiratory chain enzyme activities were performed in different tissues, the whole muscle mitochondrial DNA of the patient was sequenced, and the novel mutation was confirmed by PCR-RFLP. Transmitochondrial cybrids were constructed to confirm the pathogenicity of the mutation, and assembly/stability studies were carried out in fibroblasts and cybrids by means of mitochondrial translation inhibition in combination with blue native gel electrophoresis.Principal Findings
Biochemical analyses revealed a decrease in respiratory chain complex III activity in patient''s skeletal muscle, and a combined enzyme defect of complexes III and IV in fibroblasts. Mutant transmitochondrial cybrids restored normal enzyme activities and steady-state protein levels, the mutation was mildly conserved along evolution, and the proband''s mother and maternal aunt, both clinically unaffected, also harboured the homoplasmic mutation. These data suggested a nuclear genetic origin of the disease. However, by forcing the de novo functioning of the OXPHOS system, a severe delay in the biogenesis of the respiratory chain complexes was observed in the mutants, which demonstrated a direct functional effect of the mitochondrial genetic background.Conclusions
Our results point to possible pitfalls in the detection of pathogenic mitochondrial mutations, and highlight the role of the genetic mtDNA background in the development of mitochondrial disorders. 相似文献350.
Before we can realize our long term goal of engineering lung tissue worthy of clinical applications, advances in the identification and utilization of cell sources, development of standardized procedures for differentiation of cells, production of matrix tailored to meet the needs of the lung and design of methods or techniques of applying the engineered tissues into the injured lung environment will need to occur. Design of better biomaterials with the capacity to guide stem cell behavior and facilitate lung lineage choice as well as seamlessly integrate with living lung tissue will be achieved through advances in the development of decellularized matrices and new understandings related to the influence of extracellular matrix on cell behavior and function. We have strong hopes that recent developments in the engineering of conducting airway from decellularized trachea will lead to similar breakthroughs in the engineering of distal lung components in the future.Key words: adult and embryonic stem cells, tissue engineered lung, lung stem cells, lung matrices, lung development 相似文献