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971.
Justin J. J. van der Hooft Jacques Vervoort Raoul J. Bino Ric C. H. de Vos 《Metabolomics : Official journal of the Metabolomic Society》2012,8(4):691-703
The identification of large series of metabolites detectable by mass spectrometry (MS) in crude extracts is a challenging task. In order to test and apply the so-called multistage mass spectrometry (MS n ) spectral tree approach as tool in metabolite identification in complex sample extracts, we firstly performed liquid chromatography (LC) with online electrospray ionization (ESI)?CMS n , using crude extracts from both tomato fruit and Arabidopsis leaf. Secondly, the extracts were automatically fractionated by a NanoMate LC-fraction collector/injection robot (Advion) and selected LC-fractions were subsequently analyzed using nanospray-direct infusion to generate offline in-depth MS n spectral trees at high mass resolution. Characterization and subsequent annotation of metabolites was achieved by detailed analysis of the MS n spectral trees, thereby focusing on two major plant secondary metabolite classes: phenolics and glucosinolates. Following this approach, we were able to discriminate all selected flavonoid glycosides, based on their unique MS n fragmentation patterns in either negative or positive ionization mode. As a proof of principle, we report here 127 annotated metabolites in the tomato and Arabidopsis extracts, including 21 novel metabolites. Our results indicate that online LC?CMS n fragmentation in combination with databases of in-depth spectral trees generated offline can provide a fast and reliable characterization and annotation of metabolites present in complex crude extracts such as those from plants. 相似文献
972.
Respiratory Syncytial Virus (RSV) is a major cause of viral brochiolitis in infants and young children and is also a significant problem in elderly and immuno-compromised adults. To date there is no efficacious and safe RSV vaccine, partially because of the outcome of a clinical trial in the 1960s with a formalin-inactivated RSV vaccine (FI-RSV). This vaccine caused enhanced respiratory disease upon exposure to the live virus, leading to increased morbidity and the death of two children. Subsequent analyses of this incident showed that FI-RSV induces a Th2-skewed immune response together with poorly neutralizing antibodies. As a new approach, we used reconstituted RSV viral envelopes, i.e. virosomes, with incorporated monophosphoryl lipid A (MPLA) adjuvant to enhance immunogenicity and to skew the immune response towards a Th1 phenotype. Incorporation of MPLA stimulated the overall immunogenicity of the virosomes compared to non-adjuvanted virosomes in mice. Intramuscular administration of the vaccine led to the induction of RSV-specific IgG2a levels similar to those induced by inoculation of the animals with live RSV. These antibodies were able to neutralize RSV in vitro. Furthermore, MPLA-adjuvanted RSV virosomes induced high amounts of IFNγ and low amounts of IL5 in both spleens and lungs of immunized and subsequently challenged animals, compared to levels of these cytokines in animals vaccinated with FI-RSV, indicating a Th1-skewed response. Mice vaccinated with RSV-MPLA virosomes were protected from live RSV challenge, clearing the inoculated virus without showing signs of lung pathology. Taken together, these data demonstrate that RSV-MPLA virosomes represent a safe and efficacious vaccine candidate which warrants further evaluation. 相似文献
973.
Santos-Filho OA Forge D Hoelz LV de Freitas GB Marinho TO Araújo JQ Albuquerque MG de Alencastro RB Boechat N 《Journal of molecular modeling》2012,18(9):4061-4072
Pneumocystis carinii is typically a non-pathogenic fungus found in the respiratory tract of healthy humans. However, it may cause P. carinii pneumonia (PCP) in people with immune deficiency, affecting mainly premature babies, cancer patients and transplant recipients, and people with acquired immunodeficiency syndrome (AIDS). In the latter group, PCP occurs in approximately 80% of patients, a major cause of death. Currently, there are many available therapies to treat PCP patients, including P. carinii dihydrofolate reductase (PcDHFR) inhibitors, such as trimetrexate (TMX), piritrexim (PTX), trimethoprim (TMP), and pyrimethamine (PMT). Nevertheless, the high percentage of adverse side effects and the limited therapeutic success of the current drug therapy justify the search for new drugs rationally planned against PCP. This work focuses on the study of pyrimidine inhibitors of PcDHFR, using both CoMFA and CoMSIA 3D-QSAR methods. 相似文献
974.
A retained intron in the 3′‐UTR of Calm3 mRNA mediates its Staufen2‐ and activity‐dependent localization to neuronal dendrites 下载免费PDF全文
975.
Thomas?B?rnerEmail authorView authors OrcID profile
return OK on get 《Protoplasma》2017,254(5):1845-1855
DNA and machinery for gene expression have been discovered in chloroplasts during the 1960s. It was soon evident that the chloroplast genome is relatively small, that most genes for chloroplast-localized proteins reside in the nucleus and that chloroplast membranes, ribosomes, and protein complexes are composed of proteins encoded in both the chloroplast and the nuclear genome. This situation has made the existence of mechanisms highly probable that coordinate the gene expression in plastids and nucleus. In the 1970s, the first evidence for plastid signals controlling nuclear gene expression was provided by studies on plastid ribosome deficient mutants with reduced amounts and/or activities of nuclear-encoded chloroplast proteins including the small subunit of Rubisco, ferredoxin NADP+ reductase, and enzymes of the Calvin cycle. This review describes first models of plastid-to-nucleus signaling and their discovery. Today, many plastid signals are known. They do not only balance gene expression in chloroplasts and nucleus during developmental processes but are also generated in response to environmental changes sensed by the organelles. 相似文献
976.
Bezerra EH Rocha BA Nagano CS Bezerra Gde A Moura TR Bezerra MJ Benevides RG Sampaio AH Assreuy AM Delatorre P Cavada BS 《Biochemical and biophysical research communications》2011,(4):566-570
Diocleinae lectins are highly homologous in their primary structure which features metal binding sites and a carbohydrate recognition domain (CRD). Differences in the biological activity of legume lectins have been widely investigated using hemagglutination inhibition assays, isothermal titration microcalorimetry and co-crystallization with mono- and oligosaccharides. Here we report a new lectin crystal structure (ConBr) extracted from seeds of Canavalia brasiliensis, predict dimannoside binding by docking, identify the α-aminobutyric acid (Abu) binding pocket and compare the CRD of ConBr to that of homologous lectins. Based on the hypothesis that the carbohydrate affinity of lectins depends on CRD configuration, the relationship between tridimensional structure and endothelial NO synthase activation was used to clarify differences in biological activity. Our study established a correlation between the position of CRD amino acid side chains and the stimulation of NO release from endothelium. 相似文献
977.
Rantala A Rizzi E Castiglioni B de Bellis G Sivonen K 《Environmental microbiology》2008,10(3):653-664
We developed a new tool to detect and identify hepatotoxin-producing cyanobacteria of the genera Anabaena , Microcystis , Planktothrix , Nostoc and Nodularia . Genus-specific probe pairs were designed for the detection of the microcystin ( mcyE ) and nodularin synthetase genes ( ndaF ) of these five genera to be used with a DNA-chip. The method couples a ligation detection reaction, in which the polymerase chain reaction (PCR)-amplified mcyE / ndaF genes are recognized by the probe pairs, with a hybridization on a universal microarray. All the probe pairs specifically detected the corresponding mcyE / ndaF gene sequences when DNA from the microcystin- or nodularin-producing cyanobacterial strains were used as template in the PCR. Furthermore, the strict specificity of detection enabled identification of the potential hepatotoxin producers. Detection of the genes was very sensitive; only 1–5 fmol of the PCR product were needed to produce signal intensities that exceeded the set background threshold level. The genus-specific probe pairs also reliably detected potential microcystin producers in DNA extracted from six lake and four brackish water samples. In lake samples, the same microcystin producers were identified with quantitative real-time PCR analysis. The specificity, sensitivity and ability of the DNA-chip in simultaneously detecting all the main hepatotoxin producers make this method suitable for high-throughput analysis and monitoring of environmental samples. 相似文献
978.
de Melo Reis RA Ventura AL Schitine CS de Mello MC de Mello FG 《Neurochemical research》2008,33(8):1466-1474
Müller cells represent the main type of glia present in the retina interacting with most, if not all neurons in this tissue.
Müller cells have been claimed to function as optic fibers in the retina delivering light to photoreceptors with minimal distortion
and low loss [Franze et al (2007) Proc Natl Acad Sci 104:8287–8292]. Most of the mediators found in the brain are also detected
in the retinal tissue, and glia cells are active players in the synthesis, release, signaling and uptake of major mediators
of synaptic function. Müller glia trophic factors may regulate many different aspects of neuronal circuitry during synaptogenesis,
differentiation, neuroprotection and survival of photoreceptors, Retinal Ganglion Cells (RGCs) and other targets in the retina.
Here we review the role of several transmitters and trophic factors that participate in the neuron-glia loop in the retina.
Special issue article in honor of Dr. Ricardo Tapia. 相似文献
979.
D'Avolio A Sciandra M Siccardi M Baietto L de Requena DG Bonora S Di Perri G 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2007,848(2):374-378
A simple method for the quantification of tipranavir, the first non-peptidic HIV protease inhibitor, was developed and validated. Quinoxaline, as internal standard, was added to 50 microl of plasma before a liquid-liquid extraction by 600 microl of protein precipitation solution. The extracts were diluted before being injected in the chromatographic system. Chromatographic separation was made on a C18 column using potassium phosphate buffer (pH 3.2) and acetonitrile with gradient. Detection was performed by an UV detector at 260 nm. Relative error at three control quality concentrations ranged from -1.81 to 1.72%. Intra-day (CV%) and inter-day (CV%) precision ranged from 0.94 to 2.55% and from 3.07 to 4.24%, respectively. LOQ and LOD were 0.090 microg/ml and 0.035 microg/ml, respectively. Mean recovery was 87.1%+/-2.4%. Calibration curve was linear up to 180 microg/ml. Concentration range when optimized (0.703-180 microg/ml) proved to be adequate to measure tipranavir concentration in HIV-1-positive patients, therefore this method could be suitable for therapeutic drug monitoring of this drug. 相似文献
980.
López-Gatius F Hunter RH Garbayo JM Santolaria P Yániz J Serrano B Ayad A de Sousa NM Beckers JF 《Theriogenology》2007,67(8):1324-1330
The present study was designed to establish whether plasma pregnancy-associated glycoprotein-1 (PAG-1) measurements during the early fetal period can be associated with early fetal loss. Blood samples were obtained and ultrasound controls performed on days 35, 42, 49, 56, and 63 of gestation or until pregnancy loss from 98 lactating dairy cows. Radioimmunoassay systems were used to determine PAG-1 and progesterone concentrations. Of the 98 pregnancies investigated 18 (18.4%) suffered early fetal loss: 15 (18.5%) in cows carrying singletons, and 3 (16.7%) in twin pregnancies. In cows suffering pregnancy loss, all living embryos registered on day 35 seemed normal in size and development in all weekly ultrasound controls before fetal expulsion. Using analysis of variance, plasma PAG-1 and progesterone values were not different between no loss and fetal loss groups for every gestation period. Based on the odds ratio, and considering only PAG-1 values obtained on day 35 of gestation, the risk of fetal loss was 10 and 6.8 times more likely in cows with low (<2.5 ng/ml) and high (>4 ng/ml) PAG-1 values, respectively, than in cows with medium PAG-1 values, used as reference. Of the 10 inseminating bulls included in the study, one was related to increased fetal loss by odds ratio of 21.7, whereas one bull was attributed fetal loss rate reduced by odds ratio of 12.5 (1/0.08) These findings can have a clear clinical application: PAG-1 measurements from one single sample taken on day 35 of gestation provided more useful information than a series of values obtained from day 35 to 63 of gestation, and can be indicators of subsequent fetal loss. 相似文献