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951.
Pelletier JP Boileau C Boily M Brunet J Mineau F Geng C Reboul P Laufer S Lajeunesse D Martel-Pelletier J 《Arthritis research & therapy》2005,7(5):R1091-R1102
This study sought to evaluate the levels of mRNA expression and protein synthesis of MMP-13, cathepsin K, aggrecanase-1 (ADAMTS-4),
aggrecanase-2 (ADAMTS-5) and 5-lipoxygenase (5-LOX) in cartilage in the experimental anterior cruciate ligament (ACL) dog
model of osteoarthritis (OA), and to examine the effects of treatment with licofelone, a 5-lipoxygenase (LOX)/cyclooxygenase
(COX) inhibitor, on the levels of these catabolic factors. Sectioning of the ACL of the right knee was performed in three
experimental groups: group 1 received no active treatment (placebo group); and groups 2 and 3 received therapeutic concentrations
of licofelone (2.5 or 5.0 mg/kg/day orally, respectively) for 8 weeks, beginning the day following surgery. A fourth group
consisted of untreated dogs that were used as normal controls. Specimens of cartilage were selected from lesional areas of
OA femoral condyles and tibial plateaus, and were processed for real-time quantitative PCR and immunohistochemical analyses.
The levels of MMP-13, cathepsin K, ADAMTS-4, ADAMTS-5 and 5-LOX were found to be significantly increased in OA cartilage.
Licofelone treatment decreased the levels of both mRNA expression and protein synthesis of the factors studied. Of note was
the marked reduction in the level of 5-LOX gene expression. The effects of the drug were about the same at both tested dosages.
In vivo treatment with therapeutic dosages of licofelone has been found to reduce the degradation of OA cartilage in experimental
OA. This, coupled with the results of the present study, indicates that the effects of licofelone are mediated by the inhibition
of the major cartilage catabolic pathways involved in the destruction of cartilage matrix macromolecules. Moreover, our findings
also indicate the possible auto-regulation of 5-LOX gene expression by licofelone in OA cartilage. 相似文献
952.
Sergio?EncarnaciónEmail author Magdalena?Hernández Gabriel?Martínez-Batallar Sandra?Contreras María?del?Carmen?Vargas Jaime?Mora 《Biological procedures online》2005,7(1):117-135
We propose two-dimensional gel electrophoresis (2-DE) and mass spectrometry to define the protein components of regulons and
stimulons in bacteria, including those organisms where genome sequencing is still in progress. The basic 2-DE protocol allows
high resolution and reproducibility and enables the direct comparison of hundreds or even thousands of proteins simultaneously.
To identify proteins that comprise stimulons and regulons, peptide mass fingerprint (PMF) with matrix-assisted laser desorption
ionization/time-of-flight mass spectrometry (MALDI-TOF-MS) analysis is the first option and, if results from this tool are
insufficient, complementary data obtained with electrospray ionization tandem-MS (ESI-MS/MS) may permit successful protein
identification. ESI-MS/MS and MALDI-TOF-MS provide complementary data sets, and so a more comprehensive coverage of a proteome
can be obtained using both techniques with the same sample, especially when few sequenced proteins of a particular organism
exist or genome sequencing is still in progress. 相似文献
953.
954.
Horak P Pils D Haller G Pribill I Roessler M Tomek S Horvat R Zeillinger R Zielinski C Krainer M 《Molecular cancer research : MCR》2005,3(6):335-343
Dysregulation of apoptosis may support tumorigenesis by allowing cells to live beyond their normally intended life span. The various receptors for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) are located on chromosome 8p21.2, a region frequently deleted in ovarian cancer. Lack of expression of TRAIL receptor 1 (death receptor 4, DR4) correlates with resistance to TRAIL-induced apoptosis in ovarian cancer cells. Reconstitution of DR4 in the TRAIL-resistant A2780 ovarian cancer cell line was investigated with the demethylating agent 5-aza-2'-deoxycytidine and transient gene transfer. Regulation of other genes in the TRAIL pathway by 5-aza-2'-deoxycytidine was assessed in DNA GeneChip experiments. Primary ovarian cancers were analyzed by methylation-specific PCR and immunohistochemical analysis of a tissue microarray. Regulation of DR4 expression by demethylation or transient transfection is of functional relevance for TRAIL resistance in an ovarian cancer cell line. Hypermethylation of the DR4 promoter could be found in 10 of 36 (27.7%) DNAs isolated from ovarian cancer tissue. In an independent set of 68 ovarian cancer cases, a complete loss or down-regulation of DR4 protein expression was observed 10.3% and 8.8% patients, respectively. A significant (P = 0.019) majority of these patients was below 50 years of age. Our findings show a functional relevance of the level of DR4 expression in ovarian cancer and suggest a substantial contribution of DR4 hypermethylation and consequent loss of DR4 expression to ovarian cancer pathogenesis, particularly in premenopausal patients. 相似文献
955.
Gailus-Durner V Fuchs H Becker L Bolle I Brielmeier M Calzada-Wack J Elvert R Ehrhardt N Dalke C Franz TJ Grundner-Culemann E Hammelbacher S Hölter SM Hölzlwimmer G Horsch M Javaheri A Kalaydjiev SV Klempt M Kling E Kunder S Lengger C Lisse T Mijalski T Naton B Pedersen V Prehn C Przemeck G Racz I Reinhard C Reitmeir P Schneider I Schrewe A Steinkamp R Zybill C Adamski J Beckers J Behrendt H Favor J Graw J Heldmaier G Höfler H Ivandic B Katus H Kirchhof P Klingenspor M Klopstock T Lengeling A 《Nature methods》2005,2(6):403-404
956.
Vieira AR Avila JR Daack-Hirsch S Dragan E Félix TM Rahimov F Harrington J Schultz RR Watanabe Y Johnson M Fang J O'Brien SE Orioli IM Castilla EE Fitzpatrick DR Jiang R Marazita ML Murray JC 《PLoS genetics》2005,1(6):e64
Nonsyndromic or isolated cleft lip with or without cleft palate (CL/P) occurs in wide geographic distribution with an average birth prevalence of 1/700. We used direct sequencing as an approach to study candidate genes for CL/P. We report here the results of sequencing on 20 candidate genes for clefts in 184 cases with CL/P selected with an emphasis on severity and positive family history. Genes were selected based on expression patterns, animal models, and/or role in known human clefting syndromes. For seven genes with identified coding mutations that are potentially etiologic, we performed linkage disequilibrium studies as well in 501 family triads (affected child/mother/father). The recently reported MSX1 P147Q mutation was also studied in an additional 1,098 cleft cases. Selected missense mutations were screened in 1,064 controls from unrelated individuals on the Centre d'Étude du Polymorphisme Humain (CEPH) diversity cell line panel. Our aggregate data suggest that point mutations in these candidate genes are likely to contribute to 6% of isolated clefts, particularly those with more severe phenotypes (bilateral cleft of the lip with cleft palate). Additional cases, possibly due to microdeletions or isodisomy, were also detected and may contribute to clefts as well. Sequence analysis alone suggests that point mutations in FOXE1, GLI2, JAG2, LHX8, MSX1, MSX2, SATB2, SKI, SPRY2, and TBX10 may be rare causes of isolated cleft lip with or without cleft palate, and the linkage disequilibrium data support a larger, as yet unspecified, role for variants in or near MSX2, JAG2, and SKI. This study also illustrates the need to test large numbers of controls to distinguish rare polymorphic variants and prioritize functional studies for rare point mutations. 相似文献
957.
Springer J Ruth P Beuerlein K Palus S Schipp R Westermann B 《Journal of molecular histology》2005,36(5):345-353
Summary Biogenic amines (serotonin and catecholamines), play an important role in the control of the blood flow not only in vertebrates,
but also in invertebrates such as cephalopods. In contrast to the well investigated hearts of the ȁ8modern,ȁ9 coleoid cephalopods,
the innervation of the heart of the archaic Nautilus pompilius L. has not been studied in detail. In this study the distribution and effects of biogenic amines in the Nautilus heart were investigated. Serotonin and catecholamines were visualised by the glyxoylic acid induced fluorescence. High performance
liquid chromatotography analysis was performed to discriminate between the catecholamines, which showed a high content of
noradrenaline in the 4 auricles, the aorta and the ventricle, whereas the ventricle showed a high dopamine content. Adrenaline
was found at a very low concentration in the ventricle. Serotonin and dopamine were also immunohistochemically localised to
larger nerves and throughout the heart, respectively. In organ bath experiments, the auricles showed little spontaneous activity.
After adding serotonin, they displayed rhythmical contractions, which were accelerated dose-dependently by noradrenaline.
In summary, these data suggest an important role for biogenic amines in the control of the heart of Nautilus pompilius L., with serotonin possibly stimulating excitatory nerve fibres, whereas noradrenaline is likely to influence the muscle
contraction itself. 相似文献
958.
Kuhn K Prinz T Schäfer J Baumann C Schärfke M Kienle S Schwarz J Steiner S Hamon C 《Proteomics》2005,5(9):2364-2368
Comparative proteome profiling using stable isotope peptide labelling and mass spectrometry has emerged as a promising strategy. Here, we show the broad potential of our proprietary protein sequence tag (PST) technology. A special feature of PST is its ability to detect a wide variety of proteins including the pharmaceutically relevant membrane and nuclear proteins. This procedure addresses a similar number of proteins, compared to the multidimensional protein identification technology approach, but offers additionally a quantitative analysis with its recently developed quantitative PST version. 相似文献
959.
Mitochondrial diseases may be caused by numerous mutations that alter proteins of the respiratory chain and of other metabolic pathways in the mitochondrium. For clinicians this disease group poses a considerable diagnostic challenge due to ambiguous genotype-phenotype relationships. Until now, only 30% of the mitochondriopathies can be diagnosed at the molecular level. We therefore need a new diagnostic tool that offers a wide view on the mitochondrial proteins. Here, we present a method to generate a high-resolution, large-gel two-dimensional gel electrophoretic (2-DE) map of a purified fraction of mitochondrial proteins from Epstein-Barr virus-immortalized lymphoblastoid cell line (LCL). LCLs can be easily obtained from patients and control subjects in a routine clinical setting. They often express the biochemical phenotype and can be cultured to high cell numbers, sufficient to gain enough purified material for 2-DE. In total we identified 166 mitochondrial proteins. Thirteen proteins were earlier not known to be of mitochondrial origin. Thirty-nine proteins were associated with human diseases ranging from respiratory chain enzyme deficiencies to disorders of beta-oxidation and amino acid metabolism. This 2-DE map is intended to be the first step to diagnose mitochondrial diseases at the proteomic level. 相似文献
960.
Phosphoproteomic analysis using immobilized metal ion affinity chromatography on the basis of cellulose powder 总被引:2,自引:0,他引:2
Feuerstein I Morandell S Stecher G Huck CW Stasyk T Huang HL Huber LA Bonn GK 《Proteomics》2005,5(1):46-54
Detailed characterization of phosphoproteins as well as other post-translationally modified proteins such as glycoproteins, is required to fully understand protein function and regulatory events in cells and organisms. Therefore, an experimental strategy for the isolation of phosphoproteins using a new immobilized metal ion affinity chromatograph (IMAC) material on the basis of cellulose has been developed and characterized. Different approaches have been used to test the material. Recovery rates were determined by 32P labelling of a myelin basic protein fragment and by reversed-phase high-performance liquid chromatography-electrospray ionization mass spectrometry using a tryptic digest of the model protein bovine beta-casein. Selectivity was demonstrated by enrichment and separation of phosphopeptides from different samples, such as from a digest of horse myoglobin as well as from a digest of in vitro phosphorylated extracellular signal regulates kinase 2 (ERK2) mixed with synthetic phosphopeptides, phosphorylated on different amino acid residues. Furthermore, simplification and optimization of sample pretreatment was achieved by combining the separating (IMAC) and desalting (C18) step during preparative high performance liquid chromatography. The comparison between our material and a commercially available IMAC system (POROS 20 MC; Perspective BioSystems) emphasizes the competitiveness of the cellulose. Confirmed by the obtained data, the cellulose material performed as well as the commercially available sorbent, however with the advantage, that it can be produced rather easily and at very low cost. 相似文献