全文获取类型
收费全文 | 1634篇 |
免费 | 135篇 |
国内免费 | 2篇 |
出版年
2022年 | 13篇 |
2021年 | 23篇 |
2020年 | 7篇 |
2019年 | 12篇 |
2018年 | 24篇 |
2017年 | 22篇 |
2016年 | 41篇 |
2015年 | 72篇 |
2014年 | 71篇 |
2013年 | 87篇 |
2012年 | 130篇 |
2011年 | 107篇 |
2010年 | 82篇 |
2009年 | 71篇 |
2008年 | 123篇 |
2007年 | 101篇 |
2006年 | 105篇 |
2005年 | 102篇 |
2004年 | 98篇 |
2003年 | 90篇 |
2002年 | 78篇 |
2001年 | 32篇 |
2000年 | 11篇 |
1999年 | 26篇 |
1998年 | 24篇 |
1997年 | 21篇 |
1996年 | 17篇 |
1995年 | 17篇 |
1994年 | 19篇 |
1993年 | 19篇 |
1992年 | 16篇 |
1991年 | 13篇 |
1990年 | 8篇 |
1989年 | 5篇 |
1988年 | 6篇 |
1987年 | 7篇 |
1986年 | 9篇 |
1985年 | 7篇 |
1983年 | 4篇 |
1982年 | 5篇 |
1981年 | 4篇 |
1979年 | 3篇 |
1978年 | 4篇 |
1977年 | 5篇 |
1976年 | 8篇 |
1975年 | 2篇 |
1974年 | 3篇 |
1973年 | 2篇 |
1968年 | 4篇 |
1965年 | 2篇 |
排序方式: 共有1771条查询结果,搜索用时 15 毫秒
141.
Awad AB Fink CS Trautwein EA Ntanios FY 《The Journal of nutritional biochemistry》2005,16(11):650-655
Previous studies from our laboratory on tumor cells suggest that phytosterols stimulate ceramide production, which was associated with cell growth inhibition and stimulation of apoptosis. The objective of the present study was to examine the effect of phytosterols on ceramide metabolism in small intestinal cells that represent the first cells in contact with dietary phytosterols. Caco(2) cells, an accepted model for human intestinal epithelial cells, were used in this study. Ceramide and ceramide-containing lipids were examined by labeling the ceramide pool with (3)H-serine. Cells were supplemented with 16 microM of sterols (cholesterol, beta-sitosterol or campesterol) for 16 days postconfluence and continued to differentiate. Of the two phytosterols, beta-sitosterol, but not campesterol, induced more than double the serine labeling when compared with cholesterol. This increase was uniform in sphingomyelin (SM), ceramide and sphingosine labeling. Sterols had no effect on SM concentration in the cells. In addition, sterol had no effect on the activity of SM synthase or sphingomyelinases. There was an inhibition of ceramidases with campesterol supplementation. These data suggest that the observed increases in SM and sphingosine labeling were due to an increase in ceramide turnover. The increase in ceramide turnover with beta-sitosterol supplementation was not associated with growth inhibition but was with increases in ceramide glycosylation products such as cerebrosides and gangliosides. It was concluded that beta-sitosterol has no effect on differential Caco(2), a model of normal small intestinal cells. The increase in the glycosylated ceramide products may offer a means to protect the cells from the harmful effect of ceramide by excreting them with lipoproteins. 相似文献
142.
Maria?C.?Romero Elke?HammerEmail author Renate?Hanschke Angelica?M.?Arambarri Frieder?Schauer 《World journal of microbiology & biotechnology》2005,21(2):101-106
The filamentous fungusTalaromyces helicus , isolated from oil-contaminated sludge, oxidizes biphenyl via 4-hydroxybiphenyl to the dihydroxylated derivatives 4,4-dihydroxybiphenyl and 3,4-dihydroxybiphenyl, which, to a certain extent, are converted to glycosyl conjugates. The sugar moiety of the conjugate formed from 4,4-dihydroxybiphenyl was identified as glucose. Further metabolites: 2-hydroxybiphenyl, 2,5-dihydroxylated biphenyl, and the ring cleavage product 4-phenyl-2-pyrone-6-carboxylic acid accumulated only in traces. From these results the main pathway for biotransformation of biphenyl in T. helicus could be proposed to be the excretion of dihydroxylated derivatives (>75%) and their glucosyl conjugates (<25%). 相似文献
143.
The dynamics of the nucleobase and the ribose moieties in a 14-nt RNA cUUCGg hairpin-loop uniformly labeled with 13C and 15N were studied by 13C spin relaxation experiments. R1, R1ρ and the 13C-{1H} steady-state NOE of C6 and C1′ in pyrimidine and C8 and C1′ in purine residues were obtained at 298 K. The relaxation data were analyzed by the model-free formalism to yield dynamic
information on timescales of pico-, nano- and milli-seconds. An axially symmetric diffusion tensor with an overall rotational
correlation time τc of 2.31±0.13 ns and an axial ratio of 1.35±0.02 were determined. Both findings are in agreement with hydrodynamic calculations.
For the nucleobase carbons, the validity of different reported 13C chemical shift anisotropy values (Stueber, D. and Grant, D. M., 2002 J. Am. Chem. Soc. 124, 10539–10551; Fiala et al., 2000 J. Biomol. NMR 16, 291–302; Sitkoff, D. and Case, D. A., 1998 Prog. NMR Spectroscopy 32, 165–190) is discussed. The resulting dynamics are in agreement with the structural features of the cUUCGg motif in that
all residues are mostly rigid (0.82 < S2 < 0.96) in both the nucleobase and the ribose moiety except for the nucleobase of U7, which is protruding into solution (S2 = 0.76). In general, ribose mobility follows nucleobase dynamics, but is less pronounced. Nucleobase dynamics resulting from
the analysis of 13C relaxation rates were found to be in agreement with 15N relaxation data derived dynamic information (Akke et al., 1997 RNA 3, 702–709).
Electronic supplementary material Electronic supplementary material is available for this article at
and accessible for authorised users. 相似文献
144.
From dormant to germinating spores of Streptomyces coelicolor A3(2): new perspectives from the crp null mutant 总被引:1,自引:0,他引:1
Piette A Derouaux A Gerkens P Noens EE Mazzucchelli G Vion S Koerten HK Titgemeyer F De Pauw E Leprince P van Wezel GP Galleni M Rigali S 《Journal of proteome research》2005,4(5):1699-1708
The complete understanding of the morphological differentiation of streptomycetes is an ambitious challenge as diverse sensors and pathways sensitive to various environmental stimuli control the process. Germination occupies a particular position in the life cycle as the good achievement of the process depends on events occurring both during the preceding sporulation and during germination per se. The cyclic AMP receptor protein (crp) null mutant of Streptomyces coelicolor, affected in both sporulation and germination, was therefore presented as a privileged candidate to highlight new proteins involved in the shift from dormant to germinating spores. Our multidisciplinary approach-combining in vivo data, the analysis of spores morphological properties, and a proteome study-has shown that Crp is a central regulatory protein of the life cycle in S. coelicolor; and has identified spores proteins with statistically significant increased or decreased expression that should be listed as priority targets for further investigations on proteins that trigger both ends of the life cycle. 相似文献
145.
Rat Sertoli cells express epithelial but also mesenchymal genes after immortalization with SV40 总被引:2,自引:0,他引:2
Konrad L Munir Keilani M Cordes A Völck-Badouin E Laible L Albrecht M Renneberg H Aumüller G 《Biochimica et biophysica acta》2005,1722(1):6-14
A new immortal Sertoli cell line from pubertal rat testis was established and characterized. We have generated the clonal line SCIT-C8 expressing established markers for Sertoli cells (SC) like transferrin, clusterin and steel factor/stem cell factor (SCF). Additionally, the immortalized cells express afadin, a protein which is a member of tight and adherens junctions, therefore the cells may be useful for studies of the blood-testis barrier (BTB) in vitro. In contrast to primary SC, the immortalized cells lost expression of androgen receptor and responsiveness to androgens and follicle-stimulating hormone. Surprisingly, we found mRNA expression and protein secretion of the mesenchymal markers, fibronectin and entactin-1, which we also observed for the immortalized SC lines, ASC-17D and 93RS2. In comparison to primary SC, the immortalized cells demonstrated enhanced adhesion in vitro. This correlated with the expression of entactin-1 because adhesion was strongly reduced by antibody perturbation experiments. Additionally, we found the alternatively spliced and primarily muscle cell-specific long variant of TGF-beta2 not only in peritubular cells (PC), but also in the primary and immortalized SC. Furthermore, all immortalized cell lines secreted higher amounts of TGF-beta2 than primary SC. In conclusion, the immortalized SC lines from different developmental stages showed a similar pattern of epithelial and mesenchymal markers. 相似文献
146.
In this report we analyze the UL104 open reading frame of human cytomegalovirus (HCMV) genome that encodes the putative portal protein. An affinity-purified monospecific antiserum directed against a GST-UL104 fusion protein identified proteins of approximate M(r) 73000 and 145000 in HCMV-infected cells and purified virions. Furthermore, using an in vitro assay the ability of pUL104 to bind double-stranded DNA was shown. Analysis under native conditions of pUL104 revealed that the monomeric and dimeric forms of the protein also form high molecular weight complexes upon sucrose gradient centrifugation. The protein has been purified from recombinant baculovirus UL104 infected cells. The quaternary structure of rpUL104 was investigated by gel permeation chromatography and electron microscopy. The purified rpUL104 was found to assemble into high molecular weight complexes, a prerequisite of portal proteins which form channels for DNA import into capsids. 相似文献
147.
Verleyen P Clynen E Huybrechts J Van Lommel A Vanden Bosch L De Loof A Zdarek J Schoofs L 《Developmental biology》2004,273(1):38-47
Thirty-five years ago, Zdarek and Fraenkel demonstrated that nervous tissue extracts influenced development by accelerating pupariation in the grey flesh fly, Neobellieria bullata. We have now identified this pupariation factor as SVQFKPRLamide, designated Neb-pyrokinin-2 (Neb-PK-2). To achieve this, the central nervous system of N. bullata wandering stage larvae, that is, preceding pupariation, were dissected and extracted before HPLC separation. Chromatographic fractions were screened with a bioassay for pupariation accelerating activity. Only one fraction showed huge pupariation activity. Mass spectrometry revealed the presence of a pyrokinin, whose primary sequence could not be unequivocally determined by tandem mass spectrometry. However, this Neb-pyrokinin appeared to be very prominent in the ring gland from which it was subsequently purified and identified. Synthetic Neb-PK-2 accelerates pupariation with a threshold dose of only 0.2 pmol and therefore, Neb-pyrokinin is considered to be the genuine pupariation factor. The immunohistochemical distribution pattern of Neb-PK-2 is very similar to that of Drosophila pyrokinin-2, from which it differs by only one amino acid residue. Hence, the recently identified G-protein coupled receptors (CG8784, CG8795) for Drosophila pyrokinin-2 might play an important role in puparium formation. 相似文献
148.
Kikkawa Y Yu H Genersch E Sanzen N Sekiguchi K Fässler R Campbell KP Talts JF Ekblom P 《Experimental cell research》2004,300(1):94-108
The presence of many laminin receptors of the beta1 integrin family on most cells makes it difficult to define the biological functions of other major laminin receptors such as integrin alpha6beta4 and dystroglycan. We therefore tested the binding of a beta1 integrin-null cell line GD25 to four different laminin variants. The cells were shown to produce dystroglycan, which based on affinity chromatography bound to laminin-1, -2/4, and -10/11, but not to laminin-5. The cells also expressed the integrin alpha6Abeta4A variant. GD25 beta1 integrin-null cells are known to bind poorly to laminin-1, but we demonstrate here that these cells bind avidly to laminin-2/4, -5, and -10/11. The initial binding at 20 min to each of these laminins could be inhibited by an integrin alpha6 antibody, but not by a dystroglycan antibody. Hence, integrin alpha6Abeta4A of GD25 cells was identified as a major receptor for initial GD25 cell adhesion to three out of four tested laminin isoforms. Remarkably, cell adhesion to laminin-5 failed to promote cell spreading, proliferation, and extracellular signal-regulated kinase (ERK) activation, whereas all these responses occurred in response to adhesion to laminin-2/4 or -10/11. The data establish GD25 cells as useful tools to define the role integrin alpha6Abeta4A and suggest that laminin isoforms have distinctly different capacities to promote cell adhesion and signaling via integrin alpha6Abeta4A. 相似文献
149.
Terminases are a class of proteins which catalyze the generation of unit-length genomes during DNA packaging. These essential proteins are conserved throughout the herpesviruses and many double-stranded DNA bacteriophages. We have determined the structure of the large terminase subunit pUL56 of human cytomegalovirus, a highly pathogenic virus, to 2.6 nm resolution. Image analysis of purified pUL56 suggests that the molecule exists as a dimer formed by the association of two ring-like structures positioned on top of each other and connected by a pronounced density on one side. The 3D reconstruction of pUL56 provides first structural insights into the active protein. 相似文献
150.
Moina micrura Kurz, 1874 (Crustacea: Anomopoda), commonly regarded as a cosmopolitan cladoceran species, can be found almost all over the world except for arctic and cold-temperate regions. M. micrura has been recorded from virtually all types of limnetic habitats, including temperate permanent ponds, lakes, ephemeral desert pools, and tropical brackish fishponds. Its high morphological and ecological variability has so far been considered a result of intraspecific diversity. We tested the hypothesis that genetically isolated groups exist within M. micrura with crossing experiment and DNA sequence analysis. We compared two clones originating from populations from Central Europe (area of the type locality) and Australia. As there was extremely low production of hybrid eggs, the results of the crossing experiment strongly suggest the existence of reproductive isolation. The sequence divergence of mitochondrial gene for 12S rRNA (7.9%) was comparable to intraspecific differences within Daphnia pulex group. The sequence divergence of the clones of Moina macrocopa from Europe and Central Africa was much lower (1.4%). Our results suggest that at least two sibling species exist within M. micrura complex, which is in agreement with other data questioning the cosmopolitanism of various cladoceran species. 相似文献