全文获取类型
收费全文 | 624495篇 |
免费 | 66225篇 |
国内免费 | 412篇 |
专业分类
691132篇 |
出版年
2018年 | 5881篇 |
2016年 | 7938篇 |
2015年 | 9945篇 |
2014年 | 11851篇 |
2013年 | 16674篇 |
2012年 | 18649篇 |
2011年 | 19560篇 |
2010年 | 13415篇 |
2009年 | 12281篇 |
2008年 | 17560篇 |
2007年 | 18215篇 |
2006年 | 17144篇 |
2005年 | 16421篇 |
2004年 | 16335篇 |
2003年 | 15605篇 |
2002年 | 15299篇 |
2001年 | 26761篇 |
2000年 | 26763篇 |
1999年 | 21118篇 |
1998年 | 7254篇 |
1997年 | 7523篇 |
1996年 | 7160篇 |
1995年 | 6708篇 |
1994年 | 6507篇 |
1993年 | 6456篇 |
1992年 | 17425篇 |
1991年 | 17221篇 |
1990年 | 16977篇 |
1989年 | 16567篇 |
1988年 | 15646篇 |
1987年 | 14987篇 |
1986年 | 13744篇 |
1985年 | 13871篇 |
1984年 | 11458篇 |
1983年 | 9941篇 |
1982年 | 7449篇 |
1981年 | 6773篇 |
1980年 | 6433篇 |
1979年 | 10907篇 |
1978年 | 8516篇 |
1977年 | 7879篇 |
1976年 | 7615篇 |
1975年 | 8382篇 |
1974年 | 9218篇 |
1973年 | 9154篇 |
1972年 | 8365篇 |
1971年 | 7678篇 |
1970年 | 6792篇 |
1969年 | 6523篇 |
1968年 | 6309篇 |
排序方式: 共有10000条查询结果,搜索用时 11 毫秒
61.
We have previously shown that replacing the P1-site residue (Ala) of chicken ovomucoid domain 3 (OMCHI3) with a Met or Lys results in the acquisition of inhibitory activity toward chymotrypsin or trypsin, respectively. However, the inhibitory activities thus induced are not strong. In the present study, we introduced additional amino acid replacements around the reactive site to try to make the P1-site mutants more effective inhibitors of chymotrypsin or trypsin. The amino acid replacement Asp-->Tyr at the P2' site of OMCHI3(P1Met) resulted in conversion to a 35000-fold more effective inhibitor of chymotrypsin with an inhibitor constant (K(i)) of 1. 17x10(-11) M. The K(i) value of OMCHI3(P1Met, P2'Ala) indicated that the effect on the interaction with chymotrypsin of removing a negative charge from the P2' site was greater than that of introducing an aromatic ring. Similarly, enhanced inhibition of trypsin was observed when the Asp-->Tyr replacement was introduced into the P2' site of OMCHI3(P1Lys). Two additional replacements, Asp-->Ala at the P4 site and Arg-->Ala at the P3' site, made the mutant a more effective inhibitor of trypsin with a K(i) value of 1. 44x10(-9) M. By contrast, Arg-->Ala replacement at the P3' site of OMCHI3(P1Met, P2'Tyr) resulted in a greatly reduced inhibition of chymotrypsin, and Asp-->Ala replacement at the P4 site produced only a small change when compared with a natural variant of OMCHI3. These results clearly indicate that not only the P1-site residue but also the characteristics, particularly the electrostatic properties, of the amino acid residues around the reactive site of the protease inhibitor determine the strength of its interactions with proteases. Furthermore, amino acids with different characteristics are required around the reactive site for strong inhibition of chymotrypsin and trypsin. 相似文献
62.
Z Zhong A Toukdarian D Helinski V Knauf S Sykes J E Wilkinson C O'Bryne T Shea C DeLoughery R Caspi 《Applied and environmental microbiology》2001,67(12):5771-5779
An agar-degrading marine bacterium identified as a Microscilla species was isolated from coastal California marine sediment. This organism harbored a single 101-kb circular DNA plasmid designated pSD15. The complete nucleotide sequence of pSD15 was obtained, and sequence analysis indicated a number of genes putatively encoding a variety of enzymes involved in polysaccharide utilization. The most striking feature was the occurrence of five putative agarase genes. Loss of the plasmid, which occurred at a surprisingly high frequency, was associated with loss of agarase activity, supporting the sequence analysis results. 相似文献
63.
Binding of the cationic tetra(tributylammoniomethyl)-substituted hydroxoaluminum phthalocyanine (AlPcN4) to bilayer lipid membranes was studied by fluorescence correlation spectroscopy (FCS) and intramembrane field compensation (IFC) methods. With neutral phosphatidylcholine membranes, AlPcN4 appeared to bind more effectively than the negatively charged tetrasulfonated aluminum phthalocyanine (AlPcS4), which was attributed to the enhancement of the coordination interaction of aluminum with the phosphate moiety of phosphatidylcholine by the electric field created by positively charged groups of AlPcN4. The inhibitory effect of fluoride ions on the membrane binding of both AlPcN4 and AlPcS4 supported the essential role of aluminum-phosphate coordination in the interaction of these phthalocyanines with phospholipids. The presence of negative or positive charges on the surface of lipid membranes modulated the binding of AlPcN4 and AlPcS4 in accord with the character (attraction or repulsion) of the electrostatic interaction, thus showing the significant contribution of the latter to the phthalocyanine adsorption on lipid bilayers. The data on the photodynamic activity of AlPcN4 and AlPcS4 as measured by sensitized photoinactivation of gramicidin channels in bilayer lipid membranes correlated well with the binding data obtained by FCS and IFC techniques. The reduced photodynamic activity of AlPcN4 with neutral membranes violating this correlation was attributed to the concentration quenching of singlet excited states as proved by the data on the AlPcN4 fluorescence quenching. 相似文献
64.
J R Silkensen A P Skubitz K M Skubitz M E Rosenberg 《The journal of peptide research》1999,54(5):449-457
Expression of the glycoprotein clusterin is markedly increased following tissue injury. One function of clusterin is to promote cell interactions which are perturbed in these pathologic settings. Clusterin causes cell aggregation and adhesion in vitro yet the molecular mechanism for this effect is not known. In order to identify the active site(s) of clusterin, 34 peptides, each 15 amino acid residues in length, were synthesized from hydrophilic regions of human clusterin. When studied individually, none of the peptides caused aggregation of LLC-PK1 cells, a porcine renal epithelial cell line. However, two out of the 34 peptides inhibited clusterin-induced cell aggregation in a dose-dependent manner. Scrambled versions of these two 'active' peptides did not inhibit cell aggregation. Seven peptides promoted cell adhesion. In conclusion, these findings provide evidence for novel amino acid sequences mediating clusterin-induced renal cell interactions. 相似文献
65.
66.
I. N. Semenchuk L. A. Taranova A. A. Kalenyuk P. V. Il’yasov A. N. Reshetilov 《Applied Biochemistry and Microbiology》2000,36(1):69-72
The operating and storage stability of a receptor element of an amperometric biosensor based on thePseudomonas rathonis strain T capable of degrading surfactants was tested. Microbial cells were immobilized by incorporation in gels (agar, agarose,
and calcium-alginate), polyvinyl alcohol membrane, adhesion to Chromatographic paper GF/A, or by cross-linking induced by
glutaric aldehyde. Incorporation of microbial cells in agar gel provides long-standing conservation of their activity and
viability during measurements of high concentrations of surfactants and allows the receptor element of the biosensor to be
rapidly recovered after measurements. 相似文献
67.
68.
Annie Arguello XiaoYong Yang Daniel Vogt Amelia Stanco John L. R. Rubenstein Benjamin N. R. Cheyette 《PloS one》2013,8(6)
Synaptogenesis has been extensively studied along with dendritic spine development in glutamatergic pyramidal neurons, however synapse development in cortical interneurons, which are largely aspiny, is comparatively less well understood. Dact1, one of 3 paralogous Dact (Dapper/Frodo) family members in mammals, is a scaffold protein implicated in both the Wnt/β-catenin and the Wnt/Planar Cell Polarity pathways. We show here that Dact1 is expressed in immature cortical interneurons. Although Dact1 is first expressed in interneuron precursors during proliferative and migratory stages, constitutive Dact1 mutant mice have no major defects in numbers or migration of these neurons. However, cultured cortical interneurons derived from these mice have reduced numbers of excitatory synapses on their dendrites. We selectively eliminated Dact1 from mouse cortical interneurons using a conditional knock-out strategy with a Dlx-I12b enhancer-Cre allele, and thereby demonstrate a cell-autonomous role for Dact1 during postsynaptic development. Confirming this cell-autonomous role, we show that synapse numbers in Dact1 deficient cortical interneurons are rescued by virally-mediated re-expression of Dact1 specifically targeted to these cells. Synapse numbers in these neurons are also rescued by similarly targeted expression of the Dact1 binding partner Dishevelled-1, and partially rescued by expression of Disrupted in Schizophrenia-1, a synaptic protein genetically implicated in susceptibility to several major mental illnesses. In sum, our results support a novel cell-autonomous postsynaptic role for Dact1, in cooperation with Dishevelled-1 and possibly Disrupted in Schizophrenia-1, in the formation of synapses on cortical interneuron dendrites. 相似文献
69.
D S Faffe G H Silva P M Kurtz E M Negri V L Capelozzi P R Rocco W A Zin 《Journal of applied physiology》2001,90(4):1400-1406
The dynamic mechanical properties of lung tissue and its contents of collagen and elastic fibers were studied in strips prepared from mice instilled intratracheally with saline (C) or silica [15 (S15) and 30 days (S30) after instillation]. Resistance, elastance, and hysteresivity were studied during oscillations at different frequencies on S15 and S30. Elastance increased from C to silica groups but was similar between S15 and S30. Resistance was augmented from C to S15 and S30 and was greater in S30 than in S15 at higher frequencies. Hysteresivity was higher in S30 than in C and S15. Silica groups presented a greater amount of collagen than did C. Elastic fiber content increased progressively along time. This increment was related to the higher amount of oxytalan fibers at 15 and 30 days, whereas elaunin and fully developed elastic fibers were augmented only at 30 days. Silicosis led not only to pulmonary fibrosis but also to fibroelastosis, thus assigning a major role to the elastic system in the silicotic lung. 相似文献
70.
S. I. Tkachenko A. R. Mingaleev V. M. Romanova A. E. Ter-Oganes’yan T. A. Shelkovenko S. A. Pikuz 《Plasma Physics Reports》2009,35(9):734-753
Distribution of matter in the discharge channel formed upon a nanosecond electrical explosion of a single wire in air and
vacuum was studied experimentally. Simultaneous use of optical, UV, and X-ray diagnostics made it possible to distinguish
qualitatively different regions of the discharge channel, such as the current-carrying layers and the region occupied by a
weakly conducting cold plasma. Several series of experiments with 25-μm-diameter 12-mm-long wires made of different materials
were performed. The charging voltage and the current amplitude were varied in the ranges of U
0 = 10–20 kV and I
max ∼ 5–10 kA, respectively. Explosion regimes with a current pause and with and without current interruption, as well as with
wire preheating in air and vacuum, were studied. Shadow and schlieren images of the discharge channel were obtained using
optical probing at the second harmonic of a YAG: Nd+3 laser (λ = 0.532 μm, τ ∼ 10 ns). In the experiments carried out in vacuum, X-ray images of the discharge channel were also
obtained using an X-pinch as a point source of probing radiation and UV images were recorded using a four-frame MCP camera. 相似文献