全文获取类型
收费全文 | 6803篇 |
免费 | 320篇 |
国内免费 | 283篇 |
专业分类
7406篇 |
出版年
2024年 | 11篇 |
2023年 | 79篇 |
2022年 | 151篇 |
2021年 | 160篇 |
2020年 | 222篇 |
2019年 | 317篇 |
2018年 | 318篇 |
2017年 | 205篇 |
2016年 | 211篇 |
2015年 | 171篇 |
2014年 | 430篇 |
2013年 | 686篇 |
2012年 | 202篇 |
2011年 | 413篇 |
2010年 | 235篇 |
2009年 | 290篇 |
2008年 | 297篇 |
2007年 | 320篇 |
2006年 | 267篇 |
2005年 | 279篇 |
2004年 | 250篇 |
2003年 | 223篇 |
2002年 | 186篇 |
2001年 | 113篇 |
2000年 | 86篇 |
1999年 | 95篇 |
1998年 | 86篇 |
1997年 | 109篇 |
1996年 | 110篇 |
1995年 | 84篇 |
1994年 | 69篇 |
1993年 | 71篇 |
1992年 | 67篇 |
1991年 | 54篇 |
1990年 | 29篇 |
1989年 | 43篇 |
1988年 | 27篇 |
1987年 | 35篇 |
1986年 | 31篇 |
1985年 | 34篇 |
1984年 | 77篇 |
1983年 | 44篇 |
1982年 | 56篇 |
1981年 | 31篇 |
1980年 | 32篇 |
1979年 | 31篇 |
1978年 | 11篇 |
1977年 | 19篇 |
1976年 | 12篇 |
1974年 | 15篇 |
排序方式: 共有7406条查询结果,搜索用时 15 毫秒
41.
《Bioscience, biotechnology, and biochemistry》2013,77(4):726-728
Streptomyces sp. strain No. 6288 produces S-Hemolysin, which is a unique phospholipase C with a high substrate specificity for sphingomyelin. Moreover, the strain also produced two kinds of phospholipase inhibitors, designated as SHI and S-PLI, in different phases of cultivation. The purified SHI was shown to be a basic substance containing an amino group and glycoside moiety, and it was a more effective inhibitor of S-Hemolysin and sphingomyelinase from Streptomyces sp. with a higher substrate specificity for sphingomyelin than α-toxin from Clostridium perfringens. 相似文献
42.
Thomas Vedvick Richard G. Buckholz Michael Engel Mary Urcan Jennifer Kinney Sally Provow Robert S. Siegel Gregory P. Thill 《Journal of industrial microbiology & biotechnology》1991,7(3):197-201
Summary A synthetic gene encoding aprotinin (bovine pancreatic trypsin, inhibitor) was fused to theSaccharomyces cerevisiae prepro alpha mating factor leader sequence at the dibasic amino acid processing site.Pichia pastoris strains were developed to'express one or multiple copies of a methanol-inducible expression cassette containing the gene fusion.P. pastoris containing a single copy of the vector secreed approximately 150 mg/l of immunoreactive protein. A construct bearing five copies of the expression cassette secreted 930 mg/l of aprotinin. The purified aprotinin molecule was equipoten with the native molecule in a trypsin inhibition assay. Protein sequence analysis showed that the alpha factor-aprotinin fusion was not processed at the basic amino acid residues Lys-Arg. Instead, recombinant aprotinin had additional N-terminal amino acids derived from prepro alpha factor. The N-terminal extension was variably 11 or 4 amino acids. Inclusion of the spacer DNA sequence encoding Glu and Ala between aprotinin and the Lys-Arg processing site led to the secretion of a biologically active aprotinin containing only a Glu-Ala N-terminal extension. 相似文献
43.
Functional genomic studies and drug candidate testing both require high throughput, parallel experimentation strategies to screen for variable cellular behaviors. In this article we describe the use of an impedance sensing electrode array that is capable of sensing cell "presence" as well as the extent of cell (focal) attachment to the substrate. The signals provided by mouse fibroblasts on a sensing structure containing four different sized electrodes are reported. In the absence of cells, each electrode's impedance was found to depend as expected on electrode size and frequency. The impedance increased by several-fold when fibroblasts attached and spread out over time. More notably, the sensors also detected the cellular response to the protein kinase C inhibitor, H-7. H-7 inhibits actomyosin contractility; thereafter, the loss of focal adhesion complexes occurs. The sensors, in turn, detected an impedance decrease after H-7 addition and an increase in impedance after H-7 removal. 相似文献
44.
Umesh C. Haldar Sanat K. Saha Ronald C. Beavis Nirmal K. Sinha 《Journal of Protein Chemistry》1996,15(2):177-184
Two trypsin inhibitors, LA-1 and LA-2, have been isolated from ridged gourd (Luffa acutangula Linn.) seeds and purified to homogeneity by gel filtration followed by ion-exchange chromatography. The isoelectric point is atpH 4.55 for LA-1 and atpH 5.85 for LA-2. The Stokes radius of each inhibitor is 11.4 å. The fluorescence emission spectrum of each inhibitor is similar to that of the free tyrosine. The biomolecular rate constant of acrylamide quenching is 1.0×109 M–1 sec–1 for LA-1 and 0.8 × 109 M–1 sec–1 for LA-2 and that of K2HPO4 quenching is 1.6×1011 M–1 sec–1 for LA-1 and 1.2×1011M–1 sec–1 for LA-2. Analysis of the circular dichroic spectra yields 40%-helix and 60%-turn for La-1 and 45%-helix and 55%-turn for LA-2. Inhibitors LA-1 and LA-2 consist of 28 and 29 amino acid residues, respectively. They lack threonine, alanine, valine, and tryptophan. Both inhibitors strongly inhibit trypsin by forming enzymeinhibitor complexes at a molar ratio of unity. A chemical modification study suggests the involvement of arginine of LA-1 and lysine of LA-2 in their reactive sites. The inhibitors are very similar in their amino acid sequences, and show sequence homology with other squash family inhibitors. 相似文献
45.
天然产物中透明质酸酶抑制剂的研究 总被引:4,自引:0,他引:4
透明质酸酶抑制剂是对透明质酸酶的激活有抑制作用的物质。透明质酸酶是透明质酸的特异性裂解酶,而透明质酸在人体许多发育和调控过程中起重要作用,抑制透明质酸酶的活性可使透明质酸不被分解,维持正常的生理功能,笔者对天然产物中透明质酸酶抑制的研究开发进行了概述,并探讨了透明质酸酶抑制在医药及保健食品工业中的应用潜力。 相似文献
46.
47.
《Chronobiology international》2013,30(5):352-360
We report a comparison of fibrinolytic variables between 10 Caucasians on a predominantly European diet and 10 Greenland Eskimos on a traditional Inuit diet containing a substantial amount of fish and sea animals. We studied the diurnal variation in tissue type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) antigens and activities during a 24-h period. Blood samples were taken every 4 h. The variations of the sinusoidal curves were evaluated by the Friedman χ2 test. t-PA and PAI-1 antigen in plasma fluctuated significantly during the 24 h (Eskimos p < 0.000007 and p < 0.0007; Caucasians p < 0.00003 and p < 0.02), with a peak in the early morning and a nadir in the afternoon. This also held true for PA1 activity (Eskimos p < 0.0008; Caucasians p < O.Ol), whereas t-PA activity showed an inverse but still significant pattern (Eskimos p < 0.006; Caucasians p < 0.0008). Amplitudes, areas underneath, and overall medians of the sinusoidal curves did not deviate between the two groups with respect to t-PA and PAL In contrast to the significant variation of t-PA and PAI, the plasma concentrations of fibrin degradation products (D-Dimer), a measure of effective fibrinolysis, remained constant during the 24 h, and the absolute differences between groups did not reach statistical significance. These findings suggest that circadian variation of fibrinolytic activators and inhibitors is a basic biologic phenomenon, which is not affected by life-style, dietary habits, or ethnic differences. Furthermore, the lack of diurnal variation in D-Dimer raises the question of whether there is a causal relationship between low morning activities of t-PA and the frequent onset of myocardial infarction at that time of day, as suggested by several authors. 相似文献
48.
Structural features and molecular evolution of Bowman-Birk protease inhibitors and their potential application 总被引:3,自引:0,他引:3
The Bowman-Birk inhibitors (BBIs) are well-studied serine protease inhibitors that are abundant in dicotyledonous and monocotyledonous plants. BBIs from dicots usually have a molecular weight of 8k and are double-headed with two reactive sites, whereas those from monocots can be divided into two classes, one approximately 8 kDa in size with one reactive site (another reactive site was lost) and the other approximately 16 kDa in size with two reactive sites. The reactive site is located at unique exposed surfaces formed by a disulfide-linked β-sheet loop that is highly conserved, rigid and mostly composed of nine residues. The structural features and molecular evolution of inhibitors are described, focusing on the conserved disulfide bridges. The sunflower trypsin inhibitor-1 (SFTI-1), with 14 amino acid residues, is a recently discovered bicyclic inhibitor, and is the most small and potent naturally occurring Bowman-Birk inhibitor.Recently, BBIs have become a hot topic because of their potential applications. BBIs are now used for defense against pathogens and insects in transgenic plants, which has advantages over using toxic and polluting insecticides. BBIs could also be applied in the prevention of cancer, Dengue fever, and inflammatory and allergic disorders, because of their inhibitory activity with respect to the serine proteases that play apivotal role in the development and pathogenesis of these diseases. The canonical nine-residue loop of BBIs/STH-1 provides an ideal template for drug design of specific inhibitors to target their respective proteases. 相似文献
49.
Beth A. Bouchard Matthew T. GisselMatthew F. Whelihan Kenneth G. MannSaulius Butenas 《Biochimica et Biophysica Acta (BBA)/General Subjects》2014
Background
Expression of tissue factor (TF) antigen and activity in platelets is controversial and dependent upon the laboratory and reagents used. Two forms of TF were described: an oxidized functional form and a reduced nonfunctional form that is converted to the active form through the formation of an allosteric disulfide. This study tests the hypothesis that the discrepancies regarding platelet TF expression are due to differential expression of the two forms.Methods
Specific reagents that recognize both oxidized and reduced TF were used in flow cytometry of unactivated and activated platelets and western blotting of whole platelet lysates. TF-dependent activity measurements were used to confirm the results.Results
Western blotting analyses of placental TF demonstrated that, in contrast to anti-TF#5, which is directed against the oxidized form of TF, a sheep anti-human TF polyclonal antibody recognizes both the reduced and oxidized forms. Flow cytometric analyses demonstrated that the sheep antibody did not react with the surface of unactivated platelets or platelets activated with thrombin receptor agonist peptide, PAR-1. This observation was confirmed using biotinylated active site-blocked factor (F)VIIa: no binding was observed. Likewise, neither form of TF was detected by western blotting of whole platelet lysates with sheep anti-hTF. Consistent with these observations, no FXa or FIXa generation by FVIIa was detected at the surface of these platelets. Similarly, no TF-related activity was observed in whole blood using thromboelastography.Conclusion and significance
Platelets from healthy donors do not express either oxidized (functional) or reduced (nonfunctional) forms of TF. 相似文献50.
Stephen J Peroutka 《Current opinion in biotechnology》1997,8(6):688-691
The long anticipated ‘genetic revolution’ in neuropsychiatry has yet to have an impact on the practice of clinical medicine. Excitement in the 1980s over major genetic breakthroughs in schizophrenia and manic depression, for example, has been replaced in the late 1990s by the sobering realization that most common neuropsychiatric disorders are multifactorial. Despite considerable effort and resources, no ‘causative’ genetic variation has been identified that plays a definitive major role in any common neuropsychiatric disorder. 相似文献