全文获取类型
收费全文 | 769篇 |
免费 | 75篇 |
国内免费 | 2篇 |
出版年
2024年 | 1篇 |
2023年 | 3篇 |
2022年 | 2篇 |
2021年 | 17篇 |
2020年 | 8篇 |
2019年 | 11篇 |
2018年 | 20篇 |
2017年 | 16篇 |
2016年 | 33篇 |
2015年 | 61篇 |
2014年 | 55篇 |
2013年 | 39篇 |
2012年 | 81篇 |
2011年 | 59篇 |
2010年 | 43篇 |
2009年 | 36篇 |
2008年 | 59篇 |
2007年 | 38篇 |
2006年 | 49篇 |
2005年 | 31篇 |
2004年 | 38篇 |
2003年 | 27篇 |
2002年 | 35篇 |
2001年 | 8篇 |
2000年 | 11篇 |
1999年 | 9篇 |
1998年 | 2篇 |
1997年 | 3篇 |
1996年 | 6篇 |
1995年 | 3篇 |
1994年 | 5篇 |
1993年 | 2篇 |
1992年 | 3篇 |
1991年 | 7篇 |
1988年 | 1篇 |
1987年 | 6篇 |
1986年 | 3篇 |
1985年 | 1篇 |
1984年 | 1篇 |
1980年 | 1篇 |
1979年 | 1篇 |
1978年 | 4篇 |
1977年 | 1篇 |
1976年 | 4篇 |
1972年 | 1篇 |
1962年 | 1篇 |
排序方式: 共有846条查询结果,搜索用时 15 毫秒
51.
Jong Seok Lee Jae Wan Wee Hye Young Lee Hyo Sil An Eock Kee Hong 《Biotechnology and Bioprocess Engineering》2010,15(3):453-459
Hericium erinaceus is a well known edible and medicinal mushroom used in East-Asia. Recently, H. erinaceus has attracted a lot of attention owing to its antitumor, immuno-modulatory, and cytotoxic effect. It has been postulated that the fruiting body of H. erinaceus contains a polysaccharide that is similar to β-D-glucan, which is known to have antitumor activity against Sarcoma 180. However, optimized liquid culture conditions for enhanced polysaccharide productivity have yet to be developed, which is a necessary step for industrial applications. Therefore, the aim of this study was to determine the optimal liquid culture conditions for maximum polysaccharide production. In shake flask cultures, the optimal concentration of ascorbic acid was found to be 2.0 g/L, which prevented the broth from changing color from yellow to black. The optimal culture conditions were determined to be 23°C, 200 rpm, and a 10% inoculum size, at an uncontrolled initial pH. In addition, the modified medium contained 20 g/L glucose, 10 g/L yeast extract, and 2.0 g/L ascorbic acid. The maximum mycelial biomass and exo-polysaccharide (EPS) production in the modified medium containing uracil was 13.43 and 1.26 g/L, respectively. 相似文献
52.
Park IS Park JU Seo MJ Kim MJ Lee HH Kim SR Kang BW Choi YH Joo WH Jeong YK 《Journal of microbiology (Seoul, Korea)》2010,48(6):836-841
A fibrinolytic enzyme of the mushroom, Schizophyllum commune was purified with chromatographic methods, including a DEAE-Sephadex A-50 ion-exchange column and gel filtrations with Sephadex
G-75 and Sephadex G-50 columns. The analysis of fibrin-zymography and SDS-PAGE showed that the enzyme was a monomeric subunit
that was estimated to be approximately 17 kDa in size. The fibrinolytic activity of the enzyme in plasminogen-rich and plasminogen-free
fibrin plates was 1.25 and 0.44 U/ml, respectively. The N-terminal amino acid sequence of the purified enzyme was identified
as HYNIXNSWSSFID, which was highly distinguished from known fibrinolytic enzymes. The relative activity of the purified enzyme
with an addition of 5 mM EDTA, Phosphoramidon, and Bestatin was about 76, 64, and 52%, respectively, indicating that it is
a metalloprotease. The optimum temperature for the purified enzyme was approximately 45°C, and over 87% of the enzymatic activity
was maintained as a stable state in a pH range from 4.0 to 6.0. Therefore, our results suggest that the potential thrombolytic
agent from S. commune is a unique type of fibrinolytic enzyme. 相似文献
53.
Stéphanie Blanchard Anthony D. William Angeline C.-H. Lee Anders Poulsen Ee Ling Teo Weiping Deng Noah Tu Evelyn Tan Kay Lin Goh Wai Chung Ong Chee Pang Ng Kee Chuan Goh Zahid Bonday Eric T. Sun 《Bioorganic & medicinal chemistry letters》2010,20(8):2443-2447
A series of alkenyl indazoles were synthesized and evaluated in Aurora kinase enzyme assays. Several promising leads were optimized for selectivity towards Aurora B. Excellent binding affinity and good selectivity were achieved with optimized compounds in isolated Aurora subfamily assays. 相似文献
54.
Fanconi anemia (FA) patients are specifically defective in the repair of interstrand DNA crosslinks (ICLs), a complex process involving at least 13 FA proteins and other repair/checkpoint proteins. Of the 13 FA proteins, FANCD1/BRCA2, FANCD2, and FANCJ were previously found to be functionally conserved in C. elegans. We have also identified C. elegans homologs of FANCM and FANCI, and determined their epistatic relationships with homologs of FANCD2, checkpoint proteins, and RAD51 upon DNA crosslinking. The counterparts of FANCM, FANCI, and three checkpoint proteins (RPA, ATR and CHK1) are required for focus formation and ubiquitination associated with FANCD2 in C. elegans. However, C. elegans FANCM affects neither RPA focus formation nor CHK1 phosphorylation induced by ICLs, unlike the reported role of human FANCM, which influences ATR-CHK1 signaling at stalled replication forks. Although focus formation by both FANCD2 and RAD51 requires ATR-CHK1 signaling, FANCD2 and RAD51 acted independently in the formation of their respective foci. Thus, the FANCD2 activation pathway involving FANCM, FANCI, and the checkpoint proteins is conserved in C. elegans but with distinct differences. 相似文献
55.
The beetle family Eucinetidae, represented by Eucinetus haemorrhoidalis (Germar), is newly recorded from southern parts of the Korean Peninsula, extending its distributional range by several hundred kilometers to the south. A habitus photo and illustrations of the diagnostic characters of species are provided. 相似文献
56.
A gene encoding an alkaline (pI of 8.67) chitinase was cloned and sequenced from Chromobacterium sp. strain C-61. The gene was composed of 1,611 nucleotides and encoded a signal sequence of 26 N-terminal amino acids and
a mature protein of 510 amino acids. Two chitinases of 54 and 52 kDa from both recombinant Escherichia coli and C-61 were detected on SDS-PAGE. Maximum chitinase activity was obtained in the culture supernatant of recombinant E. coli when cultivated in TB medium for 6 days at 37°C and was about fourfold higher than that from C-61. Chi54 from the culture
supernatants could be purified by a single step based on isoelectric point. The purified Chi54 had about twofold higher binding
affinity to chitin than to cellulose. The chi54 encoded a protein that included a type 3 chitin-binding domain belonging to group A and a family 18 catalytic domain belonging
to subfamily A. In the catalytic domain, mutation of perfectly conserved residues and highly conserved residues resulted in
loss of nearly all activity, while mutation of nonconserved residues resulted in enzymes that retained activity. In this process,
a mutant (T218S) was obtained that had about 133% of the activity of the wild type, based on comparison of K
cat values. 相似文献
57.
Yang HY Jeong DK Kim SH Chung KJ Cho EJ Yang U Lee SR Lee TH 《Biochemical and biophysical research communications》2007,359(4):1030-1036
Peroxiredoxin III (Prdx III), the mitochondrial peroxidase, was preferentially expressed in murine erythroleukemia (MEL) cells. However, the mechanisms by which Prdx III regulates erythroid differentiation are unknown. In this study, K562 cells were differentiated by Ara-C treatment, and Prdx III was dramatically increased until day 5. We also investigated Prdx III expression pattern on in vitro erythropoiesis of human CD34(+) cells. When human CD34(+) cells became proerythrocyte on day 7, Prdx III was diminished, and then augmented on day 12. We established the stable sublines of Prdx III overexpression (O/E), and dominant-negative (D/N). The intracellular ROS level of Prdx III O/E cell line was lower than D/N stable cell lines. Moreover, Prdx III O/E cell line was placed in G1-arrest, but not D/N cell lines. Finally, the expression level of beta-globin and GATA-1 was dramatically increased in Prdx III O/E cell line. 相似文献
58.
Inhibition of Fc epsilon RI-mediated mast cell responses by ES-62, a product of parasitic filarial nematodes 总被引:2,自引:0,他引:2
Melendez AJ Harnett MM Pushparaj PN Wong WS Tay HK McSharry CP Harnett W 《Nature medicine》2007,13(11):1375-1381
Atopic allergy is characterized by an increase in IgE antibodies that signal through the high-affinity Fcepsilon receptor (FcepsilonRI) to induce the release of inflammatory mediators from mast cells. For unknown reasons, the prevalence of allergic diseases has recently increased steeply in the developed world. However, this increase has not been mirrored in developing countries, even though IgE concentrations are often greatly elevated in individuals from these countries, owing to nonspecific IgE induction by universally present parasitic worms. Here we offer one explanation for this paradox based on the properties of ES-62, a molecule secreted by filarial nematodes. We found that highly purified, endotoxin-free ES-62 directly inhibits the FcepsilonRI-induced release of allergy mediators from human mast cells by selectively blocking key signal transduction events, including phospholipase D-coupled, sphingosine kinase-mediated calcium mobilization and nuclear factor-kappaB activation. ES-62 mediates these effects by forming a complex with Toll-like receptor 4, which results in the sequestration of protein kinase C-alpha (PKC-alpha). This causes caveolae/lipid raft-mediated, proteasome-independent degradation of PKC-alpha, a molecule important for the coupling of FcepsilonRI to phospholipase D and mast cell activation. We also show that ES-62 is able to protect mice from mast cell-dependent hypersensitivity in the skin and lungs, indicating that it has potential as a novel therapeutic for allergy. 相似文献
59.
An endonuclease from Xanthomonas oryzae pathovar oryzae KACC 10331, XorII, was recombinantly produced in Escherichia coli using a T7 system. XorII was purified using a combination of ion exchange and immobilized metal affinity chromatography (IMAC). An optimized washing protocol was carried out on an IMAC in order to obtain a high purity product. The final amount of purified XorII was approximately 2.5 mg/L of LB medium. The purified recombinant XorII was functional and showed the same cleavage pattern as PvuI. The enzyme activity tested the highest at 25 degrees in 50 mM NaCl, 10 mM Tris-HCl, 10 mM MgCl2, and 1 mM dithiothreitol at a pH of 7.9. 相似文献
60.
Two stable and water-soluble organometallic carbonyl cluster derivatives have been prepared and shown to enter the cell with ease. The CO stretching vibrations afford strong mid-infrared signals which have been demonstrated, for the first time, to be of utility in cell imaging via an IR microscope. 相似文献