全文获取类型
收费全文 | 8806篇 |
免费 | 831篇 |
国内免费 | 103篇 |
专业分类
9740篇 |
出版年
2023年 | 37篇 |
2022年 | 137篇 |
2021年 | 185篇 |
2020年 | 108篇 |
2019年 | 136篇 |
2018年 | 150篇 |
2017年 | 141篇 |
2016年 | 252篇 |
2015年 | 449篇 |
2014年 | 432篇 |
2013年 | 525篇 |
2012年 | 685篇 |
2011年 | 631篇 |
2010年 | 385篇 |
2009年 | 346篇 |
2008年 | 464篇 |
2007年 | 420篇 |
2006年 | 431篇 |
2005年 | 373篇 |
2004年 | 329篇 |
2003年 | 307篇 |
2002年 | 257篇 |
2001年 | 237篇 |
2000年 | 229篇 |
1999年 | 186篇 |
1998年 | 84篇 |
1997年 | 59篇 |
1996年 | 69篇 |
1995年 | 57篇 |
1994年 | 47篇 |
1993年 | 61篇 |
1992年 | 126篇 |
1991年 | 95篇 |
1990年 | 100篇 |
1989年 | 108篇 |
1988年 | 78篇 |
1987年 | 83篇 |
1986年 | 89篇 |
1985年 | 90篇 |
1984年 | 61篇 |
1983年 | 63篇 |
1982年 | 51篇 |
1981年 | 49篇 |
1980年 | 46篇 |
1979年 | 76篇 |
1978年 | 61篇 |
1977年 | 45篇 |
1976年 | 47篇 |
1974年 | 45篇 |
1973年 | 41篇 |
排序方式: 共有9740条查询结果,搜索用时 15 毫秒
131.
Sandra S. Chan Hugo P. Monteiro Guilherme P. Deucher Ronaldo L. Abud Daud Abuchalla Virginia B.C. Junqueira 《Free radical biology & medicine》1998,24(9):1411-1418
In the present work, we studied the role of polymorphonuclear leukocytes (PMN) in aged individuals and coronary heart disease (CHD)-bearing patients, two physiopathological processes associated with overproduction of reactive oxygen species (ROS). The effects of antioxidant supplementation on the functional activity of PMN from CHD patients were also determined. The function of PMNs was evaluated by measuring of phagocytosis, killing activity, and ROS production. Luminol amplified chemiluminescence (CL) was used to estimate ROS production by stimulated PMNs. Total cholesterol and the LDL-cholesterol fraction from CHD patients were found to be higher than those recommended, returning to normal levels after antioxidant therapy. PMN CL of CHD patients was found to be higher than the associated control groups. Antioxidant therapy administrated to CHD patients lead to an increase in the killing activity accompanied by a decrease in PMN CL of these subjects. The study also showed that killing activity of PMN from human subjects over 60 years was significantly lower than the activity measured in younger subjects. PMN CL produced after stimulation was found to be positively correlated with the increasing age of human subjects (r = .946, p < .01). 相似文献
132.
Lizbeth Castro-Concha Victor M. Loyola-Vargas José L. Chan Manuel L. Robert 《Plant Cell, Tissue and Organ Culture》1990,22(2):147-151
Agave tequilana stem explants were used to produce adventitious shoots under a set of different water potentials induced by different concentrations of gelrite in the medium. At high water potentials all shoots were vitrified; as the medium water potential became more negative the degree of vitrification decreased but the number of shoots per explant also diminished. The enzymes NADH and NAD-GDH (EC. 1.4.1.2) were measured along the water potential gradient. GDH activity was high in the non-vitrified tissues and decreased significantly in the vitrified ones.Abbreviations GDH
glutamate dehydrogenase
- MS
Murashige and Skoog medium
- MSO
methionine sulfoximine
- PVP
polyvinylpolypyrrolidone
- GS
glutamine synthetase
- GOGAT
glutamine: oxoglutarate amino transferase 相似文献
133.
Infection by echoviruses 1 and 8 depends on the alpha 2 subunit of human VLA-2. 总被引:4,自引:10,他引:4 下载免费PDF全文
J M Bergelson N St John S Kawaguchi M Chan H Stubdal J Modlin R W Finberg 《Journal of virology》1993,67(11):6847-6852
Anti-VLA-2 antibodies protected HeLa cells from infection by echoviruses 1 and 8 but not from infection by other echovirus serotypes. Echoviruses 1 and 8 bound to and infected nonpermissive hamster cells transfected with the alpha 2 subunit of human VLA-2. These results indicate that the human alpha 2 subunit is critical for infection by echoviruses 1 and 8 but that other echovirus serotypes must bind receptors other than VLA-2. 相似文献
134.
Wen-Tai Li Anirban Mahapatra David G. Longstaff Gang Zhao Michael K. Chan Joseph A. Krzycki 《Journal of molecular biology》2009,385(4):1156-1164
Pyrrolysine, the 22nd amino acid, is encoded by amber (TAG = UAG) codons in certain methanogenic archaea and bacteria. PylS, the pyrrolysyl-tRNA synthetase, ligates pyrrolysine to tRNAPyl for amber decoding as pyrrolysine. PylS and tRNAPyl have potential utility in making tailored recombinant proteins. Here, we probed interactions necessary for recognition of substrates by archaeal PylS via synthesis of close pyrrolysine analogs and testing their reactivity in amino acid activation assays. Replacement of the methylpyrroline ring of pyrrolysine with cyclopentane indicated that solely hydrophobic interactions with the ring-binding pocket of PylS are sufficient for substrate recognition. However, a 100-fold increase in the specificity constant of PylS was observed with an analog, 2-amino-6-((R)-tetrahydrofuran-2-carboxamido)hexanoic acid (2Thf-lys), in which tetrahydrofuran replaced the pyrrolysine methylpyrroline ring. Other analogs in which the electronegative atom was moved to different positions suggested PylS preference for a hydrogen-bond-accepting group at the imine nitrogen position in pyrrolysine. 2Thf-lys was a preferred substrate over a commonly employed pyrrolysine analog, but the specificity constant for 2Thf-lys was 10-fold lower than for pyrrolysine itself, largely due to the change in Km. The in vivo activity of the analogs in supporting UAG suppression in Escherichia coli bearing genes for PylS and tRNAPyl was similar to in vitro results, with l-pyrrolysine and 2Thf-lys supporting the highest amounts of UAG translation. Increasing concentrations of either PylS substrate resulted in a linear increase in UAG suppression, providing a facile method to assay bioactive pyrrolysine analogs. These results illustrate the relative importance of the H-bonding and hydrophobic interactions in the recognition of the methylpyrroline ring of pyrrolysine and provide a promising new series of easily synthesized pyrrolysine analogs that can serve as scaffolds for the introduction of novel functional groups into recombinant proteins. 相似文献
135.
The frequency of fulminant pneumonia due to Mycoplasma pneumoniae is relatively rare despite the high prevalence of Mycoplasma species infection in the general population. We recently encountered such a case and have reviewed the English-language literature on cases of M pneumoniae pneumonia that have resulted in respiratory failure or death. Due to host factors or on epidemiologic grounds, fulminant cases seem to be more common in young healthy adults, in males, and possibly in smokers among the 46 patients we found. An enhanced host cellular immune response may be responsible for the development of severe cases. A spectrum of small airways disease is characteristic, including cellular bronchiolitis and bronchiolitis obliterans with and without organizing pneumonia. Based largely on anecdotal experience, corticosteroid use may be salutary in patients with respiratory failure. For reasons that are not well known, the incidence of pulmonary thromboembolism is increased in fatal cases. 相似文献
136.
Robert Chan Madaline Gilbert Kristin M. Thompson H. Nicholas Marsh David M. Epstein P. Shannon Pendergrast 《Nucleic acids research》2006,34(5):e36
The specific down-regulation of gene expression in cells is a powerful method for elucidating a gene's function. A common method for suppressing gene expression is the elimination of mRNA by RNAi or antisense. Alternatively, oligonucleotide-derived aptamers have been used as protein-directed agents for the specific knock-down of both intracellular and extracellular protein activity. Protein-directed methods offer the advantage of more closely mimicking small molecule therapeutics' mechanism of activity. Furthermore, protein-directed methods may synergize with RNA-directed methods since the two methods attack gene expression at different levels. Here we have knocked down a well-characterized intracellular protein's activity, NFκB, by expressing either aptamers or small interfering RNAs (siRNAs). Both methods can diminish NFκB's activity to similar levels (from 29 to 64%). Interestingly, expression of both aptamers and siRNAs simultaneously, suppressed NFκB activity better than either method alone (up to 90%). These results demonstrate that the expression of intracellular aptamers is a viable alternative to siRNA knock-down. Furthermore, for the first time, we show that the use of aptamers and siRNA together can be the most effective way to achieve maximal knock-down of protein activity. 相似文献
137.
The core prerequisites for an efficient proteome-scale analysis of mammalian membrane proteins are effective isolation, solubilization, digestion and multidimensional liquid chromatography-tandem mass spectrometry (LC-MS/MS). This protocol is for analysis of the mammalian membrane proteome that relies on solubilization and tryptic digestion of membrane proteins in a buffer containing 60% (vol/vol) methanol. Tryptic digestion is followed by strong cation exchange (SCX) chromatography and reversed phase (RP) chromatography coupled online with MS/MS for protein identification. The use of a methanol-based buffer eliminates the need for reagents that interfere with chromatographic resolution and ionization of the peptides (e.g., detergents, chaotropes, inorganic salts). Sample losses are minimized because solubilization and digestion are carried out in a single tube avoiding any sample transfer or buffer exchange between these steps. This protocol is compatible with stable isotope labeling at the protein and peptide level, enabling identification and quantitation of integral membrane proteins. The entire procedure--beginning with isolated membrane fraction and finishing with MS data acquisition--takes 4-5 d. 相似文献
138.
Linda S. M. Ooi Wing-Shan Ho Karry L. K. Ngai Li Tian Paul K. S. Chan Samuel S. M. Sun Vincent E. C. Ooi 《Journal of biosciences》2010,35(1):95-103
A mannose-binding lectin (Narcissus tazetta lectin [NTL]) with potent antiviral activity was isolated and purified from the bulbs of the Chinese daffodil Narcissus tazetta var. chinensis, using ion exchange chromatography on diethylaminoethyl (DEAE)-cellulose, affinity chromatography on mannose-agarose and
fast protein liquid chromatography (FPLC)-gel filtration on Superose 12. The purified lectin was shown to have an apparent
molecular mass of 26 kDa by gel filtration and 13 kDa by SDS-PAGE, indicating that it is probably a dimer with two identical
subunits. The cDNA-derived amino acid sequence of NTL as determined by molecular cloning also reveals that NTL protein contains
a mature polypeptide consisting of 105 amino acids and a C-terminal peptide extension. Three-dimensional modelling study demonstrated
that the NTL primary polypeptide contains three subdomains, each with a conserved mannose-binding site. It shows a high homology
of about 60%–80% similarity with the existing monocot mannose-binding lectins. NTL could significantly inhibit plaque formation
by the human respiratory syncytial virus (RSV) with an IC50 of 2.30 μg/ml and exhibit strong antiviral properties against influenza A (H1N1, H3N2, H5N1) and influenza B viruses with
IC50 values ranging from 0.20 μg/ml to 1.33 μg/ml in a dose-dependent manner. It is worth noting that the modes of antiviral action
of NTL against RSV and influenza A virus are significantly different. NTL is effective in the inhibition of RSV during the
whole viral infection cycle, but the antiviral activity of NTL is mainly expressed at the early stage of the viral cycle of
influenza A (H1N1) virus. NTL with a high selective index (SI=CC50/IC50≥141) resulting from its potent antiviral activity and low cytotoxicity demonstrates a potential for biotechnological development
as an antiviral agent. 相似文献
139.
酵母甘露聚糖(Mannan,简称Man)是能参与生物信息流影响生物体、特别是在糖基化方面起着重要调控作用的生物信息大分子.它是否能在抑制生物机体中肿瘤生长方面具有重要作用?研究结果表明:酵母甘露聚糖既能使患S-180瘤鼠的体质增强的同时又有抑制其体内所患S-180瘤生长的功效.Man抑制患鼠机体内的S-180瘤生长的功效(抑瘤率)随用量的增加而提高,当Man用量达360 mg(40 mg/Kg/d·9d)时,其抑瘤率达98.4%的最高水平,此时鼠体重增加1.66倍.Man的抑瘤功效有最佳适用量并存在性别敏感性,通常是雄性鼠的抑瘤率高于雌性鼠.Man抑制鼠S-180肿瘤生长的作用优于市售5-氟尿嘧啶的作用. 相似文献
140.
Liver microsomal membrane fluidity and lipid characteristics in vitamin A-deficient rats. 总被引:2,自引:0,他引:2 下载免费PDF全文
Rat liver microsomes (microsomal fraction) were isolated from vitamin A-deficient and -sufficient rats and analysed for membrane lipid characteristics. Membrane fluidity was found to be significantly decreased in microsomes from the vitamin A-deficient rats, but not in liposomes prepared from lipid extracts. Microsomes from vitamin A-deficient animals showed a significant decrease in C18:2, omega 6 and an increase in C22:5, omega 6 fatty acids. 相似文献