全文获取类型
收费全文 | 1151篇 |
免费 | 49篇 |
国内免费 | 1篇 |
出版年
2023年 | 6篇 |
2022年 | 20篇 |
2021年 | 38篇 |
2020年 | 25篇 |
2019年 | 21篇 |
2018年 | 48篇 |
2017年 | 21篇 |
2016年 | 44篇 |
2015年 | 34篇 |
2014年 | 67篇 |
2013年 | 114篇 |
2012年 | 108篇 |
2011年 | 84篇 |
2010年 | 71篇 |
2009年 | 47篇 |
2008年 | 48篇 |
2007年 | 44篇 |
2006年 | 49篇 |
2005年 | 44篇 |
2004年 | 45篇 |
2003年 | 33篇 |
2002年 | 36篇 |
2001年 | 12篇 |
2000年 | 15篇 |
1999年 | 15篇 |
1998年 | 12篇 |
1997年 | 8篇 |
1996年 | 7篇 |
1995年 | 9篇 |
1994年 | 8篇 |
1993年 | 3篇 |
1992年 | 5篇 |
1991年 | 2篇 |
1989年 | 4篇 |
1988年 | 3篇 |
1987年 | 5篇 |
1986年 | 3篇 |
1985年 | 8篇 |
1984年 | 3篇 |
1983年 | 3篇 |
1981年 | 3篇 |
1980年 | 3篇 |
1979年 | 4篇 |
1978年 | 3篇 |
1977年 | 3篇 |
1972年 | 2篇 |
1964年 | 1篇 |
1963年 | 1篇 |
1961年 | 1篇 |
1960年 | 1篇 |
排序方式: 共有1201条查询结果,搜索用时 15 毫秒
31.
Vivek Kumar Mukesh M. Mudgal Nidhi Rani Amrita Jha Manu Jaggi Anu T. Singh 《Journal of enzyme inhibition and medicinal chemistry》2013,28(3):763-770
A new series of functionalized amino acid derivatives N-substituted 1-N-(tert-butoxycarbonyl)-2,2-dimethyl-4-phenyl-5-oxazolidine carboxamide (1-17) and 1-N-substituted-3-amino-2-hydroxy-3-phenylpropane-1-carboxamide (18-34) were synthesized and evaluated for their in vitro cytotoxicity against human cancer cell lines. Compound 6 has shown interesting cytotoxicity (IC50 = 5.67 μm) in ovarian cancer, while compound 10 exhibited promising cytotoxicity in ovarian (IC50 = 6.1 μm) and oral (IC50 = 4.17 μm) cancers. These compounds could be of use in designing new anti-cancer agents. 相似文献
32.
Rinky Rajput Ved Vrat Verma Vishal Chaudhary Rani Gupta 《Extremophiles : life under extreme conditions》2013,17(1):29-41
??-Glutamyl transpeptidase of a thermo-acidophilic archaeon Picrophilus torridus was cloned and expressed using E. coli Rosetta-pET 51b(+) expression system. The enzyme was expressed at 37 °C/200 rpm with ??-GT production of 1.99 U/mg protein after 3 h of IPTG induction. It was improved nearby 10-fold corresponding to 18.92 U/mg protein in the presence of 2 % hexadecane. The enzyme was purified by Ni2+-NTA with a purification fold of 3.6 and recovery of 61 %. It was synthesized as a precursor heterodimeric protein of 47 kDa with two subunits of 30 kDa and 17 kDa, respectively, as revealed by SDS-PAGE and western blot. The enzyme possesses hydrolase activity with optima at pH 7.0 and 55 °C. It was thermostable with a t 1/2 of 1 h at 50 °C and 30 min at 60 °C, and retained 100 % activity at 45 °C even after 24 h. It was inhibited by azaserine and DON and PMSF. Pt??-GT shared 37 % sequence identity and 53 % homology with an extremophile ??-GT from Thermoplasma acidophilum. Functional residues identified by in silico approaches were further validated by site-directed mutagenesis where Tyr327 mutated by Asn327 introduced significant transpeptidase activity. 相似文献
33.
Shrey Kohli Aastha Chhabra Astha Jaiswal Yashika Rustagi Manish Sharma Vibha Rani 《PloS one》2013,8(10)
Background
Extracellular matrix (ECM) remodeling facilitates biomechanical signals in response to abnormal physiological conditions. This process is witnessed as one of the major effects of the stress imposed by catecholamines, such as epinephrine and norepinephrine (NE), on cardiac muscle cells. Matrix metalloproteinases (MMPs) are the key proteases involved in degradation of the ECM in heart.Objectives
The present study focuses on studying the effect of curcumin on Gelatinase B (MMP-9), an ECM remodeling regulatory enzyme, in NE-induced cardiac stress. Curcumin, a bioactive polyphenol found in the spice turmeric, has been studied for its multi-fold beneficial properties. This study focuses on investigating the role of curcumin as a cardio-protectant.Methods
H9c2 cardiomyocytes were subjected to NE and curcumin treatments to study the response in stress conditions. Effect on total collagen content was studied using Picrosirus red staining. Gelatinase B activity was assessed through Gel-Diffusion Assay and Zymographic techniques. RT-PCR, Western Blotting and Immunocytochemistry were performed to study effect on expression of gelatinase B. Further, the effect of curcumin on the localization of NF-κB, known to regulate gelatinase B, was also examined.Results
Curcumin suppressed the increase in the total collagen content under hypertrophic stress and was found to inhibit the in-gel and in-situ gelatinolytic activity of gelatinase B. Moreover, it was found to suppress the mRNA and protein expression of gelatinase B.Conclusions
The study provides an evidence for an overall inhibitory effect of curcumin on Gelatinase B in NE-induced hypertrophic stress in H9c2 cardiomyocytes which may contribute in the prevention of ECM remodeling. 相似文献34.
35.
V. Haridas Kullampalayam Shanmugam Rajgokul Sandhya Sadanandan Tanvi Agrawal Vats Sharvani M. V. S. Gopalakrishna M. B. Bijesh Kanhaiya Lal Kumawat Anirban Basu Guruprasad R. Medigeshi 《PLoS neglected tropical diseases》2013,7(1)
Background
Japanese encephalitis virus (JEV) is a major cause of viral encephalitis in South and South-East Asia. Lack of antivirals and non-availability of affordable vaccines in these endemic areas are a major setback in combating JEV and other closely related viruses such as West Nile virus and dengue virus. Protein secondary structure mimetics are excellent candidates for inhibiting the protein-protein interactions and therefore serve as an attractive tool in drug development. We synthesized derivatives containing the backbone of naturally occurring lupin alkaloid, sparteine, which act as protein secondary structure mimetics and show that these compounds exhibit antiviral properties.Methodology/Principal Findings
In this study we have identified 3,7-diazabicyclo[3.3.1]nonane, commonly called bispidine, as a privileged scaffold to synthesize effective antiviral agents. We have synthesized derivatives of bispidine conjugated with amino acids and found that hydrophobic amino acid residues showed antiviral properties against JEV. We identified a tryptophan derivative, Bisp-W, which at 5 µM concentration inhibited JEV infection in neuroblastoma cells by more than 100-fold. Viral inhibition was at a stage post-entry and prior to viral protein translation possibly at viral RNA replication. We show that similar concentration of Bisp-W was capable of inhibiting viral infection of two other encephalitic viruses namely, West Nile virus and Chandipura virus.Conclusions/Significance
We have demonstrated that the amino-acid conjugates of 3,7-diazabicyclo[3.3.1]nonane can serve as a molecular scaffold for development of potent antivirals against encephalitic viruses. Our findings will provide a novel platform to develop effective inhibitors of JEV and perhaps other RNA viruses causing encephalitis. 相似文献36.
Reed canary grass (RCG, Phalaris arundinacea L.) is a suitable energy crop for cultivation in northern peatlands. However, the atmospheric impact of RCG cultivation as influenced by harvest frequency and fertilization is not clear. Here, we compared the biomass yield and greenhouse gas (GHG) balance for RCG cultivation in peatlands affected by cutting frequency and fertilizer managements. The managements included one-cut (OC) and two-cut (TC) systems that were either fertilized (TC-F) or unfertilized (TC-U) after the first cut in summer. Biomass yield of OC, TC-F and TC-U were 12, 16 and 11 Mg dry biomass per hectare per year, respectively. GHG fluxes of CO2, N2O and CH4 were measured with closed chamber techniques in the period between first and second (final) harvest of the TC managements, i.e. from 15 June to 23 September 2011. In the GHG monitoring period of 100 days, all systems were net sources of CO2 corresponding to 64?±?3, 217?±?15 and 50?±?23 g?CO2-C?m?2 (mean?±?standard error, n?=?3) from the OC, TC-F and TC-U systems, respectively. In the same period, emissions of N2O from TC-F were ten times higher as compared to OC and TC-U. Emissions of CH4 were negligible from all systems. The TC systems could not improve the GHG balance during cultivation (271, 663 and 210 g CO2e-C?m?2 emissions from the OC, TC-F and TC-U systems, respectively), but in a broader GHG life cycle perspective, the increased biomass yield by TC-F could replace more fossil fuel and offset at least some of the higher emissions from the system. 相似文献
37.
Jagriti Narang Nidhi Chauhan Poonam Rani C. S. Pundir 《Bioprocess and biosystems engineering》2013,36(4):425-432
A method is described for construction of an amperometric triglyceride (TG) biosensor based on covalent co-immobilization of lipase, glycerol kinase and glycerol-3-phosphate oxidase onto gold polypyrrole nanocomposite decorated poly indole-5-carboxylic acid electrodeposited on the surface of a gold electrode. The enzyme electrode was characterized by transmission electron microscopy, scanning electron microscopy, electrochemical impedance studies, Fourier transform infrared spectroscopy and cyclic voltammetry. Biosensor showed optimum response within 4 s at pH 6.5 and 35 °C, when polarized at +0.1 V against Ag/AgCl. There was a linear relationship between sensor response and triolein concentration in the range 50–700 mg/dl. Biosensor was employed for determination of TG in serum. Detection limit of the biosensor was 20 mg/dl. Biosensor was evaluated with 91–95 % recovery of added triolein in sera and 4.14 and 5.85 % within and between batch coefficients of variation, respectively. There was a good correlation (r = 0.99) between sera TG values by standard method (Enzymic colorimetric) and the present method. The biosensor was unaffected by a number of serum substances at their physiological concentration. Biosensor lost 50 % of its initial activity after its 100 uses over 7 months, when stored at 4 °C. 相似文献
38.
39.
Swati Kaushik Eshita Mutt Ajithavalli Chellappan Sandhya Sankaran Narayanaswamy Srinivasan Ramanathan Sowdhamini 《PloS one》2013,8(2)
Background
Development of sensitive sequence search procedures for the detection of distant relationships between proteins at superfamily/fold level is still a big challenge. The intermediate sequence search approach is the most frequently employed manner of identifying remote homologues effectively. In this study, examination of serine proteases of prolyl oligopeptidase, rhomboid and subtilisin protein families were carried out using plant serine proteases as queries from two genomes including A. thaliana and O. sativa and 13 other families of unrelated folds to identify the distant homologues which could not be obtained using PSI-BLAST.Methodology/Principal Findings
We have proposed to start with multiple queries of classical serine protease members to identify remote homologues in families, using a rigorous approach like Cascade PSI-BLAST. We found that classical sequence based approaches, like PSI-BLAST, showed very low sequence coverage in identifying plant serine proteases. The algorithm was applied on enriched sequence database of homologous domains and we obtained overall average coverage of 88% at family, 77% at superfamily or fold level along with specificity of ∼100% and Mathew’s correlation coefficient of 0.91. Similar approach was also implemented on 13 other protein families representing every structural class in SCOP database. Further investigation with statistical tests, like jackknifing, helped us to better understand the influence of neighbouring protein families.Conclusions/Significance
Our study suggests that employment of multiple queries of a family for the Cascade PSI-BLAST searches is useful for predicting distant relationships effectively even at superfamily level. We have proposed a generalized strategy to cover all the distant members of a particular family using multiple query sequences. Our findings reveal that prior selection of sequences as query and the presence of neighbouring families can be important for covering the search space effectively in minimal computational time. This study also provides an understanding of the ‘bridging’ role of related families. 相似文献40.
Neal N. Padte Mar Boente-Carrera Chasity D. Andrews Jenny McManus Brooke F. Grasperge Agegnehu Gettie Jordana G. Coelho-dos-Reis Xiangming Li Douglass Wu Joseph T. Bruder Martha Sedegah Noelle Patterson Thomas L. Richie Chi-Huey Wong David D. Ho Sandhya Vasan Moriya Tsuji 《PloS one》2013,8(10)
A key strategy to a successful vaccine against malaria is to identify and develop new adjuvants that can enhance T-cell responses and improve protective immunity. Upon co-administration with a rodent malaria vaccine in mice, 7DW8-5, a recently identified novel analog of α-galactosylceramide (α-GalCer), enhances the level of malaria-specific protective immune responses more strongly than the parent compound. In this study, we sought to determine whether 7DW8-5 could provide a similar potent adjuvant effect on a candidate human malaria vaccine in the more relevant non-human primate (NHP) model, prior to committing to clinical development. The candidate human malaria vaccine, AdPfCA (NMRC-M3V-Ad-PfCA), consists of two non-replicating recombinant adenoviral (Ad) vectors, one expressing the circumsporozoite protein (CSP) and another expressing the apical membrane antigen-1 (AMA1) of Plasmodium falciparum. In several phase 1 clinical trials, AdPfCA was well tolerated and demonstrated immunogenicity for both humoral and cell-mediated responses. In the study described herein, 25 rhesus macaques received prime and boost intramuscular (IM) immunizations of AdPfCA alone or with an ascending dose of 7DW8-5. Our results indicate that 7DW8-5 is safe and well-tolerated and provides a significant enhancement (up to 9-fold) in malaria-specific CD8+ T-cell responses after both priming and boosting phases, supporting further clinical development. 相似文献