全文获取类型
收费全文 | 566篇 |
免费 | 49篇 |
国内免费 | 25篇 |
专业分类
640篇 |
出版年
2024年 | 1篇 |
2023年 | 18篇 |
2022年 | 44篇 |
2021年 | 67篇 |
2020年 | 44篇 |
2019年 | 37篇 |
2018年 | 50篇 |
2017年 | 19篇 |
2016年 | 28篇 |
2015年 | 41篇 |
2014年 | 34篇 |
2013年 | 45篇 |
2012年 | 39篇 |
2011年 | 28篇 |
2010年 | 17篇 |
2009年 | 12篇 |
2008年 | 13篇 |
2007年 | 14篇 |
2006年 | 14篇 |
2005年 | 8篇 |
2004年 | 10篇 |
2003年 | 6篇 |
2002年 | 4篇 |
2001年 | 4篇 |
2000年 | 6篇 |
1999年 | 6篇 |
1998年 | 2篇 |
1997年 | 6篇 |
1996年 | 7篇 |
1995年 | 1篇 |
1994年 | 3篇 |
1993年 | 1篇 |
1992年 | 3篇 |
1991年 | 3篇 |
1989年 | 1篇 |
1988年 | 2篇 |
1982年 | 2篇 |
排序方式: 共有640条查询结果,搜索用时 0 毫秒
51.
Bladder cancer-associated protein gene (BLCAP) is a novel candidate tumor suppressor gene identified from the human bladder
carcinoma. Our previous studies have shown that BLCAP overexpression could inhibit cell growth by inducing apoptosis in HeLa
cells [Zuo Z, Zhao M, Liu J, Gao G, Wu X: Tumor Biol 27: 221–226, 2006]. Such evidence suggests the alterations in BLCAP may
play an important role in tumorigenesis. To further study the biological function of the BLCAP gene, we constructed a recombinant
retroviral vector encoding BLCAP cDNA. Overexpressed BLCAP, via stable infection of exogenous BLCAP, resulted in growth inhibition
of the human tongue cancer cell line Tca8113 in vitro, accompanied by S phase cell cycle arrest and apoptosis. The growth inhibition was correlated with up-regulation of p21WAF1/CIP1 expression and down-regulation of Bcl-XL and Bcl-2 expressions. However, p53 expression and NF-κB activity remained unchanged
post infection. Furthermore, no changes in p53 phosphorylation at Ser46 and nuclear localization, which are critical to p53
function, were observed in BLCAP-overexpressed cells. Taken together, BLCAP may play a role not only in regulating cell proliferation
but also in coordinating apoptosis and cell cycle via a novel way independent of p53 and NF-κB.
Jun Yao and Li Duan contributed equally to this work. 相似文献
52.
53.
Background
Protein transduction is safer than viral vector-mediated transduction for the delivery of a therapeutic protein into a cell. Fusion proteins with an arginine-rich cell-penetrating peptide have been produced in E. coli, but the low solubility of the fusion protein expressed in E. coli impedes the large-scale production of fusion proteins from E. coli.Results
Expressed protein ligation is a semisynthetic method to ligate a bacterially expressed protein with a chemically synthesized peptide. In this study, we developed expressed protein ligation-based techniques to conjugate synthetic polyarginine peptides to Cre recombinase. The conjugation efficiency of this technique was higher than 80%. Using this method, we prepared semisynthetic Cre with poly-L-arginine (ssCre-R9), poly-D-arginine (ssCre-dR9) and biotin (ssCre-dR9-biotin). We found that ssCre-R9 was delivered to the cell to a comparable level or more efficiently compared with Cre-R11 and TAT-Cre expressed as recombinant fusion proteins in E. coli. We also found that the poly-D-arginine cell-penetrating peptide was more effective than the poly-L-arginine cell-penetrating peptide for the delivery of Cre into cell. We visualized the cell transduced with ssCre-dR9-biotin using avidin-FITC.Conclusions
Collectively, the results demonstrate that expressed protein ligation is an excellent technique for the production of cell-permeable Cre recombinase with polyarginine cell-penetrating peptides. In addition, this approach will extend the use of cell-permeable proteins to more sophisticated applications, such as cell imaging.Electronic supplementary material
The online version of this article (doi:10.1186/s12896-015-0126-z) contains supplementary material, which is available to authorized users. 相似文献54.
Xiangyang Wu Yong Wu Ruijuan Zheng Fen Tang Lianhua Qin Detian Lai Lu Zhang Lingming Chen Bo Yan Hua Yang Yang Wang Feifei Li Jinyu Zhang Fei Wang Lin Wang Yajuan Cao Mingtong Ma Zhonghua Liu Jianxia Chen Xiaochen Huang Jie Wang Ruiliang Jin Peng Wang Qin Sun Wei Sha Liangdong Lyu Pedro MouraAlves Anca Dorhoi Gang Pei Peng Zhang Jiayu Chen Shaorong Gao Felix Randow Gucheng Zeng Chang Chen XinShan Ye Stefan H E Kaufmann Haipeng Liu Baoxue Ge 《EMBO reports》2021,22(7)
Mycobacterial arabinogalactan (AG) is an essential cell wall component of mycobacteria and a frequent structural and bio‐synthetical target for anti‐tuberculosis (TB) drug development. Here, we report that mycobacterial AG is recognized by galectin‐9 and exacerbates mycobacterial infection. Administration of AG‐specific aptamers inhibits cellular infiltration caused by Mycobacterium tuberculosis (Mtb) or Mycobacterium bovis BCG, and moderately increases survival of Mtb‐infected mice or Mycobacterium marinum‐infected zebrafish. AG interacts with carbohydrate recognition domain (CRD) 2 of galectin‐9 with high affinity, and galectin‐9 associates with transforming growth factor β‐activated kinase 1 (TAK1) via CRD2 to trigger subsequent activation of extracellular signal‐regulated kinase (ERK) as well as induction of the expression of matrix metalloproteinases (MMPs). Moreover, deletion of galectin‐9 or inhibition of MMPs blocks AG‐induced pathological impairments in the lung, and the AG‐galectin‐9 axis aggravates the process of Mtb infection in mice. These results demonstrate that AG is an important virulence factor of mycobacteria and galectin‐9 is a novel receptor for Mtb and other mycobacteria, paving the way for the development of novel effective TB immune modulators. 相似文献
55.
A three-step electrodeposition method has been successfully adopted to fabricate morphology-controlled novel Au microspheres on self-doped polyaniline nanofibers (nanoSPAN) modified glassy carbon electrode. The deposition conditions, such as HAuCl(4) concentration and deposition step, have significant influences on the morphologies and electrochemical properties of the resulted Au microspheres. Well hierarchical and homogeneously dispersed flower-like Au microspheres (HHFAu) were obtained under optimal conditions by the three-step electrodeposition strategy in 5.0mM HAuCl(4) solution. HHFAu possess large surface area, excellent electron transfer ability and good biocompatibility. The DNA probe could be effectively attached to HHFAu and thus a high-performance DNA biosensor was constructed by using electrochemical impedance spectroscopy as detection method. A gene fragment of the cauliflower mosaic virus 35S gene, which is related to one of the screening genes for the transgenically modified plants, has been satisfactorily detected. The linear range was from 1.0 × 10(-13)M to 1.0 × 10(-6)M and the detection limit was 1.9 × 10(-14)M. This HHFAu/nanoSPAN-based impedance biosensing platform holds great promise for the detection of other biological and chemical molecules. 相似文献
56.
Qingxin Zhou Xinxing Lv Xi Zhang Xiangfeng Meng Guanjun Chen Weifeng Liu 《World journal of microbiology & biotechnology》2011,27(8):1905-1910
Swollenin is a novel plant expansin-like protein that has been proposed to have a cellulose disruption activity. In this study,
the recombinant swollenin (SWO2) from Trichoderma pseudokoningii S38 was successfully produced and purified in Aspergillus niger with a final yield of up to 10 mg of purified protein from 1 l of fermentation supernatant. The recombinant protein was found
to exhibit very low level of endoglucanase activity and caused a slight increase in the crystallinity when treating cellulose.
Simultaneous incubation of SWO2 with low-dose cellulases resulted in a significant synergistic activity in cellulose hydrolysis.
Specifically, an even greater increase in the synergistic activity was obtained when cellulose was pretreated with swollenin
followed by cellulase hydrolysis. Our results, therefore, provide a novel approach for the potential application of swollenin
in the efficient saccharification of cellulosic materials. 相似文献
57.
58.
Background
The DNase I hypersensitive sites (DHSs) are associated with the cis-regulatory DNA elements. An efficient method of identifying DHSs can enhance the understanding on the accessibility of chromatin. Despite a multitude of resources available on line including experimental datasets and computational tools, the complex language of DHSs remains incompletely understood.Methods
Here, we address this challenge using an approach based on a state-of-the-art machine learning method. We present a novel convolutional neural network (CNN) which combined Inception like networks with a gating mechanism for the response of multiple patterns and longterm association in DNA sequences to predict multi-scale DHSs in Arabidopsis, rice and Homo sapiens.Results
Our method obtains 0.961 area under curve (AUC) on Arabidopsis, 0.969 AUC on rice and 0.918 AUC on Homo sapiens.Conclusions
Our method provides an efficient and accurate way to identify multi-scale DHSs sequences by deep learning.59.
Xinxing Lu Qiuling Fan Li Xu Lin Li Yuan Yue Yanyan Xu Yan Su Dongcheng Zhang Lining Wang 《PloS one》2015,10(2)
ObjectiveTo investigate the effect of ursolic acid on autophagy mediated through the miRNA-21-targeted phosphoinositide 3 kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) pathway in rat mesangial cells cultured under high glucose (HG) conditions.MethodsRat glomerular mesangial cells were cultured under normal glucose, HG, HG with the PI3K inhibitor or HG with ursolic acid conditions. Cell proliferation and hypertrophy were assayed using an MTT assay and the ratio of total protein to cell number, respectively. The miRNA-21 expression was detected using RT-qPCR. The expression of phosphatase and tensin homolog (PTEN)/AKT/mTOR signaling signatures, autophagy-associated protein and collagen I was detected by western blotting and RT-qPCR. Autophagosomes were observed using electron microscopy.ResultsCompared with mesangial cells cultured under normal glucose conditions, the cells exposed to HG showed up-regulated miRNA-21 expression, down-regulated PTEN protein and mRNA expression, up-regulated p85PI3K, pAkt, pmTOR, p62/SQSTMI, and collagen I expression and down-regulated LC3II expression. Ursolic acid and LY294002 inhibited HG-induced mesangial cell hypertrophy and proliferation, down-regulated p85PI3K, pAkt, pmTOR, p62/SQSTMI, and collagen I expression and up-regulated LC3II expression. However, LY294002 did not affect the expression of miRNA-21 and PTEN. Ursolic acid down-regulated miRNA-21 expression and up-regulated PTEN protein and mRNA expression.ConclusionsUrsolic acid inhibits the glucose-induced up-regulation of mesangial cell miRNA-21 expression, up-regulates PTEN expression, inhibits the activation of PI3K/Akt/mTOR signaling pathway, and enhances autophagy to reduce the accumulation of the extracellular matrix and ameliorate cell hypertrophy and proliferation. LY294002相似文献
60.
Whole‐field macro‐ and micro‐deformation characteristic of unbound water‐loss in dentin hard tissue 下载免费PDF全文
Zhenning Chen Bobby Nadeau Kevin Yu Xinxing Shao Xiaoyuan He M. Cynthia Goh Anil Kishen 《Journal of biophotonics》2018,11(9)
High‐resolution deformation measurements in a functionally graded hard tissue such as human dentin are essential to understand the unbound water‐loss mediated changes and their role in its mechanical integrity. Yet a whole‐field, 3‐dimensional (3D) measurement and characterization of fully hydrated dentin in both macro‐ and micro‐scales remain to be a challenge. This study was conducted in 2 stages. In stage‐1, a stereo‐digital image correlation approach was utilized to determine the water‐loss and load‐induced 3D deformations of teeth in a sagittal section over consecutively acquired frames, from a fully hydrated state to nonhydrated conditions for a period up to 2 hours. The macroscale analysis revealed concentrated residual deformations at the dentin‐enamel‐junction and the apical regions of root in the direction perpendicular to the dentinal tubules. Significant difference in the localized deformation characteristics was observed between the inner and outer aspects of the root dentin. During quasi‐static loadings, further increase in the residual deformation was observed in the dentin. In stage‐2, dentin microstructural variations induced by dynamic water‐loss were assessed with environmental scanning electron microscopy and atomic force microscopy (AFM), showing that the dynamic water‐loss induced distention of dentinal tubules with concave tubular edges, and concurrent contraction of intertubular dentin with convex profile. The findings from the current macro‐ and micro‐scale analysis provided insight on the free‐water‐loss induced regional deformations and ultrastructural changes in human dentin. 相似文献