Hydrogen Sulfide and Cellular Redox Homeostasis
作者：
Xie, Zhi-Zhong* ;Liu, Yang;Bian, Jin-Song*
期刊：
Oxidative Medicine and Cellular Longevity ,2016年2016:6043038 ISSN：1942-0900
通讯作者：
Xie, Zhi-Zhong;Bian, Jin-Song
作者机构：
[Xie, Zhi-Zhong; Liu, Yang] Univ South China, Inst Pharm & Pharmacol, Hengyang 421001, Hunan, Peoples R China.;[Xie, Zhi-Zhong; Liu, Yang] Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang 421001, Hunan, Peoples R China.;[Bian, Jin-Song] Natl Univ Singapore, Yong Loo Lin Sch Med, Dept Pharmacol, Singapore 117597, Singapore.
通讯机构：
[Xie, Zhi-Zhong] Univ South China, Inst Pharm & Pharmacol, Hengyang 421001, Hunan, Peoples R China.;[Xie, Zhi-Zhong] Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang 421001, Hunan, Peoples R China.;[Bian, Jin-Song] Natl Univ Singapore, Yong Loo Lin Sch Med, Dept Pharmacol, Singapore 117597, Singapore.
摘要：
Intracellular redox imbalance is mainly caused by overproduction of reactive oxygen species (ROS) or weakness of the natural antioxidant defense system. It is involved in the pathophysiology of a wide array of human diseases. Hydrogen sulfide (H2S) is now recognized as the third "gasotransmitters" and proved to exert a wide range of physiological and cytoprotective functions in the biological systems. Among these functions, the role of H2S in oxidative stress has been one of the main focuses over years. However, the underlying mechanisms for the antioxidant effect of H2S are still poorly comprehended. This review presents an overview of the current understanding of H2S specially focusing on the new understanding and mechanisms of the antioxidant effects of H2S based on recent reports. Both inhibition of ROS generation and stimulation of antioxidants are discussed. H2S-induced S-sulfhydration of key proteins (e.g., p66Shc and Keap1) is also one of the focuses of this review.
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Simultaneous determination of flavanones, hydroxycinnamic acids and alkaloids in citrus fruits by HPLC-DAD-ESI/MS
作者：
He, Dongxiu;Shan, Yang;Wu, Yuehui;Liu, Guozhu;Chen, Bo* ;Yao, Shouzhuo
期刊：
Food Chemistry ,2011年127(2):880-885 ISSN：0308-8146
通讯作者：
Chen, Bo
作者机构：
[Liu, Guozhu; He, Dongxiu; Chen, Bo; Yao, Shouzhuo] Hunan Normal Univ, Minist Educ, Key Lab Chem Biol & Tradit Chinese Med Res, Changsha 410081, Hunan, Peoples R China.;[He, Dongxiu] Univ S China, Sch Life Sci & Technol, Dept Pharm, Hengyang 421001, Peoples R China.;[Wu, Yuehui; Shan, Yang] Hunan Agr Prod Proc Inst, Changsha 410081, Hunan, Peoples R China.
通讯机构：
[Chen, Bo] Hunan Normal Univ, Minist Educ, Key Lab Chem Biol & Tradit Chinese Med Res, Changsha 410081, Hunan, Peoples R China.
关键词：
Flavanones;Hydroxycinnamic acids;Alkaloids;HPLC-DAD–ESI/MS;Citrus hybrids;Citrus pulp;Citrus peel
摘要：
A simple and accurate method has been developed to simultaneously separate and determine 10 bioactive compounds in citrus fruits by high-performance liquid chromatography coupled with diode array detection and electrospray ionisation mass spectrometry (HPLC-DAD-ESI/MS). This HPLC assay was performed on a reversed-phase C18 column with acetonitrile and 0.1% (v/v) aqueous formic acid as mobile phase. DAD has been performed at 273, 283 and 324 nm for quantification of the alkaloids, flavanones and hydroxycinnamic acids. MS was also employed to identify the each analyte. Ten analytes (naringin, hesperidin, neohesperidin, ferulic acid, p-coumaric acid, chlorogenic acid, caffeic acid, octopamine, synephrine and tyramine) demonstrated good linearity (r ≥0.9990) in a relatively wide concentration range. The method revealed high average recovery (range, 92.1-97.9%) and good precision with interday and intraday variations with less than 4.71%. The limits of detection (LODs) ranged from 0.02 to 0.11 μg/ml, while the limits of quantification (LOQs) were defined in the range of 0.08-0.39 μg/ml. The proposed method has been successfully applied to analyse three types of bioactive constituents in eight citrus hybrids pulps and eight citrus hybrids peels, which has been successfully cultivated in China. ©2011 Elsevier Ltd. All rights reserved.
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英文
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Tristetraprolin-dependent post-transcriptional regulation of inflammatory cytokine mRNA expression by apolipoprotein A-I: Role of ATP-binding membrane cassette transporter a1 and signal transducer and activator of transcription 3
作者：
Yin, Kai;Deng, Xiang;Mo, Zhong-Cheng;Zhao, Guo-Jun;Jiang, Jin;Cui, Li-Bao;Tan, Chun-Zhi;Wen, Ge-Bo;Fu, Yuchang;Tang, Chao-Ke*
期刊：
Journal of Biological Chemistry ,2011年286(16):13834-13845 ISSN：0021-9258
通讯作者：
Tang, Chao-Ke
作者机构：
[Zhao, Guo-Jun; Tan, Chun-Zhi; Tang, Chao-Ke; Wen, Ge-Bo; Yin, Kai; Mo, Zhong-Cheng; Jiang, Jin; Cui, Li-Bao] Univ S China, Affiliated Hosp 1, Key Lab Atherosclerol Hunan Prov, Life Sci Res Ctr,Inst Cardiovasc Res, Hengyang 421001, Peoples R China.;[Wen, Ge-Bo; Yin, Kai] Univ S China, Affiliated Hosp 1, Dept Diagnost, Coll Med, Hengyang 421001, Peoples R China.;[Wen, Ge-Bo; Deng, Xiang] Univ S China, Affiliated Hosp 1, Dept Endocrinol & Metab, Hengyang 421001, Peoples R China.;[Fu, Yuchang] Univ Alabama, Dept Nutr Sci, Birmingham, AL 35294 USA.
通讯机构：
[Tang, Chao-Ke] Univ S China, Affiliated Hosp 1, Key Lab Atherosclerol Hunan Prov, Life Sci Res Ctr,Inst Cardiovasc Res, Hengyang 421001, Peoples R China.
摘要：
Atherosclerosis is an inflammatory disease characterized by the accumulation of macrophages in the arterial intima. The activated macrophages secreted more pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α, which promote the development of the disease. Apolipoprotein A-I (apoA-I), the major component of high density lipoprotein, is involved in reverse cholesterol transport of lipid metabolism. Recently, it has been found that apoA-I suppresses inflammation via repression of inflammatory cytokine expression;the mechanisms of the apoA-I-suppressive action, however, are not yet well characterized. In this study, we have for the first time found that apoA-I suppresses the expression of some inflammatory cytokines induced by lipopolysaccharide via a specific post-transcriptional regulation process, namely mRNA destabilization, in macrophages. Our further studies have also shown that AU-rich elements in the 3′-untranslated region of TNF-αmRNA are responsive to the apoA-I-mediated mRNA destabilization. The apoA-I-induced inflammatory cytokine mRNA destabilization was associated with increased expression of mRNA-destabilizing protein tristetraprolin through aJAK2/STAT3 signaling pathway-dependent manner. When blocking interaction of apoA-Iwith ATP-binding membrane cassette transporter A1 (ABCA1), a major receptor for apoA-Iin macrophages, it would almost totally abolish the effect of apoA-I on tristetraprolin expression. These results present not only a novel mechanism for the apoA-I-mediated inflammation suppression in macrophages but also provide new insights for developing strategies for modulating vascular inflammation and atherosclerosis. ©2011 by The American Society for Biochemistry and Molecular Biology, Inc.
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英文
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A generic approach for expanding homolog-targeted residue screening of sulfonamides using a fast matrix separation and class-specific fragmentation-dependent acquisition with a hybrid quadrupole-linear ion trap mass spectrometer
作者：
Huang, Chunlin;Guo, Bin* ;Wang, Xiaoying;Li, Jie;Zhu, Weitao;Chen, Bo;Ouyang, Shan;Yao, Shouzhuo
期刊：
Analytica Chimica Acta ,2012年737:83-98 ISSN：0003-2670
通讯作者：
Guo, Bin
作者机构：
[Li, Jie; Huang, Chunlin] Univ S China, Sch Pharm & Life Sci, Dept Biochem & Mol Biol, Hengyang 421001, Peoples R China.;[Guo, Bin; Wang, Xiaoying; Zhu, Weitao; Chen, Bo; Yao, Shouzhuo] Hunan Normal Univ, Minist Educ China, Key Lab Chem Biol & Tradit Chinese Med Res, Changsha 410081, Hunan, Peoples R China.;[Ouyang, Shan] Shenzhen Entry Exit Inspect & Quarantine Bur Peop, Food Inspect & Quarantine Ctr, Shenzhen 518067, Peoples R China.
通讯机构：
[Guo, Bin] Hunan Normal Univ, Minist Educ China, Key Lab Chem Biol & Tradit Chinese Med Res, Changsha 410081, Hunan, Peoples R China.
关键词：
Sulfonamides;Multi-residue;Dispersive solid-phase extraction;Animal tissues;Triple-quadrupole linear ion trap mass spectrometer;Semi-targeted screening
摘要：
A generic and efficient homolog-targeted approach was used to expand screening and detection of target class of sulfonamides and structural analogs, based on a fast single-tube extraction/partitioning-multifunction adsorption cleanup (SEP/MAC) for class-specific fragmentation-dependent acquisition with a liquid chromatography-hybrid triple-quadrupole linear ion trap mass spectrometer (LC-QqLIT). By combining the two-stage process conducted in a single tube as one-pot protocol, the straightforward SEP/MAC procedure was optimized to offer clean extracts with reasonable recovery (71-109% with RSDs < 20%) and decreased matrix interferences (-9 to 19%) of multiresidual sulfonamide extraction from different tissue samples. The novel use of neutral loss scan of 66 Da (NLS) or precursor ion scanning of m/z 108 (PreS) in positive ion mode was found to achieve more comprehensive coverage of protonated molecular ions of a wide array of sulfonamides including N-4-acetyl and hydroxylamine metabolites plus their possible dimers. Moreover, the PreS-triggered automatically enhanced product ion spectral acquisition enabled simultaneous screening, profiling and confirmation of an unlimited number of analytes belonging to the sulfonamide class within a single analysis. The validation and application results of the generic SEP/MAC-based LC-QqLIT strategy consistently demonstrated favorable performances with acceptable accuracy (67-116%), precision (RSDs < 25%), and sensitivity (LOQs <= 7.5 ng g(-1)) to meet the acceptance criteria for all the sulfonamide-tissue combinations. Thus, the integration of the matrix-independent SEP/MAC procedure and the multiparameter matching algorithm with the unit-resolution LC-QqLIT instrument can serve as a valuable semi-targeted discovery strategy for rapid screening and reliable quantitative/confirmatory analysis of real samples. (C) 2012 Elsevier B.V. All rights reserved.
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英文
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Design and Synthesis of Fluorinated Amphiphile as 19F MRI/Fluorescence Dual-Imaging Agent by Tuning the Self-Assembly
作者：
Bo, Shaowei;Song, Cong;Li, Yu;Yu, Weijiang;Chen, Shizhen;Zhou, Xin;Yang, Zhigang;Zheng, Xing* ;Jiang, Zhong-Xing
期刊：
Journal of Organic Chemistry ,2015年80(12):6360-6366 ISSN：0022-3263
通讯作者：
Zheng, Xing
作者机构：
[Bo, Shaowei; Zheng, Xing] Univ South China, Inst Pharm Pharmacol, Hengyang 421001, Peoples R China.;[Li, Yu; Yu, Weijiang; Jiang, Zhong-Xing; Bo, Shaowei; Song, Cong; Yang, Zhigang] Wuhan Univ, Minist Educ, Key Lab Combinatorial Biosynth & Drug Discovery, Wuhan 430071, Peoples R China.;[Li, Yu; Yu, Weijiang; Jiang, Zhong-Xing; Bo, Shaowei; Song, Cong; Yang, Zhigang] Wuhan Univ, Sch Pharmaceut Sci, Wuhan 430071, Peoples R China.;[Zhou, Xin; Chen, Shizhen] Chinese Acad Sci, Wuhan Inst Phys & Math, Natl Ctr Magnet Resonance Wuhan, Key Lab Magnet Resonance Biol Syst,State Key Lab, Wuhan 430071, Peoples R China.;[Jiang, Zhong-Xing; Bo, Shaowei; Zheng, Xing] Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang 421001, Peoples R China.
通讯机构：
[Zheng, Xing] Univ South China, Inst Pharm Pharmacol, Hengyang 421001, Peoples R China.
摘要：
Both<sup>19</sup>F MRI and optical imaging are powerful noninvasive molecular imaging modalities in biomedical applications. To integrate these two complementary imaging modalities, the design and synthesis of a novel<sup>19</sup>F MRI/fluorescence dual-modal imaging agent is reported herein. Through Sonogashira coupling reaction between the fluorinated phenylacetylene and 1,2,4,5-tetraiodobenzene, a fluorophore with 48 symmetrical fluorines at its periphery was constructed with high efficacy. High aqueous solubility was achieved by PEGylation of the fluorophore with monodisperse PEGs. However, an unexpected self-assembly of the PEGylated amphiphilic fluorophore in water "turned off" the<sup>19</sup>F NMR signal. However, hydrogenation of the triple bonds or introduction of branched monodisperse PEGs was able to efficiently tune the self-assembly, resulting in the "turning on" of the<sup>19</sup>F NMR signal. One of these amphiphiles combines the advantages of label-free fluorescence, high<sup>19</sup>F MRI sensitivity, biocompatibility, and excellent aqueous solubility. The results demonstrate the great potential of such amphiphiles for real-time<sup>19</sup>F MRI and fluorescence dual-modality imaging. ©2015 American Chemical Society.
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英文
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Design and Synthesis of Fluorinated Dendrimers for Sensitive 19F MRI
作者：
Yu, Weijiang;Yang, Yuqi;Bo, Shaowei;Li, Yu;Chen, Shizhen;Yang, Zhigang;Zheng, Xing;Jiang, Zhong-Xing* ;Zhou, Xin
期刊：
Journal of Organic Chemistry ,2015年80(9):4443-4449 ISSN：0022-3263
通讯作者：
Jiang, Zhong-Xing
作者机构：
[Li, Yu; Yu, Weijiang; Jiang, Zhong-Xing; Yang, Zhigang] Wuhan Univ, Key Lab Combinatorial Biosynth & Drug Discovery, Minist Educ, Wuhan 430071, Peoples R China.;[Li, Yu; Yu, Weijiang; Jiang, Zhong-Xing; Yang, Zhigang] Wuhan Univ, Sch Pharmaceut Sci, Wuhan 430071, Peoples R China.;[Yang, Yuqi; Zhou, Xin; Chen, Shizhen] Chinese Acad Sci, Wuhan Inst Phys & Math, Natl Ctr Magnet Resonance Wuhan, Key Lab Magnet Resonance Biol Syst,State Key Lab, Wuhan 430071, Peoples R China.;[Jiang, Zhong-Xing; Bo, Shaowei; Zheng, Xing] Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hunan Prov Educ Dept, Hengyang 421001, Peoples R China.;[Bo, Shaowei; Zheng, Xing] Univ South China, Inst Pharm & Pharmacol, Hengyang 421001, Peoples R China.
通讯机构：
[Jiang, Zhong-Xing] Wuhan Univ, Key Lab Combinatorial Biosynth & Drug Discovery, Minist Educ, Wuhan 430071, Peoples R China.
摘要：
To achieve high sensitivity for <sup>19</sup>F MRI, a class of novel dendritic molecules with multiple pseudosymmetrical fluorines was designed and efficiently synthesized. Through iterative bromination and Williamson ether synthesis under mild conditions, a fluorinated dendrimer with 540 pseudosymmetrical fluorines was conveniently prepared without performing the group protection in a convergent way. The dendrimer is characterized by a strong <sup>19</sup>F NMR peak and short relaxation times. Eventually, an appreciably enhanced <sup>19</sup>F MRI at an extremely low concentration (18.5 μM) was achieved, which demonstrated the potential utility of such dendritic molecules in highly sensitive <sup>19</sup>F MRI. (Chemical Equation Presented). ©2015 American Chemical Society.
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英文
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Cyclodextrin-functionalized polymers as drug carriers for cancer therapy
作者：
Wei, Hua* ;Yu, Cui-yun
期刊：
Biomaterials Science ,2015年3(7):1050-1060 ISSN：2047-4830
通讯作者：
Wei, Hua
作者机构：
[Wei, Hua] Lanzhou Univ, Key Lab Nonferrous Met Chem & Resources Utilizat, Lanzhou 730000, Gansu, Peoples R China.;[Wei, Hua] Lanzhou Univ, Dept Chem & Chem Engn, Lanzhou 730000, Gansu, Peoples R China.;[Yu, Cui-yun] Univ South China, Dept Pharm, Inst Pharm & Pharmacol, Hengyang 421001, Peoples R China.
通讯机构：
[Wei, Hua] Lanzhou Univ, Key Lab Nonferrous Met Chem & Resources Utilizat, Lanzhou 730000, Gansu, Peoples R China.
摘要：
Cyclodextrins (CDs) represent the most extensively investigated cyclic molecules due to their wide availability, facile functionalization, unique amphiphilicity and inclusion capacity. The marriage of CD chemistry with polymer science has generated novel biomaterials by integrating supermolecular host-guest chemistry and state-of-the-art polymer chemistry techniques. The current mini-review focuses on the recent progress in CD-functionalized polymers as drug carriers for cancer therapy. CD-functionalized polymers with different structures are summarized. Their application as drug carriers for cancer therapy is then highlighted. In the end, the future directions of this rapidly developing research field are discussed. This journal is ©The Royal Society of Chemistry.
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英文
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Assessment of heavy metal contamination and bioaccumulation in soybean plants from mining and smelting areas of southern Hunan Province, China
作者：
Zhou, Hang;Zeng, Min;Zhou, Xin;Liao, Bo-Han* ;Liu, Jun;Lei, Ming;Zhong, Qian-Yun;Zeng, Hui
期刊：
Environmental Toxicology and Chemistry ,2013年32(12):2719-2727 ISSN：0730-7268
通讯作者：
Liao, Bo-Han
作者机构：
[Zhou, Hang] Hunan Agr Univ, Coll Biosci & Technol, Changsha, Hunan, Peoples R China.;[Zhong, Qian-Yun; Liao, Bo-Han; Zeng, Hui; Zeng, Min; Zhou, Xin; Lei, Ming; Zhou, Hang] Cent South Univ Forestry & Technol, Inst Environm Sci & Engn, Changsha, Hunan, Peoples R China.;[Liu, Jun] Univ South China, Coll Pharm & Life Sci, Hengyang, Peoples R China.
通讯机构：
[Liao, Bo-Han] Cent South Univ Forestry & Technol, Inst Environm Sci & Engn, Changsha, Hunan, Peoples R China.
关键词：
Assessment;Geoaccumulation index;Heavy metal accumulation;Soil contamination;Soybean (Glycine max)
摘要：
Soybean is one of most important dicotyledonous food crops and is widely planted in Hunan Province, China. However, mining activity causes contamination of the soil in which soybean grows. To assess the impact of mining-induced soil contamination on soybean plants, a geoaccumulation index (I<inf>geo</inf>) was used to evaluate 20 soil samples from the mining and smelting areas of southern Hunan Province. The results indicated that Zn ranged from uncontaminated to a moderately contaminated level (I<inf>geo</inf><1), Pb was at a strongly contaminated level (I<inf>geo</inf>>3), and Cd was at an extremely contaminated level (I<inf>geo</inf>>5) across the whole study area. All of the studied soybean plants were affected by heavy metal Pb and Cd contamination, and the mean concentrations in seeds were 13.9mg/kg and 2.95mg/kg, respectively. The estimated bioconcentration factor and translocation factor showed that the soybean roots had a strong Cd bioconcentration capability and the stems had a strong translocation capability in terms of Pb, Cd, and Zn, with preferential transference of metals to the soybean leaves. The bioavailable fraction in the soil was characterized by the exchangeable fraction of heavy metals. In the present study, the bioavailable fractions of Pb, Cd, and Zn were significantly positively correlated with the concentration of these metals in soybean tissues (roots, stems, leaves, husks, and seeds). ©2013 SETAC.
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英文
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Target-based metabolomics for the quantitative measurement of 37 pathway metabolites in rat brain and serum using hydrophilic interaction ultra-high-performance liquid chromatography-tandem mass spectrometry
作者：
Chen, Jiahui;Hou, Waner;Han, Bo;Liu, Guanghui;Gong, Jin;Li, Yemeng;Zhong, Danmin;Liao, Qiongfeng* ;Xie, Zhiyong*
期刊：
Analytical and Bioanalytical Chemistry ,2016年408(10):2527-2542 ISSN：1618-2642
通讯作者：
Liao, Qiongfeng;Xie, Zhiyong
作者机构：
[Chen, Jiahui; Gong, Jin; Liao, Qiongfeng; Liu, Guanghui; Hou, Waner] Guangzhou Univ Chinese Med, Sch Chinese Mat Med, Guangzhou 510407, Guangdong, Peoples R China.;[Han, Bo] Shihezi Univ, Sch Pharm, Shihezi 832000, Peoples R China.;[Zhong, Danmin; Li, Yemeng; Xie, Zhiyong] Sun Yat Sen Univ, Sch Pharmaceut Sci, Guangzhou 510006, Guangdong, Peoples R China.;[Xie, Zhiyong] Univ South China, Cooperat Innovat Ctr Mol Target New Drug Study, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Liao, Qiongfeng] Guangzhou Univ Chinese Med, Sch Chinese Mat Med, Guangzhou 510407, Guangdong, Peoples R China.;[Xie, Zhiyong] Sun Yat Sen Univ, Sch Pharmaceut Sci, Guangzhou 510006, Guangdong, Peoples R China.;[Xie, Zhiyong] Univ South China, Cooperat Innovat Ctr Mol Target New Drug Study, Hengyang 421001, Hunan, Peoples R China.
关键词：
Target-based metabolomics;Quantitative;HILIC–MS;Amino acids;neurotransmitters;purines and pyrimidines
摘要：
Amino acids, neurotransmitters, purines, and pyrimidines are bioactive molecules that play fundamental roles in maintaining various physiological functions. Their metabolism is closely related to the health, growth, development, reproduction, and homeostasis of organisms. Most recently, comprehensive measurements of these metabolites have shown their potential as innovative approaches in disease surveillance or drug intervention. However, simultaneous measurement of these metabolites presents great difficulties. Here, we report a novel quantitative method that uses hydrophilic interaction ultra-high-performance liquid chromatography-tandem mass spectrometry (HILIC-UPLC-MS/MS), which is highly selective, high throughput, and exhibits better chromatographic behavior than existing methods. The developed method enabled the rapid quantification of 37 metabolites, spanning amino acids, neurotransmitters, purines, and pyrimidines pathways, within 6.5 min. The compounds were separated on an ACQUITY UPLC®BEH Amide column. Serum and brain homogenate were extracted by protein precipitation. The intra- and interday precision of all of the analytes was less than 11.34 %, and the accuracy was between -11.74 and 11.51 % for all quality control (QC) levels. The extraction recoveries of serum ranged from 84.58 % to 116.43 % and those of brain samples from 80.80 % to 119.39 %, while the RSD was 14.61 % or less for all recoveries. This method was used to successfully characterize alterations in the rat brain and, in particular, their dynamics in serum. The following study was performed to simultaneously test global changes of these metabolites in a serotonin antagonist p-chlorophenylalanine (PCPA)-induced anxiety and insomnia rat model to understand the effect and mechanism of PCPA. Taken together, these results show that the method is able to simultaneously monitor a large panel of metabolites and that this protocol may represent a metabolomic method to diagnose toxicological and pathophysiological states. ©Springer-Verlag Berlin Heidelberg 2016.
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英文
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Pyrrolidine Dithiocarbamate Inhibits NF-KappaB Activation and Upregulates the Expression of Gpx1, Gpx4, Occludin, and ZO-1 in DSS-Induced Colitis
作者：
Yin, Jie;Wu, Miaomiao;Duan, Jielin;Liu, Gang* ;Cui, Zhijie;Zheng, Jie;Chen, Shuai;Ren, Wenkai;Deng, Jinping;Tan, Xiangwen;Al-Dhabi, Naif Abdullah;Duraipandiyan, Veeramuthu;Liao, Peng;Li, Tiejun;Yin Yulong
期刊：
Applied Biochemistry and Biotechnology ,2015年177(8):1716-1728 ISSN：0273-2289
通讯作者：
Liu, Gang
作者机构：
[Liu, Gang; Cui, Zhijie; Chen, Shuai; Li, Tiejun; Liao, Peng; Yin, Jie; Ren, Wenkai; Wu, Miaomiao; Duan, Jielin; Yin Yulong] Chinese Acad Sci, Sci Observing & Expt Stn Anim Nutr & Feed Sci Sou, Hunan Prov Engn Res Ctr Hlth Livestock,Minist Agr, Key Lab Agroecol Proc Subtrop Reg,Inst Subtrop Ag, Changsha 410125, Hunan, Peoples R China.;[Cui, Zhijie; Yin, Jie; Wu, Miaomiao; Duan, Jielin] Univ Chinese Acad Sci, Beijing 100039, Peoples R China.;[Zheng, Jie; Deng, Jinping] Hunan Agr Univ, Coll Anim Sci, Changsha 410128, Hunan, Peoples R China.;[Tan, Xiangwen] Univ South China, Dept Lab Anim Sci, Hengyang 421001, Peoples R China.;[Duraipandiyan, Veeramuthu; Al-Dhabi, Naif Abdullah] King Saud Univ, Coll Sci, Addiriyah Chair Environm Studies, Dept Bot & Microbiol, Riyadh 11451, Saudi Arabia.
通讯机构：
[Liu, Gang] Chinese Acad Sci, Sci Observing & Expt Stn Anim Nutr & Feed Sci Sou, Hunan Prov Engn Res Ctr Hlth Livestock,Minist Agr, Key Lab Agroecol Proc Subtrop Reg,Inst Subtrop Ag, Changsha 410125, Hunan, Peoples R China.
关键词：
PDTC;IBD;Antioxidant gene;Tight junction;NF-κB
摘要：
Inflammatory bowel disease (IBD) correlates with oxidative stress, inflammation, and alteration in several signal pathways, including nuclear transcription factor-kappaB (NF-κB). Pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB, has been widely demonstrated to exhibit an antioxidant and anti-inflammatory function. This study aimed to test the hypothesis that NF-κB inhibitor PDTC confers a beneficial role in a colitis model induced by dextran sodium sulfate (DSS) in mouse. The results showed that DSS decreased daily weight gain, induced colonic inflammation, suppressed the expression of antioxidant enzymes and tight junctions, and activated NF-κB and nuclear factor erythroid 2-related factor 2/Kelch-like ECH-associated protein 1 (Nrf2/Keap1) signaling pathways. PDTC significantly upregulated (P < 0.05) Gpx1, Gpx4, occludin, and ZO-1 expressions in the DSS-induced colitis model. Meanwhile, PDTC reversed (P < 0.05) the activation of NF-κB signal pathway caused by DSS treatment. In conclusion, PDTC could serve as an adjuvant therapy for the patient with IBD. ©2015, Springer Science+Business Media New York.
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英文
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Fabrication of galactosylated chitosan-5-fluorouracil acetic acid based nanoparticles for controlled drug delivery
作者：
Yu, Cui-Yun* ;Li, Na-Mei;Yang, Sa;Ning, Qian;Huang, Can;Huang, Wen;He, Zi-Ning;He, Dong-Xiu;Tan, Xiang-Wen;Sun, Li-Chun
期刊：
Journal of Applied Polymer Science ,2015年132(40):- ISSN：1097-4628
通讯作者：
Yu, Cui-Yun
作者机构：
[He, Zi-Ning; Huang, Wen; Huang, Can; He, Dong-Xiu; Tan, Xiang-Wen; Li, Na-Mei; Sun, Li-Chun; Yang, Sa; Yu, Cui-Yun; Ning, Qian] Univ South China, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang 421001, Peoples R China.;[Yu, Cui-Yun] Univ South China, Inst Pharm & Pharmacol, Learning Key Lab Pharmacoprote Hunan Prov, Hengyang 421001, Peoples R China.;[Sun, Li-Chun] Tulane Univ, Dept Med, Hlth Sci Ctr, New Orleans, LA 70112 USA.
通讯机构：
[Yu, Cui-Yun] Univ South China, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang 421001, Peoples R China.
关键词：
biomaterials;biomedical applications;drug-delivery systems;nanostructured polymers
摘要：
In this study, a novel type of macromolecular prodrug, N-galactosylated chitosan (GC)-5-fluorouracil acetic acid (FUA) conjugate based nanoparticles, was designed and synthesized as a carrier for hepatocellular carcinoma drug delivery. The GC-FUA nanoparticles were produced by an ionic crosslinking method based on the modified ionic gelation of tripolyphosphate with GC-FUA. The structure of the as-prepared GC-FUA was characterized by Fourier transform infrared and H-1-NMR analyses. The average particle size of the GC-FUA nanoparticles was 160.1 nm, and their drug-loading content was 21.22 +/- 2.7% (n=3). In comparison with that of the freshly prepared nanoparticles, this value became larger after 7 days because of the aggregation of the GC-FUA nanoparticles. An in vitro drug-release study showed that the GC-FUA nanoparticles displayed a sustained-release profile compared to 5-fluorouracil-loaded GC nanoparticles. All of the results suggest that the GC-FUA nanoparticles may have great potential for anti-liver-cancer applications. (c) 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 42625.
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英文
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Highly selective and sensitive fluorescence chemosensor for the detection of palladium species based on Tsuji-Trost reaction
作者：
Xu, Zhong-Yong;Li, Jing;Guan, Su;Zhang, Lei* ;Dong, Chang-Zhi
期刊：
Spectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy ,2015年148:7-11 ISSN：1386-1425
通讯作者：
Zhang, Lei
作者机构：
[Guan, Su; Xu, Zhong-Yong; Li, Jing; Zhang, Lei] South China Univ, Sch Biosci & Bioengn, Guangzhou 510006, Guangdong, Peoples R China.;[Dong, Chang-Zhi] Univ Paris Diderot, Sorbonne Paris Cite, CNRS, UMR 7086,ITODYS, F-75205 Paris 13, France.;[Zhang, Lei] S China Univ Technol, Sch Biosci & Bioengn, Guangzhou 510006, Guangdong, Peoples R China.
通讯机构：
[Zhang, Lei] S China Univ Technol, Sch Biosci & Bioengn, Guangzhou 510006, Guangdong, Peoples R China.
关键词：
Palladium;Fluorescence sensor;NBD;Tsuji–Trost reaction
摘要：
Abstract A new chemosensor 7-nitro-2,1,3-benzoxadiazole-4-allyl-N-(thiophen-2-ylmethyl)carbamate (NBDTC) was synthesized and utilized for palladium detection based on the Tsuji-Trost reaction. NBDTC displayed specific and ratiometric fluorescent responses toward palladium species. The chemosensor showed more than 50-fold enhancement in fluorescence intensity with the presence of PEG400 and palladium because NBDTC can be transformed to NBDT under palladium-catalyzing Tsuji-Trost reaction. NBDTC displayed high selectivity and sensitivity for palladium species with the detection limit of 1.13 ×10<sup>-9</sup>M. ©2015 Published by Elsevier B.V.
语种：
英文
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A near-infrared BODIPY-based fluorescent probe for the detection of hydrogen sulfide in fetal bovine serum and living cells
作者：
Zhang, Jian;Zhou, Junliang;Dong, Xiaochun* ;Zheng, Xing;Zhao, Weili
期刊：
RSC Advances ,2016年6(56):51304-51309 ISSN：2046-2069
通讯作者：
Dong, Xiaochun
作者机构：
[Dong, Xiaochun; Zhang, Jian; Zhao, Weili] Fudan Univ, Sch Pharm, Shanghai 200120, Peoples R China.;[Zhou, Junliang; Zheng, Xing] Univ South China, Inst Pharm & Pharmacol, Hengyang 421001, Peoples R China.
通讯机构：
[Dong, Xiaochun] Fudan Univ, Sch Pharm, Shanghai 200120, Peoples R China.
摘要：
A near-infrared (NIR) "off-on" fluorescent probe BDP-680 was developed by taking advantage of the Michael addition reaction of unsaturated double bonds in conformationally restricted BODIPY to allow detection of H<inf>2</inf>S. The new probe possesses a highly selective and sensitive response to H<inf>2</inf>S. Such a NIR probe has been applied successfully to detect H<inf>2</inf>S in 10% deproteinized fetal bovine serum (FBS). The detection limit of the probe for NaSH in 10% deproteinized FBS sample was calculated to be about 1 μM which is consistent with the value obtained from serum free solutions (0.5 μM). The probe is of low toxicity and has been successfully used to detect H<inf>2</inf>S in living cells. ©2016 The Royal Society of Chemistry.
语种：
英文
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Design, synthesis and biological evaluation of flavonoid salicylate derivatives as potential anti-tumor agents
作者：
Deng, Xiangping;Wang, Zhe;Liu, Juan;Xiong, Shujuan;Xiong, Runde;Cao, Xuan;Chen, Yanming;Zheng, Xing;Tang, Guotao*
期刊：
RSC Advances ,2017年7(61):38171-38178 ISSN：2046-2069
通讯作者：
Tang, Guotao
作者机构：
[Wang, Zhe; Deng, Xiangping; Liu, Juan; Xiong, Shujuan; Tang, Guotao; Xiong, Runde; Cao, Xuan; Zheng, Xing] Univ South China, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Inst Pharm & Pharmacol, Hengyang 421001, Hunan, Peoples R China.;[Chen, Yanming] Mudanjiang Youbo Pharmaceut Co Ltd, Mudanjiang, Peoples R China.
通讯机构：
[Tang, Guotao] Univ South China, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Inst Pharm & Pharmacol, Hengyang 421001, Hunan, Peoples R China.
摘要：
A series of flavonoid salicylate derivatives containing trimethoxybenzene and a series of chrysin salicylate derivatives were synthesized for use as anti-tumor agents, and evaluated for antiproliferative activity using three human tumor cells: MCF-7 (breast carcinoma cells), HepG2 (liver carcinoma cells), MGC-803 (gastric carcinoma cells) and the mice tumor cells MFC (forestomach carcinoma cells). A substituent group of a suitable size and the trimethoxybenzene had a certain influence on the bioactivity of the flavonoid salicylate derivatives. Compound 2 and its salicylate derivatives 7a-7g containing the trimethoxybenzene exhibited more antiproliferative activity. Among them, compound 7g displayed the most potent antiproliferative activity against MGC-803 cells and MFC cells with the concentration causing 50% inhibition of cell growth (IC<inf>50</inf>) values of 11.05 ±1.58 μM and 13.73 ±2.04 μM, respectively. The flow cytometry results showed that compound 7g caused the cell cycle to be arrested in the G0/G1 phase and induced apoptosis of MFC cells in a dose-dependent manner. Furthermore, compound 7g showed good anti-tumor activity in vivo. These results suggested that compound 7g could be a new, potent anti-tumor candidate which should be optimized and evaluated further. ©2017 The Royal Society of Chemistry.
语种：
英文
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Hydrogen sulfide alleviates uranium-induced acute hepatotoxicity in rats: Role of antioxidant and antiapoptotic signaling
作者：
Yuan, Yan;Zheng, Jifang* ;Zhao, Tingting;Tang, Xiaoqing;Hu, Nan
期刊：
Environmental Toxicology ,2017年32(2):581-593 ISSN：1520-4081
通讯作者：
Zheng, Jifang
作者机构：
[Zheng, Jifang; Yuan, Yan; Zhao, Tingting] Univ South China, Sch Pharmaceut & Biol Sci, Inst Biol, Dept Biol, Changsheng West Rd 28, Hengyang City 421001, Hunan, Peoples R China.;[Tang, Xiaoqing] Univ South China, Coll Med, Inst Neurosci, Dept Physiol, Changsheng West Rd 28, Hengyang City 421001, Hunan, Peoples R China.;[Hu, Nan] Univ South China, Key Discipline Lab Natl Def Biotechnol Uranium Mi, Changsheng West Rd 28, Hengyang City 421001, Hunan, Peoples R China.
通讯机构：
[Zheng, Jifang] Univ South China, Sch Pharmaceut & Biol Sci, Inst Biol, Dept Biol, Changsheng West Rd 28, Hengyang City 421001, Hunan, Peoples R China.
关键词：
hydrogen sulfide;uranium;hepatotoxicity;oxidative stress;apoptosis
摘要：
As an endogenous gaseous mediator, H<inf>2</inf>S exerts antioxidative, antiapoptotic, and cytoprotective effects in livers. This study was designed to investigate the protective role of H<inf>2</inf>S against uranium-induced hepatotoxicity in adult SD male rats after in vivo effect of uranium on endogenous H<inf>2</inf>S production was determined in livers. The levels of endogenous H<inf>2</inf>S and H<inf>2</inf>S-producing enzymes (CBS and CSE) were measured in liver homogenates from uranium -intoxicated rats. In rats injected intraperitoneally (i.p.) with uranyl acetate or NaHS (an H<inf>2</inf>S donor) alone or in combination, we examined biochemical parameters to assess liver function, revealed hepatic histopathological alteration, investigated oxidative stress markers, and explored apoptotic signaling in liver homogenates. The results suggest that uranium-intoxication in rats decreased CBS and CSE protein expression, H<inf>2</inf>S synthesis capacity, and endogenous H<inf>2</inf>S generation. NaHS administration in uranium-intoxicated rats produced amelioration in liver biochemical indices and histopathological effects, decreased MDA content, and increased GSH level and antioxidative enzymes activities like SOD, CAT, GPx, and GST. NaHS administration in uranium-intoxicated rats attenuated uranium-activated phosphorylation state of JNK. NaHS treatment in uranium-intoxicated rats increased antiapoptotic Bcl-2 but decreased pro-apoptotic Bax, resulting in the rise of Bcl-2/Bax ratio. NaHS treatment in uranium-intoxicated rats reduced the apoptosis mediator caspase-3 and cytochrome c release and elevated ATP contents. Taken together, these data implicate that H<inf>2</inf>S can afford protection to rat livers against uranium-induced adverse effects mediated by up-regulation of antioxidant and antiapoptotic signaling. The anti-apoptotic property of H<inf>2</inf>S may be involved, at least in part, in inhibiting JNK signaling. ©2016 Wiley Periodicals, Inc. Environ Toxicol 32: 581–593, 2017. ©2016 Wiley Periodicals, Inc.
语种：
英文
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Direct Imaging of Single Plasmonic Metal Nanoparticles in Capillary with Laser Light-Sheet Scattering Imaging
作者：
Cao, Xuan;Feng, Jingjing;Pan, Qi;Xiong, Bin;He, Yan* ;Yeung, Edward S.
期刊：
Analytical Chemistry ,2017年89(5):2692-2697 ISSN：0003-2700
通讯作者：
He, Yan
作者机构：
[Xiong, Bin; He, Yan; Cao, Xuan] Hunan Univ, Coll Chem & Chem Engn, Coll Biol, State Key Lab Chemobiosensing & Chemometr, Changsha 410082, Hunan, Peoples R China.;[Pan, Qi; He, Yan; Feng, Jingjing] Tsinghua Univ, Dept Chem, Beijing 100084, Peoples R China.;[Cao, Xuan] Univ South China, Inst Pharm & Pharmacol, Hengyang 421001, Hunan, Peoples R China.;[Yeung, Edward S.] Iowa State Univ, Dept Chem, Ames, IA 50011 USA.
通讯机构：
[He, Yan] Hunan Univ, Coll Chem & Chem Engn, Coll Biol, State Key Lab Chemobiosensing & Chemometr, Changsha 410082, Hunan, Peoples R China.;[He, Yan] Tsinghua Univ, Dept Chem, Beijing 100084, Peoples R China.
摘要：
Understanding the heterogeneous distribution of the physical and chemical properties of plasmonic metal nanoparticles is fundamentally important to their basic and applied research. Traditionally, they are obtained either indirectly via bulk spectroscopic measurements plus electron microscopic characterizations or through single molecule/particle imaging of nanoparticles immobilized on planar substrates. In this study, by using light-sheet scattering microscopy with a supercontinuum white laser, highly sensitive imaging of individual metal nanoparticles (MNPs) flowing inside a capillary, driven by either pressure or electric field, was achieved for the first time. We demonstrate that single plasmonic nanoparticles with different size or chemical modification could be differentiated through their electrophoretic mobility in a few minutes. This technique could potentially be applied to high throughput characterization and evaluation of single metal nanoparticles as well as their dynamic interactions with various local environments.
语种：
英文
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Diastereoselective Synthesis of CF3-Containing Vicinal Diamines
作者：
Huang, Qiu-xia;Zheng, Qu-tong;Duan, Yaya;Lin, Jin-Hong* ;Xiao, Ji-Chang* ;Zheng, Xing*
期刊：
Journal of Organic Chemistry ,2017年82(15):8273-8281 ISSN：0022-3263
通讯作者：
Xiao, Ji-Chang;Zheng, Xing;Lin, Jin-Hong
作者机构：
[Xiao, Ji-Chang; Zheng, Xing; Huang, Qiu-xia; Zheng, Qu-tong] Univ South China, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Inst Pharm & Pharmacol, 28 Western Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.;[Lin, JH; Xiao, Ji-Chang; Zheng, Xing; Huang, Qiu-xia; Duan, Yaya; Lin, Jin-Hong; Zheng, Qu-tong] Univ Chinese Acad Sci, Chinese Acad Sci, Shanghai Inst Organ Chem, Key Lab Organofluorine Chem, 345 Lingling Rd, Shanghai 200032, Peoples R China.
通讯机构：
[Xiao, JC; Zheng, X] Univ South China, Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Inst Pharm & Pharmacol, 28 Western Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.;[Lin, JH; Xiao, JC; Zheng, X] Univ Chinese Acad Sci, Chinese Acad Sci, Shanghai Inst Organ Chem, Key Lab Organofluorine Chem, 345 Lingling Rd, Shanghai 200032, Peoples R China.
摘要：
The highly diastereoselective synthesis of CF3-containing vicinal diamines by a convenient two-step procedure without the need to isolate the intermediate products is described.
语种：
英文
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Nanohydroxyapatite/cellulose nanocrystals/silk fibroin ternary scaffolds for rat calvarial defect regeneration
作者：
Chen, Xiaoming;Zhou, Runmei;Chen, Bin;Chen, Jianting*
期刊：
RSC Advances ,2016年6(42):35684-35691 ISSN：2046-2069
通讯作者：
Chen, Jianting
作者机构：
[Chen, Jianting; Chen, Xiaoming] Southern Med Univ, Nanfang Hosp, Dept Orthoped Spinal Surg, Guangzhou 510515, Guangdong, Peoples R China.;[Chen, Xiaoming] Univ South China, Affiliated Hosp 2, Dept Orthoped Spinal Surg, Hengyang 421001, Peoples R China.;[Zhou, Runmei] Univ South China, Affiliated Hosp 2, Dept Pharm, Hengyang 421001, Peoples R China.;[Chen, Bin] Chenzhou 1 Peoples Hosp, Dept Orthoped Spinal Surg, Chenzhou 423000, Peoples R China.
通讯机构：
[Chen, Jianting] Southern Med Univ, Nanfang Hosp, Dept Orthoped Spinal Surg, Guangzhou 510515, Guangdong, Peoples R China.
摘要：
The purpose of this study was to design and characterise a novel biomimetic scaffold for the repair of critical size calvarial defects. In this study, we developed a new hydroxyapatite/cellulose nanocrystals/silk fibroin (HA/CNC/SF) scaffold by mixing a solution of SF with HA and CNC nanoparticles. The scaffold was fabricated by freeze-drying. The average pore size and porosity of the HA/CNC/SF scaffolds were 110 ±7.3 μm and 90 ±6.2%, respectively. The thermostability and mechanical properties of the scaffolds were significantly better than those of either SF, CNC/SF or HA/SF scaffolds. Additionally, the composite scaffold exhibited excellent biocompatibility and superior osteoconductivity. Hence, the efficacy of the HA/CNC/SF scaffold was evaluated for bone regeneration in the rat calvarial defect model. The results demonstrated that the rat calvarial defect healed with new-formed bone within 12 weeks of implantation and the degradation rate of the scaffold was a good match to the bone regeneration rate. Therefore, the HA/CNC/SF scaffold is considered to be a strong candidate for the repair of bone defects in bone tissue engineering. ©The Royal Society of Chemistry 2016.
语种：
英文
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Simultaneous fluorescence analysis of the different carbohydrates expressed on living cell surfaces using functionalized quantum dots
作者：
He, Dongxiu* ;Wang, Danxia;Shi, Xiaoxin;Quan, Wenjie;Xiong, Runde;Yua, Cui-yun;Huang, Honglin
期刊：
RSC Advances ,2017年7(20):12374-12381 ISSN：2046-2069
通讯作者：
He, Dongxiu
作者机构：
[Huang, Honglin; Yua, Cui-yun; He, Dongxiu; Wang, Danxia] Univ South China, Inst Pharm & Pharmacol, Hengyang, Hunan, Peoples R China.;[Quan, Wenjie; Shi, Xiaoxin; Yua, Cui-yun; He, Dongxiu; Xiong, Runde] Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang, Hunan, Peoples R China.
通讯机构：
[He, Dongxiu] Univ South China, Inst Pharm & Pharmacol, Hengyang, Hunan, Peoples R China.;[He, Dongxiu] Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang, Hunan, Peoples R China.
摘要：
The aberrant expression of carbohydrates has been associated with the occurrence, growth, progression and metastasis of tumors. Tumor-associated carbohydrates may have great potential as tumor markers for the early diagnosis of hepatocellular carcinoma (HCC). Therefore, characterizing HCC-associated carbohydrate expression is of great importance to assist the early diagnosis of HCC. A fluorescence method for characterizing carbohydrates expressed on both normal human cells (LO2 cells and endothelial cells) and HCC cells (HepG2 cells) using functionalized quantum dots (QDs) has been proposed in this study. The QDs were successfully fabricated and covalently conjugated with Datura stramonium agglutinin (DSA) or Lens culinaris agglutinin (LCA) via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) coupling reaction. The formed functionalized QDs (lectin-QDs conjugates) were characterized using ultraviolet and fluorescence spectra, agarose gel electrophoresis, hemagglutination activity tests, lectin competitive-binding assay and carbohydrate inhibition assays. The functionalized QDs were found to retain stable fluorescence and carbohydrate recognition abilities. Significant differences of carbohydrates expressed between on the normal cells and the HepG2 cells were evaluated by fluorescence imaging and flow cytometric analysis. The experimental results illustrate that the functionalized QDs could be used as promising tools for monitoring in situ cell surface carbohydrate expression and evaluating the differences in the carbohydrates expressed on normal cells and tumor cells surfaces, which is very important for helping the early diagnosis of HCC. ©The Royal Society of Chemistry.
语种：
英文
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Discrimination of A1555G and C1494T point mutations in the mitochondrial 12S rRNA Gene by On/Off switch
作者：
Guo, Zi-Fen;Guo, Wu-Shuang;Xiao, Li;Gao, Guo-Qiang;Lan, Fen;Lu, Xue-Guan;Li, Kai* ;Liao, Duan-Fang
期刊：
Applied Biochemistry and Biotechnology ,2012年166(1):234-242 ISSN：0273-2289
通讯作者：
Li, Kai
作者机构：
[Guo, Zi-Fen; Lan, Fen; Gao, Guo-Qiang; Guo, Wu-Shuang; Liao, Duan-Fang] Univ S China, Inst Pharm & Pharmacol, Hengyang 421001, Hunan, Peoples R China.;[Li, Kai] Soochow Univ, Coll Med, Dept Pharmacol, Suzhou 215123, Jiangsu, Peoples R China.;[Lu, Xue-Guan; Xiao, Li] Soochow Univ, Affiliated Hosp 2, Mol Med Ctr, Suzhou 215004, Jiangsu, Peoples R China.;[Liao, Duan-Fang] Hunan Univ Chinese Med, Sch Pharm, Dept Tradit Chinese Diognot, Changsha 410208, Hunan, Peoples R China.
通讯机构：
[Li, Kai] Soochow Univ, Coll Med, Dept Pharmacol, Suzhou 215123, Jiangsu, Peoples R China.
关键词：
Mitochondrial 12S rRNA;Exo(+) polymerase;Phosphorothioate modification;Aminoglycoside antibiotics
摘要：
The objective of this study was to apply the "on/off" switch consisting of 3' phosphorothioate-modified allele specific primers and exo(+) polymerase in single base discrimination of A1555G and C1494T mutations in the highly conserved sites of the mitochondrial 12S rRNA. The two point mutations are the hotspot mutations associated with either aminoglycoside antibiotics induced deafness or inherited nonsyndromic hearing loss. The PCR products of mitochondrial DNA (mtDNA) 12S rRNA gene were inserted into the pMD19-T vector for transformation into Escherichia coli JM109 competent cells for preparing wild-type pMD19-T/mt vector. Inverse PCR was carried out for mtDNA 12S rRNA gene C1494T and A1555G mutagenesis and DpnI endonuclease degradating methylated pMD19-T/mt vector existing in the inverse PCR products was carried out to construct the mutation-type pMD19-T/mtM vector. These constructed vectors were confirmed by DNA sequencing. Allelic specific primers targeting wild-type and mutation-type templates were designed with 3' terminal phosphorothioate modification. Two-directional primer extension was performed using Pfu polymerases. Amplified by exo(+) polymerase, allelic specific primers perfectly matching wild-type allele were extended while no products were produced from primers targeting point-mutated deafness-related allele. Similarly, allelic specific primers perfectly matching point-mutated deafness-related mutation-type allele were extended and no products were yielded from primers targeting wild-type allele. No specific product was observed in the primer extension reaction mediated by on/off switch in screening the mtDNA 12S rRNA gene harboring either C1494T or A1555G mutation in 40 healthy volunteers tested. These data suggest that the "off switch" mediated by exo(+) polymerase is highly reliable in the diagnosis of monogenic diseases and the novel "on/off" switch has enormous applications in systematic and extended screening of the12S rRNA gene A1555G and C1494T mutations. The established assay can be widely used not only for hearing loss patients but also for normal subjects before the use of aminoglycoside antibiotics.
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英文
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