LIGHT 信号通路在鼠衣原体生殖道感染中的作用
作者：
陈丽丽;孙源彬;徐莎;符玺宗;周洲;...
期刊：
微生物学报 ,2015年55(4):492-500 ISSN：0001-6209
作者机构：
[陈丽丽; 周洲; 陆春雪; 吴移谋] 南华大学病原生物学研究所;[孙源彬; 徐莎; 符玺宗] 南华大学公共卫生学院;[杨菲; 许桂莲] 第三军医大学免疫学研究所
关键词：
鼠衣原体;与单纯疱疹病毒糖蛋白D 竞争结合疱疹病毒侵入介体的淋巴毒素类似物;泌尿生殖道感染;病理损伤
摘要：
【目的】初步探讨与单纯疱疹病毒糖蛋白D 竞争结合疱疹病毒侵入介体的淋巴毒素类似物(lymphotoxin-like inducible protein that competes with glycoprotein D for herpesvirus entry on T cells,LIGHT)在抗衣原体感染免疫及介导衣原体生殖道病理损伤过程的作用。【方法】用1 × 10~4 IFUs 的MoPn 经生殖道感染野生型(wild type,wt) 、LIGHT KO 小鼠,每组一半小鼠于感染后49d,再次感染相同剂量的MoPn。每隔3 - 4 d取生殖道分泌物,测定其中衣原体包涵体的数量。初次感染后80d,处死小鼠,眼眶取血,分离血清,用间接免疫荧光法测定其中抗体类型及效价; 同时分离生殖道,肉眼观察其输卵管、子宫角水肿程度,然后甲醛固定、切片,H&E 染色后,显微镜下观察各组织炎性浸润程度和管腔水肿程度。分离小鼠脾细胞,体外用衣原体EB 刺激,测定上清中IL-4、IL-5、IL-17和IFN-γ 等细胞因子水平。【结果】LIGHT KO 小鼠阴道带菌时间与wt 组相当,大部分小鼠均在原发感染后28d 左右完全清除感染,且均产生对再次感染的免疫力。LIGHT KO 和wt 小鼠子宫角和输卵管均出现一定程度的病变,但差异无统计学意义。两组小鼠在原发和继发感染MoPn 后,均产生高效价的特异性抗MoPn IgG 抗体,总抗体及各IgG 抗体亚类效价差异均无统计学意义(P > 0. 05),且IgG2a /IgG1比值均大于1。和wt 小鼠一样,LIGHT KO 小鼠脾淋巴细胞经衣原体再次刺激后均可产生较高水平的IFN-γ 和IL-17,且未能检测到IL-4和IL-5。【结论】小鼠抗MoPn 生殖道感染及MoPn 引起的生殖道病理损伤不依赖于LIGHT 信号通路。
语种：
中文
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Insulin receptor binding motif tagged with igG4 Fc (Yiminsu) works as an insulin sensitizer to activate Akt signaling in hepatocytes
作者：
Wang, J.;Zou, T.;Yang, H. X.;Gong, Y. Z.;Xie, X. J.;...
期刊：
Genetics and Molecular Research ,2015年14(3):8819-8828 ISSN：1676-5680
通讯作者：
Liu, H. Y.
作者机构：
[Yang, H. X.; Wang, J.] Univ South China, Key Lab Atherosclerol Hunan Prov, Inst Cardiovasc Res, Hengyang, Hunan, Peoples R China.;[Yang, H. X.; Wang, J.] Univ South China, Sch Publ Hlth, Hengyang, Hunan, Peoples R China.;[Zou, T.] Univ South China, Affiliated Hosp 1, Dept Cardiovasc Med, Hengyang, Hunan, Peoples R China.;[Gong, Y. Z.; Xie, X. J.; Liao, D. F.] Hunan Univ Chinese Med, State Key Lab Chinese Med Powder & Med Innovat Hu, Div Stem Cell Regulat & Applicat, Changsha, Hunan, Peoples R China.;[Liu, H. Y.] Metammune LLC, Morrisville, NC 27560 USA.
通讯机构：
[Liu, H. Y.] M;Metammune LLC, Morrisville, NC 27560 USA.
关键词：
IgG4 Fc;Insulin sensitizer;Type 2 diabetes;Yiminsu
摘要：
Insulin resistance is a key feature of obesity and type 2 diabetes mellitus (T2DM). Interaction of insulin with the insulin receptor (IR) leads to both its auto-phosphorylation and phosphorylation of tyrosine residues on the IR substrate (IRS) proteins, initiating the activation of intracellular signaling cascades. The metabolic effects of IRS are known to be mediated through pathways involving phosphatidyl-inositol 3-kinase (PI-3K), which result in the activation of Akt signaling. The C-terminal region of the IR ectodomain is required to facilitate the conformational changes that are required for high-affinity binding to insulin. Furthermore, the CH2 and CH3 domains in the Fc fragments of immunoglobulins are responsible for their binding to the Fc receptor, which triggers transcytosis. In this study, we created a fusion peptide of the C-terminal end of the human IR ectodomain with the IgG4 Fc fragment, including an intervening polyG fragment to ensure enough space for insulin binding. We named this new peptide “Yiminsu”, meaning an insulin sensitizer. The results of our analyses show that Yiminsu significantly facilitates insulin signaling via the activation of Akt in hepatocytes in a dose- and time-dependent manner. Further studies are required to determine whether Yiminsu can act as an insulin sensitizer. © FUNPEC-RP.
语种：
英文
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Silver nanoparticles activate endoplasmic reticulum stress signaling pathway in cell and mouse models: The role in toxicity evaluation
作者：
Huo, Lingling;Chen, Rui;Zhao, Lin;Shi, Xiaofei;Bai, Ru;...
期刊：
Biomaterials ,2015年61:307-315 ISSN：0142-9612
通讯作者：
Chen, Chunying
作者机构：
[Chang, Yan-Zhong; Huo, Lingling] Hebei Normal Univ, Key Lab Anim Physiol Biochem & Mol Biol Hebei Pro, Key Lab Mol & Cellular Biol, Minist Educ,Coll Life Sci, Shijiazhuang 050016, Peoples R China.;[Shi, Xiaofei; Zhao, Yuliang; Zhao, Lin; Chen, Chunying; Bai, Ru; Chen, Rui; Huo, Lingling] Natl Ctr Nanosci & Technol China, CAS Key Lab Biomed Effects Nanomat & Nanosafety, Beijing 100190, Peoples R China.;[Shi, Xiaofei; Long, Dingxin; Chen, Feng; Zhao, Lin] Univ South China, Sch Publ Hlth, Hengyang 421001, Peoples R China.;[Chen, Chunying] Natl Ctr Nanosci & Technol China, 11 Beiyitiao, Beijing 100190, Peoples R China.
通讯机构：
[Chen, Chunying] N;Natl Ctr Nanosci & Technol China, 11 Beiyitiao, Beijing 100190, Peoples R China.
关键词：
Apoptosis;Cytotoxicity;Intratracheal instillation;Signaling pathway;Silver
摘要：
Silver nanoparticles (AgNPs) attract considerable public attention both for their antimicrobial properties and their potential adverse effects. In the present study, endoplasmic reticulum (ER) stress was used as a sensitive and early biomarker to evaluate the toxic potential of AgNPs in three different human cell lines in vitro and in vivo in mice. In 16HBE cells, the activation of ER stress signaling pathway was observed by upregulated expression including xbp-1s, chop/. DDIT3, TRIB3, ADM2, BIP, Caspase-12, ASNS and HERP at either the mRNA and/or protein levels. However, these changes were not observed in HUVECs or HepG2 cells. Furthermore, mice experiments showed that different tissues had various sensitivities to AgNPs following intratracheal instillation exposure. The lung, liver and kidney showed significant ER stress responses, however, only the lung and kidney exhibited apoptosis by TUNEL assay. The artery and tracheal tissues had lower ER stress and apoptosis after exposure. The lowest observable effect concentrations (LOEC) were proposed based on evaluation of AgNP induced ER stress response in cell and mouse models. In summary, preliminary evaluation of AgNP toxicity by monitoring the ER stress signaling pathway provides new insights toward the understanding the biological impacts of AgNPs. The adverse effects of exposure to AgNPs may be avoided by rational use within the safe dose. © 2015 Elsevier Ltd.
语种：
英文
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Single Nanoparticle to 3D Supercage: Framing for an Artificial Enzyme System
作者：
Cai, Ren;Yang, Dan;Peng, Shengjie;Chen, Xigao;Huang, Yun;...
期刊：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY ,2015年137(43):13957-13963 ISSN：0002-7863
通讯作者：
Tan, Weihong
作者机构：
[Yang, Shengyuan; Liu, Yuan; Liu, Zhenbao; Chen, Xigao; Hou, Weijia; Huang, Yun; Cai, Ren; Tan, Weihong] Univ Florida, Dept Chem, Ctr Res Bio Nano Interface, Gainesville, FL 32611 USA.;[Yang, Shengyuan; Liu, Yuan; Liu, Zhenbao; Chen, Xigao; Hou, Weijia; Huang, Yun; Cai, Ren; Tan, Weihong] Univ Florida, UF Genet Inst, Hlth Canc Ctr, Dept Physiol & Funct Genom, Gainesville, FL 32611 USA.;[Yang, Shengyuan; Liu, Yuan; Liu, Zhenbao; Chen, Xigao; Hou, Weijia; Huang, Yun; Cai, Ren; Tan, Weihong] Univ Florida, McKnight Brain Inst, Gainesville, FL 32611 USA.;[Tan, Weihong] Hunan Univ, Coll Biol, Coll Chem & Chem Engn, Mol Sci & Biomed Lab,State Key Lab Chemo Bio Sen, Changsha 410082, Hunan, Peoples R China.;[Tan, Weihong] Hunan Univ, Collaborat Res Ctr Mol Engn Theranost, Changsha 410082, Hunan, Peoples R China.
通讯机构：
[Tan, Weihong] U;Univ Florida, Dept Chem, Ctr Res Bio Nano Interface, Gainesville, FL 32611 USA.
摘要：
A facile strategy has been developed to fabricate Cu(OH)2 supercages (SCs) as an artificial enzyme system with intrinsic peroxidase-mimic activities (PMA). SCs with high catalytic activity and excellent recyclability were generated via direct conversion of amorphous Cu(OH)2 nanoparticles (NPs) at room temperature. More specifically, the process that takes a single nanoparticle to a 3D supercage involves two basic steps. First, with addition of a copper-ammonia complex, the Cu2+ ions that are located on the surface of amorphous Cu(OH)2 NPs would evolve into a fine lamellar structure by coordination and migration and eventually convert to 1D nanoribbons around the NPs. Second, accompanied by the migration of Cu2+, a hollow cavity is generated in the inner NPs, such that a single nanoparticle eventually becomes a nanoribbon-assembled 3D hollow cage. These Cu(OH)2 SCs were then engineered as an artificial enzymatic system with higher efficiency for intrinsic PMA than the peroxidase activity of a natural enzyme, horseradish peroxidase. © 2015 American Chemical Society.
语种：
英文
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STAT1-Mediated Down-Regulation of Bcl-2 Expression Is Involved in IFN-gamma/TNF-alpha-Induced Apoptosis in NIT-1 Cells
作者：
Cao, Zhao-hui;Zheng, Quan-you;Li, Gui-qing;Hu, Xiao-bo;Feng, Shao-long;...
期刊：
PLOS ONE ,2015年10(3):e0120921 ISSN：1932-6203
通讯作者：
Xu, Gui-lian
作者机构：
[Zheng, Quan-you; Zhang, Ke-qin; Cao, Zhao-hui] Third Mil Med Univ, Daping Hosp, Inst Surg Res, Dept Urol, Chongqing 400042, Peoples R China.;[Li, Gui-qing; Xu, Gui-lian] Third Mil Med Univ, Dept Immunol, Chongqing, Peoples R China.;[Cao, Zhao-hui; Hu, Xiao-bo] Univ South China, Sch Publ Hlth, Sch Pharm & Biol Sci, Dept Biochem & Mol Biol, Hengyang 421001, Peoples R China.;[Feng, Shao-long] Univ South China, Sch Publ Hlth, Dept Hlth Lab Technol, Hengyang 421001, Peoples R China.
通讯机构：
[Xu, Gui-lian] T;Third Mil Med Univ, Dept Immunol, Chongqing, Peoples R China.
关键词：
Apoptosis;Mitochondria;Scanning confocal microscopy;Cytokines;Cytokine therapy;Small interfering RNA;Flow cytometry;Lasers
摘要：
Tumor necrosis factor (TNF)-α and interferon (IFN)-γ are the major pro-inflammatory cytokines involved in beta-cell destruction. The fate of islet beta-cells in the cytokine-induced intrinsic mitochondrial apoptotic pathway is determined by the interaction between members of the Bcl-2 family. However, the mechanism through which beta-cell apoptosis is regulated remains unclear. In this study, we treated the murine beta-cell line NIT-1 with TNF-α and IFN-γ and then investigated the regulation of signal transducer and activator of transcription-1 (STAT-1) and expression of the members of the Bcl-2 family in this apoptotic pathway. Results showed that TNF-α and IFN-γ synergistically reduced NIT-1 cell viability. In addition, the decrease in cell growth was due to apoptosis as shown by apoptotic body formation, detected by confocal laser microscope, and a significant increase in Annexin-Vup+ cell percentage, detected by flow cytometry. Combination treatment with TNF-α and IFN-γ caused a remarkable increase in the release of cytochrome c, and in the activation of caspase-9 and caspase-3, as well as, an obvious enhancement in STAT-1 phosphorylation; the treatment, however, resulted in the down-regulation in Bcl-2 expression. The enhancement in STAT-1 activity and a down-regulation in Bcl-2 expression was also observed in MIN6 cells, another murine beta-cell derived line, after cells exposure to the combination of TNF-α and IFN-γ treatment. Knockdown of STAT-1 gene expression by siRNA or inhibition of STAT-1 activation with fludarabine reversed Bcl-2 down-expression and led to a significant decrease in apoptosis in TNF-α- and IFN-γ-treated NIT-1 cells. Taken together, our results suggest that STAT1-mediated down-regulation of Bcl-2 is involved in NIT-1 cell apoptosis induced by combination treatment with TNF-α and IFN-γ.
语种：
英文
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Characterization and complete genome sequence analysis of Staphylococcus aureus bacteriophage JS01
作者：
Jia, Hongying;Dong, Wenyang;Yuan, Lvfeng;Ma, Jiale;Bai, Qinqin;...
期刊：
Virus Genes ,2015年50(2):345-348 ISSN：0920-8569
通讯作者：
Yao, Huochun
作者机构：
[Yuan, Lvfeng; Pan, Zihao; Dong, Wenyang; Yao, Huochun; Ma, Jiale; Jia, Hongying; Lu, Chengping] Nanjing Agr Univ, Coll Vet Med, Nanjing 210095, Jiangsu, Peoples R China.;[Yuan, Lvfeng; Pan, Zihao; Dong, Wenyang; Yao, Huochun; Ma, Jiale; Jia, Hongying; Lu, Chengping] Minist Agr, Key Lab Anim Bacteriol, Nanjing 210095, Jiangsu, Peoples R China.;[Bai, Qinqin] Univ South China, Coll Publ Hlth, Hengyang 421000, Peoples R China.
通讯机构：
[Yao, Huochun] M;Minist Agr, Key Lab Anim Bacteriol, Nanjing 210095, Jiangsu, Peoples R China.
关键词：
Bovine mastitis;Staphylococcus aureus phage;Genome sequence analysis
摘要：
Staphylococcus aureus is a primary pathogen that causes bovine mastitis resulting in serious economic losses and herd management problems in dairy cows. A novel bacteriophage, JS01, specifically infecting bovine S. aureus, was isolated from milk of mastitis-affected cattle. TEM observation showed that it belonged to the family Siphovirus. The JS01 strain demonstrated a broad host range. The prediction result of PHACTS suggested that the JS01 strain was temperate phage. The JS01 genome is 43,458 bp long, with a GC content of 33.32 % and no tRNAs. Annotation and functional analysis of the predicted ORFs revealed six functional groups: structure and morphology, DNA replication and regulation, packaging, lysogeny, lysis, and pathogenicity. Comparative analysis between JS01, S. aureus MSSA476, and S. aureus prophage PVL was also performed. The characterization and genomic analysis of JS01 provide a better understanding of S. aureus-targeting bacteriophages and useful information for the development of phage-based biocontrol agents against S. aureus.
语种：
英文
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Neuropathy Target Esterase Is Degraded by the Ubiquitin-Proteasome Pathway with ARA54 as the Ubiquitin Ligase
作者：
Long, Ding-Xin;Wang, Pan;Sun, Ying-Jian;Chen, Rui;Wu, Yi-Jun*
期刊：
BIOCHEMISTRY ,2015年54(50):7385-7392 ISSN：0006-2960
通讯作者：
Wu, Yi-Jun
作者机构：
[Sun, Ying-Jian; Wang, Pan; Wu, Yi-Jun; Long, Ding-Xin; Chen, Rui] Chinese Acad Sci, Inst Zool, State Key Lab Integrated Management Pest Insects, Lab Mol Toxicol, Beijing 100101, Peoples R China.;[Long, Ding-Xin] Univ South China, Sch Publ Hlth, Hengyang 421001, Peoples R China.;[Sun, Ying-Jian] Beijing Agr Coll, Dept Vet Med & Anim Sci, Beijing 102206, Peoples R China.;[Wu, Yi-Jun] Chinese Acad Sci, Inst Zool, 1-5 Beichenxilu Rd, Beijing 100101, Peoples R China.
通讯机构：
[Wu, Yi-Jun] C;Chinese Acad Sci, Inst Zool, 1-5 Beichenxilu Rd, Beijing 100101, Peoples R China.
摘要：
Neuropathy target esterase (NTE) is an endoplasmic reticulum membrane-associated phospholipase B, which is essential for embryonic and nervous system development. However, the regulation of NTE at the protein level had not been thoroughly investigated. Our previous study showed that NTE was degraded not only by the macroautophagy-lysosome pathway but also by the ubiquitin-proteasome pathway. Here we further reveal that androgen receptor-associated protein 54 (ARA54) regulated the ubiquitin-proteasome degradation of NTE. We find that deletion of the regulatory domain of NTE, which possesses a putative destruction box and thus is essential for its degradation by the proteasome, prevented its degradation by the proteasome. In addition, we demonstrate that ARA54, which has a RING finger domain and E3 ligase activity, interacts directly with NTE. Overexpression of ARA54 downregulates the protein level of NTE, and knockdown of ARA54 inhibits the degradation of NTE. The mutation in the RING domain of ARA54 blocks the degradation of NTE by ARA54, which indicates that the RING domain is essential for ARA54's E3 activity. These findings suggest that ARA54 acts as the ubiquitin ligase to regulate the ubiquitin-proteasome degradation of NTE. © 2015 American Chemical Society.
语种：
英文
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Ultrasensitive detection of uranyl by graphene oxide-based background reduction and RCDzyme-based enzyme strand recycling signal amplification
作者：
Li, Ming-Hui;Wang, Yong-Sheng* ;Cao, Jin-Xiu;Chen, Si-Han;Tang, Xian;...
期刊：
Biosensors and Bioelectronics ,2015年72:294-299 ISSN：0956-5663
通讯作者：
Wang, Yong-Sheng
作者机构：
[Chen, Si-Han; Li, Ming-Hui; Tang, Xian; Cao, Jin-Xiu; Zhu, Yu-Feng; Wang, Yong-Sheng; Huang, Yan-Qin; Zhu, Xiao-Feng] Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.
通讯机构：
[Wang, Yong-Sheng] U;Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.
关键词：
Enzyme strand recycling amplification;G-quadruplexes;Graphene oxide;N-methyl- mesoporphyrin IX;RNA-cleaving chimera DNAzyme;Zero-background signal
摘要：
We proposed a novel strategy which combines graphene oxide-based background reduction with RCDzyme-based enzyme strand recycling amplification for ultrahigh sensitive detection of uranyl. The RCDzyme is designed to contain a guanine (G)-rich sequence that replaces the partial sequence in an uranyl-specific DNAzyme. This multifunctional probe can act as the target recognition element, DNAzyme and the primer of signal amplification. The presence of UO22+ can induce the cleavage of the substrate strands in RCDzyme. Then, each released enzyme strand can hybridize with another substrate strands to trigger many cycles of the cleavage by binding uranyl, leading to the formation of more G-quadruplexes by split guanine-rich oligonucleotide fragments. The resulting G-quadruplexes could bind to N-methyl-mesoporphyrin IX (NMM), causing an amplified detection signal for the target uranyl. Next, graphene oxide-based background reduction strategy was further employed for adsorbing free ssDNA and NMM, thereby providing a proximalis zero-background signal. The combination of RCDzyme signal amplification and proximalis zero-background signal remarkably improves the sensitivity of this method, achieving a dynamic range of two orders of magnitude and giving a detection limit down to 86. pM, which is much lower than those of related literature reports. These achievements might be helpful in the design of highly sensitive analytical platform for wide applications in environmental and biomedical fields. © 2015 Elsevier B.V.
语种：
英文
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Determination of leucomalachite green, leucocrystal violet and their chromic forms using excitation–emission matrix fluorescence coupled with second-order calibration after dispersive liquid–liquid microextraction
作者：
Ju, Saiqin;Deng, Jian* ;Cheng, Jianlin;Xiao, Ni;Huang, Kaihui;...
期刊：
Food Chemistry ,2015年185:479-487 ISSN：0308-8146
通讯作者：
Deng, Jian
作者机构：
[Xiao, Ni; Huang, Kaihui; Hu, Canhui; Zhao, Haiqing; Deng, Jian; Ju, Saiqin; Xie, Jin] Univ South China, Coll Chem & Chem Engn, Hengyang 421001, Hunan, Peoples R China.;[Cheng, Jianlin] Univ South China, Coll Publ Hlth, Hengyang 421001, Hunan, Peoples R China.;[Zhan, Xiaozhu] Sanming Supervis & Testing Ctr Qual & Safety Agro, Sanming 365000, Fujian, Peoples R China.
通讯机构：
[Deng, Jian] U;Univ South China, Coll Chem & Chem Engn, Hengyang 421001, Hunan, Peoples R China.
关键词：
Alternating trilinear decomposition;Crystal violet;Dispersive liquid-liquid microextraction;Leucocrystal violet;Leucomalachite green;Malachite green;Parallel factor analysis
摘要：
A novel spectrofluorimetric method has been developed for the simultaneous determination of leucomalachite green (LMG), leucocrystal violet (LCV), malachite green (MG) and crystal violet (CV) by combining the sensitivity of molecular fluorescence and the selectivity of the second-order calibration. Residues of LMG, LCV, MG and CV were simultaneously extracted from fish and shrimp muscle with acetonitrile. The non-fluorescent CV and MG were then reduced to the corresponding fluorescent LMG and LCV by reacting with sodium borohydride. After preconcentration with dispersive liquid-liquid microextraction technique, the extracts were analyzed by using excitation-emission matrix fluorescence coupled with second-order calibration methods based on parallel factor analysis (PARAFAC) and alternating trilinear decomposition (ATLD) algorithms. The limits of detection obtained were 2.21-2.65 ng g-1 by PARAFAC and 2.30-2.86 ng g-1 by ATLD, respectively. The developed method was successfully applied to simultaneous determination of the four analytes in grass carp and shrimp samples with recoveries of 90.53-103.03% for PARAFAC and 90.40-102.75% for ATLD. The accuracy of this novel method was also verified by high performance liquid chromatography. © 2015 Elsevier Ltd.
语种：
英文
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LncRNA在糖尿病及其并发症中的作用
作者：
阳际花;奉水东;杨丝丝;朱责梅;凌宏艳
期刊：
生理科学进展 ,2015年46(5):371-374 ISSN：0559-7765
作者机构：
[阳际花; 杨丝丝; 朱责梅; 凌宏艳] 南华大学医学院生理学教研室;[奉水东] 南华大学公共卫生学院社会医学与卫生事业管理学教研室
关键词：
糖尿病;糖尿病并发症
摘要：
长链非编码 RNA（long non-coding RNA，LncRNA）是一类转录本长度超过200个核苷酸、无蛋白质编码功能的 RNA 分子，起初认为它是转录过程中的副产物，无生物学功能。随着研究的不断推进，人们发现 LncRNA 不但在细胞增殖、分化和代谢中发挥重要的调节作用，而且还参与癌症、糖尿病、神经退行性疾病的病理进程。本文主要就LncRNA在糖尿病及其并发症中的作用作一综述。
语种：
中文
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基于T-Hg~(2+)-T结构的非标记共振光散射法检测汞离子
作者：
Yang Sheng-yuan;Xu Xiao-na* ;Yu Jun-hui;Yang Hui-xian;Hu Cheng-lan
期刊：
光谱学与光谱分析 ,2015年35(12):3471-3474 ISSN：1000-0593
通讯作者：
Xu Xiao-na
作者机构：
[Yu Jun-hui; Yang Hui-xian; Yang Sheng-yuan; Xu Xiao-na; Hu Cheng-lan] Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.
通讯机构：
[Xu Xiao-na] U;Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.
关键词：
核酸探针;共振光散射
摘要：
根据目标物诱导核酸分子构象发生变化,导致体系共振光散射强度增大的原理建立了检测Hg~(2+)的新方法。当溶液中有Hg~(2+)存在时,基于"T-Hg~(2+)-T"特异性结合作用,促使Hg~(2+)特异核酸探针的构象发生变化,由单链状态变为刚性双链结构,使体系共振光散射强度增大。在波长566 nm处,当汞离子浓度在7.2 ×10~(-9)~9×10~(-8)mol? L~(-1)范围内时,体系的共振光散射增强程度△I与汞离子的浓度(c)具有良好的线性关系。其线性方程为△I=5.12c+3.55(r=0.999 5)。将该方法用于环境水样中汞离子的测定,相对标准偏差(RSD)小于1.9%;样品加标回收率为99.4%~104.3%。该方法以Hg~(2+)的高度特异性核酸探针作为识别元件,通过控制Hg~(2+)浓度的变化调节共振光散射强度的变化,将Hg~(2+)的检测转化为对DNA分子构象变化的检测,该转化有利于增强体系共振光散射强度,提高方法灵敏度,该共振光散射检测方法能检测到浓度为2.16×10~(-9)mol?L~(-1)的Hg~(2+)。同时,由于"T-T"错配碱基对对Hg~(2+)的特异结合能力,显著提高了该分析方法对Hg~(2+)的选择性。另外,该方法操作简便、不需标记。
语种：
中文
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Urinary metal concentrations in relation to semen quality: A cross-sectional study in China
作者：
Zeng, Qiang;Feng, Wei;Zhou, Bin;Wang, Yi-Xin;He, Xiao-Sheng;...
期刊：
Environmental Science & Technology ,2015年49(8):5052-5059 ISSN：0013-936X
通讯作者：
Lu, Wen-Qing
作者机构：
[Wang, Yi-Xin; Yang, Pan; Feng, Wei; Lu, Wen-Qing; You, Ling; Zeng, Qiang; He, Xiao-Sheng] Huazhong Univ Sci & Technol, Tongji Med Coll, Sch Publ Hlth, Dept Occupat & Environm Hlth, Wuhan 430030, Hubei, Peoples R China.;[Wang, Yi-Xin; Yang, Pan; Feng, Wei; Lu, Wen-Qing; You, Ling; Zeng, Qiang; He, Xiao-Sheng] Huazhong Univ Sci & Technol, Sch Publ Hlth, Tongji Med Coll, Key Lab Environm & Hlth,Minist Educ, Wuhan 430030, Hubei, Peoples R China.;[Wang, Yi-Xin; Yang, Pan; Feng, Wei; Lu, Wen-Qing; You, Ling; Zeng, Qiang; He, Xiao-Sheng] Huazhong Univ Sci & Technol, Sch Publ Hlth, Tongji Med Coll, Minist Environm Protect, Wuhan 430030, Hubei, Peoples R China.;[Wang, Yi-Xin; Yang, Pan; Feng, Wei; Lu, Wen-Qing; You, Ling; Zeng, Qiang; He, Xiao-Sheng] Huazhong Univ Sci & Technol, Sch Publ Hlth, Tongji Med Coll, State Key Lab Environm Hlth Incubating, Wuhan 430030, Hubei, Peoples R China.;[Zhou, Bin] Univ South China, Coll Publ Hlth, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Lu, Wen-Qing] H;Huazhong Univ Sci & Technol, Tongji Med Coll, Sch Publ Hlth, Dept Occupat & Environm Hlth, Wuhan 430030, Hubei, Peoples R China.
摘要：
Exposure to metals, including essential and nonessential elements, is widespread and may be associated with altered semen quality. This study aimed to examine the association between urinary metal concentrations and semen quality in a Chinese population. We measured semen quality parameters (sperm concentration, count, motility, normal morphology, and abnormal head) and 13 metals [arsenic (As), cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), lead (Pb), manganese (Mn), molybdenum (Mo), mercury (Hg), nickel (Ni), selenium (Se), and zinc (Zn)] in the urine of 394 men from an infertility clinic. Multivariable logistic and linear regressions were used to assess the relationship between the creatinine-adjusted urinary metal concentrations and semen quality parameters. We found a significant trend for decreased odds ratios (ORs) for below-reference sperm count with increasing Se quartiles (p for trend = 0.04) and a significant trend for increased sperm percent abnormal head with increasing Ni quartiles (p for trend = 0.03). These associations persisted, even when considering multiple metals. Our results suggest that Ni exposure may be associated with deteriorated sperm morphology and that Se exposure may be associated with better semen quality. However, our findings warrant further studies in a larger and general population. © 2015 American Chemical Society.
语种：
英文
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PFOS Disturbs BDNF-ERK-CREB Signalling in Association with Increased MicroRNA-22 in SH-SY5Y Cells
作者：
Li, Wu;He, Qing-zhi;Wu, Cheng-qiu;Pan, Xiao-yuan;Wang, Jing;...
期刊：
BioMed Research International ,2015年2015:302653 ISSN：2314-6133
通讯作者：
Zeng, Huai-cai
作者机构：
[Tan, Yan; Li, Wu; Wu, Cheng-qiu; Pan, Xiao-yuan; Shan, Xiao-yun; Zeng, Huai-cai; Wang, Jing] Univ South China, Sch Publ Hlth, Dept Prevent Med, Hengyang 421001, Peoples R China.;[He, Qing-zhi; Zeng, Huai-cai] Hunan Prov Cooperat Innovat Ctr Mol Target New Dr, Hengyang 421001, Peoples R China.;[Zeng, Huai-cai] Univ South China, Sch Publ Hlth, Dept Prevent Med, 28 Western Changsheng Rd, Hengyang 421001, Peoples R China.
通讯机构：
[Zeng, Huai-cai] U;Univ South China, Sch Publ Hlth, Dept Prevent Med, 28 Western Changsheng Rd, Hengyang 421001, Peoples R China.
摘要：
Perfluorooctane sulfonate (PFOS), a ubiquitous environmental pollutant, is neurotoxic to mammalian species. However, the underlying mechanism of its neurotoxicity was unclear. We hypothesized that PFOS suppresses BDNF expression to produce its neurotoxic effects by inhibiting the ERK-CREB pathway. SH-SY5Y human neuroblastoma cells were exposed to various concentrations of PFOS to examine the role of the BDNF-ERK-CREB signalling pathway in PFOS-induced apoptosis and cytotoxicity. Furthermore, to ascertain the mechanism by which PFOS reduces BDNF signalling, we examined the expression levels of miR-16 and miR-22, which potentially regulate BDNF mRNA translation at the posttranscriptional level. Results indicated that PFOS significantly decreased cell viability and induced apoptosis in SH-SY5Y cells. In addition, BDNF and pERK protein levels decreased after PFOS treatment; however, pCREB protein levels were significantly elevated in PFOS treated groups. TrkB protein expression increased in the 10 M and 50 M PFOS groups and significantly decreased in the 100 M PFOS group. Our results demonstrated that PFOS exposure decreased miR-16 expression and increased miR-22 expression, which may represent a possible mechanism by which PFOS decreases BDNF protein levels. PFOS may inhibit BDNF-ERK-CREB signalling by increasing miR-22 levels, which may, in part, explain the mechanism of PFOS neurotoxicity. © 2015 Wu Li et al.
语种：
英文
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A highly sensitive fluorescence probe for metallothioneins based on tiron–copper complex
作者：
Xiao, Xilin* ;Xue, Jinhua;Liao, Lifu;Huang, Mingyang;Zhou, Bin;...
期刊：
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY ,2015年145:85-89 ISSN：1386-1425
通讯作者：
Xiao, Xilin
作者机构：
[Zhou, Bin; Huang, Mingyang; Xiao, Xilin; Liao, Lifu; Xue, Jinhua; He, Bo] Univ South China, Inst Pathogen Biol, Coll Publ Hlth, Coll Chem & Chem Engn, Hengyang City 421001, Hunan Province, Peoples R China.
通讯机构：
[Xiao, Xilin] U;Univ South China, Inst Pathogen Biol, Coll Publ Hlth, Coll Chem & Chem Engn, Hengyang City 421001, Hunan Province, Peoples R China.
关键词：
Metallothioneins;Tiron-copper complex;Fluorescence probe
摘要：
The fabrication of tiron-copper complex as a novel fluorescence probe for the sensitive directly detection of metallothioneins at nanomolar levels was demonstrated. In Britton-Robinson (B-R) buffer (pH 7.50), the interaction of bis(firon)copper(II) complex cation [Cu(tiron)(2)](2+) and metallothioneins enhanced the fluorescence intensity of the system. The fluorescence enhancement at 347 nm was proportional to the concentration of metallothioneins. The mechanism was studied and discussed in terms of the fluorescence spectra. Under the optimal experimental conditions, at 347 nm, there was a linear relationship between the fluorescence intensity and the concentration of the metallothioneins in the range of 8.80 x 10(-9)-7.70 x 10(-7) mol L-1, with a correlation coefficient of r = 0.995 and detection limit 2.60 x 10(-9) mol L-1. The relative standard deviation was 0.77% (n = 11), and the average recovery 94.4%. The method proposed was successfully reliable, selective and sensitive in determining of trace metallothioneins in fish visceral organ samples with the results in good agreement with those obtained by HPLC. (C) 2015 Elsevier B.V. All rights reserved.
语种：
英文
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An ultrasensitive label-free assay of 8-hydroxy-2′-deoxyguanosine based on the conformational switching of aptamer
作者：
Wang, Jia-Cheng;Wang, Yong-Sheng;Xue, Jin-Hua;Zhou, Bin;Qian, Qiu-Mei;...
期刊：
Biosensors and Bioelectronics ,2014年58:22-26 ISSN：0956-5663
通讯作者：
Wang, Yong-Song
作者机构：
[Zhou, Bin; Wang, Yong-Song; Yin, Ji-Cheng; Qian, Qiu-Mei; Zhao, Hui; Liu, Hui; Liu, Shan-Du; Xue, Jin-Hua; Wang, Yong-Sheng; Wang, Jia-Cheng] Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.;[Wang, Jia-Cheng] Dali Univ, Coll Publ Hlth, Dali 671000, Peoples R China.
通讯机构：
[Wang, Yong-Song] U;Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.
关键词：
8-hydroxy-2'-deoxyguanosine;Aptamer;Conformational switching;Gold nanoparticles;Resonance light scattering;Ultrasensitive assay
摘要：
We developed a highly sensitive label-free assay of 8-hydroxy-2'-deoxyguanosine (8-OHdG) using 8-OHdG-aptamer (Apt) as the recognition element. The Apt was adsorbed onto the surface of gold nanoparticles (AuNPs), which prevents them from aggregating under high-salt conditions. Upon addition of 8-OHdG, the conformation of the Apt changes to form a G-quadruplex structure, which leads to the aggregation of the AuNPs along with the increase of the resonance light scattering intensity. The mechanism of 8-OHdG that induces Apt to form G-quadruplexes structure was studied by circular dichroism. The response signals linearly correlated with the concentration of 8-OHdG ranging from 90.8. pM to 14.1. nM with a detection limit of 27.3. pM, which is much lower than that obtained by other methods. This method does not need any label steps and sophisticated equipment. The application for detection of 8-OHdG in real samples further demonstrated its reliability. This strategy would be helpful for developing a universal analytical method by simply replacing corresponding aptamers for various target molecules. © 2014 Elsevier B.V.
语种：
英文
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A colorimetric aptasensor for the highly sensitive detection of 8-hydroxy-2′-deoxyguanosine based on G-quadruplex–hemin DNAzyme
作者：
Liu, Hui;Wang, Yong-Sheng* ;Wang, Jia-Cheng;Xue, Jin-Hua;Zhou, Bin;...
期刊：
Analytical Biochemistry ,2014年458:4-10 ISSN：0003-2697
通讯作者：
Wang, Yong-Sheng
作者机构：
[Zhou, Bin; Chen, Si-Han; Li, Ming-Hui; Tang, Xian; Cao, Jin-Xiu; Zhao, Hui; Liu, Hui; Liu, Shan-Du; Xue, Jin-Hua; Wang, Yong-Sheng; Wang, Jia-Cheng] Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.;[Wang, Jia-Cheng] Dali Univ, Coll Publ Hlth, Dali 671000, Peoples R China.
通讯机构：
[Wang, Yong-Sheng] U;Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.
关键词：
8-Hydroxy-2'-deoxyguanosine;Colorimetric aptasensor;Conformational switching;G-quadruplex;Hemin
摘要：
A highly sensitive, low-cost colorimetric aptasensor was developed for the determination of 8-hydroxy-2′-deoxyguanosine (8-OHdG) in human urine. The method is based on a conformational switching of the 8-OHdG aptamer to form a G-quadruplex structure in the presence of 8-OHdG. The resulting G-quadruplex assembles into a peroxidase-like DNAzyme with hemin, which effectively catalyzes the oxidation of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS2-) by H2O2 to ABTS +, resulting in an increase in the absorption signal at 416 nm along with a color change of the solution. The response signals linearly correlated with the concentration of 8-OHdG, ranging from 466 pM to 247 nM with a detection limit of 141 pM. The relative standard deviation and the recovery were 1.97-3.47% (n = 11) and 98.8-100.2%, respectively. The proposed method avoids the label and derivatization steps in common methods and allows direct analysis of the samples by the naked eye without costly instruments, which is reliable, inexpensive, and sensitive. © 2014 Elsevier Inc. All rights reserved.
语种：
英文
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稳定沉默 Beclin1的人成神经瘤SH-SY5Y细胞系的构建
作者：
魏传杰;肖双;江岚;谭琰;黄波;...
期刊：
生物医学工程学杂志 ,2014年31(5):1085-1089 ISSN：1001-5515
作者机构：
南华大学 公共卫生学院,衡阳421001;[龙鼎新; 肖双; 谭琰; 江岚; 黄波; 魏传杰] 南华大学
关键词：
G418筛选;细胞克隆
摘要：
为了构建pGenesil-1-Beclin1真核表达载体,并建立Beclin1稳定沉默的人成神经瘤SH-SY5Y细胞系,将合成的shRNA片段连接pGenesil-1质粒,构建成重组质粒pGenesil-1-Beclin1,转化其至感受态大肠杆菌JM109,挑取阳性克隆进行双酶切和DNA测序鉴定.常规培养SH-SY5Y细胞,将重组质粒pGenesil-1-Beclin1和空白对照质粒pGenesil-1转染SH-SY5Y细胞,G418抗性筛选,在荧光显微镜下挑取单细胞克隆并扩大培养至稳定细胞株,经RT-PCR和Western blot检测Beclin1基因的表达.经酶切和DNA测序鉴定重组真核表达载体构建正确,成功筛选出两株细胞系.和对照组转染pGenesil-1的细胞系相比,转染pGenesil-1-Beclin1细胞系的Beclin1 mRNA表达下降71.28％±1.45％ (P＜0.05),Beclin1蛋白表达下降75.50％±2.63％ (P＜0.05).提示已经成功构建pGenesil-1-Beclin1真核表达载体并建立了Beclin1基因稳定沉默的SH-SY5Y细胞系,为研究Beclin1功能提供了细胞模型.
语种：
中文
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Induction of autophagy in human neuroblastoma SH-SY5Y cells by tri-ortho-cresyl phosphate
作者：
Long, Ding-Xin;Hu, Dan;Wang, Pan;Wu, Yi-Jun*
期刊：
Molecular and Cellular Biochemistry ,2014年396(1-2):33-40 ISSN：0300-8177
通讯作者：
Wu, Yi-Jun
作者机构：
[Wang, Pan; Wu, Yi-Jun; Long, Ding-Xin] Chinese Acad Sci, Inst Zool, Mol Toxicol Lab, State Key Lab Integrated Management Pest Insects, Beijing 100101, Peoples R China.;[Hu, Dan; Long, Ding-Xin] Univ South China, Sch Publ Hlth, Hengyang 421001, Peoples R China.;[Wu, Yi-Jun] Chinese Acad Sci, Inst Zool, Mol Toxicol Lab, State Key Lab Integrated Management Pest Insects, 1-5 Beichenxi Rd, Beijing 100101, Peoples R China.
通讯机构：
[Wu, Yi-Jun] C;Chinese Acad Sci, Inst Zool, Mol Toxicol Lab, State Key Lab Integrated Management Pest Insects, 1-5 Beichenxi Rd, Beijing 100101, Peoples R China.
关键词：
Tri-ortho-cresyl phosphate;Autophagy;Programmed cell death;LC3;Beclin 1
摘要：
Tri-ortho-cresyl phosphate (TOCP) is an organophosphorus ester and has been widely used in industry. It is found that TOCP induced delayed neurotoxicity in humans and sensitive animal species. However, the mechanism of TOCP-induced neural cytotoxicity remains unclear. In this study, we studied whether autophagy is involved in TOCP-induced neural cytotoxicity in human neuroblastoma SH-SY5Y cells. We found that 0.5 and 1.0 mM TOCP treatment significantly increased the ectopic accumulation of microtubule-associated protein 1 light chain 3 (LC3)-immunopositive puncta, Beclin 1, and LC3-II/LC3-I levels in SH-SY5Y cells in a dose-dependent manner. Notably, by monodansylcadaverine staining method, we found abundant punctate fluorescent acidic vesicular organelles in TOCP-treated cells. Furthermore, ultrastructural observation under the transmission electron microscope indicated that the cytoplasm was occupied by autophagosomes in TOCP-treated SH-SY5Y cells. Thus, these results suggest that TOCP may induce autophagy, and autophagy may be involved in the development of TOCP-induced neural cytotoxicity.
语种：
英文
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Fluorescence quenching determination of metallothioneins using 8-hydroxyquinoline-5-sulphonic acid-Cd(II) chelate
作者：
Qian, Qiu-Mei;Wang, Yong-Sheng;Zhou, Bin;Xue, Jin-Hua;Li, Le;...
期刊：
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY ,2014年118:992-998 ISSN：1386-1425
通讯作者：
Wang, Yong-Song
作者机构：
[Zhou, Bin; Wang, Yong-Song; Yin, Ji-Cheng; Qian, Qiu-Mei; Li, Le; Zhao, Hui; Liu, Shan-Du; Liu, Hui; Xue, Jin-Hua; Wang, Yong-Sheng; Wang, Jia-Cheng] Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.
通讯机构：
[Wang, Yong-Song] U;Univ South China, Coll Publ Hlth, Hengyang 421001, Peoples R China.
关键词：
Metallothioneins;8-Hydroxyquinoline-5-sulphonic acid;Cadmium;Fluorescence quenching
摘要：
A novel method for the determination of metallothioneins (MTs) in urine was developed by fluorescence quenching strategy. The response signals linearly correlated with the concentration of MTs in the ranges of 3.12 ×10 <sup>-8</sup>-1.23 ×10<sup>-6</sup> mol L<sup>-1</sup>, and the limit of detection (LOD) was 9.36 ×10<sup>-9</sup> mol L<sup>-1</sup>. The proposed method avoids the label and derivatization steps in common methods, and is reliable, inexpensive and sensitive. Furthermore, the interaction of MTs and 8-hydroxyquinoline-5-sulphonic acid (HQS)-Cd(II) chelate was investigated, and a static quenching mode was proposed to be primarily responsible for the fluorescence quenching event. It could provide a promising potential for the detection of the biomacromolecules which have no native fluorescence, and be benefit to extend the application of fluorescence strategy. ©2013 Elsevier B.V. All rights reserved.
语种：
英文
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Predictors of urinary trichloroacetic acid and baseline blood trihalomethanes concentrations among men in China
作者：
Zeng, Qiang;Zhou, Bin;Cao, Wen-Cheng;Wang, Yi-Xin;You, Ling;...
期刊：
Science of The Total Environment ,2014年493:806-811 ISSN：0048-9697
通讯作者：
Lu, Wen-Qing
作者机构：
[Wang, Yi-Xin; Yang, Pan; Cao, Wen-Cheng; Huang, Yue-Hui; Liu, Ai-Lin; Lu, Wen-Qing; You, Ling; Zeng, Qiang] Huazhong Univ Sci & Technol, Tongji Med Coll, Dept Occupat & Environm Hlth, Sch Publ Hlth, Wuhan 430074, Hubei, Peoples R China.;[Wang, Yi-Xin; Yang, Pan; Cao, Wen-Cheng; Huang, Yue-Hui; Liu, Ai-Lin; Lu, Wen-Qing; You, Ling; Zeng, Qiang] Huazhong Univ Sci & Technol, Tongji Med Coll, Minist Educ, Key Lab Environm & Hlth,Sch Publ Hlth, Wuhan 430074, Hubei, Peoples R China.;[Wang, Yi-Xin; Yang, Pan; Cao, Wen-Cheng; Huang, Yue-Hui; Liu, Ai-Lin; Lu, Wen-Qing; You, Ling; Zeng, Qiang] Huazhong Univ Sci & Technol, Tongji Med Coll, Minist Environm Protect, Sch Publ Hlth, Wuhan 430074, Hubei, Peoples R China.;[Wang, Yi-Xin; Yang, Pan; Cao, Wen-Cheng; Huang, Yue-Hui; Liu, Ai-Lin; Lu, Wen-Qing; You, Ling; Zeng, Qiang] Huazhong Univ Sci & Technol, Tongji Med Coll, State Key Lab Environm Hlth Incubating, Sch Publ Hlth, Wuhan 430074, Hubei, Peoples R China.;[Zhou, Bin] Univ South China, Coll Publ Hlth, Hengyang, Hunan, Peoples R China.
通讯机构：
[Lu, Wen-Qing] H;Huazhong Univ Sci & Technol, Tongji Med Coll, Dept Occupat & Environm Hlth, Sch Publ Hlth, Wuhan 430074, Hubei, Peoples R China.
关键词：
Biomarker;Disinfection by-products;Drinking water;Trichloroacetic acid;Trihalomethanes
摘要：
Urinary trichloroacetic acid (TCAA) and baseline blood trihalomethanes (THMs) have been measured as biomarkers of exposure to drinking water disinfection by-products (DBPs) that have been associated with increased risk of cancers and adverse reproductive outcomes. This study aimed to identify predictors of urinary TCAA and baseline blood THMs among men in China. Urine samples, blood samples, and information on socio-demographic factors and water-use activities were collected from 2216 men who participated in a cross-sectional study of exposure to drinking water DBPs and reproductive health during 2011 to 2012. Urinary TCAA and baseline blood THMs including chloroform (TCM), bromodichloromethane (BDCM), dibromochloromethane (DBCM), and bromoform (TBM) were analyzed. Multivariable linear regression was used to evaluate predictors of urinary TCAA and baseline blood THM concentrations. Tap water consumption was significantly associated with creatinine-adjusted urinary TCAA concentration (beta = 0.23 mu g/g creatinine per log10 unit; 95% CI: 0.12, 035). Men with surface water source had 0.13 (95% CI: 0.00, 027) higher mean creatinine-adjusted urinary TCAA concentrations than those with ground water source. Smoking was associated with lower concentration of creatinine-adjusted urinary TCAA. Age was significantly associated with baseline blood Br-THM (sum of BDCM, DBCM, and TBM) concentration (beta = 0.01 ng/L per unit; 95% CI: 0.00, 0.02). Increased household income was associated with decreased concentrations of baseline blood BDCM and Br-THMs. Our results suggest that tap water consumption, water source, smoking, age, and household income as the primary determinants of exposure to drinking water DBPs should be considered in exposure assessment. (C) 2014 Elsevier B.V. All rights reserved.
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英文
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