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Identification and Preliminary Analysis of Several Centromere-associated Bacterial Artificial Chromosome Clones from a Diploid Wheat Library

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Journal of Integrative Plant Biology 2006年第3期48卷 348-358页

作者： Zhao Liu Wei yue yu-shen dong Xue-Yong Zhang Key Laboratory of Crop Germplasm and Biotechnology Institute of Crop Sciences Chinese Academy of Agricultural Sciences Beijing 100081 China State Key Laboratory of Crop Genetics and Germplasm Enhancement Nanjing Agricultural University Nanjing 210095 China

Although the centromeres of some plants have been investlgated prevlously, our knowledge of the wheat centromere Is still very llmlted. To understand the structure and functlon of the wheat centromere, we used two centromeric repeats （RCS1 and CCS1-5ab） to obtain some centromere-assoclated bacterial artificial chromosome （BAC） clones in 32 RCS1-related BAC clones that had been screened out from a diploid wheat （Triticum boeoticum Boiss.; 2n=2x=14） BAC library. Southern hybridization results indicated that, of the 32 candidates, there were 28 RCS1-positive clones. Based on gel blot patterns, the frequency of RCS1 was approximately one copy every 69.4 kb in these 28 RCS1-positive BAC clones. More bands were detected when the same filter was probed with CCS1-5ab. Furthermore, the CCS1 bands covered all the bands detected by RCS1, which suggests that some CCS1 repeats were distributed together with RCS1. The frequency of CCS1 families was once every 35.8 kb, nearly twice that of RCS1. Fluorescence in situ hybridization （FISH） analysis Indicated that the five BAC clones containing RCS1 and CCS1 sequences all detected signals at the centromerlc regions in hexaplold wheat, but the signal intensities on the A-genome chromosomes were stronger than those on the B- and/or Dgenome chromosomes. The FISH analysis among nine Triticeae cereals indicated that there were A-genomespecific （or rich） sequences dispersing on chromosome arms in the BAC clone TbBACS. In addition, at the interphase cells, the centromeres of diploid species usually clustered at one pole and formed a ring-like allocation In the period before metaphase.

关键词： centromere fluorescence in situ hybridization (FISH) repeat Southern hybridization Triticum boeoticum.

Design and efficient synthesis of novel haptens and complete antigens for the AOZ,a toxic metabolite of furazolidone

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Chinese Chemical Letters 2007年第12期18卷 1490-1492页

作者： yu dong shen yu Wang Shi Wei Zhang Zhi LiXiao yuan Ming Sun Xian Zhang Bu Lian Quan Gu Food Quality and Safety Key Laboratory of Department of Education of Guangdong Province/Institute of Food Quality and Safety South China Agricultural University Guangzhou 510642 China School of Pharmaceutical Sciences Sun Yat-Sen University Guangzhou 510080 China

A good strategy was brought forward for designing efficient haptens and complete antigens for 3-amino-2-oxazolidinone （AOZ）. Haptens designed newly were achieved facilely in good yield by using LiCI-N（Et）3 as new catalysis system, the structure of which was elucidated by spectroscopy analysis, such as NMR and MS. Novel antigens for AOZ were prepared successfully by convenient active ester method. The ratios of haptens 3 and 4 to carrier proteins were proven respectively as 41 ： 1 （5a）, 39：1 （6a）, 11：1 （5b） and 9：1 （6b） by trinitrobenzene sulfonic acid （TNBS） method. The results of indirect competitive ELISA （ic-ELISA） of antiserums indicated that the haptens with a short unsaturated side chain can evoke specific immune response effectively.

关键词： Furazolidone AOZ Antigen Hapten Immunoassay

Unexpected Synthesis of 2,2'-[1-(4-Methoxyphenoxy)propane-2,2-diyl]bis(5-methoxy-3-methylbenzofuran)

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Chinese Chemical Letters 2005年第12期16卷 1581-1583页

作者： yu dong shen Hai Qiang WU Lin Kun AN Zhi Shu HUANG Xian Zhang BU Lian Quan GU School of Chemistry and Chemical Engineering Sun Yat-sen University Guangzhou 510275 School of Pharmaceutical Sciences Sun Yat-sen University Guangzhou 510080

A new bisbenzofuran analogue Ⅶ was achieved unexpectedly in one step procedure from 1-（4-methoxyphenoxy）acetone Ⅰ by using Amberlyst 15 resin as catalyst in excellent yield. The structure was elucidated by spectro... 详细信息

关键词： Benzofuran Amberlyst 15 resin cyclization

Pathogenesis and treatment to postoperative bile leakage: report of 38 cases

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Hepatobiliary & Pancreatic Diseases International 2005年第3期4卷 441-444页

作者： Jia-Min Zhang,Shi-An yu,Wei shen and Zhang-dong Zheng Jinhua Centre Hospital, Jinhua 321000 , China Jinhua Centre Hospital Jinhua 321000 China. zhangjiamin 600323@.***

Bile leakage remains a serious complication after biliary surgery. The aim of this study was to assess the etiology, diagnosis and treatment of postoperative biliary leakage. METHODS:Thirty-eight patients with biliary leakage we treated in recent 8 years were analyzed retrospectively. Among them, 8 patients had bilioenterostomy leakage, 7 accessory bile duct leakage, 7 cholecyst bed leakage, 6 leakage after removal of T-tube, 5 leakage after laparoscopic cholecystectomy, 3 leakage around T-tube, and 2 leakage caused by choledochal damage. Drainage was performed in 17 patients, reoperarion in 13, drainage plus percutaneous transhepatic cholangio drainage (PTCD) , endoscopic retrograde cholangiography (ERCP) , endoscopic nasobiliary drainage(ENBD) and endostenting in 5, and drainage plus growth hormone in 3. RESULTS; In this series, 37 patients were cured, and 1 died of multiple organ dysfunction syndrome (MODS). These patients were hospitalized for 2 weeks to 8 weeks. The drainage group was hospitalized shorter than the undrain-age group. CONCLUSIONS; A piece of white gauze can be used to touch surgical area in detecting biliary leakage intraopera-tively. Mucous to mucous suture of the bile duct and appropriate time for removal of T-tube are recommended to prevent biliary leakage. Reoperation is essential to acute peritonitis. Drainage can be used if leakage don t diffuse or it occurs after pulling out T-tube. Drainage plus ERCP, ENBD,PTCD and drainage are effective.

关键词： etiology treatment complication biliary leakage

Production and Identification of High Affinity Monoclonal Antibodies Against Pesticide Carbofuran

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Agricultural Sciences in China 2007年第9期6卷 1082-1088页

作者： YANG Jin-yi WU Qing WANG Hong PAN Ke LEI Hong-tao HU Da-yin shen yu-dong XIAO Zhi-li ZHENG Xiao-feng SUN yuan-ming Key Laboratory of Food Quality and Safety Education Department of Guangdong Province/Research Institute of Food Quality and Safety South China Agricultural University Guangzhou 510642 P.R. China Livzon Group Reagent Factory Zhuhai 519060 P.R.China

To produce high-affinity monoclonal antibodies against pesticide carbofuran, and the develop immunochemical assays for people＇s health and environmental protection, the hapten 4-[[（2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy） carbonyl]- amino]-butanoic acid （BFNB） of carbofuran was synthesized and Balb/c mice were immunized by the hapten-carrier （BFNB-bovine serum albumin, BFNB-BSA） conjugates. The splenocytes of immunized mice were fused with Sp2/0 cells and the cultural supernatants of hybridoma cells were screened by the indirect enzyme-linked immunoabsorbent assay （ELISA）, based on BFNB-ovoalbumin conjugates （BFNB-OVA）. Purified monoclonal antibody （McAb） was obtained from fluids of ascites, deposited by octanoic acid and ammonium sulfate. The affinity and the specificity of McAb were characterized by ELISA or indirect competitive ELISA. A hybridoma cell line （5D3） secreting anti-carbofuran McAb had been established. The titer of culture medium and ascites was up to 1：2.048 × 10^3 and 1：1.024 × 10^6, respectively, and the subtype of the McAb was IgG1. The affinity constant of the McAb was about 2.54 × 10×9 L mol^-1, with an IC50 value of 1.18 ng mL^-1 and a detection limit of 0.01 ng mL^-1. Cross-reactivity studies showed that the McAb was quiet specific for carbofuran, as among the four analogous compounds, they were all hardly recognized （4.59 × 10^-4% for 2,3-dihydro-2,2- dimethyl-7-benzofuranol and less than 3.0 × 10^4% for others）. The prepared McAb had a very high affinity and specificity, and it could be used to develop ELISA for rapid determination of carbofuran.

关键词： carbofuran monoclonal antibodies immunochemical assays

酚醛树脂包覆氧化天然石墨作为锂离子电池负极材料

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物理化学学报 2011年第9期27卷 2129-2134页

作者：高文超黄桃沈宇栋余爱水复旦大学化学系新能源研究院上海市分子催化和功能材料重点实验室上海200438 无锡东恒新能源材料有限公司江苏无锡214037

天然石墨经过浓硫酸氧化处理,酚醛树脂包覆并高温碳化后形成具有核壳结构的碳包覆氧化天然石墨复合材料.采用扫描电子显微镜(SEM),透射电子显微镜(TEM),X射线衍射(XRD),激光显微拉曼光谱(Raman)等检测技术对氧化处理以及酚醛树脂热解碳... 详细信息

天然石墨经过浓硫酸氧化处理,酚醛树脂包覆并高温碳化后形成具有核壳结构的碳包覆氧化天然石墨复合材料.采用扫描电子显微镜(SEM),透射电子显微镜(TEM),X射线衍射(XRD),激光显微拉曼光谱(Raman)等检测技术对氧化处理以及酚醛树脂热解碳包覆前后天然石墨材料的结构与形貌进行分析与表征.结果表明,氧化处理与适量的酚醛树脂热解碳包覆有效修复了天然石墨表面的一些缺陷结构,使其表面更为光滑.电化学测试结果显示,经过氧化处理与酚醛树脂热解碳包覆后天然石墨材料电化学性能得到明显提高.酚醛树脂包覆量为9%时,复合材料表现出最好的电化学性能,其首次放电比容量为434.0mAh·g-1,40次循环后,放电比容量保持在361.6mAh·g-1,而未经处理的天然石墨放电比容量仅为332.3mAh·g-1.该改性方法有效提高了天然石墨材料的比容量,对其进一步应用具有重要意义.

关键词：负极材料天然石墨酚醛树脂氧化包覆

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纳米抗体的稳定性及其结构基础研究进展

生物化学与生物物理进展 2022年第6期49卷 1004-1017页

作者：何晓婷董洁娴沈兴王弘沈玉栋徐振林华南农业大学食品学院广东省食品质量安全重点实验室广州510642 深圳清华大学研究院抗肿瘤创新药物研发中心深圳518057

驼类纳米抗体结构简单、易于改造,且具有低免疫原性、高稳定性、高特异性、高亲和力等特点,因而具有广泛的应用前景。纳米抗体的优势之一在于其具有较高的稳定性,比常规抗体更易于储藏和运输,甚至在高温、化学和压力等极端条件下变性后... 详细信息

驼类纳米抗体结构简单、易于改造,且具有低免疫原性、高稳定性、高特异性、高亲和力等特点,因而具有广泛的应用前景。纳米抗体的优势之一在于其具有较高的稳定性,比常规抗体更易于储藏和运输,甚至在高温、化学和压力等极端条件下变性后仍可有效地重折叠并恢复其抗原亲和力。本文综述了纳米抗体稳定性与其结构基础方面的研究进展,阐述了纳米抗体氨基酸序列、二硫键、结构域等与其稳定性的关系,揭示了高度稳定性的纳米抗体普遍具有的结构特征。基于这些结构特征,讨论了几种纳米抗体的稳定性优化策略,包括共有序列驱动的序列修复、替换易于修饰的氨基酸、净蛋白质电荷的改变、非天然二硫键的引入以及CDR超变区的移植。预期对纳米抗体的稳定性调控提供理论指导,以拓展其作为治疗药物、诊断试剂和生物传感器等方面的应用。

关键词：纳米抗体稳定性结构序列二硫键

孔雀石绿单克隆抗体制备和酶联免疫检测方法的建立

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分析化学 2016年第9期44卷 1385-1393页

作者：王宇杨金易徐振林戚平沈玉栋广州市食品检验所华南农业大学食品学院

针对孔雀石绿三苯环特征结构,设计合成1种高质量半抗原。通过偶联载体蛋白、动物免疫和细胞融合,制备出一株高质量的单克隆抗体MG-DA4-C7,亚类为Ig G1,轻链为κ型。通过对包被原浓度、抗体浓度、二抗浓度的优化,建立了孔雀石绿间接竞争... 详细信息

针对孔雀石绿三苯环特征结构,设计合成1种高质量半抗原。通过偶联载体蛋白、动物免疫和细胞融合,制备出一株高质量的单克隆抗体MG-DA4-C7,亚类为Ig G1,轻链为κ型。通过对包被原浓度、抗体浓度、二抗浓度的优化,建立了孔雀石绿间接竞争酶联免疫分析方法。方法半抑制浓度(IC;)为0.96μg/L,线性检测范围(IC;)为0.1;.1μg/L,LOD(IC;)为0.05μg/L,回归方程y=-0.3274lgx+0.4698（R;=0.9891）。本方法特异性高,与代谢物隐孔雀石绿交叉反应小于0.1%,与结晶紫、灿烂绿交叉反应率分别为18.1%和26.5%;实际样品添加回收率为87.3%;07.3%。本方法测定结果经HPLC-MS/MS方法确证,二者相关系数达0.999。本方法可用于鱼类等水产品中孔雀石绿残留的实际检测。

关键词：孔雀石绿半抗原单克隆抗体抗原识别酶联免疫分析

氟甲喹单克隆抗体制备、鉴定及间接竞争酶联免疫吸附分析法

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分析化学 2010年第3期38卷 313-317页

作者：王宇沈玉栋徐振林雷红涛王弘孙远明广东省食品质量安全重点实验室华南农业大学食品学院

以氟甲喹(FLU)为原料,合成4个碳原子手臂的半抗原(FLUABA),采用活泼酯法与牛血清白蛋白(BSA)偶联制备免疫抗原,通过免疫Balb/c小鼠及细胞融合,获得1株稳定分泌抗氟甲喹单克隆抗体的杂交瘤细胞株DB6-E7,其抗体亚类为IgG1,亲和力常数(KA)... 详细信息

以氟甲喹(FLU)为原料,合成4个碳原子手臂的半抗原(FLUABA),采用活泼酯法与牛血清白蛋白(BSA)偶联制备免疫抗原,通过免疫Balb/c小鼠及细胞融合,获得1株稳定分泌抗氟甲喹单克隆抗体的杂交瘤细胞株DB6-E7,其抗体亚类为IgG1,亲和力常数(KA)为8.19×108L/mol。将氟甲喹、FLUABA及6个碳原子手臂的半抗原FLUACA分别与卵清白蛋白(OVA)偶联作为包被抗原,研究异源包被对间接竞争ELISA灵敏度的影响。结果表明,异源包被可显著提高ELISA方法的灵敏度。基于最佳异源包被(FLU-OVA)的酶联免疫吸附分析法的IC50为26.33μg/L,检出限为4μg/L,定量检测范围为8.0～114μg/L(IC20～IC80)。与喹诺酮类药物及结构类似物几乎不存在交叉反应,特异性高。此方法可满足畜禽产品中氟甲喹残留的快速筛查。

关键词：氟甲喹单克隆抗体异源包被酶联免疫吸附分析法