Purification and Determination of Vitellogenin for Application to the Joint Toxicity of Diethylstilbestrol and 17α-Ethynylestradiol

卵黄蛋白原（Vitellogenin,Vtg）是卵生动物中卵黄蛋白的前体，通常只存在于性成熟的雌性个体中。但是如果雄性或幼体暴露于环境雌激素环境，其体内也能产生Vtg。因此，Vtg被认为是一种理想的环境雌激素蛋白质生物标志物，在研究各种化合物的雌激素效应中被广泛应用。 Vtg的纯化多采用离子交换与凝胶渗透两步骤法，耗时长，回收率低、Vtg容易降解。因此，有必要开发一种简单、快速、回收率高的Vtg纯化方法。目前Vtg的检测主要采用特异性的免疫分析方法，方法灵敏，选择性强；但耗时长，操作繁琐。建立快速、简便、可靠的Vtg检测方法成为深入开展Vtg作为环境雌激素标志物研究的关键前提。在环境雌激素毒...Vitellogenin (Vtg) is the egg yolk protein precursor in the oviparous animals. It is usually produced in sexually active female individuals. However, Vtg can also be detected in the male and the juvenile exposed to estrogen or estrogen mimic chemicals. Therefore, Vtg in male serum or plasma is generally considered to be a candidate of protein biomarker for environmental estrogens, thus widely used...学位：理学博士院系专业：海洋与环境学院环境科学与工程系_环境科学学号：B20033401

Study on estrogenic effects of 17α-ethynylestradiol on juvenile Pagrosomus major

采用静态暴露方式研究17α-乙炔基雌二醇(EE2)对真鲷(PAgrOSOMuS MAJOr)幼鱼的雌激素效应。当真鲷幼鱼暴露于0.01、0.1、0.5μg/lEE2,42 d后,真鲷幼鱼的肥满度极显著下降;血浆中卵黄蛋白原(VTg)被诱导产生,肝胰脏指数(HSI)和血浆蛋白总量极显著升高;血浆雌二醇(E2)和睾酮(T)水平显著降低,其中血浆雌二醇受EE2的干扰更显著,降低的幅度更大。结果提示,真鲷幼鱼的肥满度、肝胰脏指数、血浆蛋白总量、血浆性激素水平等可作为评估EE2等环境内分泌干扰物毒性效应的生物指标;真鲷幼鱼血浆中的VTg具有作为生物标志物,用于监测海洋水体中EE等环境内分泌干扰物的潜力。The estrogenic effects of 17α-ethynylestradiol(EE2) on the juvenile Pagrosomus major were studied via a static exposure experiment.After exposed to EE2 at doses of 0.01,0.1,0.5 μg/L for 42 d,the condition factors were significantly decreased,and plasma vitellogenins were induced,hepatopancreas somatic index(HSI) and total plasma protein were significantly increased.The levels of plasma estradiol(E2) and testosterone(T) were significantly decreased,and E2 were more affected.The results showed that the condition factor,HSI,total plasma protein and plasma sex hormone levels of juvenile Pagrosomus major may be used a biological index for evaluating the toxic effects of environmental endocrine disruptors such as EE2.The vitellogenin in the plasma of juvenile Pagrosomus major is potential to be a biomarker for monitoring the marine environmental endocrine disruptors such as EE2.国家自然科学基金(20277030

Determination of bisphenol A in environmental water samples by stir bar sorptive extraction based on molecularly imprinted polymer

以双酚A(bPA)为单体,利用整体材料“原位“聚合技术制备以分子印迹聚合物为涂层的吸附萃取搅拌棒(MIP-SbSE),然后与高效液相色谱(HPlC)-二极管阵列检测器联用,探讨其对环境水样bPA的选择萃取性能。优化萃取过程中吸附和解吸时间、解吸液种类以及基底PH值和离子强度对目标化合物的选择吸附性能。在最佳条件下,MIP-SbSE可对模板分子进行有效的选择吸附,线性范围为1.0~200μg/l,检出限(S/n=3)和定量限(S/n=10)分别为0.28μg/l和0.94μg/l。在实际水样分析中,具有良好的加标回收率,其值为96.0%~108.7%。研究结果表明,所建立的方法具有简便、灵敏和环境友好等特点。A new stir bar sorptive extraction (SBSE) based on molecularly imprinted polymer (MIP) with bisphenol A as template was prepared.Based on it,a simple,selective and sensitive method for the determination of bisphenol A (BPA) was developed combined with high performance liquid chromatography (HPLC) with diode array detection.To achieve the optimum extraction performance,several main parameters,including extraction and desorption time,the nature of desorption solvent,pH value and contents of inorganic salt in the sample matrix,were investigated.The optimized parameters for the extraction of BPA from water matrix with the MIP-SBSE are as follows: extraction and desorption time were 120 min and 10 min,respectively; the pH value of matrix was 10.0; using acetonitrile (including 1% acetic acid) as desorption solvent; no salt was added in the matrix.Under the optimized experimental conditions,the method showed good linearity between 1.0-200 μg/L.The detection limit (S/N=3) and quantification limit (S/N=10) of the proposed method for the BPA was 0.28 μg/L and 0.94 μg/L,respectively.The proposed method was successfully applied to the determination of the target compound in water samples.The recoveries of spiked target compound in real samples ranged from 96.0%-108.7%.The results indicated that the developed method possessed the advantages such as high sensitivity,simplicity,low cost and environmental friendliness.国家自然科学基金项目(No.20805039);福建省青年科技人才创新项目(No.2006F3117

Valence Subband Structure and Optical Gain of InGaAs/InP Quantum Wire

采用有效质量理论 6带模型 ,计算了 In0 .53Ga0 .4 7As/ In P量子线的光学性质 ,具体计算了In0 .53Ga0 .4 7As/ In P量子线的能带结构、态密度、载流子浓度、光学跃迁矩阵元和光学增益谱 ,并把量子线的光学增益谱和量子阱的光学增益谱作了比较。福建省自然科学基金资助课题 (No.E9910 0 0 5

红花组分HSYA对人胃腺癌BGC-823移植瘤裸鼠VEGF蛋白、KDR与缺氧诱导因子表达的影响

目的:研究红花组分羟基红花黄色素A(HSYA)对人胃腺癌皮下移植瘤裸鼠血管内皮生长因子(VEGF)蛋白、含激酶插入区受体(KDR)和缺氧诱导因子(HIF-1α)mRNA与蛋白表达的影响。方法:采用BALB/C nu/nu裸小鼠接种人胃腺癌细胞株BGC-823于右前肢腋部皮下建立裸鼠人癌移植瘤模型,随机分为模型组、对照组、HSYA高、低剂量组(0.056g/L、0.028g/L),观察抑瘤作用,酶联免疫吸附测定(ELISA)法检测瘤组织VEGF与HIF-1α蛋白以及血清VEGF蛋白表达;蛋白印迹(Western blotting)法测定瘤组织KDR磷酸化蛋白的表达;实时荧光定量PCR(RTFQ-PCR)法检测瘤组织KDR及HIF-1αmRNA表达。结果:HSYA低剂量组抑瘤明显,该组瘤组织及血清VEGF蛋白表达降低,瘤组织KDR磷酸化蛋白表达减弱,与模型组比较差异明显(P<0.05,P<0.01);HIF-1α蛋白表达较模型组瘤组织明显减少(P<0.01)。KDR mRNA表达与模型组相比减弱,差异有统计学意义(P<0.05)。结论:一定浓度的HSYA抑制肿瘤生长的可能机制与抑制VEGF、HIF-1α蛋白的表达,减弱KDR蛋白磷酸化及其基因表达,从而抑制内皮细胞活化阻碍肿瘤血管新生以及降低肿瘤缺氧微环境对血管生成的诱导有关

羟基红花黄色素A对人胃癌移植瘤裸鼠瘤组织bFGF蛋白及MMP-9表达的影响

目的:研究羟基红花黄色素A(hydroxy safflor yellow A,HSYA)对人胃癌裸鼠皮下移植瘤碱性成纤维生长因子(basic fibroblast growth factor,bFGF)蛋白以及基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)mRNA及蛋白表达的影响。方法:采用BALB/C nu/nu裸小鼠接种人胃腺癌细胞株BGC-823右前肢腋部皮下建立裸鼠人癌移植瘤模型,随机分为模型组、阳性对照环磷酰胺组(2 g.L-1)、HSYA组(0.056,0.028 g.L-1),观察抑瘤作用,并采用实时荧光定量PCR(real time-fluorescent quantitation PCR,RTFQ-PCR)检测瘤组织中MMP-9 mRNA表达;酶联免疫吸附测定法(enzyme linked immunosorbentassay,ELISA)检测瘤组织中bFGF,MMP-9蛋白表达。结果:HSYA0.028 g.L-1组抑瘤效果明显,且其MMP-9 mRNA表达及bFGF与MMP-9蛋白表达与模型组比,均明显降低(P<0.05)。结论:一定浓度的HSYA抑制肿瘤生长的机制之一可能与抑制bFGF,MMP-9的表达,减少瘤组织血管基底膜的降解,阻抑血管移行以减少血管生成有关

商业方法专利保护问题研究——以网络时代银行业金融创新为载体

一、商业方法专利概述(一)内涵界定人类社会的活动离不开各种各样的“方法“,但各国对于何谓“商业方法“并没有统一的定义。美国专利分类第705类商业方法的定义是:装置及定义的方法,用于商业运作、行政、企业管理或财务资料报表的产生,其能使资料在经过处理后有显著的改变或完成运算操作,装置及对应的方法,用于货物或服务之提供改变时的资料处理或运算操作。欧洲

An on-line automatic titration device for measuring organic phosphorus in water

采用TEFLON MICROPUMP恒量泵,研制了滴定法在线自动检测装置,用于对工业循环冷却水水质稳定剂的监测与控制。对水体中HEDP含量的分辨率可达0 .0 8mg L。改变滴定剂配方与检测波长,装置可应用于其他需要采用滴定分析法进行在线自动检测的场合。An on-line automatic measurent instrument based on titrimetry has been developed using TEFLON MICRO PUMP precision constant pump. The device is used for monitoring and controlling water quality stabilizer in industrial recycling cooling water. The detection limit for HEDP in recycled water is 0.08mg/L. The device can be used in other locations where on-line automatic detection based on titrimetry is needed

In situ Enrichment of Labile Heavy Metals in Seawater with Diffusive Gradients in Thin Films

研究表明对生物体有直接作用的金属并非溶解态的全部,而是其中的活性部分.薄膜扩散梯度技术(dgT)作为一种原位富集有效态金属的新技术,被广泛应用于自然水体里重金属的分析.本研究设计了一种能同时原位富集海水中的Mn、CO、nI、Cu、zn、Cd、Pb的dgT装置;使用自制的凝胶扩散系数测定装置获得了金属离子在扩散相凝胶中的扩散系数;考察了PH、离子强度等对富集效果的影响,当PH为5.6~8.6、离子强度在10~700MMOl/l范围内,富集效果不受影响.在厦门近岸海域成功地进行为期7d的海水中重金属的原位富集,实验结果显示,dgT能够富集该海域海水中6.672%~45.33%的溶解态重金属,即获得dgT有效态.平行样测定的相对标准偏差均小于10%,该方法可用于海水中痕量有效态重金属的分析.Heavy metals are introduced into marine environments mostly by human activities,causing adverse effects on marine ecosystems.Nowadays scientists have recognized that the bioavailability of heavy metals controls the potential of adverse effects rather than the total or filterable concentrations.Diffusive gradients in thin films(DGT)is a recently developed technique that is capable of in situ enrichment of labile metal species in aqueous systems for quantitative measurements.In this study,DGT device was built for the measurement of heavy metals in seawaters.The molecular diffusion coefficient(D)of various metals through a polyacrylamide gel had been determined.Using Chelex-100 as the ion-exchange resin,pH in the range of 5.6-8.6and the ionic strength between 10mmol/L to 700mmol/L would not influence the enrichment.The DGT devices had been applied for the pre-concentration of Mn,Co,Ni,Cu,Zn,Cd and Pb in coastal seawaters of Xiamen for seven days.The result showed that the DGT-labile part in the fraction of0.45μm filterable portion varied from 6.672%-45.33%for these metals.And the relative standard deviation was all below 10%(n=3).DGT had been confirmed to be a very promising tools for preconcentrating trace metals in waters and better understanding their bioavailability.厦门大学近海海洋环境科学国家重点实验室自主课题(MELRI1001;MELRI1202

Determination of Siderophores in Seawater by High Performance Liquid Chromatography-Tandem Mass Spectrometry Coupled with Solid Phase Extraction

铁载体是一种由海洋菌类合成并分泌、能特异性络合海水中铁的有机配体。本研究建立了固相萃取预处理、高效液相色谱-串联质谱测定海水中铁载体化合物的分析方法。海水样品经0.22μM滤膜过滤,其中铁载体由EnVI-18萃取柱萃取、甲醇洗脱后得到富集净化后的试样;采用Sb-C18反向色谱柱以0.1%(V/V)甲酸溶液和甲醇溶液梯度洗脱分离,在质谱的多反应监测正离子模式下检测。3种铁载体化合物标准物质PyOVErdInES-fE,fErrICHrOME,fErrIOXAMInE E的检测线性范围分别为0.001~3.0μg/M l,0.005~15.0μg/M l,0.001~3.0μg/M l,相关系数r2>0.99;仪器检出限分别为0.08,1.76和1.36 ng/M l;定量限分别0.27,5.87和4.53 ng/M l。在海水样品中添加铁载体化合物混合标准溶液,3种目标物测定值的相对标准偏差 0.99) was obtained for Pyoverdines-Fe,Ferrichrome,Ferrioxamine E at the concentrations of 0.001- 3.00 μg / m L,0.005- 15.00 μg / m L,0.001-3.00 μg / m L,respectively.The instrumental detection limits and limits of quantification for the three analytes were 0.08,1.76 and 1.36 ng / m L; 0.27,5.87 and 4.53 ng / m L,respectively.The relative standard deviations were lower than 12%,while the recoveries were 12.1%- 18.6% for Pyoverdines-Fe,82.0%-97.7% for Ferrichrome,and 70.0%- 98.3% for Ferrioxamine E.国家自然科学基金(No.41176075)资助~