Study of Phthalic Acid Esters Environmental Incretion in Aquicolous Propagation and of Sudan Dyes in Food by GC-EI-MS

科技与社会的进步使得人们在接触和使用大量化学物质的同时，也在向环境排放大量的垃圾，造成了严重的生态环境问题，其中环境激素问题的研究进展尤其引人注目。邻苯二甲酸酯（PhthalicAcidEsters，PAEs）类环境激素作为增塑剂广泛存在于塑料制品中，随着塑料的大量生产和应用而释放到环境中，构成对生态的严重破坏和对人类的巨大威胁，有人将PAEs称为“第二个全球性PCBs污染物”。本论文致力于水生动、植物体内PAEs含量的分析，建立了丙酮超声提取、Florisil硅藻土净化预分离和GC-EI-MS定性与定量的分析方法，分析了多种水生动植物体内的17种PAEs类环境激素。内容分为四章：第一章简要地...With the development of science and technology, people touched and used lot of chemical materials, and emitted a mass of wastes at the same time, which made great harm to the entironment especially the problem of Environmental Incretion. Phthalic Acid Esters which are a kind of Environmental Incretion consist extensively in the plastic materials as the additive. Along with the production and appli...学位：理学硕士院系专业：化学化工学院化学系_分析化学学号：20032502

Pluralinstic-supplied Reseaarch on Pension Services Against an Aging Background——Taking Xiaoshan District of Hangzhou City as an Example

养老是每位老年人都要经历的人生阶段，养老服务关乎老年人在人生最后一个阶段的生存状态与生活品质。中国改革开放以来，老年群体对服务内容多样化、品种等级层次化水平的要求越来越高，政府作为供给的主体，单独承担养老服务供给的全部责任是不现实的，需要其他供给主体来共同承担这份重担。这就要求我们要加快养老服务社会化体系建设，构建和谐社会的整体环境，围绕这个目标要求我们不仅仅要保障孤老优抚对象的老年生活，更要通过不断提升服务标准和服务质量，来改善和提高全体老年人养老服务条件，提供全方位的服务。 在人口老龄化背景下,本文对养老服务多元供给的发展进行了研究和探索，采集和整理了杭州市萧山区老年人口规模、公办与民办...Old age is a necessary life stage that everyone has to go through. And services on the old people are closely related to their living qualities in the last stage of life. The demand in improving pension service diversification and level is growing up since China's reform and opening up. As the main body of the supply of pension services, government is difficult to assume full responsibility. We ne...学位：公共管理硕士院系专业：公共事务学院_公共管理硕士学号：1392012115036

肾透明细胞癌差异表达基因的筛选及其核心基因相应miRNA和lncRNA预测分析

【目的】筛选肾透明细胞癌(ccRCC)中差异表达基因并其相关miRNA和lncRNA,寻找可作为ccRCC诊断及治疗的生物标志物。【方法】通过从TCGA数据库和GEO数据库筛选ccRCC差异表达基因并取交集,对差异表达基因进行GO功能分析和KEGG通路富集分析,使用MCODE软件筛选出差异表达基因中的核心基因,并利用mirDIP数据库和STARBASE数据库对核心基因上游的miRNA和lncRNA进行预测。【结果】共筛选出427个差异表达基因,这些差异表达基因在GO功能分析上主要与催化活性、受体活性和细胞黏附分子活性等功能相关,KEGG信号通路富集结果则表明其主要与PPAR、Rap1以及细胞因子受体相互作用等信号通路相关。从这些差异表达基因中筛选出11个核心基因,并预测到134个相应miRNA,接着对5个与ccRCC总体生存率相关的miRNA在STARBASE中共预测到6个相应lncRNA。最后将lncRNA、miRNA、核心基因以及相应信号通路在CYTOSCAPE中构建出一个互作网络。【结论】利用生物信息学技术将TCGA数据库与GEO数据库结合起来,筛选出ccRCC中差异表达基因,并对其中的核心基因进行miRNA与lncRNA预测分析,为后续找到可用于ccRCC临床诊断的靶标与治疗的靶点提供了有力帮助。国家自然科学基金(81570748);;福州总医院杰出青年培养专项(2017Q05

肾透明细胞癌Caki-1细胞系差异表达基因的生物信息学分析

目的比较肾透明细胞癌Caki-1细胞系与正常肾上皮细胞系ASE-5063中的差异表达基因(DEGs),寻找潜在的肾透明细胞癌特异性分子标志物。方法利用GEO数据库自带的GEO2R在线分析工具分析基因芯片GSE78179,将筛选出的DEGs分别导入Metascape、STRING以及Cytoscape进行综合分析并筛选出核心基因。最后使用Fun Rich等软件对筛选出的核心基因进行GO和KEGG富集分析。结果共筛选出562个DEGs,其中上调基因345个,下调基因217个。进一步使用MCODE筛选出36个关键基因,GO功能分析发现这些基因与细胞粘附分子活性、趋化因子活性、细胞通讯和信号转导等密切相关;KEGG通路富集结果则表明差异基因主要集中在趋化因子信号通路、TNF信号通路以及NF-κB信号通路等多种与肿瘤相关的通路上。结论运用生物信息学方法筛选出肾透明细胞癌Caki-1细胞系中DEGs,其中数个核心基因广泛参与多种肿瘤的病理进程,但尚未在肾透明细胞癌有相关研究报道,提示其可能是治疗肾透明细胞癌的潜在靶点。国家自然科学基金(81570748);;福州总医院杰出青年培养专项(2017Q05

基于数据挖掘分析PEG3基因在肾透明细胞癌中的表达及预后意义

目的:阐述印迹基因PEG3在肾细胞癌中的表达及意义。方法:利用Oncomine数据库及GEPIA分析PEG3在肾透明细胞癌组织中的表达情况;利用KM-Plotter做PEG3与肾透明细胞癌患者生存期的相关性分析,经Meth HC分析PEG3启动子区甲基化水平,String-DB数据库分析PEG3相互作用蛋白;通过The Human Protein Atlas分析PEG3蛋白在肾透明细胞癌中的蛋白表达情况。结果 :在mRNA水平上,PEG3在肾透明细胞癌中较正常肾组织显著低表达,表达水平与肾癌预后呈负相关(Log-rank P=0.001 1),PEG3蛋白在肾透明细胞癌中也较正常肾脏组织低表达,且其启动子甲基化水平则显著增高。与PEG3相关蛋白有NLRP7、MEST、SIAH1、PEG10、CDC25 B等,可能参与细胞有丝分裂周期调节及遗传印迹等细胞功能。结论 :通过肿瘤基因数据库信息挖掘发现,PEG3在肾透明细胞癌组织中呈低表达,且与患者生存期有关,将为后续的肿瘤研究提供重要的理论依据。国家自然科学基金(编号:81570748,81370948

基于数据挖掘分析ATP1A1基因在肾透明细胞癌中的表达及意义

目的:通过数据挖掘分析ATP1A1在肾透明细胞癌中的表达及意义。方法:通过Oncomine数据库检索关于ATP1A1的mRNA信息,采用The Human Protein Atlas分析ATP1A1蛋白在正常肾组织和肾透明细胞癌中表达情况,GEPIA网站中TCGA数据对ATP1A1低表达的肾透明细胞癌患者进行生存分析,Meth HC数据库分析ATP1A1甲基化水平和蛋白相互作用,利用String-DB数据分析ATP1A1与上下游蛋白的相互作用。结果:肾透明细胞癌(Clear cell Renal Cell Carcinoma,cc RCC)组织中ATP1A1的mRNA表达水平较正常对照组明显降低。免疫组化结果证实ATP1A1蛋白质表达变化与mRNA相似。TCGA数据中得出ATP1A1低表达患者的总体生存期明显短于高表达组患者。此外,ATP1A1基因启动子区在肾透明细胞癌中的甲基化水平明显高于正常肾组织中。同时,ATP1A1与ATP1B1、FXYD2、ATP1B2等蛋白可能存在相互作用。结论:大数据分析结果表明ATP1A1在肾透明细胞癌中低表达,并与其发生发展相关,可能作为其潜在的治疗靶点。国家自然科学基金项目(81370948;81570748

GC-EI-MS internal standard analysis of phthalic acid esters in fish

The fish samples were cleaned up by Florisil, then analyzed by GC-EI-MS to determine eight phthalic acid esters, diethyl phthalate (DEP), dipropyl phthalate (DPrP), dibutyl phthalate (DBP), butyl benzyl phthalate (BBP), dipentyl phthalate (DPeP), dicyclohexyl phthalate (DCHP), dihexyl phthalate (DHP) and diethyl hexyl phthalate (DEHP). The external standard and internal standard method were compared, and the pretreatment method was optimized. Especially, the problem of analytical blank control in this experiment was discussed carefully. The linear range of the method was 50.0 similar to 800 mu g.L-1, and the linear cor relation coefficient (R) was higher than 0.99986. The relative standard deviation of the method was less than 12.7% with the recovery values ranging from 74% to 113%. The limit of detection was lower than 3.66 mu g.L-1. The linear range, accuracy, precision and limit of detection of this method can satisfy the request of simultaneous analysis of PAEs in fish by GC-EI-MS

Promotion of proliferation of luminal B breast cancer cells by mesenchymal stem cells and its underlying molecular mechanisms

目的分析人脐带间充质干细胞(hUC-MSCs)对Luminal B型乳腺癌细胞生长增殖的影响,并初步探讨其可能的分子机理。方法绿色荧光蛋白(GFP)和荧光素酶共表达慢病毒感染人Luminal B型乳腺癌细胞BT474,并经嘌呤霉素筛选两周后,于荧光显微镜下观察GFP的表达情况,IVIS Kinetic成像系统拍照以观察和记录慢病毒感染后BT474细胞荧光素酶的表达情况;荧光显微镜下直接观察,结合MTS实验分析hUC-MSCs共培养或其浓缩上清处理对GFP和荧光素酶共表达BT474细胞生长增殖的影响;Western blot法检测hUC-MSCs浓缩上清处理对BT474细胞Akt和MAPK信号通路激活情况以及下游细胞周期调控蛋白Cyclin D1表达的影响;常规RT-PCR法检测hUC-MSCs中NRG-1、NRG-2、IGF-Ⅰ、IGF-Ⅱ和EGF等配体的表达。荧光素酶表达强度与细胞数量的相关性经由Excel软件行统计学分析,MTS实验数据则经由SPSS13.0统计软件行统计学分析。结果荧光显微镜和IVIS Kinetic成像系统的观察结果分别证实,GFP和荧光素酶经慢病毒载体系统的介导可在BT474细胞中成功地共表达,且荧光素酶的表达强度与细胞数量呈直线相关。MSCs共培养或其浓缩上清处理均可显著促进Luminal B型乳腺癌细胞BT474的生长增殖,其细胞存活比例分别为各自对照组的148.06%(P<0.005)和147.99%(P<0.001);MSCs浓缩上清处理同时激活BT474细胞内Akt和MAPK信号通路,并上调细胞周期调控蛋白Cyclin D1表达。此外,RT-PCR结果显示,hUC-MSCs中NRG-1和EGF的mRNAs水平呈高表达,而NRG-2、IGF-Ⅰ和IGF-Ⅱ等配体的mRNAs表达也可见。结论 MSCs可通过表达并分泌NRG-1等配体,从而激活Luminal B型乳腺癌细胞BT474的下游Akt和MAPK信号转导通路以上调细胞周期调控蛋白Cyclin D1的表达,进而促进其生长增殖。Objective To investigate the effect of human umbilical cord mesenchymal stem cells(hUC-MSCs) on proliferation of luminal B breast cancer cells and its underlying molecular mechanisms. Methods Human luminal B breast cancer cells BT474 were infected with GFP and luciferase co-expressing lentiviruses and then subjected for selection with Puromycin for 2 weeks. The expression of GFP and luciferase was detected by fluorescent microscopy and IVIS Kinetic image system, respectively. The effect of coculture or treatment with conditioned medium of hUCMSCs on proliferation of BT474 was analyzed with MTS assay. Western blot was carried out to detect the effect of treatment with conditioned medium of hUC-MSCs on the activation of both Akt and MAPK signalings in BT474, as well as the expression of downstream cell cycle regulator Cyclin D1. Regular RT-PCR was applied to analyze the mRNAs expression of ligands such as NRG-1, NRG-2, IGF-Ⅰ, IGF-Ⅱ, and EGF in hUC-MSCs. The correlation between relative luciferase activity and cell number was analyzed with Excel software, while MTS assay data was statistically analyzed with SPSS 13.0 software. Results The co-expression of GFP and luciferase in BT474 via lentiviral expression system was visualized by fluorescent microscopy and IVIS Kinetic image system. The linear correlation between relative luciferase activity and cell number was determined by curve fitting analysis. Coculture or treatment with conditioned medium of hUC-MSC significantly promoted the proliferation of BT474, with survival rates being 148.06 %(P < 0.005)and 147.99 %(P < 0.001)of control, respectively. In addition, treatment with conditioned medium of hUC-MSC was shown to induce activation of both Akt and MAPK signalings, which further upregulated the expression of Cyclin D1. Moreover, high mRNAs expression levels of both NRG-1 and EGF, as well as moderate mRNAs expression levels NRG-2, IGF-Ⅰ, and IGF-Ⅱ were showed by RT-PCR. Conclusion Our results here demonstrated that MSCs may promote the proliferation of luminal B breast cancer cells through paracrine of ligands such as NRG-1, which in turn results in the activation of both Akt and MAPK signalings and upregulation of the expression of Cyclin D1.国家自然科学基金面上项目(81272922);; 福建省自然科学基金面上项目(2016J01577);; 福州总医院院内课题国际合作研究专项(2015G01

Multiresidue Determination of Organophosphorus Pesticide Residues in Vegetables and Fruit by Gas Chromatography-Negative Ion Chemical Ionization-Mass Spectrometry

将气相色谱-负离子化学电离质谱法(GC-NC I-M S)应用于蔬菜水果中9种有机磷农药残留的分析测定,初步解析了这些农药的NC I-M S特征阴离子结构和断裂机理,并初步探讨了GC-NC I-M S分析蔬菜水果中有机磷农药残留时基体效应的影响。采用空白样品基体匹配校准曲线法(M C)进行定量分析,有效地降低了基体效应的影响。蔬菜水果样品用乙酸乙酯超声提取,以乙硫磷为内标物,采用GC-NC I-M S的选择离子监测方式(S IM)进行定性和定量分析。9种有机磷农药的方法检测限为0.12~1.0μg/kg。在方法的检测限与1 000μg/kg范围内,线性相关系数都大于0.999 3。当空白蔬菜水果(西红柿)样品的加标水平为100,400,800μg/kg时,平均加标回收率为78%~126%,相对标准偏差为0.58%~14.7%。An analytical method of gas chromatography coupled with negative ion chemical ionization-mass spectrometry for simultaneous determination of nine organophosphorus pesticide residues in vegetables and fruit has been developed,and the negative ions structure and partition mechanism of the nine organophosphorus pesticides were interpreted.Meanwhile,the matrix effect for sample analysis was discussed,and matrix-matched calibration for quantification was introduced to reduce the matrix effect in this method.Pesticides were extracted from sample with ethyl acetate in an ultrasonic bath,then determined by gas chromatography-mass spectrometry operated in negative chemical ionization mode and quantified in selective ion monitoring mode,and ethion was used as an internal standard.The detection limits of the method were 0.12-1.0 μg/kg for the nine organophosphorus pesticides,and the relative coefficients were higher than(0.9993).A blank sample(tomato) was spiked at 100,400,800 μg/kg for each pesticide,and the recoveries were determined to be from 78% to 126% with relative standard deviations from 0.58% to 14.7% for the pesticides

Determination of Multiple Pesticide Residues in Honey Using Gas Chromatography-Mass Spectrometry

开展了蜂蜜中23种农药残留的气相色谱-电子轰击离子源质谱(GC-E I/M S)分析方法的研究,并对其中3种农药的E I/M S碎片离子的断裂机理与结构进行了初步解析。探讨了蜂蜜试样前处理条件的优化与选择。将蜂蜜试样用乙酸乙酯提取剂超声提取、F lo ris il硅藻土色谱柱净化和正己烷-乙酸乙酯(体积比为7∶3)混合洗脱剂洗脱后,以PCB103为内标物,采用选择离子监测(S IM)方式下的GC-E I/M S分析。当试样的加标浓度为50,100和200μg/kg时,加标回收率为82%~120%,相对标准偏差小于11.0%。23种农药的检测限都小于10.0μg/kg,线性范围为10~500μg/kg,相关系数都大于0.995。此分析方法已成功地应用于蜂蜜中23种痕量农药残留的分析。An analytical method was developed for the simultaneous determination of 23 pesticide residues in various commercial honeys.Meanwhile,the characteristic ions and fragmentation mechanism of three pesticides in the process of electron ionization mass spectrometry(EI/MS) were evaluated.After the optimization of different parameters such as the extraction solvent,pesticides were extracted from honey with ethyl acetate in an ultrasonic bath,cleaned up on a Florisil column by an elution of mixture of hexane and ethyl acetate(7∶ 3,v/v), and analyzed by gas chromatography-electron ionization mass spectrometry(GCEI/MS) in the selected ion monitoring mode(SIM) with PCB103 as internal standard.Recovery studies were performed at 50,100 and 200 μg/kg fortification levels for each pesticide,and the recoveries ranged from 82% to 120% with relative standard deviations between 1.8% and 11.0% for different pesticides.The limit of detection was less than 10.0 μg/kg for all the pesticides.The developed method was linear in the range of 10-500 μg/kg,with correlation coefficients larger than 0.995.Finally,the developed analytical method has been successfully applied to the determination of pesticide residues in several honey samples