Fast detection of thanatophoric dysplasia type I p.R248C mutation hot spots and rapid prenatal diagnosis of three TD type I high-risk fetuses

目的针对致死性侏儒症I型（thanatophoric dysplasia typeⅠ,TD-Ⅰ）FGFR3基因的突变热点＂p.R248C＂,建立快速特异的酶切鉴定法（restriction endonuelease testing,RE）和扩增受阻突变系统（amplification refractory mutation system,ARMS）/RE法,并应用于后续3例疑似TD-I高危胎儿的快速产前诊断,以及时防止患胎出生,同时为今后的胚胎植入前遗传学诊断（preimplantation genetic diagnosis,PGD）打下良好基础。方法首先,针对突变热点p.R248C突变前后的序列特点,选择合适的内切酶＂Afe I＂,建立酶切鉴定法。其次,设计双错配碱基特异引物结合Apa LI酶切,建立ARMS/RE双重特异鉴定法。对阴性和阳性结果均再用普通引物扩、测的结果进行验证。结果用Afe I酶切FGFR3基因exons（6-7）普通引物的PCR产物（535 bp）,正常对照及非p.R248C突变的TD病例均能被切成255 bp和280 bp 2个片段,而胎1~胎3均切出255 bp、280 bp和535 bp 3个片段。用特异引物E7（p.R248C）扩增,正常对照和非p.R248C突变的TD病例均扩增阴性,无法进一步做酶切鉴定;而p.R248C突变均扩增阳性,当再用Apa LI酶切PCR产物（365 bp）时,胎1~胎3均切出22 bp和343 bp 2个片段。通过引物扩、测结果显示：胎1~胎3均是p.R248C杂合突变。结论该法快速特异、准确可靠,可用于p.R248C突变热点的快速检测及含该突变高危胎儿的快速产前诊断。该法还可用于含p.R248C突变TD-I型家系的PGD。胎1~胎3都是TD-I患胎,建议尽早终止妊娠。Objective To build up the specific rapid methods of RE and ARMS/RE for mutation hotspot＂p.R248C＂in the FGFR3 gene of thanatophoric dysplasia type I（TD-I）,then use the method for rapid prenatal diagnosis of 3 follow-up high-risk fetuses of TD-I to stop the birth of suffering fetuses,at the same time,lay a good foundation of preimplantation genetic diagnosis（PGD） for the future.Methods First,according to the characteristics of the sequence of the mutation hot spot p.248 C before and after mutation,the appropriate enzyme＂Afe I＂was selected to establish the identification method of enzyme digestion.After that,specific primers of double mismatch bases were devised,combining the method of digestion of Apa LI（ARMS/RE） to achieve the double identification.In the end,the sequences obtained by common primers were used to verify the negative and positive results through DNA sequencing.Results For the normal control and the patient without p.R248 C mutation of TD-I,PCR products（535 bp） of exons（6-7） of FGFR3 gene,amplified by common primers,could be digested into 2 fragments（255 bp and 280 bp） by Afe I.However,for fetuses 1~3,the PCR products could be digested into 3 fragments（255 bp,280 bp and 535 bp）.The normal control and the patient without p.R248 C mutation of TD-I were negative using the specific primers E7（p.R248C）,which could not be further identified by enzyme digestion.For the fetuses with p.R248 C mutation of TD-I,PCR products（365 bp） of exon 7 of FGFR3 gene,amplified by specific primers,could be digested by Apa LI and all the digested products were 22 bp and 343 bp.Sequencing results of common primers＇ products indicated：fetuses 1~3 were p.R248 C heterozygous mutation.Conclusions The method is fast,accurate and reliable,which can be available for the fast detection of mutation hot spot p.R248 C and the rapid prenatal diagnosis of high-risk fetus with p.R248 C mutation.This method can also be applied to PGD of high-risk fetus of TD-I with the mutation＂p.R248C＂.Fo闽粤合作科研基金（71010025

基于立即早期蛋白IE62建立水痘-带状疱疹病毒感染性滴度检测方法

目的:基于水痘-带状疱疹病毒(VZV)立即早期蛋白IE62与酶联免疫斑点检测技术(ELISpot)建立一种新型的VZV感染性滴度的快速检测方法。方法:应用生物信息学方法设计并合成VZV-IE62蛋白的多肽抗原,牛血清白蛋白偶联后免疫BALB/c小鼠,筛选和制备抗VZV-IE62单克隆抗体,应用Western blot和免疫荧光法等开展抗体性能评价,继而应用酶联免疫斑点技术(ELISpot)结合生物素-亲和素放大法建立新型的VZV感染性滴度检测方法,并与经典空斑计数法进行比较。结果:获得抗VZV-IE62的单克隆抗体1B7,应用于ELISpot方法可特异识别VZV感染后的细胞,以之为基础建立了新型的VZV感染性滴度检测方法。相比经典空斑计数法,该方法可显著缩短检测时间(从5至7 d缩短至32 h)检测结果具有较好的一致性。结论:本研究建立了一种新型的基于VZV立即早期蛋白IE62与酶联免疫斑点技术的VZV感染性滴度检测方法(VZV-IE62 ELISpot),具有潜在的转化应用前景,可为VZV的防治研究提供支持。国家自然科学基金项目(81601762)资

Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli

旨在建立基于大肠杆菌表达系统的高效可溶性表达人鳞状上皮细胞癌抗原(SCCAg)方法,获得具有较好活性的重组SCCAg抗原并应用于建立抗原检测方法; 。基于pGEX-6P-l载体和大肠杆菌E. coli; ER2566菌株开展重组SCCAg抗原可溶性表达纯化方法研究,评价纯化抗原活性,筛选特异性单克隆抗体,初步建立并评价SCCAg抗原检测方法。结果; 显示,pGEX-6P-l载体和E coli; ER2566菌株可用于建立较高效的可溶性表达和纯化SCCAg抗原的方法,获得了具有较高纯度和活性的重组SCCAg抗原,筛选获得特异性单克隆抗体并; 初步建立了 SCCAg管式化学发光检测方法。建立了有效的基于大肠杆菌表达系统的可溶性表达和纯化SCCAg的方法。The aims of this study are to establish a method for efficient soluble; expression of human squamous cell carcinoma antigen(SCCAg ) based on; Escherichia coli expression system and obtain the recombinant SCCAg; antigen in fine activity, then apply it in the detection method; establishment of antigen. The study on the method of soluble expression; and purification of recombinant SCCAg antigen was conducted based on; pGEX-6P-l vector and E. coli ER2566 strain. The activity of the purified; antigen was evaluated by Abbott Kit and the specific monoclonal antibody; was screened by indirect ELISA. It was proved that PGEX-6P-1 vector and; E. coli strain ER2566 could be used to establish efficient soluble; expression and purification method for recombinant SCCAg antigen.; Moreover, the recombinant SCCAg antigen was proved to be in high purity; and activity. Thus,the SCCAg detection method of chemical luminous tube; was established with the specific monoclonal antibodies. In conclusion,; an effective method for the expression and purification of SCCAg, which; is based on the E. coli expression system, is established.国家高技术研究发展计划(863计划

水痘-带状疱疹病毒主要衣壳蛋白ORF40单克隆抗体的制备及初步应用

目的:制备抗水痘-带状疱疹病毒(VZV)主要衣壳蛋白ORF40的单克隆抗体,对该蛋白在VZV感染细胞内的表达、分布以及在VZV病毒颗粒中的包含情况进行初步探究。方法:设计并合成VZV ORF40多肽片段,偶联...国家自然科学基金项目(81601762)资

Near-atomic cryo-electron microscopy structures of varicella-zoster virus capsids

VZV是一种广泛存在并且具有高度传染性的人类α-疱疹病毒。初次感染VZV可导致水痘，人群普遍易感（感染率约为61%～100%）。该病毒可在背根神经节潜伏感染，持续终生。夏宁邵教授团队长期开展VZV相关基础与新型疫苗研究，通过系统和精细探索建立了高效的VZV规模化培养和病毒颗粒纯化技术体系，成功获得高质量的VZV颗粒样品。首次揭示了疱疹病毒α家族的水痘-带状疱疹病毒（VZV）不同类型核衣壳的近原子分辨率结构，阐明了VZV核衣壳不同组成蛋白的相互作用网络与衣壳装配机制，可为进一步开展新型载体疫苗设计及抗病毒药物等研究提供重要支持。 我校博士后王玮、高级工程师郑清炳、博士生潘德全和俞海副教授为该论文共同第一作者，我校夏宁邵教授、程通副教授、李少伟教授以及美国罗格斯大学朱桦（Hua Zhu）教授、加利福尼亚大学洛杉矶分校周正洪（Z. Hong Zhou）教授为该论文的共同通讯作者。【Abstract】Varicella-zoster virus (VZV) is a medically important human herpesvirus that causes chickenpox and shingles, but its cell-associated nature has hindered structure studies. Here we report the cryo-electron microscopy structures of purified VZV A-capsid and C-capsid, as well as of the DNA-containing capsid inside the virion. Atomic models derived from these structures show that, despite enclosing a genome that is substantially smaller than those of other human herpesviruses, VZV has a similarly sized capsid, consisting of 955 major capsid protein (MCP), 900 small capsid protein (SCP), 640 triplex dimer (Tri2) and 320 triplex monomer (Tri1) subunits. The VZV capsid has high thermal stability, although with relatively fewer intra- and inter-capsid protein interactions and less stably associated tegument proteins compared with other human herpesviruses. Analysis with antibodies targeting the N and C termini of the VZV SCP indicates that the hexon-capping SCP—the largest among human herpesviruses—uses its N-terminal half to bridge hexon MCP subunits and possesses a C-terminal flexible half emanating from the inner rim of the upper hexon channel into the tegument layer. Correlation of these structural features and functional observations provide insights into VZV assembly and pathogenesis and should help efforts to engineer gene delivery and anticancer vectors based on the currently available VZV vaccine.This research was supported by grants from the National Science and Technology Major Projects for Major New Drugs Innovation and Development (no. 2018ZX09711003-005-003), the National Science and Technology Major Project of Infectious Diseases (no. 2017ZX10304402), the National Natural Science Foundation of China (no. 81871648, 81601762), the Research Unit of Frontier Technology of Structural Vaccinology of Chinese Academy of Medical Sciences (no. 2019RU022) and the US National Institutes of Health (DE025567/028583). 该研究获得了国家自然科学基金、新药创制国家科技重大专项和传染病防治国家科技重大专项等资助

Key Technologies of Detecting Depression with Voice Features

抑郁症是一种以抑郁症状为核心并伴随大量其他症状的精神疾病。目前诊断以主观为主，而客观的评估工具对促进抑郁症的更加快速和准确的治疗尤为重要。语音数据临床容易获取，但是语音与抑郁症二者之间还存在以下问题:语音特征是否显著预测抑郁症，纳人混淆变量一一人口学信息后，语音对抑郁症预测的贡献大小;语音特征能否区分是否抑郁;二者关联是否跨情境跨情绪稳定;以及语音特征是否能够在复杂临床诊断情境中保持高鉴别力。 研究一通过二元逻辑回归模型调查语音特征与抑郁症之间的关联是否显著。并纳人人口统计学信息，将其对预测是否抑郁的贡献作为基线水平。本研究收集584抑郁症患者和548名健康人的语音数据。结果发现，有四种语音特征对抑郁症预测起到了主要贡献:PC1 (OR=0.58, P &lt;0.0001) , PC6 (OR=1.57, P &lt;0.001) ,PC17 ( OR = 1. 53 , P &lt;0.0001)和PC24 ( OR = 1.45 , P &lt;0. 05 )。语音特征对抑郁症的单独贡献达到了35.65% (Nagelkerke&#39;s R2)。 研究二设立三种分类模型:单独基于语音的模型;单独基于人口学变量的模型;基于语音与人口学变量的模型。同时该研究纳人了其他数据集作为测试集以便说明模型的泛化能力。本研究包含三个语音数据集，数据集一同研究一，用于分类模型构建。数据集二包含500名抑郁症患者，404名健康人。数据集三包含45名抑郁症患者与58名健康人。结果发现，与以人口学变量建立的抑郁症分类预测模型相比，包含语音的模型(单独基于语音的模型;基于语音和人口学变量建立的模型)一致的达到了较高的分类准确性(F-measure)。在其他数据集上进行测试，得到的结果也是一致的。在该研究中，语音特征单独预测模型在不同测试集上的分类准确性均达到80% 。 研究三收集了45名抑郁症患者与58名健康人的语音数据。研究采用了3(情绪状态:正性，中性，负性)*3(任务类型:语言问答，文本朗读，图片描述)的实验设计，运用机器学习分类算法一一逻辑回归(Logistic Regression, LR)来构建抑郁识别模型。实验结果表明，语音对不同情境下不同情绪状态下的AUC值均在0.6以上(65.7-80.9)，语音的抑郁识别准确性可以达到82.9% o 研究四设定了三种不同的分类任务:1)对健康与非健康组进行分类;2)对健康组与各种精神疾病进行分类;3)对精神疾病两两分类。匹配后有32名躁郁症患者，抑郁症患者106例，健康患者114例，精神分裂症患者20例。从语音中提取MFCC特征并抽取i-vectors。逻辑回归模型评估结果显示:分类抑郁症和双相障碍的模型AUC值为0.5 (F-score=0.44 )。对于其他分类任务，AUC值均在0.75到0.92之间(F-score:0.73~0.91)。在模型性能的比较上，差异检验发现，抑郁症和双相障碍分类模型的性能(AUC )显著差于针对双相障碍与精神分裂症的分类模型(corrected P &lt; 0.05 )。其他分类任务模型好坏差异不显著。而语音特征对抑郁症和双相障碍的分类效果不理想。 本研究对语音特征与抑郁症的关系进行了系统的探讨，说明了以下几点:(1)语音特征能够显著预测抑郁症，语音对抑郁症具有可观的贡献;(2)语音特征能够实际预测抑郁症，模型具有一定的泛化能力;(3)语音的预测作用是跨情境跨情绪稳定的;(4)语音能够在精神疾病临床诊断的复杂情境下具有较高的鉴别能力。这些关键技术研究为进一步探究语音作为临床抑郁症诊断工具的可能性奠定了坚实的基础。</p

A COMPARATIVE STUDY ON TRANSLATION AND INTERPRETING AS PROFESSION

Translation, in general, refers to the faithful representation of what is written or spoken in one language into another. Professionally “translation” is confined to the written, and “interpretation” to the spoken language. As the profession of translator / interpreter has already established an important status in the interlingual communication in today’s world, it is thus of great significance t...学位：文学硕士院系专业：外文学院外文系_外国语言学及应用语言学学号：1993271

An Interview to Professor R.A.Marcus at the 46th ISE Meeting

马库斯教授采访录（第４６届ＩＳＥ年会，１９９５．９．１）ＡｎＩｎｔｅｒｖｉｅｗｔｏＰｒｏｆｅｓｓｏｒＲ．Ａ．Ｍａｒｃｕｓａｔｔｈｅ４６ｔｈＩＳＥＭｅｅｔｉｎｇ按１９９２年诺贝尔学奖获得者，美国加州理工学院Ｒ．Ａ．Ｍａｒｃｕｓ（马库斯）教授应邀参加第４..

新疆沙冬青和蒙古沙冬青叶片及其生境土壤中15种无机元素的含量比较

采用单道扫描等离子体发射光谱仪(ICP)和微波消解仪测定了新疆沙冬青和蒙古沙冬青两种珍稀植物的叶片及生境土壤环境中15种无机元素(Al、B、Ca、Cu、Fe、Mg、Mn、Na、Ba、La、Co、Ni、Pb、Rb和Ti)的含量,其中两种沙冬青叶片及生境土壤中含量最大的元素均为Ca,最小的均为Co.应用两个独立样本检验(Independent-samples T Test)中的曼-惠特尼U检验法(Mann-Whitney U)分析15种无机元素的差异性,结果显示,两种沙冬青的土壤环境相似;叶片的无机元素除了5种元素(B、Fe、Co、Ni和Ti)呈现显著性差异以外,其余元素含量差异不显著;通过比较叶片和土壤的元素含量发现,15种元素均呈现显著性差异;通过叶片内无机元素的浓度与生境土壤中该元素浓度的比值比较,发现Pb和Ti两种元素的叶片内浓度与生境土壤中浓度比值存在显著差异.图1表7参2