Immobilization of Antibody on Polystyrene Microbeads and Application for Immunoassay in Human Serum

摘要 液相芯片是一种多功能新型生物芯片平台，与传统的固相芯片相比，液相芯片具有灵活性好、检测通量高、反应分析时间短、操作方便等明显的优势，广泛应用于核酸杂交分析、免疫学或者基于免疫学原理进行的分析、高通量药物筛选等研究。在液相芯片中，其核心元件功能化微珠承担着分子识别和信号表达的重要任务，微珠表面生物探针分子的固定化效果直接影响芯片的灵敏度、检测限等，因此，研究微珠表面生物分子的固定化方法十分重要。 本论文基于构建液相芯片敏感元件并用于血清中肿瘤标志蛋白检测的目的，重点研究了液相芯片敏感元件聚苯乙烯微珠的活化及表面抗体分子固定化的方法，优化了分析条件，并用于血清样品的检测。研究工作主要有以...Abstract Liquid chip is a versatile biochip platform. Compared to conventional solid chip, it has some advantages such as good flexibility, high throughput, short response time and easy operation, and so is widely used in nucleic acid hybridization analysis, immunological analysis and high throughput drug screening. Functional microbeads which were core components of liquid chip undertake the imp...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：2052007115097

Construction of expression vector of HCV NS4B and its effect on cell apoptosis

目的 构建丙型肝炎病毒（HCV）2a型非结构蛋白4B（NS4B）的真核表达载体,并观察其对骨肉瘤细胞U2OS凋亡的影响。方法 以质粒PJFH1为模板,通过PCR反应扩增4目的基因片段,采用同源重组法与PCMV-tag2b载体相连,转化大肠杆菌感受态DH5α,筛选正确的克隆。以脂质体为介导转染U2OS细胞,通过Western blot检测NS4B蛋白的表达,免疫荧光检测细胞的凋亡。结果 构建pCMV-tag2b-NS4B重组质粒,经测序其与NCBI公布的序列完全一致,并可在U2OS细胞中表达,DAPI检测显示细胞凋亡率为（17.25±2.95）%,对照组凋亡率为（6.53±2.36）%。结论 成功构建NS4B的真核表达载体,并可在U2OS细胞中表达,诱导细胞凋亡。山西省自然科学基金（No：2014021037-9）; 山西省高等学校大学生创新创业训练项目（No：2014052）; 山西医科大学汾阳学院科技发展基金（No：2016B02

Quantitative Analysis of Surface-Immobilized Hydrazide Groups by Spectrophotometry

建立了一种测定固相表面酰肼基团含量的方法。运用fE(Ⅲ)-邻二氮菲显色体系,通过固相表面具有较强还原性的酰肼基团将fE(Ⅲ)还原为fE(Ⅱ),生成的fE(Ⅱ)可与邻二氮菲生成稳定的桔红色络合物,从而利用分光光度法测定溶液体系吸光度来表征固相表面酰肼基团的含量。该方法在0~0.02μMOl/Ml范围内呈良好线性关系,线性相关系数r=0.9975,检出限为5.4x10-4μMOl/Ml。用该方法测出微珠固相表面酰肼基团的活化效率为72%。方法可以很好地应用于固相载体表面酰肼基团的测定。A measurement approach of hydrazide groups on solid-phase surface by spectrophotometry was reported.By reducing Fe(Ⅲ) to Fe(Ⅱ) in solutions with microbead surface-conjugate hydrazide groups,a stable orange-red complex of Fe(Ⅱ)-orthophenanthroline was formed.The content of hydrazide groups on the bead surface was then characterized by spectrophotometric measurement.This method has advantages of high sensitivity,simplicity and practicability.The linear range is from 0 to 0.02 μmol/mL with the linear correlation coefficient of 0.9975 and detection limit of 5.4×10-4 μmol/mL(S/N=3).In this method,72% of the activation efficiency of the hydrazide groups activated on the carboxylated microbead surface was detected.It is promising to apply the approach to the quantitative assessment of hydrazide groups on various solid-phase surfaces.国家自然科学基金(No.20775065;No.20835005);教育部高校博士点基金(No.20070384023);国家基础科学人才培养基金(No.J1030415

场景主旨加工的上级水平优势：来自注意瞬脱范式的证据

目的：考察场景主旨层级加工的优先性及其对注意资源的需求。 方法：利用注意瞬脱范式，在相同的T1任务（识别面孔）条件下，记录被试在四种时间延迟条件下完成T2任务（识别不同层级的场景）的正确率。 结果：与基本水平相比，上级水平的场景主旨识别正确率更高、瞬脱量更小、瞬脱期更长；识别不同层级的场景主旨对识别面孔的正确率具有调节作用。 结论：与基本水平相比，上级水平的场景主旨识别对注意资源的需求量较小，支持场景主旨加工的上级水平优势观点。</p

Retrospect and prospect of research on C4 photosynthesis in diatoms

Diatoms（Bacillariophyta） is one of the main groups of marine phytoplankton and plays an important role in global carbon cycle. The emergence of C4 photosynthesis is a major stride in plant evolution, and also affects the global carbon cycle. However, the understanding of C4 photosynthesis of diatoms is still limited. Since the 1970 s, scientists have argued whether C4 is involved in diatoms. In this paper, the discovery history of diatom C4 photosynthesis, the diatom carbon dioxide concentration mechanism（CCM） and the comparison of C4 photosynthesis between diatom and higher plants were reviewed. New methodology in the study of diatom’s C4 photosynthesis is prospected to know better about the role of diatoms in global carbon cycle

Retrospect and prospect of research on C_4 photosynthesis in diatoms

Diatoms (Bacillariophyta) is one of the main groups of marine phytoplankton and plays an important role in global carbon cycle. The emergence of C_4 photosynthesis is a major stride in plant evolution, and also affects the global carbon cycle. However, the understanding of C_4 photosynthesis of diatoms is still limited. Since the 1970s, scientists have argued whether C_4 is involved in diatoms. In this paper, the discovery history of diatom C_4 photosynthesis, the diatom carbon dioxide concentration mechanism (CCM) and the comparison of C_4 photosynthesis between diatom and higher plants were reviewed. New methodology in the study of diatoms C_4 photosynthesis is prospected to know better about the role of diatoms in global carbon cycle

Preparation of CdSe/CdS Quantum Dots and Quantum Dots@ Polystyrene Fluorescent Microspheres

针对量子点在聚苯乙烯(PS)微球中包载率不高的问题,首先通过热循环单前驱体耦合法制备出高效发光的CdSe/CdS核壳量子点,再采用配体交换及乳液; 聚合法将制备的CdSe/CdS核壳量子点包载在PS微球中.实验结果表明:所制备的CdSe/CdS核壳量子点的荧光量子产率超过90%;经过配体交换; ,不仅提高了量子点的溶解度,而且PS微球对量子点的包载率提高至87%.该结果对于提高量子点荧光微球在荧光免疫分析中的检测灵敏度具有较重要的意义.In order to improve the encapsulation efficiency of quantum dots in; polystyrene microspheres,CdSe/CdS core-shell quantum dots were firstly; prepared by thermal cycling single precursor coupling method. The; quantum dots@ polystyrene fluorescent microspheres were then; successfully obtained by embedding the CdSe/CdS core-shell quantum dots; in polystyrene microspheres using ligand exchange and emulsion; polymerization. The results indicated that the fluorescent quantum yield; of the prepared core-shell quantum dots was more than 90%.Using the; method of ligand exchange,not only the solubility of the core-shell; quantum dots was improved,but also the encapsulation efficiency of; quantum dots in polystyrene microspheres was increased to 87%.This might; be useful in improving the detection sensitivity of quantum dots; fluorescent microspheres in immunofluorescence assay.国家自然科学基

A new mutation pattern of hepatitis B virus X gene

目的:证实乙型肝炎病毒(hepatitis Bvirus,HBV)X基因一种新的变异方式.方法:从HBV慢性感染患者血清中提取HBVDNA,扩增X基因序列,克隆入pMD19T载体,选择阳性克隆进行DNA测序,与已知HBV基因相应序列比较该患者体内HBV基因变异位点以及变异形式.结果:从21例患者中共挑选74个克隆测序,测序结果提示54个克隆X基因下游大段缺失突变,长度达234nt,位于1601-1834nt处,另有1个克隆发生245nt缺失突变.发生缺失变异的病毒株同时存在G/A1515C、G1518C和A1585T替换突变,这两种突变具有联动特征.缺失突变株HBx仅编码76aa,其第44和45位编码为LL,具有特异性.结论:观察到一种X蛋白变异方式,这种大段缺失突变导致X蛋白下游编码序列丢失,其为X因子还是X蛋白以及这种变异是否为常态形式尚需进一步研究.AIM: To identify a novel X gene mutation pattern of hepatitis B virus (HBV) in patients with chronic HBV infection.METHODS: A pair of primers was designed on the basis of nucleotide sequences of X gene. Polymerase chain reaction (PCR) was used to amplify the target region from HBV DNA sam- ples extracted from chronic hepatitis B patients in Xiamen city. After electrophoresis of the PCR products in 9 g/L agarose gel, the target regions were cut, re-purified and TA-cloned into pMD19 T vector. The inserted regions in positive clones were sequenced. Sequence comparison with HBV genome submitted in GenBank was made to find the mutation sites. RESULTS: Totally 74 strains from 21 patients with chronic HBV infection were sequenced, and the results showed that there was a charac- teristic deletion region, with a length of 234 nt (nt 1601-1834) in 54 clones, and a length of 245 nt in 1 clone. There were 3 replacement mutations bounding to deletion mutation: G/A1515C, G1518C and A1585T, which caused substitutions in the 44th and 45th amino acid site to LL. These mutant strains only coded 76 aa of up-stream HBx. CONCLUSION: A novel deletion mutation in HBV X gene is observed in patients with chronic HBV infection. The deletion mutants encode 76-aa X factor instead of X protein.厦门市首批重大疾病科研攻关资助项目,No.WKZ0501;; 厦门市卫生局医学科研立项资助项目,No.WSK0506;; 厦门大学引进人才科研启动基金资助项目,No.Z03109;; 福建省青年科技人才创新资助项目,No.2006F3127~