Research on Nonstop Cultivating Model for Doctoral Students in China——Taking an Example of X University

我国研究生教育规模的扩张，彰显着我国教育实力的增强。但与西方国家相比，我国的博士生教育发展历史较为短暂，规模的不断扩张势必带来诸多问题。博士生的培养质量的提高成为当前我们面临的主要任务，博士生培养模式的改革成为提高博士生培养质量改革的关键。“直博生”培养模式是近年来新兴起的一种博士生培养模式，它打通了硕士生和博士生两个培养阶段，这种培养模式为高校保留优质生源、提高培养效率、培养科研创新人才贡献了力量。 在系统地总结了我国“直博生”培养的历史沿革和发展状况以及发达国家“直博生”培养模式经验的基础上，以X大学为案例开展访谈调查研究。X大学是一所高水平的研究型大学，是早期开展“直博生”培养的高校之...The expansion of the scale of higher education, highlighting the enhancement of China's educational strength. However, compared with the western countries, the development history of China's doctoral education is too short,the expansion of the scale is bound to bring a lot of problems. Cultivating quality should be further improved is the main task of doctoral education, and the reform of doctoral...学位：教育学硕士院系专业：教育研究院_高等教育学学号：2572014115181

Diversity of bacterial community structure and its driving factors in three bays of Bohai Sea

[Background] Sustainable development of coastal ecosystem has become one of the most important concerns for people nowadays. Riverine output and anthropogenic interrupt have important impacts on the coastal environment. [Objective] In this study, we collected 12 samples from three transects including Bohai Bay, Liaodong Bay and Laizhou Bay to explore the microbial community and diversity in summer of 2015. [Methods] DNA was extracted from water samples by using DNA extraction kit. Samples were analyzed by Illumina HiSeq sequencing technology. We compared the differences among these three transects according to the analysis results. [Results] The diversity index and rarefaction curves showed significant differences among these three transects. The order of diversity value was Laizhou Bay>Bohai Bay>Liaodong Bay. The distribution of the dominant community was as follows: the proportion of Proteobacteria, Bacteroidetes, Cyanobacteria, Actinobacteria and Planctomycetes in the Bohai Bay was 39.8%, 25.7%, 22.4%, 5.85% and 4.38%, respectively. The dominant community proportion in Liaodong bay was Proteobacteria (37.8%), Bacteroidetes (25.7%), Cyanobacteria (17.8%), Actinobacteria (10.4%) and Planctomycetes (5.64%). While in Laizhou Bay there were only four dominant communities as follows: Proteobacteria (59.0%), Bacteroidetes (17.5%), Cyanobacteria (8.2%), Actinobacteria (7.88%). By using the principal component analysis (PCA) and Heatmap correlation analysis, we found that environmental factors were key roles in controlling the microbial diversity in the Bohai Sea. Among them, the concentration of nitrate was particularly significant according to the Mantel test analysis. [Conclusion] The microbial diversity in the three bays of Bohai Sea was very rich and multifarious. The population structure and species in the Laizhou Bay is the most complex and abundant among these three bays, and then it is Bohai Bay and Liaodong Bay. There was a significant correlation among microbial diversity, environmental factors and the spatial distribution. Above all, this study will provide a theoretical basis for further protection and ecological development of Bohai Sea

Characterizing Nitrogen Saturation of the Wuchuan Headwater Stream in the Southeast of China

以九龙江流域典型的农业源头溪流——五川溪为研究区域,开展每月一次共2年的nO3-采样用于溪流氮饱和特征的研究.结果表明,2005年和2007年溪流的nO3-浓度分别为35.5~319.5μEQl-1和5.0~353.6μEQl-1,根据STOddArd和TrAAEn提出的氮饱和划分准则,五川流域分别处于氮饱和阶段2/3和阶段2,接近氮饱和.氮饱和阶段随着nO3-浓度的增加而上升,五川溪流的氮饱和阶段存在着时间上的变化.河流生态系统中氮负荷增加,使河流达到氮饱和状态,并最终改变溪流系统硝化和反硝化等氮的生物地球化学循环过程.随着nO3-浓度的增加,五川源头溪流已成为流域内重要的nO3-源.NO3-concentrations sampled monthly in the Wuchuan stream,which is a small agricultural headwater stream in the southeast of China,within two years,were used to study the stream′s nitrogen saturation characteristics.Results showed that NO3-concentrations varied from 35.5 to 319.5 μeqL-1 in 2005 and from 5.0 to 353.6 μeqL-1 in 2007.And,according to the criteria proposed by Stoddard and Traaen,the nitrogen saturation status of the stream was at stage 2 or 3,and stage 2,respectively.Both stages were closed to saturation.It was found that the saturation stage would increase with the NO3-concentration,which caused the change of nitrogen saturation stage in the Wuchuan stream over time.The increasing of nitrogen loadings has led to the stream nitrogen saturation and eventually can alter nitrification,denitrification and other nitrogen biogeochemical cycles in the stream ecosystem.国家自然科学基金资助项目(41175130);教育部新世纪优秀人才支持计

死亡受体5胞外区域的重组、表达及活性鉴定

目的构建死亡受体5(deathreceptor5,DR5)胞外区域(eDR5)的表达载体,表达纯化重组蛋白并鉴定其生物特性。方法通过重叠PCR获得DR5胞外段编码序列,构建pET-22b(+)/DR5表达载体,转化大肠杆菌BL21(DE3),IPTG诱导表达,Ni2+柱亲和纯化,SDS-PAGE、直接ELISA鉴定纯化产物的纯度和特异性,用MTT法检测eDR5蛋白阻断DR5单克隆抗体FMU1.5和TRAIL诱导人胶质瘤细胞株U343(高表达DR5)、U373(低表达DR5)细胞凋亡的作用。结果获得了DR5胞外段编码序列,目的蛋白在上清及包涵体中都有表达,表达量占菌体总蛋白的30%以上,纯化的重组蛋白纯度达95%以上,蛋白产量达9mg/ml。ELISA结果表明所纯化蛋白为eDR5。eDR5蛋白可部分阻断FMU1.5和TRAIL诱导人胶质瘤细胞株U343细胞凋亡的作用,其阻断率与DR5表达相关。结论死亡受体5胞外段基因的成功重组、表达及纯化,为进一步的功能研究奠定了基础

Construction,expression,and purification of RGD-FasL and analysis of its activities

目的构建适于原核表达的重组蛋白RGD-FasL表达载体,并进行重组蛋白的表达纯化及抗肿瘤活性分析。方法通过重叠PCR将RGD序列插入到FasL基因的N端,获得RGD-FasL基因,构建pGEX-5X-1/RGD-FasL表达载体。转化大肠杆菌BL21(DE3),IPTG诱导表达,GST柱纯化。采用体外黏附实验、MTT比色法、流式细胞法检测融合蛋白的功能。结果通过重叠PCR获得了编码正确氨基酸序列的目的基因。目的蛋白以分泌的形式表达,表达量占菌体总蛋白的30%以上。纯化后,蛋白纯度达95%以上。体外黏附实验表明所纯化的融合蛋白可与宫颈癌Hela细胞发生特异结合。MTT比色法与流式细胞技术均表明纯化的融合蛋白能特异性地诱导肿瘤细胞发生凋亡。结论重组蛋白RGD-FasL表达载体的成功构建、表达、纯化及活性分析,为进一步的功能研究奠定了基础。 【英文摘要】 Objective To construct,express,and purify RGD-FasL gene and analysis its anti-tumor activities. Methods The RGD-FasL gene was constructed by overlapping PCR through inserting the sequence of RGD at the N-terminate of FasL gene,and then was inserted into vector pGEX-5X-1 and expressed efficiently in E.coli BL21(DE3) under optimization condition. The expressed products were purified by GST resin column. The function of recombined protein was detected by adhesion in vitro analysis,MTT colorimetric,and flow cyt..

A Study on the mechanisms of eutrophication of a shallow upstream lake in the Jiulong River Catchment

基于对九龙江上游龙潭湖富营养化水体和沉积物现状的监测结果,通过与国内富营养化深水湖库和流域下游大型富营养化浅水湖泊进行对比,深入探讨了流域上游浅水湖泊富营养化发生的原因及主导机制.流域上游浅水湖泊具有外源污染物输入较少的特点,较下游大型浅水湖泊更易受温度等气候条件和沉积物氧化还原状态的影响,以及外源输入总磷控制具有较强的滞后效应,因此对流域上游浅水湖泊富营养化的控制必须重视内源营养盐释放,特别是结合态磷的内源释放问题.Based on site monitoring chemical data of lake water and sediments during an algal bloom in the Longtan Lake,causes and mechanisms of eutrophication in shallow upstream lakes were discussed by comparing the Longtan Lake in upstream of the Jiulong River with deep eutrophic lakes and large shallow downstream eutrophic lakes in China.The shallow upstream lake is characterized as relatively simple nutrient inputs and high susceptivity to climatic variability(such as temperature),redox conditions,and strong lag effects of the external total phosphorus inputs.Therefore,attention must be paid to the sediments nutrient releases,especially,the bound phosphorus of sediments when remediating eutrophication in shallow upstream lakes.国家自然科学基金资助项目(41175130);教育部新世纪优秀人才支持计

人死亡受体5全长基因的构建及转染胶质瘤细胞

目的构建人死亡受体5(death receptor5,DR5)全长基因真核细胞表达载体pcDNA3.1/DR5,pcDNA3.1/GFP/DR5,并观察其转染胶质瘤细胞U138的效果。方法通过重叠PCR获得DR5全长编码序列,构建pcDNA3.1/GFP/DR5,pcDNA3.1/DR5表达载体,利用脂质体转染试剂盒,分别将2种质粒pcDNA3.1/GFP/DR5、pcDNA3.1/GFP共转染胶质瘤细胞U138,转染后72h,半定量逆转录聚合酶链反应(RT-PCR)检测DR5mRNA的表达,流式细胞术检测DR5的表达强度、检测Anti-DR5对胶质瘤细胞U138生长的影响。结果获得了DR5全长编码序列,成功瞬时转染U138,RT-PCR、流式细胞术检测结果表明,转染后U138细胞DR5mRNA、蛋白水平的表达明显增加,Anti-DR5可以明显抑制U138细胞的生长。结论获得了DR5全长编码序列,探索到成功转染DR5的最佳方法,为稳定筛选高表达DR5的U138细胞提供依据

Analysis of DcR3 in rat model of adjuvant arthritis

目的研究诱骗受体DcR3对佐剂型关节炎(AA)大鼠模型的作用及其机理。方法注射弗式完全佐剂建立大鼠佐剂型关节炎(AA)模型,尾静脉注射DcR3蛋白,观察大鼠关节肿胀度、间接ELISA检测血清和滑膜液中细胞因子IL-1β、TNF-α、IFN-γ的变化。RT-PCR检测滑膜和淋巴细胞中DcR3、Fas、FasL mRNA的表达以及脾脏中TGF-β、IFN-γ、TNF-α、IL-4、IL-10 mRNA的表达。Western blot分析滑膜细胞中Caspase-8、Caspase-3、Caspase-9、Bcl-2蛋白的表达。结果DcR3治疗AA大鼠后,足肿胀度降低;血液和滑膜液的IL-1β、TNF-α、IFN-γ水平下降;脾脏中TGF-β、IFN-γ、TNF-αmRNA表达下调,IL-4、IL-10 mRNA表达上调;滑膜细胞中Caspase-8、Caspase-3蛋白表达上调,Bcl-2蛋白表达下调。结论DcR3可以用于实验性大鼠AA的治疗,其治疗机制与调节滑膜细胞FasL、Fas mRNA的表达和血液淋巴细胞中Fas mRNA的表达,促进滑膜细胞和自身反应性淋巴细胞的凋亡;调节脾脏细胞Th1/Th2细胞因子... 【英文摘要】 Objective To study the immunoregulatory mechanisms of DcR3 on rats adjuvant arthritis(AA).Methods AA was induced by complete freund s adjuvant(CFA)in rats,and the rats were injected subcutaneously with DcR3.The perimeters of rat hind soles were measured before and after the injection of CFA.The pathological changes of inflammatory knee joints were observed under optical microscope.The changes of IL-1β,TNF-α,and IFN-γ in serum and synovial liquid were detected by indirect ELISA,respectively.The DcR3,Fas,and ...厦门大学科研启动基金(Z03103

Study on the prothymosin α as vaccine adjuvant of P.yoelii

目的:研究胸腺素α原(PrOTHyMOSInα,PrOTα)作为约氏疟原虫疫苗免疫佐剂的作用。方法:提取P.yOElII-17Xnl全蛋白作为抗原,用胸腺素α原作为免疫佐剂,免疫小鼠。具体方案为:昆明小鼠分为4组,每组6只,A组免疫P.yOElII-17Xnl全蛋白+PrOTα;b组免疫P.yOElII-17Xnl全蛋白;C组只注射PrOTα;d组为空白对照,以相同体积的生理盐水代替。免疫结束后感染致死的P.yOElII-17Xl,1x107个虫/只小鼠。结果:感染后的前10天A组小鼠疟原虫血症平均值要低于其他三组,且最终有3只小鼠存活下来,存活率50%,C组有一只小鼠存活,b、d组小鼠全部死亡。结论:用P.yOElII-17Xnl全蛋白做抗原,用PrOTα作为佐剂,比单独用P.yOElII-17Xnl全蛋白对小鼠有更好的免疫保护作用,提示了PrOTα可以成为一种有潜力的蛋白疫苗。Objective:To investigate the function of prothymosin α(ProTα) as vaccine adjuvant of P.yoelii.Methods:The mice were immunized with the total protein extracted from P.yoelii-17XNL as antigen,together with prothymosin α as adjuvant.Programs:Kunming mice were divided into A,B,C and D group.A group was immunized with P.yoelii-17XNL total protein and ProTα;B group was immunized with P.yoelii-17XNL;C group was only injected with ProTα;D group was the control,only injected with physiological saline.And then,the mice of each group was infected with P.yoelii-17XL,the dosage was 1×107/mice.Results:The parasitemia of A group-mice was lower than the other three groups in the first 10 days after infection,and eventually there were three mice survived in A group,the survival rate was 50%,one mouse survived in C group,all of mice in B group and D group died.Conclusion:Mice immunized with P.yoelii-17XNL total protein as antigen together with ProTα as adjuvants,had better immune protection than those immunized with P.yoelii-17XNL protein only.The present results suggest that the ProTα can act as a potential adjuvant in protein vaccine.厦门市科技项目(3502Z20083004);国家“973”项目(2007CB513103)资