检测细胞角蛋白在食管癌外周血和骨髓微转移中的意义

目的:探讨食管癌患者外周血和骨髓中CK18mRNA基因表达及临床意义。方法:采用巢式RT-PCR方法检测50例M0、10例M1的食管癌患者及10例食管良性病变患者外周血及肋骨骨髓中细胞角蛋白CK18mRNA基因的表达。结果:50例M0食管癌患者外周血、骨髓中CK18mRNA阳性表达率分别为38.0%(19/50)和42.0%(21/50);10例M1的食管癌患者外周血、骨髓中CK18mRNA阳性表达率分别为90.0%(9/10)、100%(10/10);食管良性病变患者的外周血和骨髓中K18mRNA阳性表达率均为0%(0/10),CK18mRNA阳性表达在三组间差异有显著性(P0.05),但呈正相关性(r=0.704,P<0.01)。结论:应用RT-PCR法检测食管癌患者外周血或骨髓中CK18mRNA微转移有助于食管癌的分期和预后的判断

The efect of elemene reversing the multidurg resistance of A549/DDP lung cancer cells

目的 :探讨榄香烯乳(ElEMEnE)逆转人肺腺癌耐顺铂(CISPlATIn,ddP)细胞A549/ddP的耐药性及可能的作用机制。方法 :采用MTT法检测榄香烯乳单药作用时的细胞毒作用及与ddP联合作用对A549/ddP细胞耐药的逆转作用;HOECHST 33342法进行细胞核染色,荧光显微镜下观察细胞形态的改变;fCM检测细胞凋亡率;蛋白质印迹法检测细胞质中细胞色素C(CyTOCHrOME C,CyT C)、PrO-CASPASE-3、CASPASE-3和抗凋亡因子b细胞淋巴瘤-2(b CEll lyMPHOMA-2,bCl-2)家族蛋白表达的情况。结果 :不同浓度榄香烯乳对A549/ddP细胞均有一定的抑制作用,呈时间-剂量依赖性效应,联合ddP能提高耐药细胞株A549/ddP对ddP的敏感性而逆转其耐药性。HOECHST 33342法和fCM法检测结果显示,榄香烯乳联合ddP能更大程度上诱导A549/ddP细胞的凋亡,提高耐药细胞的凋亡率,同时还上调A549/ddP细胞质中CyT C、CASPASE-3和bAd蛋白的表达,而下调PrO-CASPASE-3和bCl-2蛋白的表达。结论 :榄香烯乳能逆转A549/ddP细胞耐药性可能与其损伤线粒体膜,使线粒体释放CyT C到细胞质、同时活化CASPASE-3、上调促凋亡蛋白bAd和下调抗凋亡蛋白bCl-2表达以启动凋亡路径有关。Objective: To investigate the effect of elemene on the drug resistance of cisplatin(DDP)- resistant human lung adenocarcinoma A549/DDP cells, and its possible mechanism.Methods: The growth inhibition of A549/DDP cells treated with elemene alone or combined with different concentrations of DDP was detected by MTT assay.The morphological changes of apoptosis of A549/DDP cells treated with different concentrations of elemene(20 and 40 μg/mL) for 24 h were observed by Hoechst 33342 staing under fluorescence a microscope.The apoptosis rate of A549/DDP cells treated with different concentrations of elemene(20 and 40 μg/mL) after 24 h were detected by FCM(flow cytometry).The expressions of cytochrome C, pro-caspase-3, caspase-3 and the Bcl-2 family proteins were measured by Western blotting analysis.Results: MTT result showed that different concentrations elemene could inhibit the proliferation of A549/DDP cells in a time- and dose- dependent manner.Intriguingly, elemene plus DDP enhanced the sensitivity of A549/ DDP cells to DDP and reversed the resistance of A549/DDP cells.Elemene was also a strong inducer of apoptosis in this model system, and a synergistic effect on induction of cell death was observed when the tumor cells were treated with both agents.The result showed that elemene could enhance A549/DDP cell apoptosis.Furthermore, the combination of elemene and DDP also enhanced the protein expressions of cytochrome C, caspase-3 and Bad, and reduced the protein levels of Bcl-2 and pro-caspase-3 in the A549/DDP cells.Conclusion: The reversal of the multi-drug resistance of A549/DDP cell line by elemene may result from its effect on induction of apoptosis.Elemene treatment can impaire mitochondrial membrane, release cytochrome C into cytoplasm, activate caspase-3,up-regulate pro-apoptotic protein Bad and down-regulate anti-apoptotic protein Bcl-2, and finally caused apoptosis.福建省自然科学基金面上项目(编号:2010D014

Elemene reversed the multidurg resistance of A549 /DDP lung cancer cells via mitochondrial apoptosis pathway

目的:探讨榄香烯乳(ElEMEnE,ElE)逆转人肺腺癌耐顺铂(CISPlATIn,ddP)细胞A549/ddP的耐药性及作用机制。方法:采用MTT法检测榄香烯乳单用的细胞毒作用及与ddP合用时耐药逆转作用。荧光探针JC-1结合激光共聚焦显微镜检测线粒体膜电位的变化。dCfH-dA荧光探针结合流式细胞仪检测细胞内活性氧(rEACTIVE OXygEn SPECIES,rOS)水平。用谷胱甘肽试剂盒结合分光光度法检测计算gSH/(gSSg+gSH)比值。蛋白质印迹法检测胞质中CyTO C、PrO-CASPASE-3、CASPASE-3和bCl-2家族蛋白表达情况。结果:不同浓度榄香烯乳抑制A549/ddP细胞株生长,呈时间-剂量依赖性效应,联合顺铂能提高A549/ddP细胞株对顺铂的敏感性而逆转耐药。不同浓度榄香烯乳联合顺铂使A549/ddP细胞株线粒体膜电位下降,rOS浓度增加,gSH/(gSSg+gSH)比值降低,上调胞质中CyTO C、CASPASE-3、bAd蛋白表达,下调PrO-CASPASE-3、bCl-2蛋白表达。结论:榄香烯乳逆转A549/ddP细胞株耐药性可能与其损伤线粒体膜,活化胞内氧化还原体系,诱导线粒体凋亡路径有关。Objective: To explore the mechanism that elemene( ELE) reversed the multidurg resistance( MDR)of A549 /DDP lung adenocarcinoma cell.Methods: MTT assay was used to determine the growth inhibition of human lung adenocarcinoma A549 /DDP cells in vitro.Mitochondrial membrane potential( MMP) was monitored by JC- 1fluorescence probe with laser confocal scanning microscopy,the intercellular reactive oxygen species( ROS) level was measured by 2',7'- dichlorfluorescein- diacetate( DCFH- DA) staining and flow cytometry and the ratio of GSH /( GSSG +GSH) was calculated according to detection results of GSH kit.The expression of Cytochrome C,Caspase-3 and the Bcl- 2 family proteins and in the case of cyclosporine A and DEVD- CHO,the expression of Caspase- 3expression were measured by Western blot.Results: MTT results showed that different concentrations ELE could inhibit the proliferation of A549 /DDP cells in a time- and dose- dependent manner.Intriguingly,ELE plus cisplatin enhanced the sensitivity of A549 /DDP cells to cisplatin and reversed A549 /DDP cells dury resistance.Different concentrations ELE decreased mitochondrial membrane potential,increased intracellular ROS concentration and decreased GSH /( GSSG + GSH) ratio of A549 /DDP cells in a time- and dose- dependent manner.Furthermore,the combination with both ELE and cisplatin also enhanced the protein expression of Cytoplasmic C,Caspase- 3 and Bad,and reduced the protein levels of Bcl- 2 and Pro- caspase- 3 in the cisplatin- resistant A549 /DDP cancer cells.Conclusion: ELE reversed the MDR of A549 /DDP cell line may demage mitochondrial membrane,active intracellular redox system and induce the mitochondrial apopotosis pathway.福建省自然科学基金面上项目(编号:2010D014

苦参碱对人食管癌细胞株Eca-109增殖和凋亡的影响

目的探讨中药提取物苦参碱单体(Matrine)对人食管癌细胞株Eca-109的诱导凋亡和抑制增殖作用。方法体外培养人食管癌细胞株(Eca-109),分别给以不同浓度的苦参碱对体外培养的Eca-109细胞株进行干预。以MTT比色法、Hoechst33342染色法以及流式细胞术测定苦参碱对Eca-109细胞株的诱导凋亡及抑制增殖的作用。结果 MTT实验显示苦参碱可以明显抑制Eca-109细胞株的增殖,流式细胞术检测细胞周期显示G2期细胞明显增多,S期细胞显著减少。结论苦参碱能明显抑制Eca-109细胞株的增殖,促进其凋亡,其作用机制可能与细胞周期阻滞于G2期有关

食管癌组织中CuZn-SOD活性和表达的研究

[目的]通过对食管癌组织和癌旁组织中CuZn-SOD活性和表达检测,探讨CuZn-SOD活性和表达在食管癌发生、发展中的作用。[方法]CuZn-SOD活性和表达的检测分别采用DTNB直接法和Western Bloting方法。[结果]总SOD和CuZn-SOD的活性在食管癌组织和相应癌旁组织中的差异尚不能认为有统计学意义。30%食管癌组织中CuZn-SOD的表达明显高于癌旁组织。[结论]不同病例中表达水平结果不一致,CuZn-SOD的表达水平与肿瘤的发生或发展存在一定的联系

The effect of elemene on reversal multidurg resistance and the expression of P-glyco-protein in A549/DDP

目的:探讨榄香烯乳(ElEMEnE,ElE)逆转人肺腺癌A549/ddP细胞株耐药性及其机制。方法:采用MTT法检测榄香烯乳单用的细胞毒作用及与顺铂(CISPlATIn,ddP)合用时耐药逆转作用;采用流式细胞术检测榄香烯乳对A549/ddP细胞内罗丹明-123(rHOdAMInE-123,rH123)蓄积的影响;采用WESTErn blOT检测榄香烯乳对耐药细胞A549/ddP细胞膜上P-gP蛋白表达的影响。结果:不同浓度榄香烯对A549/ddP细胞均有一定的抑制作用,呈时间-剂量依赖性效应。单用ddP的IC50为15.46μg/Ml,合用20μg/Ml榄香烯24H,IC50为4.15μg/Ml,逆转耐药倍数为3.63。同时,榄香烯增加A549/ddP细胞内rH123的蓄集,降低细胞膜上PgP的表达,且呈剂量依赖性,表明榄香烯能减少A5 4 9/ddP细胞对药物的外排和抑制P-gP蛋白的表达。结论:榄香烯在体外逆转肿瘤细胞耐药性可能与其抑制P-gP的功能和表达有关。Objective: To investigate the elemene( ELE) reversal of human lung adenocarcinoma A549/DDP cell resistance and its mechanism.Methods: Cytotoxicity of ELE alone or in combination with DDP in A549 / DDP cells was determined by MTT assay.Based on flow cytometric technology,the effect of ELE on the uptake of rhodamine-1 2 3( Rho1 2 3) in A5 4 9 / DDP cells was detected by measuring Rho123 mean fluorescence intensity( MFI).P- gp protein expression on the A549 / DDP cell membrane were detected by Western blot.Results: Different concentration elemene inhibited the A549 / DDP cell proliferation in a time- dose- dependent manner( P < 0.05).Alonely used DDP,IC 50 was 15.46μg / ml; combined 20μg / ml elemene for 24h,DDP IC 50 was 4.15μg / ml,the reversal activity was 3.63 times higher than that of DDP.Elemene increased intracellular accumulation of Rh123 of the A549 / DDP cells, reduced the expression of P- gp on the cell membrane,in a dose- dependent manner,indicating that elemene reduce A549 / DDP cells for drug efflux and inhibition of P- gp protein pump function.Conclusion: The reversal activity of DDP is possible related to inhibition of P- gp function and expression in vitro,which lead to an increased intracellular accumulation of anticancer drugs.福建省自然科学基金面上项目(编号:2010D014

A Blended Teaching Style of Scientific Inquiry and Interactive Teaching in Application of Competition Training of the College Chemical Experiment Skills

为进一步提高实验教学质量,培养学生的创新精神、实验、实践动手能力及综合素质,本文在大学分析化学实验竞赛操作培训中构建了探究互动模式。以化学实验竞赛为动力,引领进一步深化教学改革。提出了在探究实验中互动,在互动中进行探究的教学方法,从而培养了学生的实验操作技能、创造性及创新性。In order to improve the learning and teaching quality of experiment course and cultivate students ＇innovative spirit,and experiment skills,and practical ability and comprehensive quality,a blended teaching style of scientific inquiry and interactive teaching was constructed during the college analytical chemical experiment skill training.The further teaching reform was developed using chemical experiment competition as a driving force. A new teaching method of scientific inquiry during interactive teaching and interactive teaching during scientific inquiry was put forward.Therefore,the students＇ experimental skills,creativity and innovation were improved.广西高等教育教学改革工程项目(2014JGA129);广西民族大学2103年度高等教育教学改革工程项目(2012XJGY23

Practice of the Blended Styles of Constructivism and Scientific Inquiry Teaching in Instrumental Analysis Course

为了提高《仪器分析》课程教学质量与水平，本工作在化学及应用化学两个班的教学过程中运用两种教学方法：一种是单一的讲授方法；另一种是以建构主义理论为指导，并融合探究模式，这种教学活动过程包括：课前布置任务，课堂创设问题情景，引导学生探究问题，并提出猜想或假设，对假设进行检验及演绎推理，引导学生发现规律得出结论。通过一学期的实践与比较，发现建构主义与探究式相融合的教学效果比传统讲授的教学效果要优越。n order to improve the learning and teaching quality and level of Instrumental Analysis courses, two different teaching styles were carried out in two classes of chemistry major and applied chemistry major （counterparts）, respectively.One style was to utilize traditional teacher-centered manner of lecture, the students passively listened to their teachers＆#39;lecture and take notes; another format was to be guided by constructivism, and to blend inquiry learning and teaching.This blended learning and teaching style included the following processes： subsequently, to assign homework to students before class, to design problem environments, to pilot students to inquire problem on their own, and to propose their guess or hypothesis, to test their hypothesis and conduct deductive reasoning, to motivate students to find law and finally make conclusion in classroom.The investigations found that the blended learning and teaching style of constructivism and inquiry was superior to the traditional one through a semester experiment.广西高等教育教学改革工程项目(2014GA129);广西民族大学2103年度高等教育教学改革工程项目(2012XJGY23

Applications of Scientific Inquiry Teaching in Instrumental Analysis Courses

为了提高《仪器分析》课程教学质量，该研究在环境工程与制药工程专业的《仪器分析》课程中分别采用了两种教学模式：一种是“填鸭式”的教学，老师讲学生听；另一种是探究式，引导学生对一些知识点进行探索．后者这种教学活动过程包括：设问质疑，做出猜测或假设、交流，得出初步结论．通过一学期的实践与比较，发现探究式的教学效果比传统讲授的教学效果要优越．In order to improve the learning and teaching quality of instrumental analysis courses, two different teaching styles were carried out in two classes of environmental engineering specialty （counterparts） and pharmacy engineering specialty, respectively. One style is to cramming method of teaching： students passively accept their teachers＇ lecture and take notes; another format is a scientific inquiry teaching： to guide the students to explore knowledge point. This teaching style includes the following processes： subse- quently, to assign homework to students before class, to design problem environments, to pilot students to inquire problem on their own, and to propose their guess or hypothesis, to test their hypothesis and conduct deductive reasoning, to motivate students to find law and finally make conclusion in classroom. The investi- gations found that the scientific inquiry teaching was superior to the traditional one through a semester exper- iment.广西高等教育教学改革工程项目(2014GA129);广西民族大学2103年度高等教育教学改革工程项目(2012XJGY23

建构、探究及互动混合模式在教学中的实践——以大学仪器分析课堂为例

课程教学模式的改革对于培养创新能力的人才具有重要作用。为了提高化工专业仪器分析课程教学质量,在教学过程中可运用两种教学模式:一种是传统的讲授,老师讲学生听;另一种是以建构主义理论为指导,并融合探究及互动模式,这种教学活动过程包括:课前布置任务,课堂创设问题情景,引导学生自主探究问题,并提出猜想或假设,对假设进行检验及演绎推理,引导学生发现规律得出结论。通过一学期的实践与比较,发现建构主义指导下的探究及互动混合灵活多变模式的教学效果比传统单一讲授的教学效果要优越。2014年广西高等教育教学改革工程项目(No.2014JGA129);广西民族大学2012年度高等教育教学改革工程项目(No.2012XJGY23)支