科创中国

Objective To detect the cell viability and the expressions of stem cell surface markers after chemotherapeutic drug treatment.Methods We observed the cytotoxic effects of three chemotherapeutic agents[epirubicin(Epi),fluorouracil(5-FU)and cyclophosphamide(Cyc)]in three cell lines,and the cell viabilities after removed these chemotherapeutic agents.Expressions of stem cell surface markers CD44,CD24,CD90,CD14 and aldehyde dehydrogenase1(ALDH1)in breast cancer cells were analyzed by real-time PCR.The post hoc analysis(Tukey's tests)in conjunction with one-way ANOVA was used for statistical analysis.Results The initial cytotoxic efficacy was most notable.After the treatment of the same therapeutic agents,cell viability was decreased by 64.8%35.14%,32.25%in BT-483 cells,66.4%,22.94%and 45.88%in MDA-MB-231 cells,97.1%,99.5%and 76.4%in MCF cells.The difference was significant compared with that before treatment(P=0.000).However,the inhibitory effects were diminished after chemotherapeutic agent withdrawal.Cell viabilities were increased to 167.9%,212.04%and 188.66%in MDA-MB-231 cells at48 h after withdrawal.At 72 h after withdrawal,cell viability was increased with a significant difference in three cell lines(all P values=0.000).Expressions of CD44 and ALDH1 were most prevalent for MDA-MB-231,BT-483 and MCF-7 cells.ALDH1 mRNA level was significant higher in BT-483(HER-2 overexpression cell line)than MDA-MB-231(triple negative cell line)(P=0.012).CD14 mRNA level in MCF-7 cells were significantly lower than that in MDA-MB-231 and BT-483(P=0.003,0.001).BT-483 showed significantly higher level of CD44 than MDA-MB-231 and MCF-7 cell line(P=0.013,0.020),and no significant difference was detected between MDA-MB-231 and MCF-7 breast cancer cells(P=0.955).CD90 mRNA expressions were detected in MDA-MB-231 cells and MCF-7 cells,but not in BT-483 cells.Conclusion Some malignant cells could survive in vitro and begin to proliferate again between cycles of chemotherapy.

报告1例透明细胞汗腺瘤.患者女,22岁.3年前无意中发现左下腹一核桃大褐色半透明状实性肿块,逐渐增大,现约3.2cm*1.6cm*1.3cm,有蒂,表面光滑,质地中等,无压痛感.

目的:探讨皖南蝮蛇毒抑瘤组分I(AHVAC-I)对人胃癌细胞株SGC-901生长增殖的影响及作用机制.方法:AHVAC-I处理SGC-7901细胞.CCK-8法检测其生长抑制作用,光镜观察细胞形态变化,TUNEL法检测细胞凋亡情况.结果:AHVAC-I抑制SGC-901细胞的生长增殖,并呈浓度依赖性,半数抑制浓度(IC50)为58.197μg/ml;镜下可见SGC-901细胞悬浮、胞核固缩等典型凋亡形态学变化;TUNEL法显示凋亡指数随浓度增加而增加.结论:AHVAC-I对人胃癌细胞株SGC-7901具有生长抑制作用,诱导凋亡可能是其主要的作用机制之一.

目的利用AAV介导的体细胞基因敲除技术,在人大肠癌细胞系HCTll6中对靶基因Sirt1进行敲除,达到基因沉默的目的.方法构建Sirtl打靶载体,通过病毒包装、感染、药物筛选、PCR鉴定、Westernblotting鉴定等步骤获得Sirt1基因敲除的细胞株.结果经过筛选和鉴定后获得两个Sirtl-/-阳性克隆,成功构建HCTll6Sirtl.一细胞系.结论通过体细胞敲除技术,我们成功地获得Sirtl敲除肠癌细胞株.

[目的]探讨姜黄素对自发性高血压大鼠缺血再灌注后视网膜细胞凋亡及凋亡相关基因p53表达的影响和意义.[方法]制作大鼠视网膜缺血再灌注(retinalischemiareperfusion,RIR)模型,将自发性高血压大鼠(spontaneouslYhypertensiverat,SHR)随机分为假手术组(SH)、缺血再灌注组(IR)、姜黄素处理组(CU)和对照组(sc),每组又分为再灌注后2h、6h、24h、72h和7d等5个亚组(n=9).用TUNEL(termihaldeoxynucleotidyltransferase-mediateddUTPnickendlabeling)法观察大鼠视网膜和视网膜毛细血管细胞凋亡.分光光度法检测视网膜Caspase-3酶表达活性,用免疫组织化学sP法检测再灌注后不同时段视网膜组织中p53蛋白表达变化.[结果]与IR组比较.CU组能显著减少视网膜和视网膜毛细血管的细胞凋亡数量(P〈0.01),抑制p53蛋白表达(P〈0.01).[结论]姜黄素对SHR视网膜缺血再灌注损伤有保护作用,调控凋亡相关基因p53蛋白的表达可能是其作用机制之一.

目的:研究从白屈菜乙醇提取物中分离出的白屈菜碱在诱导HeLa细胞凋亡中的作用及参与其作用的主要信号转导通路.方法:细胞先以不同浓度白屈菜碱处理48h,用噻唑蓝法分析确定半数致死量(medianlethaldose,LD50).用4',6-二脒基-2-苯基吲哚染色,追踪分析核浓染以及DNA损伤和碎片的形态学变化,并用流式细胞术分析检测活性氧(reactive oxygen species,ROS)的产生以及细胞周期阻滞和线粒体膜电位的变化.用圆二色光谱分析寻找白屈菜碱和小牛胸腺DNA可能的相互作用.用逆转录聚合酶链反应和蛋白免疫印迹法测定p38、p53、蛋白激酶B(proteinkinase B,AKT)、磷脂酰肌醇3激酶(phosphatidylinositol 3-kinases,PI3K)、Janus激酶3(Janus kinase 3,JAK3)、信号转导及转录激活因子3(signal transducer and activator of transcription 3,STAT3)等的mRNA和蛋白表达,以及E6、E7癌基因和促凋亡基因、抗凋亡基因的mRNA和蛋白表达.结果:根据白屈菜碱的LD50(30μg/mL),选定3种实验剂量,即22.5、30和37.5μg/mL.结果显示,白屈菜碱抑制了HeLa细胞增殖,诱发其细胞凋亡,表现为ROS的产生,细胞亚G1和G0/G1周期阻滞,线粒体膜电位变化和DNA碎片产生.圆二色光谱分析结果显示白屈菜碱和小牛胸腺DNA间存在有效的相互作用.信号通路的研究显示白屈菜碱通过上调p38、p53和其他促凋亡基因的表达,以及下调AKT、P13K、JAK3、STAT3、E6、E7和其他抗凋亡基因的表达,有效诱发细胞凋亡.结论:从白屈菜中分离出的自屈菜碱能通过改变p38-p53及AKT/P13激酶信号转导通路有效地诱发HeLa细胞凋亡.

The mass production of printed electronics can be achieved by roll-to-roll(R2R) printing system, so highly accurate web tension is required that can minimize the register error and keep the thickness and roughness of printed devices in limits. The web tension of a R2R system is regulated by the use of integrated load cells and active dancer system for printed electronics applications using decentralized multi-input-single-output(MISO) regularized variable learning rate backpropagation artificial neural networks. The active dancer system is used before printing system to reduce disturbances in the web tension of process span. The classical PID control result in tension spikes with the change in roll diameter of winder and unwinder rolls. The presence of dancer in R2R system shows that improved web tension control in printing span and the web tension can be enhanced from 3.75 N to 4.75 N. The overshoot of system is less than ±2.5 N and steady state error is within ±1 N where load cells have a signal noise of ±0.7 N. The integration of load cells and active dancer with self-adapting neural network control provide a solution to the web tension control of multispan roll-to-roll system.