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.

目的表皮干细胞是组织工程化皮肤的"种子细胞".文中研究碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)、胰岛素样细胞生长因子-1(insulin-like growth factor-1,IGF-1)和转化生长因子仅(transforming growth factor a,TGFa)对人表皮干细胞增殖的影响.方法采用两步酶消化法和Ⅳ型胶原差速贴壁相结合的方法获得人原代表皮干细胞,将表皮干细胞分为A组(K-SFM)、B组(K-SFM+bFGF)、C组(K-SFM+IGF-1)及D组(K-SFM+TGFot)进行培养.比较不同组之间表皮干细胞的克隆形成率、生长增殖、生长曲线和细胞周期等指标.结果B组、C组及D组培养的表皮干细胞在细胞增殖、细胞克隆率方面检测结果均明显高于A组(P〈0.05);流式细胞仪检测B组、C组与A组及D组处于G0/G1细胞比例比较差异有统计学意义(P〈0.05).结论IGF-1、bFGF及TGF-a均对表皮干细胞的增殖有促进作用,IGF-1及bFGF对表皮干细胞表型的维持有较好的作用.

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.

本研究旨在通过茎一环RT-PCR法,建立有效定量miRNA421的PCR方法,探讨该方法在定量检测miRNA方面的应用价值.利用miRBase数据库获得miRNA-421的成熟序列.设计3条miRNA421特异性反转录引物,Q-PCR上游引物及通用下游引物.通过10倍梯度稀释RNA后特异性反转录,RT-PCR分析不同引物互相配对PCR的扩增曲线及熔融曲线,鉴定出特异性好、扩增效率高的引物.为进一步鉴定引物的有效性,过表达miRNA-d21,用鉴定的成熟引物定量扩增.利用茎一环RT-PCR法设计、鉴定出miRNA-421引物:421RT7、F19,且这对引物特异性好、扩增效率高.通过设计引物组合,茎一环RT-PCR法能够很好地用于miRNA定量分析,并具有准确、快速、节约成本的优点.

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.

采用带有PVT口模的流变仪测定无定型聚合物熔体在压力0~50 MPa,温度453~563 K时的PVT(压力-体积-温度)特性曲线,分别用基团贡献修正胞腔模型(GCMCM)和胞腔模型对实测的PVT数据进行预测.结果表明:在实验中用基团贡献修正胞腔模型的比容平均预测误差较胞腔模型要大得多,其预测精度要略差于胞腔模型,这说明基团贡献修正胞腔模型的应用具有一定的局限性.同时还计算出了2种模型所对应的特征值,并分析了其特征值存在的共同点和差异的原因.