Objective： To explore and investigate the selection of effective antisense oligodeoxynuleotides with the help of computer and RNAstructure folding software. Methods： Bcl-2 gene was used as the target gene and five a...Objective： To explore and investigate the selection of effective antisense oligodeoxynuleotides with the help of computer and RNAstructure folding software. Methods： Bcl-2 gene was used as the target gene and five antisense oligodeoxynuleotides were designed to be bound to Bcl-2 mRNA optimal secondary structure regions that were predicted free from intramolecular fold or instability of free energy. The five antisense oligodeoxynucleotides were studied with experimental assay of leukemia cells, including cell grow assay with tropan blue exclusion, expression of Bcl-2 protein detected with immunochemistry and flowcytometry, Bcl-2 mRNA content detected with RT-PCR technique, as well as apoptosis observed and determined with morphonological method, electrophoresis and flowcytometry. Results： The results showed that two of the five antisense oligodeoxynucleotides were effective antisense oligodeoxynucleotides, which were able to inhibit cell growth in leukemia, to decrease the level of Bcl-2 mRNA and protein, to induce apoptosis of leukemia cells significantly. Conclusion： The computational prediction of antisense efficacy is faster than other methods and more efficient, which can potentially speed the development of sequences for both research and clinical applications.展开更多
目的:探讨A2DS2评分联合中性粒细胞百分比/白蛋白比值(neutrophil percentage/albumin ratio,NPAR)、D-二聚体对急性缺血性脑卒中(acute ischemic stroke,AIS)患者发生卒中相关性肺炎(stroke-associated pneumonia,SAP)的预测价值。方法...目的:探讨A2DS2评分联合中性粒细胞百分比/白蛋白比值(neutrophil percentage/albumin ratio,NPAR)、D-二聚体对急性缺血性脑卒中(acute ischemic stroke,AIS)患者发生卒中相关性肺炎(stroke-associated pneumonia,SAP)的预测价值。方法:回顾性分析南部战区空军医院2017年11月至2023年11月期间收治的265例AIS患者的临床资料,根据AIS患者发病1周内是否发生SAP分为SAP组和非SAP组,收集两组患者的临床资料,计算A2DS2评分、NPAR,检测两组D-二聚体水平。Spearman相关性分析A2DS2评分与NPAR、D-二聚体水平的关系。采用多因素Logistic回归分析独立危险因素。应用受试者工作特征(receiver operating characteristic,ROC)曲线评价预测价值。基于A2DS2评分、NPAR和D-二聚体的最佳截断值构建改良A2DS2(modifed A2DS2,mA2DS2)评分,应用ROC曲线分析mA2DS2评分对AIS患者发生SAP的预测价值。结果:本次研究共纳入265例AIS患者,其中78例AIS患者发病1周内发生SAP,SAP发病率为29.43%。SAP组和非SAP组在年龄、入院时美国国立卫生院研究卒中量表(national institute of health stroke scale,NIHSS)评分、吞咽困难、心房纤颤、既往卒中史、A2DS2评分、D-二聚体、纤维蛋白原水平、全身免疫炎症指数(systemic immune-inflammation index,SII)、中性粒细胞/淋巴细胞比值(neutrophil to lymphocyte ratio,NLR)、血小板/淋巴细胞的比值(platelet to lymphocyte ratio,PLR)、NPAR等方面比较有差异(P<0.05)。A2DS2评分与NPAR、D-二聚体呈正相关(P<0.05)。NPAR升高、D-二聚体升高、A2DS2评分升高是AIS患者发生SAP的危险因素(P<0.05)。A2DS2评分预测AIS患者发生SAP的曲线下面积(area under curve,AUC)为0.866,NPAR预测AIS患者发生SAP的AUC为0.815,D-二聚体预测发生SAP的AUC为0.705。ROC曲线分析结果显示,mA2DS2评分预测AIS患者发生SAP的AUC为0.898,大于A2DS2评分单独预测的AUC(Z=3.557,P<0.05)。结论:A2DS2评分、NPAR、D-二聚体水平显著升高与AIS患者发生SAP有关,A2DS2评分联合NPAR、D-二聚体检测对AIS患者发生SAP的预测效能较高。展开更多
文摘Objective： To explore and investigate the selection of effective antisense oligodeoxynuleotides with the help of computer and RNAstructure folding software. Methods： Bcl-2 gene was used as the target gene and five antisense oligodeoxynuleotides were designed to be bound to Bcl-2 mRNA optimal secondary structure regions that were predicted free from intramolecular fold or instability of free energy. The five antisense oligodeoxynucleotides were studied with experimental assay of leukemia cells, including cell grow assay with tropan blue exclusion, expression of Bcl-2 protein detected with immunochemistry and flowcytometry, Bcl-2 mRNA content detected with RT-PCR technique, as well as apoptosis observed and determined with morphonological method, electrophoresis and flowcytometry. Results： The results showed that two of the five antisense oligodeoxynucleotides were effective antisense oligodeoxynucleotides, which were able to inhibit cell growth in leukemia, to decrease the level of Bcl-2 mRNA and protein, to induce apoptosis of leukemia cells significantly. Conclusion： The computational prediction of antisense efficacy is faster than other methods and more efficient, which can potentially speed the development of sequences for both research and clinical applications.
文摘目的:探讨A2DS2评分联合中性粒细胞百分比/白蛋白比值(neutrophil percentage/albumin ratio,NPAR)、D-二聚体对急性缺血性脑卒中(acute ischemic stroke,AIS)患者发生卒中相关性肺炎(stroke-associated pneumonia,SAP)的预测价值。方法:回顾性分析南部战区空军医院2017年11月至2023年11月期间收治的265例AIS患者的临床资料,根据AIS患者发病1周内是否发生SAP分为SAP组和非SAP组,收集两组患者的临床资料,计算A2DS2评分、NPAR,检测两组D-二聚体水平。Spearman相关性分析A2DS2评分与NPAR、D-二聚体水平的关系。采用多因素Logistic回归分析独立危险因素。应用受试者工作特征(receiver operating characteristic,ROC)曲线评价预测价值。基于A2DS2评分、NPAR和D-二聚体的最佳截断值构建改良A2DS2(modifed A2DS2,mA2DS2)评分,应用ROC曲线分析mA2DS2评分对AIS患者发生SAP的预测价值。结果:本次研究共纳入265例AIS患者,其中78例AIS患者发病1周内发生SAP,SAP发病率为29.43%。SAP组和非SAP组在年龄、入院时美国国立卫生院研究卒中量表(national institute of health stroke scale,NIHSS)评分、吞咽困难、心房纤颤、既往卒中史、A2DS2评分、D-二聚体、纤维蛋白原水平、全身免疫炎症指数(systemic immune-inflammation index,SII)、中性粒细胞/淋巴细胞比值(neutrophil to lymphocyte ratio,NLR)、血小板/淋巴细胞的比值(platelet to lymphocyte ratio,PLR)、NPAR等方面比较有差异(P<0.05)。A2DS2评分与NPAR、D-二聚体呈正相关(P<0.05)。NPAR升高、D-二聚体升高、A2DS2评分升高是AIS患者发生SAP的危险因素(P<0.05)。A2DS2评分预测AIS患者发生SAP的曲线下面积(area under curve,AUC)为0.866,NPAR预测AIS患者发生SAP的AUC为0.815,D-二聚体预测发生SAP的AUC为0.705。ROC曲线分析结果显示,mA2DS2评分预测AIS患者发生SAP的AUC为0.898,大于A2DS2评分单独预测的AUC(Z=3.557,P<0.05)。结论:A2DS2评分、NPAR、D-二聚体水平显著升高与AIS患者发生SAP有关,A2DS2评分联合NPAR、D-二聚体检测对AIS患者发生SAP的预测效能较高。
