The NSC-34 cell line is a widely recognized motor neuron model and various neuronal differentiation protocols have been exploited. Under previously reported experimental conditions, only part of the cells resemble dif...The NSC-34 cell line is a widely recognized motor neuron model and various neuronal differentiation protocols have been exploited. Under previously reported experimental conditions, only part of the cells resemble differentiated neurons;however, they do not exhibit extensive and time-prolonged neuritogenesis, and maintain their duplication capacity in culture. The aim of the present work was to facilitate long-term and more homogeneous neuronal differentiation in motor neuron–like NSC-34 cells. We found that the antimitotic drug cytosine arabinoside promoted robust and persistent neuronal differentiation in the entire cell population. Long and interconnecting neuronal processes with abundant growth cones were homogeneously induced and were durable for up to at least 6 weeks in culture. Moreover, cytosine arabinoside was permissive, dispensable, and mostly irreversible in priming NSC-34 cells for neurite initiation and regeneration after mechanical dislodgement. Finally, the expression of the cell proliferation antigen Ki67 was inhibited by cytosine arabinoside, whereas the expression levels of neuronal growth associated protein 43, vimentin, and motor neuron–specific p75, Islet2, homeobox 9 markers were upregulated, as confirmed by western blot and/or confocal immunofluorescence analysis. Overall, these findings support the use of NSC-34 cells as a motor neuron model for properly investigating neurodegenerative mechanisms and prospectively identifying neuroprotective strategies.展开更多
目的探究CD5阳性弥漫大B细胞淋巴瘤(CD5-positive diffuse large B-cell lymphoma,CD5^(+)DLBCL)患者中的Ki67最佳截断值及其临床预后价值。方法选取2012年11月—2022年10月淮海淋巴瘤协作组中7家医疗中心具有完整Ki67免疫组织化学染色...目的探究CD5阳性弥漫大B细胞淋巴瘤(CD5-positive diffuse large B-cell lymphoma,CD5^(+)DLBCL)患者中的Ki67最佳截断值及其临床预后价值。方法选取2012年11月—2022年10月淮海淋巴瘤协作组中7家医疗中心具有完整Ki67免疫组织化学染色结果的CD5^(+)DLBCL且接受利妥昔单抗为基础的免疫治疗患者。选用最大选择秩统计量的方法计算Ki67的最佳截断值,并据此将患者分为高表达组和低表达组。比较2组基线信息及总生存期,并用Cox比例风险模型进行生存分析。结果共纳入231例初诊患者,平均年龄为(60.19±13.07)岁,其中男性111例(48.1%),Ann Arbor早期106例(45.9%)。Ki67的平均表达水平为70%,其最佳截断值为65%。高表达Ki67与中枢受累的比例显著升高相关(χ^(2)=6.229,P=0.013)。在校正其他混杂因素后,Cox结果表明Ki67高表达是CD5^(+)DLBCL患者预后的不良因素(HR=1.728,95%CI:1.127~2.649,P=0.012)。亚组分析结果提示,在BCL-2阳性患者中,Ki67高表达与更差的预后相关。将Ki67与IPI/NCCN-IPI联合应用后,可进一步提高对CD5^(+)DLBCL患者预后的预测准确性。结论Ki67在CD5^(+)DLBCL中的最佳截断值为65%。Ki67高表达为不良预后因素,结合IPI/NCCN-IPI可提高预后预测能力。展开更多
基金supported by FATALSDrug Project [Progetti di Ricerca@CNR SAC.AD002.173.058] from National Research Council,Italy (to CV)。
文摘The NSC-34 cell line is a widely recognized motor neuron model and various neuronal differentiation protocols have been exploited. Under previously reported experimental conditions, only part of the cells resemble differentiated neurons;however, they do not exhibit extensive and time-prolonged neuritogenesis, and maintain their duplication capacity in culture. The aim of the present work was to facilitate long-term and more homogeneous neuronal differentiation in motor neuron–like NSC-34 cells. We found that the antimitotic drug cytosine arabinoside promoted robust and persistent neuronal differentiation in the entire cell population. Long and interconnecting neuronal processes with abundant growth cones were homogeneously induced and were durable for up to at least 6 weeks in culture. Moreover, cytosine arabinoside was permissive, dispensable, and mostly irreversible in priming NSC-34 cells for neurite initiation and regeneration after mechanical dislodgement. Finally, the expression of the cell proliferation antigen Ki67 was inhibited by cytosine arabinoside, whereas the expression levels of neuronal growth associated protein 43, vimentin, and motor neuron–specific p75, Islet2, homeobox 9 markers were upregulated, as confirmed by western blot and/or confocal immunofluorescence analysis. Overall, these findings support the use of NSC-34 cells as a motor neuron model for properly investigating neurodegenerative mechanisms and prospectively identifying neuroprotective strategies.
