目的 比较使用流式细胞仪355 nm和407 nm激光器激发Hochest33342检测细胞凋亡。 方法 通过ATO药物诱导急性早幼粒白血病细胞（NB4）及血清饥饿法诱导人肺癌细胞（NCl-H292）细胞凋亡，取24、48 h时间点收集细胞，进行Hoechst33342-碘化丙啶（PI）双染，分别在配置有两种激光器的流式细胞仪上检测细胞凋亡。 结果 细胞经处理后24 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（28.20 ± 4.80）%；NCl-H292细胞凋亡率Hoechst33342+/PI-：（22.47 ± 2.78）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（25.10 ± 6.19）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：20.47 ± 1.46%。处理后48 h，355 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（33.60 ± 3.75）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（26.77 ± 1.16）%。407 nm激光器检测NB4细胞凋亡率Hoechst33342+/PI-：（29.47 ± 2.33）%。NCl-H292细胞凋亡率Hoechst33342+/PI-：（31.47 ± 3.05）%。两种细胞处理后比处理前凋亡率明显升高，但355 nm激光器与407 nm激光器检测的凋亡结果差异不明显（P＞0.05）。 结论 407 nm激光器激发Hoechst33342可检测细胞凋亡。
目的 建立急性白血病（AL）患者八色流式免疫表型分析起始管方案。 方法 用胞膜CD3（CD3）、CD19、CD10、CD34、CD45、胞浆CD79a（cCD79a）、髓过氧化物酶（MPO)和胞浆CD3（cCD3）等8种抗体建立八色流式染色方案。膜表面抗体直接染色；膜内抗体经固定破膜，再染色后上机检测。将3个血小板减少患者骨髓标本分别进行抗体的单色染色和缺一色染色；最后对17例确诊的AL初发患者标本进行检测。 结果 用单色染色来确定染色方案中各抗体的检测电压及荧光补偿；缺一色染色中，阳性细胞群较单色染色变化均＜10%，表明方案中的各抗体相互作用小。17例AL初发患者中，6例急性B淋巴细胞白血病原始细胞均为CD34和CD19阳性，5例cCD79a阳性和4例CD10阳性；4例急性T淋巴细胞白血病患者均为cCD3阳性；6例急性髓细胞白血病均为CD34和MPO阳性；1例B+T混合表型AL患者CD34、cCD3、CD19、cCD79a及CD10均为阳性，MPO和CD3为阴性，此检测方案能够确定各类AL的细胞类型。 结论 建立了AL患者八色流式免疫表型分析起始管方案，操作简便快速，适用于临床检测。
ObjectiveCD44 and CD54 are two specific biomarkers of gastric cancer stem cells and were used as targets in this study. The number of CD45–CD44+CD54+ cell subsets in peripheral blood of gastric cancer patients was detected by flow cytometry. Further, we combined these results with the clinicopathological characteristics of gastric cancer patients to analyze the significance of CD45–CD44+CD54+ cell subsets.MethodsFrom December 2016 to September 2017, 38 patients with gastric cancer in gastrointestinal surgery of West China Hospital of Sichuan University were included as the study object. The content of CD45–CD44+CD54+ cell subsets in their peripheral blood was detected by flow cytometry and its clinical significance was analyzed.ResultsThe median number of CD45–CD44+CD54+ cells were 541.9/mL (71.7–8 057.0/mL) in 38 patients and 555.9/mL (71.7–8 057.0/mL) in the group of patients with R0 resection. Patients without lymph node metastasis were found to have more CD45–CD44+CD54+ cells than patients with lymph node metastasis [941.4/mL (183.5–8 057.0)/mL vs 379.3/mL (71.7–2 269.7/mL, P=0.002], and more CD45–CD44+CD54+ cells in patients with TNM stage Ⅰ–Ⅱ than in TNM stage Ⅲ–Ⅳ [858.6/mL (183.5–8 057.0/mL) vs 364.6/mL (71.7–2 269.7/mL, P=0.015]. The patients with T3–4 stages (P= 0.025), N+ stage (P=0.009) and TNM Ⅲ–Ⅳ stage (P=0.012) had low ratios of the subgroup with high number of CD45–CD44+CD54+ cells, respectively. We made a more accurate judgment of N stage and TNM stage when we combined tumor size and the number of CD45–CD44+CD54+ cells together. However, there was no significant correlation between the number of CD45–CD44+CD54+ cells and other clinicopathological features and prognosis.ConclusionsThe number of CD45–CD44+CD54+ cell subsets is correlated with tumor progression, which might be used to predict TNM stage and N stage. However, the number of patients included in this study is too small, and the clinical significance of CD45–CD44+CD54+ subsets in gastric cancer patients needs to be further demonstrated by expanding the sample size.