Objective:Multiple myeloma(MM)is a hematologically malignant clonal plasma cell disease.This study aims to explore the association between immunophenotypes and prognosis in patients with MM,to determine whether the ex...Objective:Multiple myeloma(MM)is a hematologically malignant clonal plasma cell disease.This study aims to explore the association between immunophenotypes and prognosis in patients with MM,to determine whether the expression of CD45 and CD200 is related to the prognosis of newly diagnosed MM(NDMM)patients,and to evaluate the significance of the combined expression of CD45 and CD200 in NDMM.Methods:A total of 123 NDMM patients admitted to Shengjing Hospital of China Medical University from July 2015 to August 2019 were enrolled.Five key immunophenotypic markers(including CD38,CD138,CD45,CD56,and CD200)were screened through flow cytometry and identified using random forest analysis and univariate Cox regression analysis.Patients were divided into 3 groups:Group A,CD45 and CD200 double-positive;Group B,CD45 or CD200 single-positive;Group C,CD45 and CD200 double-negative.Kaplan-Meier curves were used to analyze overall survival(OS)and progression-free survival(PFS)across groups.Multivariate Cox regression was performed to evaluate prognostic factors,and a nomogram was constructed based on these results.Results:The OS and PFS of single-positive groups for CD38,CD138,CD45,CD56,and CD200 were all shorter than those of their respective single-negative groups(all P<0.05).Significant differences were observed in OS(P<0.001)and PFS(P=0.001)among Groups A,B,and C.Group A had shorter OS and PFS(all P=0.001)compared to the Group B+C(cases from Group B and Group C were combined).CD45 and CD200 double-positive was an independent prognostic factor for NDMM[hazard ratio(HR)=2.178,95%confidence interval(CI)1.048 to 4.529;P=0.037].The nomogram and calibration curves constructed from multivariate Cox regression analysis demonstrated good concordance(concordance index=0.706;95%CI 0.661 to 0.751).Conclusion:NDMM patients with double-positive expression of CD45 and CD200 have significantly shorter OS and PFS.Compared with the use of either marker alone,the combined assessment of CD45 and CD200 may provide better prognostic stratification for MM patients.展开更多
目的探究大鼠羊膜间充质干细胞及其外泌体的分离与鉴定方法,为其后续治疗应用奠定基础。方法①妊娠18 d SD大鼠,心脏注射400 IU/kg肝素,75%乙醇浸泡子宫,在0.9%氯化钠溶液中分离羊膜组织;②用0.75 mg/mL胶原酶和0.25%胰蛋白酶分步消化...目的探究大鼠羊膜间充质干细胞及其外泌体的分离与鉴定方法,为其后续治疗应用奠定基础。方法①妊娠18 d SD大鼠,心脏注射400 IU/kg肝素,75%乙醇浸泡子宫,在0.9%氯化钠溶液中分离羊膜组织;②用0.75 mg/mL胶原酶和0.25%胰蛋白酶分步消化、应用α-MEM培养基(添加4 ng/mL成纤维生长因子)培养;③采用形态观察、表面标志物检测和免疫荧光染色进行鉴定;④超速离心法提取外泌体,透射电镜观察形态、BCA法测定蛋白浓度和流式细胞术检测表面标志物。结果形态学观察发现干细胞贴壁生长,形态呈长梭形,细胞之间相互连接;流式细胞术检测干细胞表面抗原显示,CD29、CD90表达呈阳性,CD34、CD45表达呈阴性;细胞免疫荧光结果显示,Vimentin(波形蛋白)表达阳性。结果表明,分离得到的大鼠羊膜干细胞为间充质干细胞。透射电镜结果显示,超速离心法分离得到的外泌体形态完整、呈椭圆形,具有双分子层膜结构且大小均一,粒径平均(82.6±14.0)nm;BCA测定外泌体蛋白浓度为0.995μg/μL;流式细胞术检测表面蛋白显示CD9、CD63和CD81表达呈阳性。结论采用胶原酶和胰蛋白酶分步消化法成功分离出大鼠羊膜间充质细胞,通过超速离心法成功提取大鼠羊膜间充质干细胞外泌体,为大鼠羊膜间充质干细胞及其外泌体的研究和应用奠定基础。展开更多
基金supported by the National Natural Science Foundation,China(81870166).
文摘Objective:Multiple myeloma(MM)is a hematologically malignant clonal plasma cell disease.This study aims to explore the association between immunophenotypes and prognosis in patients with MM,to determine whether the expression of CD45 and CD200 is related to the prognosis of newly diagnosed MM(NDMM)patients,and to evaluate the significance of the combined expression of CD45 and CD200 in NDMM.Methods:A total of 123 NDMM patients admitted to Shengjing Hospital of China Medical University from July 2015 to August 2019 were enrolled.Five key immunophenotypic markers(including CD38,CD138,CD45,CD56,and CD200)were screened through flow cytometry and identified using random forest analysis and univariate Cox regression analysis.Patients were divided into 3 groups:Group A,CD45 and CD200 double-positive;Group B,CD45 or CD200 single-positive;Group C,CD45 and CD200 double-negative.Kaplan-Meier curves were used to analyze overall survival(OS)and progression-free survival(PFS)across groups.Multivariate Cox regression was performed to evaluate prognostic factors,and a nomogram was constructed based on these results.Results:The OS and PFS of single-positive groups for CD38,CD138,CD45,CD56,and CD200 were all shorter than those of their respective single-negative groups(all P<0.05).Significant differences were observed in OS(P<0.001)and PFS(P=0.001)among Groups A,B,and C.Group A had shorter OS and PFS(all P=0.001)compared to the Group B+C(cases from Group B and Group C were combined).CD45 and CD200 double-positive was an independent prognostic factor for NDMM[hazard ratio(HR)=2.178,95%confidence interval(CI)1.048 to 4.529;P=0.037].The nomogram and calibration curves constructed from multivariate Cox regression analysis demonstrated good concordance(concordance index=0.706;95%CI 0.661 to 0.751).Conclusion:NDMM patients with double-positive expression of CD45 and CD200 have significantly shorter OS and PFS.Compared with the use of either marker alone,the combined assessment of CD45 and CD200 may provide better prognostic stratification for MM patients.
