目的探讨血小板储存过程中伴随储存时间延长pri-mioroRNA-223的表达变化及意义。方法使用血细胞分析仪检测不同保存时间(1、3、5 d)的血小板质量,并进行代谢组学检测。同时,建立液滴式数字PCR(droplet digital PCR,ddPCR)方法,对其敏感...目的探讨血小板储存过程中伴随储存时间延长pri-mioroRNA-223的表达变化及意义。方法使用血细胞分析仪检测不同保存时间(1、3、5 d)的血小板质量,并进行代谢组学检测。同时,建立液滴式数字PCR(droplet digital PCR,ddPCR)方法,对其敏感性、重复性进行评价。用ddPCR检测血小板样本不同储存时间的pri-miR-223表达量,分析其对血小板储存损伤(PSL)的影响及意义。结果随着保存时间的延长,血小板不同保存时间的血小板计数(PLT)、血小板比例(P-LCR)、平均血小板体积(MPV)和血小板分布宽度(PDW)差异均无统计学意义(P>0.05)。而代谢组学研究共筛选出955种代谢产物,其中54种代谢产物上调,6种代谢产物下调。所建立的ddPCR方法特异性好,敏感性高,标准曲线表现出良好的线性相关性(Y=0.9736X+0.0832),R^(2)值为0.9984,线性动态范围灵敏度为3.3×10~0~1.9×10~5拷贝/μL。ddPCR结果显示第3、5天样本中pri-miR-223表达量比第1天明显降低。结论随着血小板体外贮存时间的延长,对PLT、P-LCR、MPV和PDW没有影响,储存期末部分代谢产物改变,且通过ddPCR检测发现pri-miR-223表达量降低,这种代谢变化可能与pri-miR-223的表达调控有关。展开更多
目的·探讨甲基转移酶3(methyltransferase like 3,METTL3)调控pri-miR-21的N^(6)-甲基腺苷(N^(6)-methyladenosine,m^(6)A)甲基化修饰在糖尿病肾病(diabetic nephropathy,DN)小鼠肾脏纤维化发病机制中的作用。方法·采用8周龄...目的·探讨甲基转移酶3(methyltransferase like 3,METTL3)调控pri-miR-21的N^(6)-甲基腺苷(N^(6)-methyladenosine,m^(6)A)甲基化修饰在糖尿病肾病(diabetic nephropathy,DN)小鼠肾脏纤维化发病机制中的作用。方法·采用8周龄雄性db/db小鼠作为DN模型小鼠,db/m小鼠作为对照,同时按照是否经尾静脉注射S-腺苷高半胱氨酸水解酶抑制剂3-脱氮腺苷(3-deazaadenosine,DAA),共随机分为4组(5只/组),分别为db/m组、db/db组、db/m+DAA组和db/db+DAA组;8周龄开始注射DAA,注射1次/5 d,共注射8次。DAA干预结束后继续饲养小鼠至19周龄,收取各组小鼠血、尿、肾脏组织标本。检测血糖、血肌酐、尿白蛋白肌酐比(albumin-to-creatinine ratio,ACR),肾脏行苏木精-伊红(H-E)染色、Masson染色及天狼星红染色观察病理变化;试剂盒检测肾脏总RNA中m^(6)A的甲基化水平;Western blotting检测肾脏METTL3及纤维化相关蛋白表达;实时定量PCR检测肾脏总pri-miR-21和成熟miR-21;使用免疫磁珠富集肾脏组织中m^(6)A甲基化RNA,并通过PCR检测其中m^(6)A甲基化的pri-miR-21。结果·相较于db/m组,db/db组小鼠血糖,血肌酐,ACR,肾脏METTL3、m^(6)A甲基化修饰水平、纤维化相关蛋白、总pri-miR-21、m^(6)A甲基化pri-miR-21和成熟miR-21表达水平均显著增加(均P<0.05),小鼠肾脏系膜基质增多、肾小球基底膜增厚、胶原纤维累积显著增加。相较于db/db组,db/db+DAA组血糖,血肌酐,ACR,肾脏m^(6)A甲基化修饰水平、纤维化相关蛋白、m^(6)A甲基化pri-miR-21和成熟miR-21表达水平均显著下降(均P<0.05),总pri-miR-21表达水平显著升高(P=0.000),METTL3蛋白表达水平未见显著变化,小鼠肾脏损伤及纤维化程度显著减轻。结论·pri-miR-21的m^(6)A甲基化修饰促进miR-21成熟,进而促进DN小鼠肾脏纤维化的发生发展;抑制METTL3可通过调控pri-miR-21的m^(6)A甲基化修饰抑制DN小鼠肾脏纤维化。展开更多
Several studies examined the impact of miR-34b/c rs4938723 polymorphism and cancer risk, but the findings are inconsistent. However, no study has been conducted to inspect the impact of miR-34b/c polymorphism on bladd...Several studies examined the impact of miR-34b/c rs4938723 polymorphism and cancer risk, but the findings are inconsistent. However, no study has been conducted to inspect the impact of miR-34b/c polymorphism on bladder cancer. This study aimed to assess possible association between rs4938723 polymorphism and bladder cancer risk.This case-control study was done on 136 pathologically proven bladder cancer patients and 144 controls. Genotyping of Pri-miR-34b/c rs4938723 polymorphism was achieved by using the polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP) method. Our findings did not show any statistically significant differences in genotype and allele frequencies between bladder cancer and controls. Larger sample sizes with diverse ethnicities are required to validate our findings.展开更多
文摘目的探讨血小板储存过程中伴随储存时间延长pri-mioroRNA-223的表达变化及意义。方法使用血细胞分析仪检测不同保存时间(1、3、5 d)的血小板质量,并进行代谢组学检测。同时,建立液滴式数字PCR(droplet digital PCR,ddPCR)方法,对其敏感性、重复性进行评价。用ddPCR检测血小板样本不同储存时间的pri-miR-223表达量,分析其对血小板储存损伤(PSL)的影响及意义。结果随着保存时间的延长,血小板不同保存时间的血小板计数(PLT)、血小板比例(P-LCR)、平均血小板体积(MPV)和血小板分布宽度(PDW)差异均无统计学意义(P>0.05)。而代谢组学研究共筛选出955种代谢产物,其中54种代谢产物上调,6种代谢产物下调。所建立的ddPCR方法特异性好,敏感性高,标准曲线表现出良好的线性相关性(Y=0.9736X+0.0832),R^(2)值为0.9984,线性动态范围灵敏度为3.3×10~0~1.9×10~5拷贝/μL。ddPCR结果显示第3、5天样本中pri-miR-223表达量比第1天明显降低。结论随着血小板体外贮存时间的延长,对PLT、P-LCR、MPV和PDW没有影响,储存期末部分代谢产物改变,且通过ddPCR检测发现pri-miR-223表达量降低,这种代谢变化可能与pri-miR-223的表达调控有关。
基金funded by a research grant (#8067) from Zahedan University of Medical Sciences
文摘Several studies examined the impact of miR-34b/c rs4938723 polymorphism and cancer risk, but the findings are inconsistent. However, no study has been conducted to inspect the impact of miR-34b/c polymorphism on bladder cancer. This study aimed to assess possible association between rs4938723 polymorphism and bladder cancer risk.This case-control study was done on 136 pathologically proven bladder cancer patients and 144 controls. Genotyping of Pri-miR-34b/c rs4938723 polymorphism was achieved by using the polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP) method. Our findings did not show any statistically significant differences in genotype and allele frequencies between bladder cancer and controls. Larger sample sizes with diverse ethnicities are required to validate our findings.
