利用阳极氧化法在纯Ti表面制得TiO2纳米管,随后用冻干法在纳米管中负载microRNA制得复合涂层。利用SEM、TEM和荧光显微镜对TiO2/microRNA复合涂层进行表征,并对microRNA的释放行为以及其进入成骨细胞后的功效进行分析。结果表明,microRNA分子成功负载在TiO2纳米管中,并在12h内持续稳定释放投递,荧光照片表明投递的microRNA成功进入成骨细胞,ALP活性测定进一步表明进入细胞中microRNA促进了成骨分化。

TiO2 nanotubes was prepared by anodic oxidation method on pure Ti surface,then the microRNA was loaded in the nanotubes by lyophilization to build composite coating.The TiO2/microRNA composite coating was characterized by SEM,TEM and fluorescence microscope.The release behavior of microRNA and its furthrer function was analysised.The results showed that microRNA successfully deposited in TiO2 nanotubes, and sustained release delivery for 1 2 h.Fluorescent images indicated that the microRNA was transfected into osteoblasts,ALP activity assay further indicated that the microRNA promoted the osteogenic differentiation.

概搜集已报道的在乳腺癌发生发展过程中有重要作用的microRNA信息，结合相关肿瘤模型的实验数据，评估microRNA在乳腺癌诊断和治疗方面的应用前景。以“癌基因”和“癌抑制基因”为分类依据，全面地总结归纳乳腺癌相关microRNA的功能和作用机制，进一步从临床诊断标记物和治疗靶点的层面分析microRNA的潜在临床应用价值。表1和2总结了microRNA参与乳腺癌发展过程的具体事件及其调控的靶基因。同时，本文还深入探讨microRNA作为临床诊断标记物及治疗靶点的可行性，揭示已有研究的不足之处，为今后的相关工作方向提供一些建议。

MicroRNAs (miRs) are smal single-stranded RNA molecules, which function as key negative regulators of post-transcriptional modulation in almost all biological processes. Abnormal expression of microRNAs has been ob-served in various types of cancer including breast cancer. Great efforts have been made to identify an association between microRNA expression profiles and breast cancer, and to understand the functional role and molecular mechanism of aberrant-expressed microRNAs. As research progressed, ‘oncogenic microRNAs’ and ‘tumor sup-pressive microRNAs’ became a focus of interest. The potential of candidate microRNAs from both intercellular (tissue) and extracel ular (serum) sources for clinical diagnosis and prognosis was revealed, and treatments involving microRNA achieved some amazing curative effects in cancer disease models. In this review, advances from the most recent studies of microRNAs in one of the most common cancers, breast cancer, are highlighted, especial

目的：探讨定向诱导造血干细胞分化为巨核细胞和血小板的分子机制。方法将脐带造血干细胞培养后，分离巨核细胞，分别提取造血干细胞和巨核细胞总RNA，进行microRNA芯片分析。结果与造血干细胞相比，巨核细胞中表达上调超过2倍的microRNA有183个，下调超过2倍的microRNA有139个。结论造血干细胞和巨核细胞microRNA表达谱存在差异，尤其是多个microRNA与定向分化巨核细胞相关。

Objective To elucidate the mechanism of megakaryocyte development and platelet production from hematopoietic stem cells. Methods CD34+ cells were isolated from umbilical cord blood and cultivated. Megakaryocytes were puriifed from cultivated CD34+cells.Total RNAs from CD34+cells and megakaryocytes were extracted.Microarray analysis was performed to determine differential expressed microRNAs.Results Differential microRNAs were defined as a statistically significant difference with 2 fold or greater change. Compared with hematopoietic stem cells,183 microRNAs showed higher expression and 139 microRNAs were down-regulated in megakaryocytes.Conclusion The differential expressed microRNAs between hematopoietic stem cells and megakaryocytes indicates that some microRNAs play an important role in regulating megakaryocytes differentiation from hematopoietic stem cells.

随着大数据时代的来临,microRNA与基因的序列数据不断增加,如何从大量的数据中挖掘有生物学意义的信息成为新的热点问题。研究表明microRNA间以协作的方式在疾病中发挥作用,并呈现出网络的结构化趋势。因此,系统分析不同microRNA间的相似性将在疾病生物学标记挖掘等研究领域起到关键的桥梁作用。而microRNA通过调节其靶基因发挥作用,所以本研究将充分利用现有靶基因数据,从功能角度分析microRNA间的相似性。研究选取前期工作所得的靶基因优化列表,利用富集分析将基因集合转化为功能节点集合,并在此基础上利用集合相似性测度计算microRNA对在不同层面的功能一致性。结果表明,相同家族的microRNA倾向于调控相同或相似的靶基因;类比于非靶基因,microRNA靶基因倾向于共享较多相似的细胞组分,而在生物学通路及生物学过程中则具有相对较低的相似性。

The numbers of microRNA and genes sequences have increased greatly with the advent of big data era. Thus how to explore useful information with biological signiifcances from massive datasets has become a new hot topic. Former researches showed that microRNAs tended to play roles in diseases in a cooperative way and the relationships could be presented in the form of network. As a result, similarity analysis for microRNAs through a system way could play an important role in the ifeld of disease biomarkers discovery. Considering that microRNAs play regulation roles by binding to their target genes, we focused on the available target gene data to analyze the similarity of microRNA pairs on functional levels. The optimization microRNA targets list generated by our former research as input were chosen and the enrichment analysis was used to map gene sets into functional term sets. The similarities between microRNAs were then calculated using similarity metrics on functional levels. Our resu

目的 筛选人退变腰椎间盘特异性表达microRNA的靶基因,并探讨JNK信号转导通路在椎间盘退变(intervertebral disc degeneration,IVDD)分子生物学机制中发挥的作用.方法 对获取的8份腰椎退行性疾病患者(男6例,女2例；44～68岁)手术髓核组织和3份腰椎骨折患者(男3例,17、18、22岁)正常髓核组织依次进行髓核细胞分离、培养、染色鉴定、提取标本总RNA;采用microRNA微阵列基因表达实验和分析技术筛选差异表达的microRNAs并采用实时qPCR技术对其中高表达者进行验证；综合MicroCosm v5、TargetScan 5.1和microRNA.org等三个数据库的靶基因信息,取交集分析预测靶基因,并分析与差异基因或靶基因功能显著相关的生物信号通路；定量PCR方法验证筛选结果.结果 退变组中microRNA-513a-5p和microRNA-494呈显著高表达,比值分别为2.222 2和2.948 5,并与验证结果相吻合；预测靶基因分别为MKK4和JunD,此两种靶基因分别位于JNK信号通路的上、下游,均参与JNK信号传导.结论 microRNA-513a-5p和microRNA-494在退变椎间盘中具有高表达,其对应的靶基因为MKK4和JunD.JNK信号传导通路在IVDD的发病机制中可能发挥重要正负反馈调节作用.

Objective To screen and validate differentially expressed microRNAs in human degenerative nucleus pulposus cells (NPCs) and to predict their target genes,and to investigate the role of JNK pathway in degenerative intervertebral disc disease.Methods Eight degenerative nucleus pulposus tissues were harvested from 8 patients (6 males and 2 females) with lumbar degenerative disease,and three normal nucleus pulposus tissues were harvested from 3 patients (3 males) with lumbar fracture,intraoperatively.Differentially expressed microRNAs were screened by microRNA microarray analysis and validated by real-time qPCR.Target genes of highly expressed microRNAs were predicted by analyzing information from data bases:MicroCosm v5,TargetScan 5.1 and microRNA.org.Signal pathways associated with the target genes were analyzed,and qPCR was used to validate the screening results.Results Twenty differentially expressed microRNAs were identified.The microRNA-513a-5p and microRNA-494 were highly expressed