BACKGROUND & AIMS: :Human coronary artery endothelial cells (HCAECs) have the potential to undergo fibrogenic endothelial-mesenchymal transition (EndMT), which results in matrix-producing fibroblasts and thereby contributes to the pathogenesis of cardiac fibrosis. Recently, the profibrotic cytokine transforming growth factor-β (TGF-β) is shown to be the crucial pathogenic driver which has been verified to induce EndMT. C-Ski is an important regulator of TGF-β signaling. However, the detailed role of c-Ski and the molecular mechanisms by which c-Ski affects TGF-β-induced EndMT in HCAECs are not largely elucidated. In the present study, we treated HCAECs with TGF-β of different concentrations to induce EndMT. We found that overexpression of c-Ski in HCAECs either blocked EndMT via hindering Vimentin, Snail, Slug, and Twist expression while enhancing CD31 expression, with or without TGF-β treatment. In contrast, suppression of c-Ski further enhanced EndMT. Currently, miRNA expression disorder has been frequently reported associating with cardiac fibrosis. By using online tools, we regarded miR-155 as a candidate miRNA that could target c-Ski, which was verified using luciferase assays. C-Ski expression was negatively regulated by miR-155. TGF-β-induced EndMT was inhibited by miR-155 silence; the effect of TGF-β on Vimentin, CD31, Snail, Slug, and Twist could be partially restored by miR-155. Altogether, these findings will shed light on the role and mechanism by which miR-155 regulates TGF-β-induced HCAECs EndMT via c-Ski to affect cardiac fibrosis, and miR-155/c-Ski may represent novel biomarkers and therapeutic targets in the treatment of cardiac fibrosis.

背景与目标： : 人冠状动脉内皮细胞 (HCAECs) 具有发生纤维化的内皮-间质转化 (EndMT) 的潜力，这导致产生基质的成纤维细胞，从而有助于心脏纤维化的发病机理。最近，纤维化细胞因子转化生长因子-β (TGF-β) 被证明是关键的致病驱动因素，已被证实可诱导EndMT。C-ski是TGF-β 信号的重要调节因子。然而，c-Ski的详细作用以及c-Ski影响HCAECs中TGF-β 诱导的EndMT的分子机制尚未得到很大的阐明。在本研究中，我们用不同浓度的TGF-β 处理HCAECs以诱导EndMT。我们发现，有或没有TGF-β 处理，HCAECs中c-Ski的过表达通过阻碍波形蛋白，蜗牛，Slug和Twist的表达来阻断EndMT，同时增强CD31的表达。相比之下，抑制c-ski进一步增强了EndMT。目前，miRNA表达紊乱经常被报道与心脏纤维化有关。通过使用在线工具，我们将miR-155视为可以靶向c-Ski的候选miRNA，使用荧光素酶分析验证了这一点。C-Ski表达受miR-155负调节。TGF-β 诱导的EndMT被miR-155沉默抑制; TGF-β 对波形蛋白，CD31，蜗牛，Slug和Twist的作用可以通过miR-155部分恢复。总之，这些发现将阐明miR-155通过c-Ski调节TGF-β 诱导的HCAECs EndMT以影响心脏纤维化的作用和机制，并且miR-155/c-Ski可能代表治疗心脏纤维化的新型生物标志物和治疗靶标。

BACKGROUND & AIMS: -2

背景与目标： -2

BACKGROUND & AIMS: :Human hair follicle mesenchymal stem cells (hHF-MSCs) are capable of differentiating into smooth muscle cells (SMCs) in response to transforming growth factor-β (TGF-β), and thus can be used for cardiovascular tissue engineering and regenerative medicine. However, the precise molecular mechanisms underlying SMC conversion of hHF-MSCs are still undefined. MicroRNAs (miRNAs) are small noncoding RNAs that modulate gene expression post-transcriptionally by binding to the complementary sequences of targeted mRNAs. Accumulating evidence indicates that miRNAs are associated with SMC differentiation in vitro andin vivo. In this study, we revealed that miR-18b was significantly downregulated during TGF-β1-induced hHF-MSCs differentiation into SMC using miRNA array profiling and quantitative RT- PCR (qRT-PCR). Over-expression of miR-18b in hHF-MSCs led to remarkable downregulation of SMC-specific markers such as SMA and calponin proteins. On the contrary, inhibition of endogenous miR-18b by its antisense oligonucleotide antagomir-18b reversed the changes of SMA and calponin proteins. We also showed that SMAD2, a key transcription regulator in TGF-β signaling which was involved in SMC differentiation, is regulated by miR-18b. miR-18b could suppress the expression of SMAD2 protein by targeting the 3'UTR of SMAD2 gene without affecting its mRNA level in hHF-MSCs. Moreover, knockdown of SMAD2 by RNA interference could block the effect of inhibition of miR-18b on SMC differentiation, indicating that SMAD2 contributed to miR-18b mediated regulation of TGF-β-induced SMC differentiation. In conclusion, this study demonstrated that miR-18b regulated the TGF-β1-induced differentiation of hHF-MSCs into SMCs by targeting SMAD2 gene, and provided novel insights into the regulatory mechanisms of TGF-β-induced SMC differentiation.

背景与目标： : 人毛囊间充质干细胞 (hhf-mscs) 能够响应转化生长因子-β (TGF-β) 分化为平滑肌细胞 (SMCs)，因此可用于心血管组织工程和再生医学。然而，hHF-MSCs的SMC转化的确切分子机制仍未确定。Microrna (mirna) 是小的非编码rna，通过与靶向mrna的互补序列结合来调节转录后的基因表达。越来越多的证据表明，mirna在体外和体内与SMC分化有关。在这项研究中，我们揭示了使用miRNA阵列谱和定量rt-pcr (qRT-PCR) 在tgf-β1诱导的hHF-MSCs分化为SMC期间miR-18b显着下调。hHF-MSCs中miR-18b的过表达导致SMC特异性标记 (例如SMA和钙蛋白) 的显着下调。相反，其反义寡核苷酸对内源性miR-18b的抑制antagomir-18b逆转了SMA和钙蛋白的变化。我们还表明，SMAD2是参与SMC分化的TGF-β 信号传导中的关键转录调节剂，受miR-18b调节。miR-18b可以通过靶向SMAD2基因的3'UTR来抑制SMAD2蛋白的表达，而不影响其在hHF-MSCs中的mRNA水平。此外，通过RNA干扰敲除SMAD2可以阻断miR-18b对SMC分化的抑制作用，表明SMAD2有助于miR-18b介导的TGF-β 诱导的SMC分化的调节。总之，本研究表明，miR-18b通过靶向SMAD2基因调控了tgf-β1诱导的hHF-MSCs向SMC的分化，并为TGF-β 诱导的SMC分化的调控机制提供了新的见解。

BACKGROUND & AIMS: OBJECTIVE:This study aimed to investigate the effect of circular RNA itchy E3 ubiquitin protein ligase on cell proliferation and apoptosis and to explore its target micro-RNAs in prostate cancer cells. METHODS:Circular RNA itchy E3 ubiquitin protein ligase expression in human prostate cancer cells and normal prostate epithelial cells was determined by real time-quantitative polymerase chain reaction assay. Circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids (circular RNA itchy E3 ubiquitin protein ligase(+) group and control overexpression plasmids group were transfected with PC-3 cells. Rescue experiment was performed by transfection of circular RNA itchy E3 ubiquitin protein ligase overexpression and micro-197 overexpression plasmids (circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids/micro RNA (+) group) into PC-3 cells. Cell Counting Kit-8 and annexin V/propidium iodide assays were conducted to evaluate cell proliferation and apoptosis, respectively. Western blot was performed to determine the expressions of apoptotic-related markers. RESULTS:Circular RNA itchy E3 ubiquitin protein ligase expression was decreased in DU 145, 22RV1, VCaP, and PC-3 cells compared to RWPE cells. In PC-3 cells, cell proliferation rate was reduced in circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids group compared to control overexpression plasmids group at 48 hours and 72 hours. Cell apoptosis rate was elevated in circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids group compared to control overexpression plasmids group at 48 hours, and Western blot showed the similar results. Micro RNA-197 but not micro RNA-31 or micro RNA-432 was the target micro-RNA of circular RNA itchy E3 ubiquitin protein ligase. In rescue experiments, cell proliferation rate was elevated, but apoptosis rate was reduced in circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids/micro RNA (+) group compared to circular RNA itchy E3 ubiquitin protein ligase overexpression plasmids group, indicating that circular RNA itchy E3 ubiquitin protein ligase upregulation inhibited cell proliferation but promoted apoptosis through downregulating micro RNA-197. CONCLUSION:Circular RNA itchy E3 ubiquitin protein ligase upregulation suppresses cell proliferation but promotes apoptosis through targeting micro RNA-197 in prostate cancer. Our study may provide a new insight for the treatment of prostate cancer.

背景与目标：

BACKGROUND & AIMS: :Neonatal hypoxic-ischemic encephalopathy (HIE) is a disorder mainly due to asphyxia during the perinatal period, and late diagnosis leads to high mortality. In this study, the expression of microRNA-199a (miR-199a) in HIE newborns was investigated, as well as its clinical significance in HIE diagnosis and prognosis. Circulating levels of S100B and NSE in HIE newborns were measured using enzyme-linked immunosorbent assay, and the expression of miR-199a was analyzed using quantitative real-time PCR. The diagnostic value of miR-199a, S100B and NSE was evaluated using the receiver operating characteristic (ROC) analysis, and their prognostic value was assessed by the evaluation of Gesell intellectual development of the HIE newborns. HIE newborns possessed significantly increased levels of S100B and NSE and decreased miR-199a (all P < 0.01). The Neonatal Behavioral Neurological Assessment (NBNA) score of HIE newborns was negatively correlated with S100B and NSE, while was positively correlated miR-199a. The ROC analysis results showed the diagnostic value of serum miR-199a, and the combined detection of miR-199a, S100B and NSE could obtained the highest diagnostic accuracy in HIE newborns. miR-199a expression was lowest in newborns with severe HIE, and it had diagnostic potential to distinguish HIE cases with different severity. Regarding the prognosis of neonatal HIE, the correlation of miR-199a, S100B, NSE with Gesell intellectual development was found in HIE newborns. The decreased miR-199a in HIE newborns serves as a potential diagnostic biomarker and may help to improve the diagnostic and prognostic value of S100B and NSE in neonatal HIE.

背景与目标： 新生儿缺氧缺血性脑病 (HIE) 是一种主要由围产期窒息引起的疾病，晚期诊断导致高死亡率。本研究探讨了microRNA-199a (miR-199a) 在HIE新生儿中的表达及其在HIE诊断和预后中的临床意义。采用酶联免疫吸附法测定HIE新生儿循环中S100B和NSE的水平，并采用实时定量PCR分析miR-199a的表达。使用接受者工作特征 (ROC) 分析评估miR-199a，S100B和NSE的诊断价值，并通过评估HIE新生儿的Gesell智力发育评估其预后价值。HIE新生儿S100B和NSE水平明显升高，miR-199a降低 (均p  <  0.01)。HIE新生儿的新生儿行为神经评估 (NBNA) 评分与S100B和NSE呈负相关，miR-199a呈正相关。ROC分析结果显示了血清miR-199a的诊断价值，miR-199a，S100B和NSE联合检测可在HIE新生儿中获得最高的诊断准确性。miR-199a在严重HIE的新生儿中表达最低，并且具有区分不同严重程度的HIE病例的诊断潜力。关于新生儿HIE的预后，发现miR-199a，S100B，NSE与HIE新生儿的Gesell智力发育相关。HIE新生儿miR-199a降低是潜在的诊断生物标志物，可能有助于提高S100B和NSE在新生儿HIE中的诊断和预后价值。

BACKGROUND & AIMS: :Exosomes are key mediators of intercellular communication and play a role in the pathogenesis and progression of cancer. Exosomes in circulating body fluids serve as molecular markers for cancer diagnosis. This study aimed to investigate the role of exosomal microRNA (miR)-1910-3p in breast cancer and determine its clinical diagnostic value. MiR-1910-3p promoted proliferation and migration of breast cancer cells in vitro and in vivo. In vitro, exosomes enriched in miR-1910-3p transferred miR-1910-3p to mammary epithelial cells and breast cancer cells, promoting proliferation and migration, inhibiting apoptosis, and inducing autophagy. In vivo, exosomes enriched in miR-1910-3p promoted the proliferation and migration of breast cancer cells. MiR-1910-3p downregulated myotubularin-related protein 3, activated the NF-κB and wnt/β-catenin signaling pathway, and promoted breast cancer progression. Serum miR-1910-3p in exosomes was an effective diagnostic marker that improved the sensitivity of breast cancer diagnosis when used in combination with the traditional tumor marker CA153. In conclusion, breast cancer cell-derived exosomes promoted the growth, metastasis, and autophagy of breast cancer cells by transferring miR-1910-3p. MiR-1910-3p in serum exosomes may serve as a novel molecular marker for breast cancer diagnosis.

背景与目标： : 外泌体是细胞间通讯的关键介质，在癌症的发病机理和进展中起作用。循环体液中的外泌体可作为癌症诊断的分子标记。本研究旨在探讨外体microRNA (miR)-1910-3p在乳腺癌中的作用，并确定其临床诊断价值。MiR-1910-3p在体外和体内促进了乳腺癌细胞的增殖和迁移。在体外，富含miR-1910-3p的外泌体miR-1910-3p转移至乳腺上皮细胞和乳腺癌细胞，促进增殖和迁移，抑制细胞凋亡，诱导自噬。在体内，富含miR-1910-3p的外泌体促进了乳腺癌细胞的增殖和迁移。MiR-1910-3p下调肌管蛋白相关蛋白3，激活NF-κ b和wnt/β-catenin信号通路，促进乳腺癌进展。外泌体中的血清miR-1910-3p是与传统肿瘤标志物ca153联合使用时提高乳腺癌诊断敏感性的有效诊断标志物。总之，乳腺癌细胞来源的外泌体通过转移miR-1910-3p促进乳腺癌细胞的生长，转移和自噬。血清外泌体中的MiR-1910-3p可作为乳腺癌诊断的新型分子标志物。

BACKGROUND & AIMS: Background:Circular RNAs (circRNAs) which are shown as a class of RNAs exhibit the importance in the regulation of gene expression and the development of biological process. However, the expression profile and molecular mechanism of circRNA ATXN7 (circATXN7) is still mostly uncertain in gastric cancer (GC). Methods:qRT-PCR analysis was performed to detect the expression of circATXN7, miR-4319 and ENTPD4 in GC tissues and cells. CCK-8, colony formation, EdU, flow cytometry, TUNEL and transwell assays were conducted to assess the effect of circATXN7 or miR-4319 on cell proliferation, apoptosis and invasion. In vivo assays were utilized to further analyze the function of circATXN7 on the tumorigenesis and progression of GC. The interaction between miR-4319 and circATXN7 (or ENTPD4) was verified using luciferase reporter and RNA pull-down assays. Results:The results showed an upregulated circATXN7 expression in GC tissues and cell lines. Besides, silenced circATXN7 hampered the proliferation and invasion as well as promoted the apoptosis in GC cells. Moreover, low expression of miR-4319 was found in GC. It was determined that circATXN7 acted as a sponge for miR-4319 and had a negative association with miR-4319. We also found that miR-4319 upregulation restrained GC cell proliferation and migration whereas enhanced apoptosis. Subsequently, ENTPD4, the target gene of miR-4319, was found overexpressed in GC. Additionally, it was negatively correlated with miR-4319 whereas positively associated with circATXN7. In vivo experiments, circATXN7 silence was confirmed to inhibit GC tumor growth. Conclusions:CircATXN7 promoted GC development through sponging miR-4319 and regulating ENTPD4, which identified circATXN7 as a new biomarker in GC.

背景与目标：

BACKGROUND & AIMS: :Multidrug resistance (MDR) is usually correlated with the poor prognosis of gastric cancer. In this study, we revealed a total of 11 microRNAs (miRNA) that regulated MDR of gastric cancer via high-throughput functional screening, and miR-508-5p reversed MDR most efficiently among these candidate miRNAs. The overexpression of miR-508-5p was sufficient to reverse cancer cell resistance to multiple chemotherapeutics in vitro and sensitize tumours to chemotherapy in vivo. Further studies showed that miR-508-5p could directly target the 3'-untranslated regions of ABCB1 and Zinc ribbon domain-containing 1 (ZNRD1), and suppress their expression at the mRNA and protein levels. Meanwhile, the suppression of ZNRD1 led to a decrease in ABCB1. These findings suggest that a miR-508-5p/ZNRD1/ABCB1 regulatory loop has a critical role in MDR in gastric cancer. In addition, miR-508-5p could be used as a prognostic factor for overall survival in gastric cancer. These data reveal an important role for miR-508-5p in the regulation of MDR in gastric cancer, and suggest the potential application of miR-508-5p in drug resistance prediction and treatment.

背景与目标： : 多药耐药 (MDR) 通常与胃癌的不良预后相关。在这项研究中，我们揭示了总共11个通过高通量功能筛选调节胃癌MDR的microrna (miRNA)，并且在这些候选miRNA中miR-508-5p最有效地逆转MDR。miR-508-5p的过表达足以在体外逆转癌细胞对多种化学疗法的抗性，并在体内使肿瘤对化学疗法敏感。进一步的研究表明，miR-508-5p可以直接靶向ABCB1和含锌带结构域1 (ZNRD1) 的3 '-非翻译区，并在mRNA和蛋白质水平上抑制它们的表达。同时，ZNRD1的抑制导致abcb1的减少。这些发现表明，miR-508-5p/ZNRD1/ABCB1调节环在胃癌MDR中具有关键作用。此外，miR-508-5p可以用作胃癌总体生存的预后因素。这些数据揭示了miR-508-5p在胃癌MDR调控中的重要作用，并提示了miR-508-5p在耐药性预测和治疗中的潜在应用。

BACKGROUND & AIMS: :Epithelial ovarian cancer (EOC) is the leading cause of death among gynecologic malignancies. Despite great efforts to improve early detection and optimize chemotherapeutic regimens, the 5-year survival rate is only 30% for patients presenting with late-stage ovarian cancer. The high mortality of this disease is due to late diagnosis in over 70% of ovarian cancer cases. A class of small noncoding RNAs, or microRNAs, was found to regulate gene expression at the post-transcriptional level. Some, but not all, of the data indicated that the miR-200 family was dysregulated in a variety of malignancies. In this study, we demonstrated that miR-200a and E-cadherin were significantly upregulated in EOC compared to benign epithelial ovarian cysts and normal ovarian tissues. However, further stratification of the subject indicated that the expression levels of miR-200a were significantly downregulated in late-stage (FIGO III+V) and grade 3 groups compared with early stage (FIGO I+II) and grade 1 to 2 groups. Similarly, relatively low levels of miR-200a were observed in the lymph compared to the node-negative group. E-cadherin expression was found to be absent in normal ovarian tissue and was frequently expressed in benign epithelial ovarian cysts, with absence or low levels observed in late-stage ovarian cancers. There was a significantly positive correlation between miR-200a and E-cadherin in EOC. The biphasic expression pattern suggested that miR-200a levels may serve as novel biomarkers for the early detection of EOC, and miR-200a and E-cadherin are candidate targets for the development of new treatment modalities against ovarian cancer.

背景与目标： : 上皮性卵巢癌 (EOC) 是妇科恶性肿瘤死亡的主要原因。尽管为改善早期检测和优化化疗方案做出了巨大努力，但对于晚期卵巢癌患者而言，5年生存率仅30%。这种疾病的高死亡率是由于超过70% 例卵巢癌病例的晚期诊断。发现一类小的非编码rna或microrna在转录后水平上调节基因表达。一些 (但不是全部) 数据表明，miR-200家族在各种恶性肿瘤中失调。在这项研究中，我们证明了与良性上皮性卵巢囊肿和正常卵巢组织相比，EOC中的miR-200a和E-cadherin显着上调。然而，对受试者的进一步分层表明，与早期 (FIGO I II) 和1至2级组相比，晚期 (FIGO III V) 和3级组的miR-200a表达水平显着下调。同样，与淋巴结阴性组相比，在淋巴中观察到相对较低的miR-200a水平。发现E-cadherin表达在正常卵巢组织中不存在，并且在良性上皮性卵巢囊肿中经常表达，在晚期卵巢癌中观察到不存在或低水平。EOC中miR-200a与E-cadherin之间存在显着正相关。双相表达模式表明，miR-200a水平可能是早期检测EOC的新型生物标志物，miR-200a和E-钙粘蛋白是开发针对卵巢癌的新治疗方式的候选靶标。

BACKGROUND & AIMS: :miRNAs are important regulators of cellular senescence yet the extent of their involvement remains to be investigated. We sought to identify miRNAs that are involved in cytokine-induced premature senescence (CIPS) in endothelial cells. CIPS was established in young human pulmonary microvascular endothelial cells (HMVEC-Ls) following treatment with a sublethal dose (20ng/ml) of tumor necrosis factor alpha (TNF-α) for 15days. In parallel, HMVEC-Ls were grown and routinely passaged until the onset of replicative senescence (RS). Differential expression analysis following miRNA microarray profiling revealed an overlapped of eight deregulated miRNAs in both the miRNA profiles of RS and TNF-α-induced premature senescence cells. Amongst the deregulated miRNAs were members of the miR 17-92 cluster which are known regulators of angiogenesis. The role of hsa-miR-20b in TNF-α-induced premature senescence, a paralog member of the miR 17-92 cluster, was further investigated. Biotin-labeled hsa-miR-20b captured the enriched transcripts of retinoblastoma-like 1 (RBL1), indicating that RBL1 is a target of hsa-miR-20b. Knockdown of hsa-miR-20b attenuated premature senescence in the TNF-α-treated HMVEC-Ls as evidenced by increased cell proliferation, increased RBL1 mRNA expression level but decreased protein expression of p16INK4a, a cellular senescence marker. These findings provide an early insight into the role of hsa-miR-20b in endothelial senescence.

背景与目标： : mirna是细胞衰老的重要调节剂，但其参与程度仍有待研究。我们试图鉴定参与细胞因子诱导的内皮细胞过早衰老 (CIPS) 的miRNAs。用亚致死剂量 (20ng/ml) 的肿瘤坏死因子 α (TNF-α) 治疗15天后，在年轻的人肺微血管内皮细胞 (HMVEC-Ls) 中建立了CIPS。同时，HMVEC-Ls生长并常规传代，直到复制性衰老 (RS) 开始。miRNA微阵列分析后的差异表达分析显示，在RS和TNF-α 诱导的早衰细胞的miRNA谱中，八个解除调节的miRNA重叠。在解除管制的mirna中，miR 17-92簇的成员是已知的血管生成调节剂。进一步研究了hsa-miR-20b在TNF-α 诱导的过早衰老 (miR 17-92簇的旁系成员) 中的作用。生物素标记的hsa-miR-20b捕获了视网膜母细胞瘤样1 (RBL1) 的富集转录本，表明RBL1是hsa-miR-20b的靶标。hsa-miR-20b的敲除减弱了TNF-α 处理的HMVEC-Ls中的过早衰老，这可以通过增加细胞增殖，增加RBL1 mRNA表达水平但降低细胞衰老标志物p16INK4a的蛋白表达来证明。这些发现提供了hsa-miR-20b在内皮衰老中的作用的早期见解。

BACKGROUND & AIMS: OBJECTIVES:MicroRNAs (miRNAs) are endogenous small RNAs of 21-25 nucleotides that can pair with sites in 3' untranslated regions in mRNAs of protein-coding genes to downregulate their expression. Recently, miR-499 and other miRNAs released in circulating blood have been reported as promising biomarkers for acute myocardial infarction (AMI). In the present study, we developed a novel reverse-transcription real-time PCR assay for human miR-499 quantification. DESIGN AND METHODS:miR-499 was reverse-transcribed with a 3' portion-specific primer into cDNAs. The cDNAs were further extended with overlap PCR. The extended cDNAs were determined by quantitative, real-time PCR. Synthetic miR-499 was put into the RT reaction over an optimal range to generate standard curves for absolute quantification of miR-499. RESULTS:In the presence of 0.0001 amol/μL to 1.0×10⁶ amol/μL of synthetic miR-499, we observed a linear correlation (R²=0.999) between the logarithm of the amount of input RNA and the CT value. The miR-499 was reliably measured at a detection limit of 0.0001 amol/μL. The miR-499 measurements in spiked plasma samples indicated excellent correlation between the novel qRT PCR and classic stem loop qRT PCR. The qRT PCR analysis demonstrated that miR-499 was detected with higher levels in plasma from the patient with AMI in acute phase (AMI) compared with those from the control groups (P<0.001). CONCLUSIONS:We developed a novel reverse-transcription real-time PCR assay for human miR-499 quantification. The good reproducibility and wide linearity range may permit more use of it in the quantification of other human miRNAs in future.

背景与目标：

BACKGROUND & AIMS: :Patients with secondary acute myeloid leukemia (sAML) arising from myelodysplastic syndromes have a poor prognosis marked by an increased resistance to chemotherapy. An urgent need exists for adjuvant treatments that can enhance or replace current therapeutic options. Here we show the potential of LB100, a small-molecule protein phosphatase 2 A (PP2A) inhibitor, as a monotherapy and chemosensitizing agent for sAML using an in-vitro and in-vivo approach. We demonstrate that LB100 decreases cell viability through caspase activation and G2/M cell-cycle arrest. LB100 enhances daunorubicin (DNR) cytotoxicity resulting in decreased xenograft volumes and improved overall survival. LB100 profoundly upregulates miR-181b-1, which we show directly binds to the 3' untranslated region of Bcl-2 mRNA leading to its translational inhibition. MiR-181b-1 ectopic overexpression further diminishes Bcl-2 expression leading to suppression of sAML cell growth, and enhancement of DNR cytotoxicity. Our research highlights the therapeutic potential of LB100, and provides new insights into the mechanism of LB100 chemosensitization.

背景与目标： : 由骨髓增生异常综合征引起的继发性急性髓细胞性白血病 (sAML) 患者的预后较差，其对化疗的抵抗力增加。迫切需要可以增强或替代当前治疗选择的辅助治疗。在这里，我们使用体外和体内方法显示了小分子蛋白磷酸酶2 a (PP2A) 抑制剂LB100作为sAML的单一疗法和化学增敏剂的潜力。我们证明LB100通过caspase激活和G2/M细胞周期阻滞降低细胞活力。LB100增强柔红霉素 (DNR) 的细胞毒性，从而减少异种移植体积并改善总生存率。LB100深刻上调miR-181b-1，我们显示其与Bcl-2 mRNA的3' 非翻译区直接结合，导致其翻译抑制。MiR-181b-1异位过度表达进一步减少Bcl-2表达，导致sAML细胞生长的抑制和DNR细胞毒性的增强。我们的研究突出了LB100的治疗潜力，并为LB100的化学增敏机制提供了新的见解。

BACKGROUND & AIMS: :Human cytomegalovirus (HCMV) has been reported to be linked to vascular disease through the induction of neovessel formation. We have previously reported that microRNA (miR)-217 and miR-199a-5p enhance endothelial angiogenesis via inhibition of sirtuin 1 (SIRT1) in HCMV-infected human umbilical vein endothelial cells (HUVECs). Here, we found that miR-138 also suppressed the expression of the SIRT1 protein and stimulated phosphorylation of signal transducer and activator of transcription 3 (p-STAT3). Moreover, the regulation of p-STAT3 expression mediated by SIRT1 was found to promote HCMV-induced angiogenesis. These findings revealed that miR-138 might promote angiogenesis of HCMV-infected HUVECs by activating the SIRT1-mediated p-STAT3 pathway, and this could provide novel insights into HCMV-induced angiogenesis.

背景与目标： : 据报道，人类巨细胞病毒 (HCMV) 通过诱导新血管形成与血管疾病有关。我们先前已经报道了microRNA (miR)-217和miR-199a-5p通过抑制HCMV感染的人脐静脉内皮细胞 (huvec) 中的sirtuin 1 (SIRT1) 来增强内皮血管生成。在这里，我们发现miR-138还抑制了SIRT1蛋白的表达并刺激了信号转导子和转录激活子3的磷酸化 (p-STAT3)。此外，发现由SIRT1介导的p-STAT3表达调节可促进HCMV诱导的血管生成。这些发现表明，miR-138可能通过激活SIRT1-mediated p-STAT3途径促进HCMV感染的HUVECs的血管生成，这可能为HCMV诱导的血管生成提供新的见解。

BACKGROUND & AIMS: :The expression pattern and role of circular RNAs (circRNAs) in the pathogenesis of gastric cancer (GC) and their underlying mechanisms remain unresolved. In this study, we identified differentially expressed circRNAs by a circRNA microarray and verified the results by quantitative reverse transcription-polymerase chain reaction using 117 clinical samples. Cell Counting Kit-8, wound healing, Transwell, and tumorsphere formation assays were conducted to assess the effects of circ-CEP85L on cell proliferation and invasion in vitro. Mouse intraperitoneal injection models were used to assess the functions of circ-CEP85L in vivo. Luciferase reporter assays, fluorescence in situ hybridization, and rescue experiments were performed to elucidate the underlying mechanism of circ-CEP85L. We found that circ-CEP85L, which has not been studied in GC, was significantly downregulated in GC tissues and that decreased circ-CEP85L expression correlated significantly with a worse prognosis. The knockdown of circ-CEP85L promoted the proliferation and invasion of GC cells, which was reversed by overexpression of circ-CEP85L. Furthermore, inhibition of circ-CEP85L promoted tumor growth in vivo. Mechanistically, circ-CEP85L was confirmed to be a direct target of miR-942-5p. In addition, rescue experiments indicated that circ-CEP85L is able to inhibit the proliferation and invasion of GC cells by sponging miR-942-5p. Finally, western blot assays verified that the downregulation of miR-942-5p efficiently reversed the inhibition of NFKBIA induced by circ-CEP85L overexpression. Therefore, we conclude that circ-CEP85L promotes NFKBIA expression by acting as a sponge of miR-942-5p; thus, inhibiting GC proliferation and invasion. circ-CEP85L is a potential target in the treatment of GC.

背景与目标： : 环状rna (circRNAs) 在胃癌 (GC) 发病中的表达模式和作用及其潜在机制仍未解决。在这项研究中，我们通过circRNA微阵列鉴定了差异表达的circRNA，并使用117临床样品通过定量逆转录-聚合酶链反应验证了结果。进行细胞计数试剂盒-8，伤口愈合，Transwell和肿瘤球形成试验，以评估circ-CEP85L对体外细胞增殖和侵袭的影响。小鼠腹腔注射模型用于评估体内circ-CEP85L的功能。进行了荧光素酶报告基因测定，荧光原位杂交和抢救实验，以阐明circ-CEP85L的潜在机制。我们发现，尚未在GC中进行研究的circ-CEP85L在GC组织中被显着下调，并且circ-CEP85L表达的降低与预后差显着相关。circ-CEP85L的敲除促进了GC细胞的增殖和侵袭，而circ-CEP85L的过表达则使其逆转。此外，抑制circ-CEP85L促进体内肿瘤生长。从机械上讲，circ-CEP85L被确认为miR-942-5p的直接目标。此外，拯救实验表明，circ-CEP85L能够通过海绵miR-942-5p抑制GC细胞的增殖和侵袭。最后，western blot测定证实了miR-942-5p的下调有效地逆转了circ-CEP85L过表达诱导的NFKBIA的抑制作用。因此，我们得出结论，circ-CEP85L通过充当miR-942-5p海绵来促进NFKBIA表达; 因此，抑制GC增殖和侵袭。circ-CEP85L是GC医治的一个潜在靶点。

BACKGROUND & AIMS: :Mirtrons are non-canonical miRNAs arising by splicing and debranching from short introns. A plethora of introns have been inferred by computational analyses as potential mirtrons. Yet, few have been experimentally validated and their functions, particularly in relation to their host genes, remain poorly understood. Here, we found that Drosophila larvae lacking either the mirtron miR-1010 or its binding site in the nicotinic acetylcholine receptor β2 (nAcRβ2) 3'UTR fail to grow properly and pupariate. Increase of cortical nAcRβ2 mediated by neural activity elevates the level of intracellular Ca2+, which in turn activates CaMKII and, further downstream, the transcription factor Adf-1. We show that miR-1010 downregulates nAcRβ2. We reveal that Adf-1 initiates the expression of SKIP, the host gene of miR-1010. Preventing synaptic potentials from overshooting their optimal range requires both SKIP to temper synaptic potentials (incoherent feedforward loop) and miR-1010 to reduce nAcRβ2 mRNA levels (negative feedback loop). Our results demonstrate how a mirtron, in coordination with its host gene, contributes to maintaining appropriate receptor levels, which in turn may play a role in maintaining homeostasis.

背景与目标： : Mirtrons是由短内含子剪接和脱支产生的非规范mirna。通过计算分析，已经推断出过多的内含子是潜在的mirtrons。然而，很少有人得到实验验证，它们的功能，特别是与宿主基因有关的功能，仍然知之甚少。在这里，我们发现果蝇幼虫在烟碱乙酰胆碱受体 β2 (nAcRβ2) 3'UTR中缺乏mirtron miR-1010或其结合位点，无法正常生长和腐烂。由神经活动介导的皮质nAcRβ2的增加会提高细胞内Ca2的水平，进而激活CaMKII，并在更下游激活转录因子Adf-1。我们显示miR-1010下调nacrβ2。我们揭示了Adf-1启动miR-1010宿主基因SKIP的表达。防止突触电位超出其最佳范围需要跳过以调节突触电位 (非相干前馈环) 和miR-1010以降低nacrβ2mrna水平 (负反馈环)。我们的结果证明了mirtron如何与其宿主基因协调，有助于维持适当的受体水平，进而可能在维持体内平衡中发挥作用。