MicroRNA 在多囊卵巢综合征中的表达及作用

非编码微小RNAs（microRNAs,miRNAs）主要在转录后水平上调控基因表达,异常miRNA表达与胰岛素抵抗、糖尿病、炎症及各种癌症形成有关。多囊卵巢综合征（polycystic ovary syndrome,PCOS）患者血清及卵泡液中miRNAs表达谱存在差异,其异常的miRNAs表达能够通过影响基因转录后水平,参与PCOS的发生发展。综述miR-93、miR-222、miR-92a/b、miR-224、miR-320、miR-9、miR-483-5p、miR-513a-3p、miR-24、miR-133b、miR-26b和miR-378在PCOS患者的胰岛素抵抗、性激素合成和分泌、颗粒细胞增生、卵泡发育异常及排卵障碍中的作用,探讨miRNA在PCOS发病机制中的作用。     

Circulating miRNAs: Potential diagnostic role for coronavirus disease 2019 (COVID-19)

Nowadays, the coronavirus disease (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) represents a major global health problem. Intensive efforts are being employed to better understand this pathology and develop strategies enabling its early diagnosis and efficient treatment. In this study, we compared the signature of circulating miRNAs in plasma of COVID-19 patients versus healthy donors. MiRCURY LNA miRNA miRNome qPCR Panels were performed for miRNA signature characterization. Individual quantitative real-time PCR (qRT-PCR) was carried out to validate miRNome qPCR results. Receiver-operator characteristic (ROC) curve analysis was applied to assess the diagnostic accuracy of the most significantly deregulated miRNA(s) as potential diagnostic biomarker(s). Eight miRNAs were identified to be differentially expressed with miR-17-5p and miR-142-5p being down-regulated whilst miR-15a-5p, miR-19a-3p, miR-19b-3p, miR-23a-3p, miR-92a-3p and miR-320a being up-regulated in SARS-CoV-2-infected patients. ROC curve analyses revealed an AUC (Areas Under the ROC Curve) of 0.815 (P = 0.031), 0.875 (P = 0.012), and 0.850 (P = 0.025) for miR-19a-3p, miR-19b-3p, and miR-92a-3p, respectively. Combined ROC analyses using these 3 miRNAs showed a greater AUC of 0.917 (P = 0.0001) indicating a robust diagnostic value of these 3 miRNAs. These results suggest that plasma miR-19a-3p, miR-19b-3p, and miR-92a-3p expression levels could serve as potential diagnostic biomarker and/or a putative therapeutic target during SARS-CoV-2-infection.     

microRNA-103a-3p confers protection against lipopolysaccharide-induced sepsis and consequent multiple organ dysfunction syndrome by targeting HMGB1

BackgroundSepsis and subsequent multiple organ dysfunction syndrome (MODS) have high global incidence and mortality rate, imposing tremendous health burden. microRNAs (miRNAs or miRs) are implicated in the pathogenesis of sepsis and MODS. The aim of this study is to explore the potential mechanisms of miR-103a-3p targeted high mobility group box 1 (HMGB1) involvement in the pathogenesis of sepsis complicated with multiple organ dysfunction syndrome (MODS).MethodsA mouse sepsis model was induced by lipopolysaccharide (LPS). Bone marrow-derived macrophages were collected and LPS was used to establish a cellular inflammation model. Targeted binding between miR-103a-3p and HMGB1 was verified by a double luciferase assay and their roles in LPS-induced sepsis were further explored using gain-of-function experiments.ResultsmiR-103a-3p was decreased while HMGB1 was increased in sepsis. In LPS-induced mouse sepsis models, the downregulation of HMGB1 was found to result in reductions in NO, TNF-α, IL-1β, IL-6, lung myeloperoxidase activity, pulmonary microvascular albumin leakage, serum alanine aminotransferase, aspartate aminotransferase activity, and lung and liver tissue apoptosis. Additionally, decreased HMGB1 blunted the inflammatory response and increased survival rate of modeled mice. Importantly, HMGB1 was confirmed to a target gene of miR-103a-3p. In cellular inflammation models, miR-103a-3p was found to alleviate LPS-induced sepsis and MODS in vitro by decreasing HMGB1.ConclusionsTaken together, our results demonstrated the inhibitory role of miR-103a-3p in sepsis via inhibiting HMGB1 expression.     

Resistance Training Diminishes the Expression of Exosome CD63 Protein without Modification of Plasma miR-146a-5p and cfDNA in the Elderly

Aging-associated inflammation is characterized by senescent cell-mediated secretion of high levels of inflammatory mediators, such as microRNA (miR)-146a. Moreover, a rise of circulating cell-free DNA (cfDNA) is also related to systemic inflammation and frailty in the elderly. Exosome-mediated cell-to-cell communication is fundamental in cellular senescence and aging. The plasma changes in exercise-promoted miR-146a-5p, cfDNA, and exosome release could be the key to facilitate intercellular communication and systemic adaptations to exercise in aging. Thirty-eight elderly subjects (28 trained and 10 controls) volunteered in an 8-week resistance training protocol. The levels of plasma miR-146a-5p, cfDNA, and exosome markers (CD9, CD14, CD63, CD81, Flotillin [Flot]-1, and VDAC1) were measured prior to and following training. Results showed no changes in plasma miR-146a-5p and cfDNA levels with training. The levels of exosome markers (Flot-1, CD9, and CD81) as well as exosome-carried proteins (CD14 and VDAC1) remained unchanged, whereas an attenuated CD63 response was found in the trained group compared to the controls. These findings might partially support the anti-inflammatory effect of resistance training in the elderly as evidenced by the diminishment of exosome CD63 protein expression, without modification of plasma miR-146a-5p and cfDNA.     

Effects of blackcurrant-based juice on atherosclerosis-related biomarkers in cultured macrophages and in human subjects after consumption of a high-energy meal

Regular consumption of fruit and vegetables may be associated with decreased CVD risk. In the present study, we investigated the effects of blackcurrant (BC) juice, rich in polyphenols and ascorbic acid, on oxidative and inflammatory biomarkers in cultured macrophages in vitro and in human subjects with an atherosclerosis-prone phenotype (after consumption of a high-energy meal). In cultured macrophages (RAW264.7), BC treatment significantly inhibited lipopolysaccharide-induced inflammation as indicated by lower mRNA levels of TNF-α, IL-1β and inducible NO synthase (iNOS) and lower nuclear p65 levels indicating decreased NF-κB activity. iNOS protein levels were lower and haem oxygenase 1 levels higher in BC-treated cells when compared with untreated controls. Subjects given a high-energy meal had elevated serum glucose and insulin levels with no significant difference between the BC-based juice and placebo treatment groups. TAG following meal ingestion tended to be attenuated after the BC treatment. Plasma ascorbic acid and radical-scavenging capacity were decreased following placebo meal consumption; however, BC significantly elevated both parameters compared with baseline and placebo ingestion. Plasma oxidised LDL, α-tocopherol and paraoxonase activity were unchanged in both treatment groups. Furthermore, production of TNF-α and IL-1β was not significantly changed by BC meal consumption. The present results suggest potential antioxidative and anti-inflammatory properties of BC in vitro in cultured macrophages. Although the observations were not directly transferable to a postprandial in vivo situation, the present results show that BC juice consumption may improve postprandial antioxidant status as indicated by higher ascorbic acid levels and free radical-scavenging capacity in plasma.     

Moderate alcohol consumption reduces plasma C-reactive protein and fibrinogen levels; a randomized,diet-controlled intervention study

OBJECTIVE: To evaluate the effect of moderate alcohol consumption on the acute phase proteins C-reactive protein and fibrinogen. DESIGN: Randomized, diet-controlled, cross-over study. SETTING: The study was performed at TNO Nutrition and Food Research, Zeist, The Netherlands. SUBJECTS: Ten middle-aged men and 10 postmenopausal women, all apparently healthy, non-smoking and moderate alcohol drinkers, were included. One women dropped out because of a treatment-unrelated cause. The remaining 19 subjects finished the experiment successfully. INTERVENTIONS: Men consumed four glasses and women consumed three glasses of beer or no-alcohol beer (control) with evening dinner during two successive periods of 3 weeks. The total diet was supplied to the subjects and had essentially the same composition during these 6 weeks. Before each treatment there was a 1 week washout period to compensate for possible carry-over effects. RESULTS: Plasma C-reactive protein and fibrinogen levels were decreased by 35% (P=0.02) and 12.4% (P< or =0.001), respectively, after 3 weeks' consumption of beer, as compared to no-alcohol beer consumption. CONCLUSIONS: Moderate alcohol consumption significantly decreased plasma C-reactive protein and fibrinogen levels. An anti-inflammatory action of alcohol may help explain the link between moderate alcohol consumption and lower cardiovascular disease risk. SPONSORSHIP: Dutch Foundation for Alcohol Research (SAR).     

肾细胞癌患者血清外泌体miR-181a-3p表达水平的临床价值

目的探讨肾细胞癌(renal cell carcinoma,RCC)患者血清外泌体微小核糖核酸-181a-3p(microRNA-181a-3p, miR-181a-3p)的表达变化,及其作为非侵入性辅助诊断RCC标志的临床价值。 方法收集2016年6月至2017年6月期间,南京总医院泌尿外科收治的未经治疗的79例RCC患者和75名年龄、性别匹配的健康体检者血清标本,分离血清外泌体并提取RNA。用实时荧光定量聚合酶链反应(quantitative real-time polymerase chai reaction,qRT-PCR)检测两组miR-181a-3p水平的2^-△Ct相对定量含量,并采用多种统计方法分析外泌体miR-181a-3p诊断RCC的效能。 结果qRT-PCR检测RCC患者血清外泌体miR-181a-3p的表达水平为0.264(0.095~0.782),较健康对照组[0.068(0.032~0.130)]明显上调(U=1022,P<0.001);同时,血清外泌体miR-181a-3p水平在早期(I/II期)RCC患者[0.202(0.105~0.543)]中也显著升高(U=699,P<0.001)。血清外泌体miR-181a-3p的受试者工作特征曲线(receiver operating characteristic curve,ROC)区分RCC与健康对照者的曲线下面积(AUC)为0.828(95%CI：0.765~0.890),敏感度为72.2%,特异度为72.0%;诊断I/II期RCC患者的AUC为0.837(95%CI：0.771~0.902),敏感度和特异度分别为75.4%和72.0%。经逻辑回归分析,血清外泌体miR-181a-3p是RCC的独立危险因素(OR=6.662,95%CI：3.294~13.474,P<0.001)。 结论RCC患者尤其早期患者血清外泌体miR-181a-3p显著升高,其可作为非侵入性辅助诊断RCC的新型分子标志。     

MiR-122–5p regulates the pathogenesis of childhood obesity by targeting CPEB1

BackgroundChildhood obesity is strongly associated with inflammation which contributes to the development of several obesity-related disorders. Accumulating evidence has suggested that microRNAs (miRNAs) are involved in the pathogenesis of multiple human diseases, including childhood obesity. MiR-122–5p was reported to be related to obesity in childhood, however, the detailed function and mechanism of miR-122–5p are still obscure.MethodsSimpson-Golabi-Behmel syndrome (SGBS) adipocytes were cocultured with macrophage cell line THP-1 or macrophage-conditioned medium (MacCM) to promote cytokine expression. Oil Red O staining was used to detect the accumulation of lipid droplets in SGBS cells. The expression of interleukin 6 (IL-6), IL-8, and monocyte chemoattractant protein 1 (MCP-1) at mRNA and protein levels was assessed by RT-qPCR and ELISA, respectively. Western blotting was used for measuring protein levels of target genes of miR-122–5p. The luciferase reporter assay was applied for detecting the binding relation between miR-122–5p and cytoplasmic polyadenylation element binding protein 1 (CPEB1).ResultsCoculture of SGBS adipocytes and THP-1 macrophages/MacCM promoted IL-6, IL-8, and MCP-1 expression at mRNA and protein levels. Overexpression of miR-122–5p inhibited IL-6, IL-8, and MCP-1 expression in SGBS adipocytes, and this inhibitory effect was rescued by CPEB1 upregulation. CPEB1 3′-untranslated region was directly targeted by miR-122–5p.ConclusionMiR-122–5p suppresses cytokine expression in SGBS adipocytes by targeting CPEB1.     

miR-146a-5p在不明原因复发性流产蜕膜组织中的表达及其意义

目的: 探讨miR-146a-5p在不明原因复发性流产（URSA）蜕膜组织中的表达变化并分析其可能作用机制。方法: 选取2019年1—12月在成都西囡妇科医院因URSA行清宫术患者（URSA组）及因无生育计划行人工流产术的正常妊娠者（正常妊娠组）各10例,收集2组所有对象的蜕膜组织。比较2组蜕膜组织中miR-146a-5p水平,利用生物信息学方法预测miR-146a-5p潜在靶基因和相关分子机制,并检测蜕膜组织中关键靶分子水平并分析其与miR-146a-5p水平之间的相关性。结果: URSA组miR-146a-5p水平低于正常妊娠组（P<0.001）。在至少3个数据库均预测到的miR-146a-5p靶基因共35个,其中IRAK1、CD80和TRAF6富集于Toll样受体（TLR）信号通路;IRAK1、SORT1和TRAF6富集于神经营养素（NT）信号通路,miR-146a-5p预测靶基因编码的功能蛋白之间存在3对相互作用关系。URSA组IRAK1、CD80、TRAF6和SORT1基因和蛋白水平均高于正常妊娠组（P<0.05）,miR-146a-5p水平与这4个分子基因和蛋白水平均呈负相关（P<0.05）。结论: 蜕膜组织中miR-146a-5p可能通过调控TLR和NT信号通路参与URSA发病机制,为后续URSA防治策略研究提供了新的思路和靶点。     

Alterations in microRNA expression patterns in liver diseases

In the past few years there has been growing interest for a type of short RNAs called microRNAs, which are involved in the regulation of gene expression mainly in a negative way. There are about 1000 known microRNA today. It has been demonstrated that expression level of microRNA may become altered from normal to diseased state, thus microRNAs could be employed as a reliable tool in the diagnosis of diseases. A liver-characteristic microRNA (miR-122) needed for functioning hepatocytes has been identified, which usually shows a decreased expression level upon liver injury. miR-122 has been suggested as a biomarker since it was downregulated in the liver tissue upon acetaminophen-induced toxicity and in turn elevated miR-122 level was detected in the plasma. Moreover, miR-122 level in the plasma was found to be more sensitive as compared with conventional assays based on the release of liver enzymes. Also, miR-122 expression tends to decrease as carcinogenesis progresses. In addition, miR-122 enhances the replication of hepatitis C virus and its level seems to influence the efficiency of interferon therapy. Nowadays, many microRNAs are known whose distinctive alterations in their specific patterns seem to characterize individual pathological processes. In this article, the major alterations in microRNA expression patterns in liver diseases such as drug- and alcohol-induced liver diseases, non-alcoholic fatty liver diseases, fibrosis, viral infections (hepatitis), cirrhosis and hepatocellular carcinoma are summarized.     

Moderate consumption of beer, red wine and spirits has counteracting effects on plasma antioxidants in middle-aged men

OBJECTIVE: To evaluate the in vivo effects of moderate consumption of red wine, beer and spirits on antioxidants, antioxidant enzymes and antioxidant capacity. DESIGN: Randomized, diet-controlled, cross-over study. SUBJECTS: Twelve apparently healthy, non-smoking middle-aged men were included; 11 of them completed the study. INTERVENTIONS: Each subject consumed four glasses of red wine, beer, spirits and water (negative control) with evening dinner during four successive periods of 3 weeks, daily at the Institute. The total diet was supplied to the subjects and had essential the same composition during these 12 weeks. RESULTS: Neither the enzyme activities of serum glutathion peroxidase, erythrocyte glutathion reductase and superoxide dismutase nor the plasma concentrations of alpha- and gamma-tocopherol, lutein, zeaxantin, beta-cryptoxanthin, lycopene and alpha-carotene were affected. Plasma beta-carotene concentrations were decreased after 3 weeks' consumption of red wine, beer and spirits (40 g alcohol/day) as compared to consumption of water, by 15% (P=0.0005), 11% (P=0.010) and 13% (P=0.003), respectively. Also, plasma ascorbic acid was decreased after beer (15%, P=0.004) and spirits (12%, P=0.030), but not after wine consumption. Serum uric acid concentrations were increased after consumption of beer (15%, P<0.0001), spirits (8%, P=0.008) and red wine (9%, P=0.003). The overall serum antioxidant capacity, assessed as Trolox equivalent antioxidant capacity (TEAC), was similar for all treatments. CONCLUSIONS: Moderate consumption of red wine, beer and spirits has counteracting effects on plasma antioxidant components, resulting in no significant effect on overall antioxidant status. The effects on antioxidant parameters are largely independent of the type of alcoholic beverage, and probably irrelevant to chronic disease risk. SPONSORSHIP: Dutch Foundation for Alcohol Research (SAR).     

Vitamin D-mediated attenuation of miR-155 in human macrophages infected with dengue virus: Implications for the cytokine response

Clinical manifestations of dengue disease rely on complex interactions between dengue virus (DENV) and host factors that drive altered immune responses, including excessive inflammation. We have recently established that vitamin D can modulate DENV-induced cytokine responses and restrict infection in human macrophages. Cytokine responses are finely regulated by several homeostatic mechanisms, including microRNAs (miRNAs) that can rapidly target specific genes involved in the control of immune signaling pathways. However, the modulation of miRNAs by vitamin D during DENV infection is still unknown. Here, using a qPCR miRNA array we profiled immune-related miRNAs induced by DENV infection in human monocyte-derived macrophages (MDM) differentiated in absence or presence of vitamin D (D₃-MDM). We found several miRNAs differentially expressed in both MDM and D₃-MDM upon DENV infection. Interestingly, from these, a set of 11 miRNAs were attenuated in D₃-MDM as compared to MDM. Gene set enrichment analysis of the predicted mRNA targets of these attenuated miRNAs suggested a predominant role of miR-155-5p in the TLR-induced cytokine responses. Indeed, validation of miR-155-5p attenuation in D₃-MDM was linked to increased expression of its target gene SOCS-1, a key component for TLR4 signaling regulation. Likewise, TLR4 activation with LPS further corroborated the same miR-155-5p/SOCS-1 negative correlation observed in D₃-MDM upon DENV exposure. Moreover, D₃-MDM differentiation induced down-regulation of surface TLR4 that was linked to less TLR4/NF-κB-derived secretion of IL-1β. These data suggest a key role of vitamin D in the control of inflammatory cytokine responses during DENV infection of human macrophages via the TLR4/NF-κB/miR-155-5p/SOCS-1 axis.     

基于TCGA数据库构建肺腺癌预后相关的微小RNAs风险模型

目的寻找肺腺癌(lung adenocarcinoma,LUAD)特异性的预后相关微小RNAs(microRNAs, miRNAs),为LUAD患者预后预测及个性化治疗方案制定提供依据。 方法下载TCGA数据库中522例LUAD患者组织标本的miRNA-Seq数据和临床病理及生存时间数据,用R语言对LUAD与癌旁组织中差异miRNAs进行分析。采用LASSO & COX回归模型在训练集(245例LUAD)中进行LUAD预后相关miRNAs筛选,并构建基于7个miRNAs表达谱的线性风险模型。根据风险值的高低,以中位风险值为界将患者分为高、低风险组,并分别在测试集(245例LUAD)和总体标本(490例LUAD)中对风险模型预测患者预后的有效性进行验证。采用COX回归分析miRNAs风险模型是否是独立的预后因子。 结果LUAD组织与癌旁组织中共有72个差异表达的miRNAs(上调45个、下调27个)。从训练集中确定miR-101-3p、miR-148a-3p、miR-192-5p、miR-193b-3p、miR-505-3p、miR-584-5p和miR-99a-5p 7个与总生存期相关的miRNAs构建预后风险模型。在训练集、测试集及总体标本中,高风险组患者与低风险组患者相比,总体生存时间均显著降低(P均<0.05)。经多因素COX回归分析,风险模型在训练集、测试集及总体样本中均是一个独立的预后因子(训练集HR=1.97,P=0.02;测试集HR=1.927,P=0.009;总体HR=1.909,P=0.001)。 结论研究确定了7个与LUAD患者预后相关的miRNAs,基于7个miRNAs构建的风险模型是1个独立的预后因子。     

Changes of body weight and plasma ghrelin levels after gastric banding and gastric bypass

OBJECTIVE: Ghrelin is an enteric peptide with strong orexigenic and adipogenic effects. Plasma ghrelin levels are decreased in obese subjects but increase after weight loss; this increase is not observed after Roux-en-Y gastric bypass (RYGB). Prospective and comparative data after adjustable silicone gastric banding (ASGB) have not been reported previously. RESEARCH METHODS AND PROCEDURES: Overnight fasting plasma ghrelin concentration was measured in morbidly obese subjects at baseline and 3, 6, 12, and 24 months after ASGB (n = 8) or RYGB (n = 5) and in nonoperated controls (n = 7). RESULTS: After RYGB, body weight (BW) decreased by 29.5 +/- 5.5 kg (mean +/- SE, p < 0.001), whereas plasma ghrelin failed to increase significantly (+167 +/- 119 pg/mL, not significant). In contrast, after ASGB, BW decreased less (by 22.8 +/- 5.9 kg; p < 0.001), and plasma ghrelin significantly increased by 377 +/- 201 pg/mL (p = 0.025). Neither BW nor plasma ghrelin changed in nonoperated controls. Plasma leptin decreased in both operated groups (similarly p < 0.05) but not in nonoperated controls. Plasma growth hormone and insulin-like growth factor 1 were not correlated with changes in plasma ghrelin concentrations. DISCUSSION: Plasma ghrelin levels failed to increase during substantial weight loss after RYGB, but did increase in response to lesser weight loss after ASGB. These findings suggest that the plasma ghrelin response after weight loss is impaired after exclusion of major parts of the stomach and the duodenum (RYGB), and the smaller long-term weight loss after ASGB compared with RYGB may be due, at least in part, to an absent increase in plasma ghrelin after RYGB.     

LncRNA ZFAS1 plays a role in regulating the inflammatory responses in sepsis-induced acute lung injury via mediating miR-193a-3p

ObjectiveTo explore the role of lncRNA ZFAS1-mediated miR-193a-3p in the regulation of inflammatory responses in rats with sepsis-induced acute lung injury (ALI).MethodsSepsis-induced ALI models were constructed by LPS induction and then injected with ZFAS1 overexpression plasmid. Thereafter, lung injury score and the W/D weight ratio were calculated. Besides, bronchoalveolar lavage fluid (BALF) was isolated from rats to perform the cell count and protein quantification, while qRT-PCR and ELISA were performed to detect the inflammatory cytokines expressions. In vitro, NR8383 cells were transfected and then treated with LPS, followed by the measurement of inflammatory cytokines, cell viability and cell apoptosis.ResultsIn comparison with the Control group, rats in the LPS group presented sharp increases in the W/D weight ratio and injury score of lung, total protein concentration and the count of neutrophils and macrophages in BALF. Besides, rats in LPS group also resulted in a decrease in ZFAS1 expression and increase in miR-193a-3p expression in lung tissues, with the increased pro-inflammatory cytokines. Dual luciferase reporter gene assay confirmed a target relation between miR-193a-3p and ZFAS1. As compared to the Blank group, NR8383 cells in the LPS group had up-regulated pro-inflammatory cytokines with declined cell viability and elevated cell apoptosis; and meanwhile, ZFAS1 and Bcl-2 were decreased but miR-193a-3p and Bax were increased. Overexpression of ZFAS1 could significantly improve LPS-induced ALI in vivo and in vitro with reduced levels of pro-inflammatory cytokines.ConclusionOverexpression of ZFAS1, possibly via targeting the expression of miR-193a-3p, could inhibit the apoptosis and ameliorate the inflammatory responses of ALI in sepsis.     

Repeated exposure to alcoholic beer does not induce long-lasting changes in alcohol self-administration and intake in Sardinian alcohol-preferring and Sardinian non-preferring rats

AIMS: Rats avidly consume non-alcoholic beer, and addition of alcohol to non-alcoholic beer may function as a medium to induce intake of large amounts of alcohol in rats. The present study investigated whether Sardinian alcohol-preferring (sP) and Sardinian non-preferring (sNP) rats, initially exposed to non-alcoholic beer, and subsequently to non-alcoholic beer containing increasing concentrations of alcohol, would develop unusually high alcohol self-administration and drinking behaviours: (i) when alcohol was added to non-alcoholic beer, and (2) once beer was withdrawn and a plain alcohol solution was made available. METHODS: In Experiment 1, rats were exposed to operant, 30-min/day self-administration sessions of non-alcoholic beer with increasing concentrations of alcohol [0, 2.5, 5, 7.5, and 10% (v/v)] for a total of 45 days. After a brief 'beer-fading' phase, the rats were exposed to self-administration sessions of a plain 10% (v/v) alcohol solution. In Experiment 2, the rats were exposed to non-alcoholic beer with increasing concentrations of alcohol [0, 2.5, 5, 7.5, and 10% (v/v)] and water under the 2-bottle choice regimen with unlimited access (24 h/day) for a total of 35 days. After a brief 'beer-fading' phase, the rats were exposed to the choice between a plain 10% (v/v) alcohol solution and water. RESULTS: sP and sNP rats did not differ in self-administration (Experiment 1) and intake (Experiment 2) of non-alcoholic beer. In Experiment 1, as alcohol content increased, the amount of self-administered alcohol increased progressively in sP rats (up to 1-1.2 g/kg) and remained stable in sNP rats (approximately 0.65 g/kg). When the plain 10% alcohol solution was available, the amount of self-administered alcohol in sP rats initially dropped, and tended to increase-up to approximately 0.6 g/kg-on continuing exposure. In sNP rats, their lever-pressing behaviour was rapidly extinguished after beer withdrawal. In Experiment 2, as alcohol content was increased, daily alcohol intake increased progressively in sP rats (up to 8-9 g/kg) and averaged approximately 2.4 g/kg in sNP rats. When the plain alcohol solution was available, daily alcohol intake in sP rats was initially low, reaching control values on continuing exposure; conversely, daily alcohol intake was completely suppressed in sNP rats. CONCLUSIONS: These results suggest that exposure to alcoholic beer resulted in unusually high intakes of alcohol in both sP and sNP rats for as long as non-alcoholic beer was added to alcohol; however, these high levels of alcohol self-administration and intake were not maintained once non-alcoholic beer was withdrawn.     

Alcohol consumption and plasma homocysteine.

A few reports show that consumption of spirits and of wine correlate with elevated plasma total homocysteine (tHcy), which is associated with the risk of cardiovascular disease. We analyzed the relation between tHcy and current daily ethanol consumption cross-sectionally in middle-aged Japanese men (n = 974, age 51-59 years). Plasma tHcy was positively associated with consumption of whiskey but not with consumption of shochu (Japanese spirits), sake, beer, or wine. Odds ratios of an increase in daily intake of 30 ml ethanol (approximately 1 standard deviation) for hyperhomocysteinemia (>14.0 micromol/l) were 2.58 (95% confidence interval, 1.29-5.14) for whiskey, 1.08 (0.78-1.50) for shochu, 0.99 (0.59-1.66) for sake, 0.98 (0.58-1.63) for beer, and 1.70 (0.31-9.50) for wine in a multivariate logistic regression analysis adjusted for the daily number of cigarettes smoked, physical activity, vegetable consumption, and serum creatinine levels. After inclusion of plasma folate and vitamin B12 in the multivariate analysis model, the association between whiskey ethanol consumption and hyperhomocysteinemia remained significant with odds ratio of 2.79 (1.36-5.72). These results suggest that whiskey consumption correlates with hyperhomocysteinemia independently of plasma folate or vitamin B12 or lifestyle factors in the population studied.     

Circulating miR-29c,miR-30c,miR-193a-5p and miR-885-5p: Novel potential biomarkers for HTLV-1 infection diagnosis

Human T-cell leukemia virus type 1 (HTLV-1) is an oncoretrovirus that infects 5–10 million people worldwide. Currently, different methods are used to test HTLV-1 infection. However, a biomarker that could enable an early and accurate diagnosis of HTLV-1 infection is still lacking. Here, we compared the serum miRNA expression profile in HTLV-1 infected patients versus healthy individuals to identify a potential biomarker for diagnosis of HTLV-1 infection.TaqMan miRNA microarray (TLDA) was carried out to compare the miRNA expression profile in infected versus healthy individuals. Quantitative real-time RT-PCR (qRT-PCR) was applied to validate TLDA results. Receiver-operator characteristic (ROC) curve analysis was performed to determine the diagnostic accuracy of the most highly and significantly identified deregulated miRNA(s) as potential biomarker(s). We identified deregulated expression for ten miRNAs with miR-127, miR-136, miR-142-3p, miR-221, and miR-423-5p being down-regulated whilst let-7b, miR-29c, miR-30c, miR-193a-5p, and miR-885-5p being up-regulated in infected individuals. ROC curve analyses showed an AUC (Areas Under the ROC Curve) of 0.875 (95% CI: 0.7819–0.9581; P = .0021), 0.861 (95% CI: 0.7596–0.9754; P = .003), 0.856 (95% CI: 0.689–0.895; P = .011), and 0.849 (95% CI: 0.678–0.855; P = .017) for miR-29c, miR-30c, miR-193a-5p, and miR-885-5p respectively. Combined ROC analyses using these 4 miRNAs showed a greater AUC of 0.907 (95% CI: 0.809–1; P = .000001) indicating a robust diagnostic value of these 4 miRNAs. Our findings highlight serum miR-29c, miR-30c, miR-193a-5p and miR-885-5p as novel potential biomarkers important for HTLV-1 diagnosis.     

肺癌诊断相关miRNA标志物的筛选和验证

目的 获得肺癌组织和正常肺部组织中差异表达的miRNA用于肺癌的早期诊断.方法 首先运用miRNA芯片比较5对来自临床的正常肺部组织和肺癌组织,筛选差异性表达的miRNA,然后用荧光实时定量PCR进行鉴定.最后用ROC曲线分析评价特定miRNA标志物区分肺癌组织与正常肺部组织的能力.结果 miR-10b-5p和miR-199a-5p在肺癌组织中的相对表达量(2.76±0.71;2.05±0.38)显著高于肺部正常组织中的相对表达量(1.00±0.17;0.96±0.18),差异有统计学意义(t=2.40、2.56,P＜0.05).ROC曲线评价miR-10b-5p和miR-199a-5p区分肺癌组织和肺部正常组织的能力,曲线下面积(AUC)分别为0.731和0.672.两个miRNA联合区分肺癌组织和肺部正常组织的AUC为0.773,灵敏度为57.9％,特异性为94.7％.结论 miR-10b-5p和miR-199a-5p联合可作为候选肿瘤标志物组合用于早期肺癌诊断.     

Circulating MicroRNAs,Polychlorinated Biphenyls,and Environmental Liver Disease in the Anniston Community Health Survey

Background: Polychlorinated biphenyl (PCB) exposures have been associated with liver injury in human cohorts, and steatohepatitis with liver necrosis in model systems. MicroRNAs (miRs) maintain cellular homeostasis and may regulate the response to environmental stress.Objectives: We tested the hypothesis that specific miRs are associated with liver disease and PCB exposures in a residential cohort.Methods: Sixty-eight targeted hepatotoxicity miRs were measured in archived serum from 734 PCB-exposed participants in the cross-sectional Anniston Community Health Survey. Necrotic and other liver disease categories were defined by serum keratin 18 (K18) biomarkers. Associations were determined between exposure biomarkers (35 ortho-substituted PCB congeners) and disease biomarkers (highly expressed miRs or previously measured cytokines), and Ingenuity Pathway Analysis was performed.Results: The necrotic liver disease category was associated with four up-regulated miRs (miR-99a-5p, miR-122-5p, miR-192-5p, and miR-320a) and five down-regulated miRs (let-7d-5p, miR-17-5p, miR-24-3p, miR-197-3p, and miR-221-3p). Twenty-two miRs were associated with the other liver disease category or with K18 measurements. Eleven miRs were associated with 24 PCBs, most commonly congeners with anti-estrogenic activities. Most of the exposure-associated miRs were associated with at least one serum hepatocyte death, pro-inflammatory cytokine or insulin resistance bioarker, or with both. Within each biomarker category, associations were strongest for the liver-specific miR-122-5p. Pathways of liver toxicity that were identified included inflammation/hepatitis, hyperplasia/hyperproliferation, cirrhosis, and hepatocellular carcinoma. Tumor protein p53 and tumor necrosis factor α were well integrated within the top identified networks.Discussion: These results support the human hepatotoxicity of environmental PCB exposures while elucidating potential modes of PCB action. The MiR-derived liquid liver biopsy represents a promising new technique for environmental hepatology cohort studies. https://doi.org/10.1289/EHP9467