凉血通瘀方干预高血压大鼠急性脑出血模型对脑组织中差异miRNA表达的影响＊

目的 研究凉血通瘀方对高血压大鼠急性脑出血模型脑组织miRNA表达的影响，对差异表达的miRNA靶基因进行分析，探索凉血通瘀方可能的药效机制。方法 将自发性高血压大鼠随机分成对照组（B）和实验组（C）。适应性饲养一周后，C组灌胃凉血通瘀方，B组灌胃等体积生理盐水，连续5天，每天1次。构建脑出血模型后收集脑组织，借助全转录组测序技术获得miRNA表达量，与miRBase数据库比对获取已知miRNA，使用miRDeep2预测新miRNA。差异分析软件为DESeq2，筛选阈值为|log₂FC| ≥1 并且P <0.05。对显著差异表达的miRNA进行靶基因预测，对靶基因进行GO功能、KEGG通路富集和PPI网络分析。结果 实验组和对照组对比，共发现21个显著差异表达的miRNA，上调有9个，下调有12个，共预测得到1243个有统计学意义的靶基因。GO富集分析发现，生物过程中突触囊泡分泌的调节、神经递质分泌的调节和神经递质运输的调节占前三位，神经元投射终点、全膜、质膜区域和细胞投射则是主要的细胞成分。分子功能分别为小GTPase绑定、底物特异性跨膜转运蛋白活性和离子跨膜转运体活性。通路分析结果显示，靶基因在癌证通路、pI3K-Akt信号通路、人类乳头瘤病毒感染、神经活性配体-受体相互作用和MAPK通路等分布广泛。采用STRING网站和Cytoscape软件，根据MCC算法筛选出ADRA2C、CASR、CCL28、CCR1、DRD2、GNAT3、GRM2、DYNC1LI1、GABBR1、GNAI1等核心靶基因。结论 凉血通瘀方对脑出血急性期鼠脑组织内miRNA的表达有重要影响；显著差异表达miRNAs可能通过靶向核心基因调控凉血通瘀方干预急性脑出血的病理过程及预后。     

microRNA在结肠癌组织的表达及其临床意义

目的 探讨结肠癌组织与对应正常结肠组织miRNA表达特征及其在结肠癌的诊断中的意义.方法 通过定量聚合酶链反应(qRT-PCR)法检测了6对结肠癌及其远端正常结肠的手术标本中200种miRNA的表达情况,并应用PAM分析法探讨miRNA在预测结肠癌病变中的作用.结果 检测6对结肠癌及其远端正常结肠的手术标本中miRNA的表达情况发现在结肠癌组织/远端正常结肠组织中存在132种miRNA,并且有95种miRNA在结肠癌发生过程中表达量发生了显著的变化(变化倍数大于1.5),其中有48种表达增加应用PAM分析方法对9对结肠癌组织/远端正常结肠组织进行了定性判断,发现预测结果 与病理切片诊断结果 一致.结论 miRNA与结肠癌的发展有着密切的联系,并且其可以作为一个有效的肿瘤标志物以进行结肠癌的早期预测和诊断.     

Effect of Dietary Fatty Acids on MicroRNA Expression Related to Metabolic Disorders and Inflammation in Human and Animal Trials

Dietary fatty acids (DFAs) play key roles in different metabolic processes in humans and other mammals. DFAs have been considered beneficial for health, particularly polyunsaturated (PUFAs) and monounsaturated fatty acids (MUFAs). Additionally, microRNAs (miRNAs) exert their function on DFA metabolism by modulating gene expression, and have drawn great attention for their potential as biomarkers and therapeutic targets. This review explicitly examined the effects of DFAs on miRNA expression associated with metabolic diseases, such as obesity, non-alcoholic fatty liver disease (NAFLD), and cardiovascular disease (CVD), as well as inflammation, published in the last ten years. DFAs have been shown to induce and repress miRNA expression associated with metabolic disease and inflammation in different cell types and organisms, both in vivo and in vitro, depending on varying combinations of DFAs, doses, and the duration of treatment. However, studies are limited and heterogeneous in methodology. Additionally, recent studies demonstrated that high fat ketogenic diets, many enriched with saturated fats, do not increase serum saturated fat content in humans, and are not associated with increased inflammation. Thus, these findings shed light on the complexity of novel treatment and DFA interventions for metabolic disease and to maintain health. Further studies are needed to advance molecular therapeutic approaches, including miRNA-based strategies in human health and disease.     

Blood biomarkers of mild traumatic brain injury: State of art

BackgroundMild traumatic brain injury (mTBI) is one of the most common causes of emergency department visits around the world. Up to 90% of injuries are classified as mTBI. Cranial computed tomography (CCT) is a standard diagnosis tool to identify intracranial complications in adults with mTBI. Alternatively, children can be admitted for inpatient observation with CCT scans performed only on those with clinical deterioration. The use of blood biomarkers is a supplementary tool for identifying patients at risk of intracerebral lesions who may need imaging.MethodWe realised a bibliographic state of art providing a contemporary clinical and laboratory framework for blood biomarker testing in mTBI management.ResultsThe S100B protein is the only biomarker that can be used today in the clinical routine for management of mTBI with appropriate evidence-based medicine. Due to its excellent negative predictive value, S100B protein is an alternative choice to CCT scanning for mTBI management with considered, consensual and pragmatic use. In this state of art, we propose points to help clinicians and clinical pathologists use serum S100B protein in the clinical routine. A state of art on the different biomarkers (GFAP, UCH-L1, NF [H or L], tau, H-FABP, SNTF, NSE, miRNAs, MBP) is also conducted. Some of these other biomarkers, used alone (GFAP, UCH-L1) or in combination (GFAP + H-FABP ± S100B ± IL10) can improve the specificity of S100B.ConclusionUsing a bibliographic state of art, we highlighted the added values of the blood biomarkers for the clinical management of mTBI.     

基于双链特异性核酸酶触发滚环扩增的microRNA-21太赫兹超材料传感方法的构建

目的:构建一种太赫兹(THz)超材料传感方法用于microRNA-21(miRNA-21)的信号放大检测。方法:首先构建THz超材料传感方法,并用聚丙烯酰胺凝胶电泳及zeta电位验证方法的可行性;对传感器检测条件进行优化之后,对不同浓度的miRNA-21以及其他不同的miRNAs进行检测,并与其他microRNA检测方法进行比较;最后,对该传感器的回收率进行了评价。结果:在最优实验条件下,通过双链特异性核酸酶(DSN)循环识别与滚环扩增(RCA)的双重信号放大策略,该THz超材料传感器对靶标miRNA-21的响应范围为10 fmol/L至10 nmol/L,检测限为8.49 fmol/L。并且该传感器具有较好的特异性,具备了从多种miRNAs中识别靶标miRNA-21的能力,并且在商业化人血清样本中的回收率可达94.33%到115.33%。结论:该THz传感器可以实现靶标miRNA-21的高灵敏、高特异性检测,具备了在miRNA相关疾病无标记诊断与早期预警的潜力。     

肿瘤相关基因NET-1的研究进展

NET-1(tspan1)是新近被证实的四次跨膜蛋白超家族(transmembrane 4 superfamily,TM4SF)的成员之一。作为一种肿瘤相关基因,与肿瘤细胞的增殖、迁移、内吞作用和诱导血管形成密切相关。NET-1蛋白的异常表达可能是引起和维持癌恶性表型的原因之一,其表达水平对癌的预后具有评估价值,该基因可能成为肿瘤基因治疗的靶点。     

糖肾方对db/db小鼠肾脏纤维化的保护作用

目的:观察糖肾方(Tang Shen Fang,TSF)对于自发性2型糖尿病肾病db/db小鼠肾脏纤维化的保护作用。方法:8周龄雄性db/db小鼠随机分为2组,分别为模型组(db/db)和观察组(db/db+TSF),每组9只;8周龄雄性db/m小鼠为正常对照组(db/m),顺应性喂养2周后,给予糖肾方干预12周,处死小鼠,分离小鼠肾脏用于后续检测。利用PAS和Masson染色观察肾脏病理;利用Western Blotting、Real-time PCR和免疫组化等技术检测肾脏组织纤维化TGF-β1/Smad3通路以及miRNA21和miRNA29b的表达。结果:糖肾方可以改善db/db小鼠尿微量白蛋白水平,减轻小鼠肾脏损伤;Western Blotting、Real-time PCR和免疫组化结果显示,糖肾方可下调db/db小鼠肾脏中纤维化标志分子TGF-β1、Smad3、CollagenⅠ和Fibronectin的表达;下调db/db小鼠肾脏miRNA 21的表达,上调miRNA 29b的表达。结论:糖肾方可作用于TGF-β1/Smad3信号通路抑制肾脏纤维化。     

血清miR-21和miR-34a与晚期胃癌患者Folfox化疗方案化疗疗效的关系研究

目的 探讨晚期胃癌患者Folfox方案化疗前后血清miR-21和miR-34a水平变化及其与化疗疗效的关系。方法 以在我院接受Folfox方案化疗的45例晚期胃癌患者为研究对象。采用Real-time PCR检测化疗前后miR-21和miR-34a水平,分析miR-21和miR-34a水平与化疗疗效和患者预后的关系,采用受试者工作曲线(ROC)评估miR-21和miR-34a对化疗疗效的评估。结果 化疗后,部分缓解6例(13.33%)、疾病稳定10例(22.22%)、疾病进展29例(64.44%),客观缓解率为13.33%,疾病控制率为35.56%,中位生存期为8.1个月。化疗后miR-21水平下调,而miR-34a水平上调,差异有统计学意义(P＜0.05)。化疗有效组miR-21水平低于无效组,而miR-34a水平高于无效组,差异有统计学意义(P＜0.05)。miR-21与化疗疗效呈负相关,miR-34a与之呈正相关(r_s=-0.431,0.504,P＜0.05)。miR-21低水平患者中位生存期7.6个月,而高水平患者为6.0个月;miR-34a低水平患者中位生存期5.5个月,而高水平患者为8.3个月,Log-Rank检验显示,miR-21低水平患者中位生存期大于高水平患者(P＜0.05),miR-34a高水平患者中位生存期大于低水平患者(P＜0.05)。miR-21和miR-34a联合评估化疗疗效的曲线下面积为0.865、灵敏度为80.9%、特异度为89.8%。结论 血清miR-21和miR-34a与晚期胃癌化疗疗效和患者中位生存期相关。     

不同海拔地区藏族胃癌患者miRNA的初步筛选

目的 筛选不同海拔地区生活的藏族患者的胃癌组织微小RNA(miRNA)差异表达,探讨高原地区外环境缺氧是否参与调控肿瘤的发生、侵袭和转移等.方法 选择5对不同海拔地区生活的藏族胃癌患者作为研究对象(其中海拔﹥3500 m者5例,海拔﹤2450 m者5例),对胃癌组织及其配对癌旁组织进行miRNA芯片检测,分析差异miRNA,查询差异miRNA-靶基因,预测的靶基因应用DAVID数据库进行功能富集分析(GO分析)和pathway富集分析.结果 在5对不同海拔地区生活的藏族胃癌患者的胃癌组织及其配对癌旁组织中找出16个差异表达的miRNA,在海拔﹥3500 m组均上调,仅有hsa-miR-1260b、hsa-miR-130b-3p、hsa-miR-3648、hsa-miR-4284预测到验证的靶基因,其中hsa-miR-1260b预测118个,hsa-miR-130b-3p预测151个.这些靶基因富集在细胞增殖、基因表达调控、转录调控、细胞周期、细胞内信号传导等生物学过程和功能上;pathway富集到癌症通路上.结论 在高原藏族胃癌患者的肿瘤生长、侵袭、转移及肿瘤耐药等过程中,hsa-miR-1260b、hsa-miR-130b-3p可能发挥着重要的作用.