Peripheral blood RNA biomarkers can predict lesion severity in degenerative cervical myelopathy

Degenerative cervical myelopathy is a common cause of spinal cord injury,with longer symptom duration and higher myelopathy severity indicating a worse prognosis.While numerous studies have investigated serological biomarkers for acute spinal cord injury,few studies have explored such biomarkers for diagnosing degenerative cervical myelopathy.This study involved 30 patients with degenerative cervical myelopathy(51.3±7.3 years old,12 women and 18 men),seven healthy controls(25.7±1.7 years old,one woman and six men),and nine patients with cervical spondylotic radiculopathy(51.9±8.6 years old,three women and six men).Analysis of blood samples from the three groups showed clear differences in transcriptomic characteristics.Enrichment analysis identified 128 differentially expressed genes that were enriched in patients with neurological disabilities.Using least absolute shrinkage and selection operator analysis,we constructed a five-gene model(TBCD,TPM2,PNKD,EIF4G2,and AP5Z1)to diagnose degenerative cervical myelopathy with an accuracy of 93.5%.One-gene models(TCAP and SDHA)identified mild and severe degenerative cervical myelopathy with accuracies of 83.3%and 76.7%,respectively.Signatures of two immune cell types(memory B cells and memory-activated CD4^(+)T cells)predicted levels of lesions in degenerative cervical myelopathy with 80%accuracy.Our results suggest that peripheral blood RNA biomarkers could be used to predict lesion severity in degenerative cervical myelopathy.

Non-coding RNAs in acute ischemic stroke:from brain to periphery

Acute ischemic stroke is a clinical emergency and a condition with high morbidity,mortality,and disability.Accurate predictive,diagnostic,and prognostic biomarkers and effective therapeutic targets for acute ischemic stroke remain undetermined.With innovations in high-throughput gene sequencing analysis,many aberrantly expressed non-coding RNAs(ncRNAs)in the brain and peripheral blood after acute ischemic stroke have been found in clinical samples and experimental models.Differentially expressed ncRNAs in the post-stroke brain were demonstrated to play vital roles in pathological processes,leading to neuroprotection or deterioration,thus ncRNAs can serve as therapeutic targets in acute ischemic stroke.Moreover,distinctly expressed ncRNAs in the peripheral blood can be used as biomarkers for acute ischemic stroke prediction,diagnosis,and prognosis.In particular,ncRNAs in peripheral immune cells were recently shown to be involved in the peripheral and brain immune response after acute ischemic stroke.In this review,we consolidate the latest progress of research into the roles of ncRNAs(microRNAs,long ncRNAs,and circular RNAs)in the pathological processes of acute ischemic stroke–induced brain damage,as well as the potential of these ncRNAs to act as biomarkers for acute ischemic stroke prediction,diagnosis,and prognosis.Findings from this review will provide novel ideas for the clinical application of ncRNAs in acute ischemic stroke.

Heme status affects human hepatic messenger RNA and microRNA expression

AIM:To assess effects of heme on messenger RNA(mRNA) and microRNA(miRNA) profiles of liver cells derived from humans.METHODS:We exposed human hepatoma cell line Huh-7 cells to excess iron protoporphyrin(heme)(10 μmol/L) or induced heme deficiency by addition of 4,6-dioxoheptanoic acid(500 μmol/L),a potent inhibitor of aminolevulinic acid dehydratase,for 6 h or 24 h.We harvested total RNA from the cells and performed both mRNA and miRNA array analyses,with use of Affymetrix chips,reagents,and instruments(human genome U133 plus 2.0 and miRNA 2.0 arrays).We assessed changes and their significance and interrelationships with Target Scan,Pathway Studios,and Ingenuity software.RESULTS:Changes in mRNA levels were most numerous and striking at 6 h after heme treatment but were similar and still numerous at 24 h.After 6 h of heme exposure,the increase in heme oxygenase 1 gene expression was 60-fold by mRNA and 88-fold by quantitative reverse transcription-polymerase chain reaction.We found striking changes,especially up-regulation by heme of nuclear erythroid-2 related factor-mediated oxidative stress responses,protein ubiquitination,glucocorticoid signaling,P53 signaling,and changes in RNAs that regulate intermediary metabolism.Fewer mRNAs were down-regulated by heme,and the fold decreases were less exuberant than were the increases.Notable decreases after 24 h of heme exposure were patatin-like phospholipase domain-containing protein 3(-6.5-fold),neuronal PAS domain protein 2(-1.93-fold),and protoporphyrinogen oxidase(-1.7-fold).CONCLUSION:Heme excess exhibits several toxic effects on liver and kidney,which deserve study in humans and in animal models of the human porphyrias or other disorders.

精神分裂症患者外周血中差异表达circRNA、miRNA、mRNA筛选及核心竞争性内源RNA网络构建

目的筛选精神分裂症患者外周血中差异表达的circRNA、miRNA、mRNA,并构建核心竞争性内源RNA(ceRNA)网络,解析精神分裂症发生发展的分子机制。方法精神分裂症患者6例记为精神分裂症组,同期体检健康的人群4例记为健康对照组。采集两组研究对象的外周血,提取总RNA后构建cDNA文库并进行高通量测序,采用R软件筛选精神分裂症患者外周血中差异表达circRNA、miRNA、mRNA。采用miRanda和qTar数据库共同预测与差异表达miRNA相互作用的circRNA和mRNA,与差异表达的circRNA、mRNA取交集后,采用R软件构建差异表达的ceRNA网络。基于DAVID数据库对精神分裂症患者差异表达ceRNA网络中差异mRNA进行GO和KEGG富集分析。采用string数据库构建精神分裂症患者差异表达ceRNA网络中差异mRNA之间的PPI网络,选用cytoscape软件中cytoHubba插件筛选影响精神分裂症发生发展的核心mRNA,选择与核心mRNA存在相互作用关系的miRNA和circRNA构建精神分裂症患者外周血中差异表达的核心ceRNA网络。结果相比健康对照组,精神分裂症组患者外周血中差异表达circRNA、miRNA、mRNA分别为31、213、2624个。构建了由差异表达的5个circRNA、4个miRNA和57个mRNA组成的ceRNA网络。精神分裂症患者差异表达ceRNA网络中mRNA主要参与化学刺激的感官知觉、多细胞生物生长的负调控等生物过程,涉及泛素-泛素连接酶活性、鸟苷酸结合、酶结合等分子功能,以及细胞质、肌动蛋白细胞骨架、轴突末端等细胞成分,ceRNA网络中mRNA显著富集于NF-κB信号通路、PI3K-Akt信号通路、Ras信号通路、泛素介导的蛋白水解等代谢通路。构建的精神分裂症患者差异表达ceRNA网络中mRNA PPI网络共包含23个节点和22条边,筛选获得影响精神分裂症发生发展的2个核心mRNA为BRCA1、GNAS。构建的精神分裂症患者外周血中差异表达的核心ceRNA网络包含5个节点和4条边。5个节点中包含2个核心mRNA:BRCA1、GNAS,1个miRNA:hsa-miR-502-5p,2个circRNA:hsa_circ_0001423、hsa_circ_0008494。4条边分别为hsa_circ_0001423/hsa-miR-502-5p/BRCA1、hsa_circ_0001423/hsa-miR-502-5p/GNAS、hsa_circ_0008494/hsa-miR-502-5p/BRCA1和hsa_circ_0008494/hsa-miR-502-5p/GNAS。结论与健康对照人群相比,精神分裂症患者外周血中有差异表达的circRNA 31个、miRNA 213个、mRNA 2624个,最终构建了包含5个节点和4条边的核心ceRNA网络。

Significance of vascular endothelial growth factor messenger RNA expression in gastric cancer

IM To study VEGF mRNA expression in gastric carcinoma and to clarify the association of its expression with the clinicopathologic features of the disease.METHODS In situ hybridization (ISH) and histochemistry were used to examine and analyze the expression of VEGF mRNA and antigen, and microvessel count (MVC) in 28 cases of gastric carcinomatous tissue in combination with clinical materials.RESULTS Ninteen of 28 gastric carcinomas were positive for VEGF mRNA. VEGF mRNA was mainly expressed in malignant cells and not in normal epithelium of gastric mucosa. Its expression was further increased in tumor cells adjacent to tumor necrosis zones, where stromal cells expressed VEGF mRNA occasionally. There was a close correlation between MVC and VEGF mRNA positivity (P<0005). High VEGF mRNA levels were significantly associated with serosal invasion, lymph node metastasis and TNM staging (P<005, respectively).CONCLUSION VEGF mRNA expression is associated with tumor invasion and metastasis by stimulating angiogenesis in gastric carcinoma.expression; endothelial growth factor.

Expression of alpha-fetoprotein messenger RNA in BEL-7404 human hepatoma cells and effect of L-4-oxalysine on the expression

AIM To investigate alpha-fetoprotein (AFP) mRNA expression in BEL-7404 human hepatoma cells and the effect of L-4-oxalysine (OXL) on the expression.METHODS BEl-7404 human hepatoma cells were maintained in RPMI 1640 media. Human AFP cDNA probe was labelled with digoxigenin-11-dUTP by the random primer labelling method. The expression of AFP mRNA in Bel-7404 cells was determined by an in situ hybridization technique with digoxigenin-labelled human AFP cDNA probe. The positive intensities of AFP mRNA in cells were analyzed by microspectrophotometer and expressed as absorbance at 470nm. For the experiment with OXL, cells were incubated with various concentrations of the agent for 72h.RESULTS Essentially all the hepatoma cells contained AFP mRNA in the cytoplasm, although in various amounts. The specificity of the hybridization reaction was confirmed by control experiments in which the use of RNase-treated BEL-7404 cells, non-AFP-producing cells (HL-60 human leukemia cells) or a nonspecific cDNA probe resulted in negative hybridization. When the cells were treated with OXL (25, 50mg/L), the content of AFP mRNA in the cytoplasm was decreased with the inhibition percentages of 34.3% and 70.1%, respectively (P＜0.05).CONCLUSION AFP mRNA was expressed in BEL-7404 human hepatoma cells and OXL suppressed AFP mRNA expression in the cells.

非编码RNA和外泌体在妊娠期糖尿病发生机制中的作用及早期诊断价值

背景:近几年有很多关于外泌体非编码RNA与妊娠期糖尿病发生机制的研究,但缺乏不同来源尤其是胎盘来源外泌体的最新系统综述。目的:文章对微小RNA、长链非编码RNA和环状RNA以及外泌体在妊娠期糖尿病中的变化情况及对疾病发生发展的可能作用进行综述,为临床妊娠期糖尿病的早期筛查与治疗提供潜在靶标。方法:从PubMed、Web of Science、中国知网、万方数据及维普数据库进行非编码RNA或外泌体非编码RNA与妊娠期糖尿病相关文献进行检索,最终纳入74篇文献进行综述。结果与结论:①非编码RNA通过调节各种生理功能参与了妊娠期糖尿病的发生发展,为妊娠期糖尿病的研究提供了新的方向。②外泌体在人体中广泛存在,红细胞、上皮细胞和胎盘细胞等均可分泌外泌体,这些不同来源外泌体中的非编码RNA被证明可以在妊娠期糖尿病的发病机制、诊断以及治疗方法中发挥一定的作用。③微小RNA与妊娠期糖尿病:外周血微小RNA在妊娠期糖尿病中的作用主要是影响滋养层细胞、胰腺β细胞功能以及妊娠期糖尿病血糖水平;胎盘微小RNA可反映妊娠期糖尿病的严重程度,并损伤滋养层细胞的功能。④长链非编码RNA与妊娠期糖尿病:外周血长链非编码RNA可以通过磷脂酰肌醇3激酶/蛋白激酶B通路诱导胰岛素抵抗并且有可能为妊娠期糖尿病的诊断与治疗提供新的思路;胎盘长链非编码RNA可以调控妊娠期糖尿病胎盘滋养层细胞增殖迁移,促进妊娠期糖尿病的发生发展。⑤环状RNA与妊娠期糖尿病:外周血和胎盘环状RNA可通过损伤胎盘滋养层细胞增殖迁移与代谢等功能诱导糖尿病的发生发展。⑥外泌体非编码RNA与妊娠期糖尿病:外周血外泌体非编码RNA可影响妊娠期糖尿病血糖水平和葡萄糖稳态,通过影响胎盘功能参与妊娠期糖尿病的发生发展。⑦非编码RNA有潜力成为妊娠期糖尿病早期诊断的标志物,此外工程化的外泌体能更好地实现妊娠期糖尿病的靶向治疗等,这些最新信息为妊娠期糖尿病的基础研究和临床转化提供了参考依据。⑧未来还需改进外周血外泌体的提取和纯化方法,并排除种族、饮食和体力活动等因素以提高结果的可重复性,而循环非编码RNA及外泌体在妊娠期糖尿病的预测和诊断中的临床应用还需要更多的前瞻性临床研究。

宫颈癌RNA分子的研究进展

信使RNA(mRNA)、微小RNA(miRNA)、长链非编码RNA(lncRNA)和环状RNA(circRNA)等在内的RNA分子之间彼此联系、互相协调,共同构成动态的分子网络,即竞争性内源RNA(ceRNA)调控网络。近年来研究揭示,ceRNA调控网络在促进宫颈癌的发生、发展和转归等过程中发挥着重要的调控作用。ceRNA调控网络中的任何一种RNA分子出现异常,均有可能通过多条信号通路导致宫颈癌细胞增殖、浸润、迁移或凋亡。现就近年来取得的研究进展,重点对mRNA、miRNA、lncRNA和circRNA等RNA分子与宫颈癌的关系进行综述,旨在深入阐明宫颈癌发病相关RNA分子机制,从而为宫颈癌的诊断和治疗提供新思路。

慢性乙型肝炎患者血清HBsAg与单个核细胞乙型肝炎病毒RNA水平对聚乙二醇干扰素治疗效果的预测价值

目的探讨慢性乙型肝炎患者外周血清HBsAg、乙型肝炎病毒(HBV)DNA与血清HBV RNA及单个核细胞(PBMC)HBV RNA的关系。方法50例慢性乙型肝炎患者,给予Peg-IFNα-2b 180μg皮下注射,每周一次,分别在初始治疗、24周和48周,检测患者血清HBV五项、肝功能、HBV DNA、HBV RNA及PBMC中HBV RNA的变化。结果患者三个时间段的生化指标ALT、AST、TBIL、AKP及GGT比较差异无统计学意义(P>0.05);免疫学标志物HBsAg、HBsAb、HBeAg、HBeAb差异有统计学意义(P<0.05);血清HBV DNA及HBV RNA与PBMC HBV RNA差异有显著统计学意义(P<0.001)。结论Peg-IFNα-2b抗病毒治疗各时间段,肝功能转氨酶无显著变化;治疗24周,血HBsAg、HBV DNA与HBV RNA及PBMC HBV RNA快速下降,有显著相关性;治疗48周,血清HBsAg、HBV DNA与HBV RNA及PBMC HBV RNA相关性减弱。因此,24周HBV RNA下降幅度优于48周,更能预测临床治愈。

参芪复方对糖尿病GK大鼠肝脏组织mRNA、lncRNA表达谱的影响

目的应用全转录组测序与实时聚合酶链锁反应(Real-time polymerase chain reaction,RT-qPCR)技术,探索参芪复方调控糖尿病GK大鼠肝脏信使RNA(messenger RNA,mRNA)、长链非编码RNA(long non-coding RNA,lncRNA)表达谱的机制研究。方法采用高脂高糖饲料建立2型糖尿病模型,将GK大鼠随机分为模型组、参芪复方组和西药组。另将10只Wistar大鼠设为空白组,予普通饲料喂养。参芪复方组大鼠予参芪复方浸膏灌胃,西药组予西格列汀混悬液灌胃,模型组及空白组灌服生理盐水。干预12周,每周检测空腹血糖(fasting blood glucose,FBG)。运用HE染色法观察大鼠肝脏组织病理形态,在空白组、模型组和参芪复方组中每组随机选取4只大鼠进行全转录组测序,构建mRNA、lncRNA差异表达谱,分析差异基因生物学功能,并进行RT-qPCR验证。结果与空白组相比,糖尿病GK大鼠FBG显著升高(P<0.05),模型组大鼠肝脏可见肝细胞排列紊乱,边界模糊,出现弥漫空泡状改变,呈中度脂肪变性,伴见炎性浸润、弥漫水肿;与模型组相比,参芪复方组大鼠FBG在12周时明显降低(P<0.05),肝脏组织排列较整齐,脂质沉积、细胞水肿及炎细胞浸润明显减轻。全转录组结果显示,与空白组相比,模型组大鼠mRNA、lncRNA表达谱存在显著差异,决定了糖尿病大鼠生理病理上的差异表现。参芪复方可广泛调控mRNA、lncRNA的差异表达,富集分析显示参芪复方通过调控多条内分泌系统及代谢过程相关信号通路改善糖尿病,包括胰岛素分泌、非酒精性脂肪性肝病、胆固醇代谢、鞘脂代谢、胰岛素抵抗等信号通路。RT-qPCR结果显示,基因Irf1、Trim30、Tmcc3、Insig1与转录组结果一致,转录组准确性较高。结论参芪复方发挥调节血糖及改善肝脏脂质沉积、水肿的作用,可能与广泛调控mRNA、lncRNA的差异表达有关。

mRNA疫苗制备技术在甲型流感病毒疫苗研究中的应用

禽流感病毒(Avian influenza virus,AIV)、H1N1和H3N2亚型人流感病毒(Influenza virus,IV)均属于甲型流感病毒(Influenza A virus,IAV)。IAV含有多种亚型,毒株变异快,因此IAV疫苗株需要根据当年的优势毒株进行相应的调整,这给疫苗的研制和生产带来巨大挑战。信使核糖核酸(messenger RNA,mRNA)疫苗制备工艺简单、生产时间短且容易规模化生产,在疾病预防方面具有广阔的应用前景。笔者对mRNA疫苗的种类、优化策略和IAV mRNA疫苗的研究现状进行了综述,旨在为IAV mRNA疫苗的研发提供理论参考。

弱精子症患者精子lncRNA与mRNA基因表达特征分析

目的:通过DNBSEQ平台测序技术构建文库,并以此来确定人类正常与弱精子症精子中lncRNA和mRNA的表达差异,并通过生物信息学方法分析其在弱精子症中的生物学意义。方法:将9份正常精液样本及9份弱精子症精液样本分组后,分离精子,提取总RNA,通过DNBSEQ测序平台检测得到RNA-seq在精子中的表达情况,通过GO富集分析和KEGG通路分析,进一步分析其相关功能情况。结果:使用DNBSEQ平台检测,每组平均产出10.64G数据,共检测到282185个RNA,包含有107009个lncRNA,其中15157个lncRNA具有表达差异,2190个lncRNA表达上调,12967个表达下调;共检测到19514个mRNA,其中13736个mRNA具有表达差异,4995个mRNA表达上调,8741个mRNA表达下调。差异基因主要富集在精子细胞膜、离子通道的功能以及精子发育、受精相关的通路上。结论:通过对弱精子症与正常精子的测序分析,鉴别出差异表达的lncRNA和mRNA,可通过胞膜离子通道对精子功能进行调控,进而导致弱精子症的发生,可为弱精子症的进一步研究提供分子基础。

POCD小鼠海马组织中差异表达miRNA的筛选、靶基因预测和生物信息学分析及其调控机制探讨

目的采用生物信息学方法分析术后认知功能障碍(POCD)小鼠海马组织中差异表达miRNA(DEMs)及其靶基因(DEGs),并构建POCD小鼠海马组织中差异表达miRNA-mRNA的调控网络图,探讨其机制。方法在GEO数据库中搜索下载POCD小鼠海马组织与正常小鼠海马组织基因表达谱芯片数据GSE95070,采用R软件lim⁃ma函数包筛选出差异表达的miRNA,即DEMs。将DEMs分别输入到miRTarBase、TargetScan、miRDB数据库中,三个数据库中皆存在的靶基因为差异表达的DEMs靶基因,即DEGs。使用R软件中的ggplot2、enrichplot函数包对DEGs进行了基因本体(GO)功能富集分析和KEGG信号通路分析。通过STRING在线工具构建DEGs蛋白互作网络(PPI),将蛋白间相互作用得分>0.4的节点输入到可视化工具Cytoscape软件中,利用cytoHubba插件筛选出节点度值排名前10的节点,即为可能参与小鼠POCD发生发展的枢纽基因。选取枢纽基因及其相应的DEMs,利用Cyto⁃scape软件构建POCD小鼠海马组织中差异表达miRNA-mRNA调控网络图。结果共筛选出19个显著差异表达的DEMs,得到448个DEGs。GO功能富集分析结果显示,DEGs在DNA结合的转录因子激活活性、特异性RNA聚合酶Ⅱ、微小GTP酶结合等分子功能中显著富集,在树突的形成以及调节、促进神经胶质细胞的增殖等生物学过程中显著富集,在突触膜的有机组成成分、细胞边缘以及囊泡运输等细胞组分中显著富集。KEGG信号通路分析结果显示,DEGs在Cushing综合征相关通路、cGMP-PKG信号通路、Hepatitis C信号通路、Apelin信号通路等通路中显著富集。小鼠POCD发生发展的10个枢纽基因分别为Gsk3β、Igf1、Cd34、Ubxn7、Smad2、Smarca4、Stat1、Grb2、Sin3a、Ncam1,相应的6个DEMs分别为mmu-miR-362-3p、mmu-miR-3065-5p、mmu-miR-592-3p、mmu-miR-28a-3p、mmu-miR-181c-5p、mmu-miR-351-5p,成功构建了POCD小鼠海马组织中差异表达miRNA-mRNA调控网络图,其中mmu-miR-362-3p调控Gsk3β的关系对、mmu-miR-3065-5p调控Igf1的关系对在调控网络中节点度最高。结论与正常小鼠海马组织相比,POCD小鼠海马组织中有19个差异表达的DEMs和448个DEGs;DEGs与树突的形成以及调节等有关,参与介导Cushing综合征相关通路、cGMP-PKG信号通路等。mmu-miR-362-3p、mmu-miR-3065-5p等差异表达的miRNA可能是通过调节Gsk3β、Igf1等靶基因的表达,影响POCD的发生发展。

基于RNA-seq不明原因复发性流产绒毛组织mRNA基因差异性表达研究

目的:构建不明原因复发性流产(unexplained recurrent spontaneous abortion,URSA)孕妇绒毛组织的mRNA差异性表达谱并进行功能分析,为阐明URSA的发生发展机制提供依据。方法:收集2022年3月于新疆医科大学第一附属医院妇科门诊收治的4例URSA(试验组)和4例选择性终止妊娠孕妇(对照组)的绒毛组织样本。应用转录组测序技术(RNA-seq)筛选2组差异表达基因(differentially expressed genes,DEGs),并对其进行GO功能注释、KEGG通路分析及蛋白质-蛋白质相互作用(PPI)网络分析,进一步研究差异表达mRNA的功能,并筛选出URSA的关键基因。结果:RNA-seq分析共筛选出3350个DEGs,其中上调基因2259个,下调基因1091个。通过PPI网络分析,筛选出与URSA有关的前10个关键基因:TYROBP、CD74、GH1、GH2、STAT5A、HLA-DRA、STAT5B、HLA-DRB1、CISH和HLA-A。通过GO和KEGG生物学功能分析,发现差异性表达的mRNA参与到多种白细胞分化相关免疫反应通路和同种异体免疫应答等生物学过程。结论:利用RNA-seq获得的URSA的mRNA差异性表达谱,筛选出可能与URSA有关的关键基因及生物学通路,为进一步研究URSA的发病机制和诊疗靶点提供了理论依据。

治疗遗传病的RNA药物研究进展

RNA药物能够通过识别互补序列靶向对应基因以抑制特定蛋白或RNA的表达,或通过翻译合成目的基因编码的蛋白来发挥遗传性疾病治疗作用。RNA药物主要分为寡核苷酸药物(包括反义寡核苷酸、小干扰RNA和RNA适配体)和信使RNA药物等。其中,反义寡核苷酸和小干扰RNA已用于临床治疗遗传病,而RNA适配体和信使RNA药物目前还处于临床试验阶段。当前主要通过对RNA药物进行化学修饰(如对信使RNA进行假尿嘧啶修饰)来降低免疫原性和提升药物疗效,以及开发纳米粒载体、细胞外囊泡和类病毒载体等递送载体来解决RNA药物的稳定性、特异靶向性和安全性等问题。本文概述了目前用于治疗遗传病的11种RNA药物的具体作用分子机制,并简单讨论了RNA药物的化学修饰及递送载体的研究现状。

妊娠期高血压病患者血清DNMT1 mRNA和LncRNA UCA1水平表达与妊娠结局的关系

目的分析妊娠期高血压疾病(hypertensive disorder complicating pregnancy,HDCP)患者血清DNA甲基转移酶1(DNA methyltransferase 1,DNMT1)信使RNA(messenger RNA,mRNA)、长链非编码RNA(long non-codingRNA,LncRNA)尿路上皮癌胚抗原1(urothelial carcinoembryonic antigen 1,UCA1)水平与妊娠结局的关系。方法选取2021年3月~2023年8月在邯郸市妇幼保健院诊治的HDCP患者195例为病例组、健康妊娠孕妇195例为对照组。收集所有孕妇临床资料并检测分娩前1天生化指标;荧光定量PCR法检测血清DNMT1 mRNA,LncRNA UCA1水平;根据病情将病例组分为妊娠期高血压(pregnancy induced hypertension,PIH)组、轻度子痫前期(preeclampsia,PE)组、重度PE组;根据HDCP患者分娩时不良妊娠结局情况分为妊娠结局良好组和妊娠结局不良组;比较对照组和病例组临床资料和生化指标、血清DNMT1 mRNA,LncRNA UCA1水平;比较不同严重程度HDCP患者血清DNMT1mRNA和LncRNA UCA1水平;比较不同妊娠结局HDCP患者临床资料和生化指标、血清DNMT1 mRNA和LncRNAUCA1水平;分析HDCP患者血清DNMT1 mRNA与LncRNA UCA1的相关性,影响HDCP患者妊娠结局的因素,血清DNMT1 mRNA和LncRNA UCA1对HDCP患者发生不良妊娠结局的预测价值。结果与对照组比较,病例组收缩压、舒张压、白细胞计数水平明显升高,血清DNMT1 mRNA(0.72±0.18 vs 1.00±0.04),LncRNA UCA1(0.61±0.16vs 1.00±0.02)水平明显降低,差异具有统计学意义(t=40.651,32.595,7.837,21.205,33.775,均P<0.001);PIH组、轻度PE组、重度PE组血清DNMT1 mRNA(0.85±0.20,0.74±0.18,0.50±0.15),LncRNA UCA1(0.77±0.18,0.58±0.16,0.43±0.13)水平依次降低,差异具有统计学意义(F=52.687,64.030,均P<0.001);HDCP患者血清DNMT1 mRNA与LncRNA UCA1呈正相关(r=0.582,P<0.001);与妊娠结局良好组比较,妊娠结局不良组HDCP严重程度较高,收缩压、舒张压、白细胞计数水平明显升高,血清DNMT1 mRNA(0.80±0.20 vs 0.59±0.15),LncRNA UCA1(0.72±0.17 vs 0.43±0.14)水平明显降低,差异具有统计学意义(χ^(2)=18.386,t=2.615~12.290,均P<0.05);重度PE[OR(95%CI)=1.708(1.193~2.445)]、收缩压[OR(95%CI)=1.495(1.090~2.049)]、舒张压[OR(95%CI)=1.621(1.076~2.442)]是影响HDCP患者发生不良妊娠结局的危险因素,DNMT1 mRNA[OR(95%CI)=0.833(0.725~0.957)],LncRNA UCA1[OR(95%CI)=0.796(0.696~0.909)]是影响HDCP患者发生不良妊娠结局的保护因素(均P<0.05);DNMT1 mRNA和LncRNA UCA1二者联合预测HDCP患者发生不良妊娠结局的曲线下面积(area under curve,AUC)大于DNMT1 mRNA及LncRNA UCA1单独预测的AUC(0.926 vs 0.832,0.844),差异具有统计学意义(Z=2.932,2.345,均P<0.05)。结论HDCP患者血清DNMT1 mRNA和LncRNA UCA1水平均较低,与病情程度、妊娠结局相关,DNMT1 mRNA联合LncRNA UCA1检测对不良妊娠结局有较佳预测效能。

长链非编码RNA-N1LR在脑缺血再灌注血脑屏障损伤的作用机制研究

目的探讨长链非编码RNA(lncRNA)-N1LR在脑缺血再灌注损伤后血脑屏障的作用机制。方法原代小鼠脑微血管内皮细胞常规培养,经氧糖剥夺/复糖复氧(OGD/R)处理模拟脑缺血再灌注损伤,实验分对照组、OGD组、lncRNA-N1LR过表达组(OGD处理后转染质粒lncRNA-N1LR过表达)、lncRNA-N1LR沉默组(OGD处理后转染质粒lncRNA-N1LR沉默)。采用逆转录聚合酶链反应检测各组中lncRNA-N1LR mRNA、紧密连接蛋白5(claudin-5)及闭合蛋白(occludin)mRNA表达水平;异硫氰酸荧光素-葡聚糖(FITC-dextran)渗透法检测血脑屏障通透性;免疫蛋白印迹法检测claudin-5、occludin蛋白表达。结果与对照组比较,OGD组lncRNA-N1LR mRNA、occludin、claudin-5 mRNA表达水平下降(0.31±0.01 vs 1.00±0.10,0.42±0.03 vs 1.01±0.13,0.38±0.03 vs 1.00±0.15,P<0.05),血脑屏障FITC-dextran通透性明显升高(58.79±3.04 vs 8.87±0.63,P<0.01)。与OGD组比较,lncRNA-N1LR过表达组lncRNA-N1LR mRNA、occludin、claudin-5 mRNA表达水平升高(0.67±0.07 vs 0.31±0.01,0.92±0.02 vs 0.42±0.03,0.70±0.08 vs 0.38±0.03,P<0.05),血脑屏障FITC-dextran通透性降低(41.57±2.43 vs 58.79±3.04,P<0.05)。与OGD组比较,lncRNA-N1LR沉默组lncRNA-N1LR mRNA、occludin、claudin-5 mRNA表达水平降低(0.21±0.02 vs 0.31±0.01,0.31±0.03 vs 0.42±0.03,0.22±0.02 vs 0.38±0.03,P<0.05),血脑屏障FITC-dextran通透性升高(72.34±1.43 vs 58.79±3.04,P<0.05)。结论LncRNA-N1LR上调可能通过降低血脑屏障通透性发挥神经保护作用。

COPD患者lncRNA MIR155HG表达与肺功能的相关性及其对AECODP的辅助诊断价值

目的 探讨慢性阻塞性肺疾病(COPD)患者长链非编码RNA(lnc RNA) MIR155HG与肺功能的相关性,及其对慢性阻塞性肺疾病急性加重(AECOPD)的辅助诊断价值。方法 选取2018年10月—2021年3月南阳市中心医院COPD患者117例,其中稳定期患者60例(稳定期组)、AECOPD患者57例(AECOPD组),以60名健康体检者作为正常对照组。收集所有研究对象一般资料,并检测肺功能[第1秒用力呼气容积(FEV1)、用力肺活量(FVC),计算FEV1/FVC%和外周血单个核细胞(PBMC)中lnc RNAMIR155HG的表达。采用Logistic回归分析评估AECOPD的危险因素。采用受试者工作特征(ROC)曲线评价各项指标诊断AECOPD的效能。结果 AECOPD组、稳定期组、正常对照组白细胞(WBC)计数、中性粒细胞百分比(NEUT%)、降钙素原(PCT)、C反应蛋白(CRP)、纤维蛋白原(Fib)、PBMC中lnc RNA MIR155HG相对表达量依次降低(P<0.05),FEV1和FEV1/FVC%依次升高(P<0.05)。稳定期组和AECOPD组PBMC中lnc RNA MIR155HG相对表达量与WBC计数、NEUT%、PCT、CRP和Fib水平均呈正相关(P<0.001),与FEV1和FEV1/FVC%均呈负相关(P<0.001)。Logistic回归分析结果显示,lnc RNAMIR155HG表达升高、FEV1降低是发生AECOPD的危险因素[比值比(OR)值分别为2.381、0.682,95%可信区间(CI)分别为1.526~3.715、0.531~0.876]。ROC曲线分析结果显示,FEV1、lnc RNA MIR155HG单项和联合检测诊断AECOPD的曲线下面积(AUC)分别为0.826、0.854、0.939。结论 COPD患者PBMC中lnc RNA MIR155HG水平升高,且与炎症指标和肺功能相关,或可作为AECOPD的早期诊断标志物。