LncRNA NEAT1 promotes proliferation of ovarian cancer cells and angiogenesis of co-incubated human umbilical vein endothelial cells by regulating FGF9 through sponging miR-365: An experimental study

Objective:To uncover the function of lncRNA NEAT1 in ovarian cancer (OC) cells and its mechanism.Methods:The expression patterns of lncRNA NEAT1 and FGF9 in human OC cells and human ovarian epithelial cells was determined. OC cells were transfected with sh-NEAT1, pcDNA3.1-NEAT1, miR-365 mimic, miR-365 inhibitor or pcDNA3.1-NEAT1 + sh-NEAT1 before cell proliferation rate and cell clone formation rate were measured. After the transfected OC cells were co-cultivated with human umbilical vein endothelial cells (HUVECs), Matrigel angiogenesis assay tested angiogenesis of HUVECs; qRT-PCR and Western blot tested the expressions of vascular endothelial growth factor (VEGF), angiogenin 1 (Ang-1) and matrix metalloproteinase 2 (MMP2). Dual-luciferase reporter assay determined the targeted binding of NEAT1 and FGF9 to miR-365.Results:LncRNA NEAT1 and FGF9 are over-expressed in OC cells. Knockdown of NEAT1 or FGF9, or over-expression of miR-365 results in decreased proliferation rate and cell clones as well as inhibited angiogenesis and down-regulated expressions of VEGF, Ang-1 and MMP2. Over-expression of NEAT1 or knockdown of miR-365 can reverse the effect caused by FGF9 knockdown. NEAT1 can down-regulate the expression of miR-365 while up-regulating that of FGF9. Dual-luciferase reporter assay determined that NEAT1 competes with FGF9 for binding to miR-365.Conclusion:LncRNA NEAT1 up-regulates FGF9 by sponging miR-365, thus promoting OC cell proliferation and angiogenesis of HUVECs.     

药物性肝损伤患者血清miR-21和miR-124a水平变化及其临床意义探讨

目的 探讨药物性肝损伤(DILI)患者血清微小RNA(miR)-21和miR-124a水平变化及其临床意义。方法 2019年3月～2022年3月我院收治的87例DILI患者和60例同期体检的健康人,采用实时荧光定量PCR法检测血清miR21和miR-124a mRNA水平,采用ELISA法检测核转录因子-κB(NF-κB)和白介素-6(IL-6)水平。结果 在DILI发生时,血清ALT、AST、GGT和TBIL峰值水平分别为(143.6±51.8)U/L、(158.2±38.8)U/L、(131.6±26.8)U/L和(41.9±9.6)μmol/L;DILI患者血清miR-21、miR124a mRNA、NF-κB和IL-6水平分别为(1.4±0.3)、(2.6±0.4)、(3615.4±526.4)pg/ml和(12.7±1.8)pg/ml,均显著高于健康人【分别为(1.0±0.1)、(1.1±0.1)、(692.2±144.6)pg/ml和(3.4±0.7)pg/ml,P<0.05】;混合型DILI患者血清NF-κB和IL-6水平分别为(3874.5±282.5)pg/ml...     

NEAT1和miR-206与急性冠脉综合征患者PCI术后心肌损伤及预后的相关性分析

目的 探究血清核富集转录体1（NEAT1）、微小RNA-206（miR-206）对急性冠脉综合征患者经皮冠状动脉介入（PCI）术后心肌损伤及心血管不良事件（MACE）的预测价值。方法 选取2017年1月-2020年1月在本院行PCI手术的急性冠脉综合征患者102例为研究对象,根据治疗6个月后是否发生心肌损伤及术后随访12个月是否发生MACE,分为心肌损伤组（32例）和心肌未损伤组（70例）,MACE组（24例）和非MACE组（78例）。采用实时荧光定量PCR法检测血清中miR-206、NEAT1水平,采用Pearson法分析miR-206水平与NEAT1水平的相关性,采用受试者工作特征（ROC）曲线评价血清NEAT1、miR-206预测急性冠脉综合征患者PCI术后心肌损伤及MACE的价值。结果 与心肌未损伤组相比,心肌损伤组血清NEAT1水平较高（P＜0.05）,血清miR-206水平较低（P＜0.05）;与非MACE组相比,MACE组术后肌钙蛋白I（cTnI）、术后肌酸激酶同工酶（CK-MB）、血清NEAT1水平较高（P＜0.05）,血清miR-206水平较低（P＜0.05）;ROC曲线显示,血清miR-206水平预测急性冠脉综合征患者PCI术后心肌损伤及MACE的曲线下面积（AUC）分别为0.813、0.824,血清NEAT1水平预测急性冠脉综合征患者PCI术后心肌损伤及MACE的AUC分别为0.851、0.876,二者联合预测急性冠脉综合征患者PCI术后心肌损伤的AUC为0.908,二者联合预测急性冠脉综合征患者PCI术后MACE的AUC为0.926;心肌损伤组及MACE组患者血清miR-206水平与NEAT1水平均呈负相关（P＜0.05）。结论 急性冠脉综合征PCI术后心肌损伤患者血清NEAT1表达上调,血清miR-206表达下调,miR-206、NEAT1可预估急性冠脉综合征患者PCI术后心肌损伤及MACE,且二者联合预测价值较高。     

支气管哮喘患儿血清miR-34、Notch1水平与气道炎症的相关性分析

目的探究微小RNA-34(miR-34)、Notch1在支气管哮喘(BA)患儿血清中的表达水平及其与气道炎症的关系。方法选取2018年10月~2020年5月本院收治的94例BA患儿为BA组,并选取同期90例体检健康儿童为健康组。比较两组一般资料;采用实时荧光定量PCR(qRT-PCR)法检测血清miR-34、Notch1 mRNA表达水平;酶联免疫吸附法(ELISA)检测血清白介素-17(IL-17)、白介素-10(IL-10)水平;采用血细胞分析仪检测嗜酸性粒细胞(EOS)水平;Pearson法分析BA患儿血清miR-34、Notch1 mRNA表达水平与IL-17、IL-10、EOS,及miR-34表达水平与Notch1 mRNA的相关性;Logistic回归分析发生BA的影响因素。结果BA组患儿血清miR-34及IL-10水平,均明显低于健康组(P<0.05),Notch1 mRNA、IL-17、EOS水平、家族哮喘史、个人过敏史、家族过敏史均明显高于健康组(P<0.05);BA患儿血清miR-34表达水平与IL-17、EOS、Notch1 mRNA呈负相关(P<0.05),与IL-10呈正相关(P<0.05);血清Notch1 mRNA表达水平与IL-17、EOS呈正相关(P<0.05),与IL-10呈负相关(P<0.05);miR-34是影响BA发生的独立保护因素(P<0.05),Notch1、IL-17、家族哮喘史、个人过敏史、家族过敏史是影响BA发生的独立危险因素(P<0.05)。结论BA患儿血清miR-34表达水平降低,Notch1表达水平升高,两者呈负相关,且均与气道炎症显著相关,检测血清miR-34、Notch1表达水平,均有助于辅助诊断BA,两者联合可提高对BA的诊断价值。     

支气管哮喘患儿血清miR-34、Notch1水平与气道炎症的相关性分析

目的探究微小RNA-34(miR-34)、Notch1在支气管哮喘(BA)患儿血清中的表达水平及其与气道炎症的关系。方法选取2018年10月~2020年5月本院收治的94例BA患儿为BA组,并选取同期90例体检健康儿童为健康组。比较两组一般资料;采用实时荧光定量PCR(qRT-PCR)法检测血清miR-34、Notch1 mRNA表达水平;酶联免疫吸附法(ELISA)检测血清白介素-17(IL-17)、白介素-10(IL-10)水平;采用血细胞分析仪检测嗜酸性粒细胞(EOS)水平;Pearson法分析BA患儿血清miR-34、Notch1 mRNA表达水平与IL-17、IL-10、EOS,及miR-34表达水平与Notch1 mRNA的相关性;Logistic回归分析发生BA的影响因素。结果BA组患儿血清miR-34及IL-10水平,均明显低于健康组(P<0.05),Notch1 mRNA、IL-17、EOS水平、家族哮喘史、个人过敏史、家族过敏史均明显高于健康组(P<0.05);BA患儿血清miR-34表达水平与IL-17、EOS、Notch1 mRNA呈负相关(P<0.05),与IL-10呈正相关(P<0.05);血清Notch1 mRNA表达水平与IL-17、EOS呈正相关(P<0.05),与IL-10呈负相关(P<0.05);miR-34是影响BA发生的独立保护因素(P<0.05),Notch1、IL-17、家族哮喘史、个人过敏史、家族过敏史是影响BA发生的独立危险因素(P<0.05)。结论BA患儿血清miR-34表达水平降低,Notch1表达水平升高,两者呈负相关,且均与气道炎症显著相关,检测血清miR-34、Notch1表达水平,均有助于辅助诊断BA,两者联合可提高对BA的诊断价值。     

Long Noncoding RNA Antisense Noncoding RNA in the INK4 Locus Correlates With Risk,Severity, Inflammation and Infliximab Efficacy in Crohn's Disease

Background

The aim of this study was to investigate the association of intestinal mucosa long noncoding RNA (lncRNA) antisense noncoding RNA in the INK4 locus (ANRIL) expression with disease risk, activity and inflammatory cytokines levels of Crohn's disease (CD).

miR-126-5p expression in the plasma of patients with sepsis-induced acute lung injury and its correlation with inflammation and immune function

Objective

This work was implemented to elucidate the miR-126-5p expression in the plasma of patients with sepsis-induced acute lung injury (ALI) and its correlation with inflammation and immune function.

Methods

The peripheral blood of patients with sepsis-induced ALI was obtained, and the levels of inflammatory factors (interleukin-6 [IL-6], C-reactive protein [CRP], and procalcitonin [PCT]) were determined. Meanwhile, T lymphocyte subsets (CD3+, CD4+, and CD8+), and immunoglobulins (IgA, IgM, and IgG) were tested. miR-126-5p and TRAF6 mRNA expression in plasma was assessed. Receiver operating characteristic (ROC) curve was performed to assess the diagnostic accuracy of miR-126-5p in sepsis without ALI and sepsis with ALI. Correlation between miR-126-5p expression and clinical indicators was analyzed. The targets of miR-126-5p were predicted using the bioinformatics method, and the direct targets were verified through investigations.

Results

miR-126-5p expression in plasma of patients with sepsis-induced ALI was reduced than that of patients with sepsis without ALI. miR-126-5p expression was negatively correlated with IL-6, CRP, and PCT but positively correlated with IgA, IgM, and IgG as well as CD3+, CD4+, and CD8+ in patients with sepsis-induced ALI. ROC curve suggested that miR-126-5p (AUC: 0.777; 95%CI: 0.689–0.866) could distinguish patients with sepsis with ALI from patients with sepsis without ALI. TRAF6 expression in patients with sepsis-induced ALI was higher than that in patients with sepsis without ALI. TRAF6 was a target gene of miR-126-5p,

Conclusion

This research highlights that miR-126-5p is reduced in the plasma of patients with sepsis-induced ALI, and miR-126-5p relates to systemic inflammation and immune function indicators.     

miR-150及其靶基因IRAK2在变应性鼻炎小鼠模型中的表达

目的探讨miR-150及其靶基因IRAK2在变应性鼻炎小鼠鼻黏膜中的表达及其作用。方法SPF级Balb/c小鼠20只,随机分为两组,变应性鼻炎模型组(AR)、对照组(NC),每组10只。模型组用卵清蛋白(OVA)致敏建立变应性鼻炎小鼠模型;对照组使用生理盐水替代。每组随机取4只小鼠,病理检查;每组剩余6只小鼠取其鼻黏膜,实时定量PCR检测小鼠鼻黏膜中miR-150及IRAK2 mRNA表达水平,蛋白印迹法(Western blot)检测IRAK2蛋白的表达,采用酶联免疫吸附法(ELISA)测定血清中OVA特异性IgE、IL-4、IL-13的含量,模型组与对照组miR-150相对表达量与IRAK2 mRNA的相对表达量、OVA特异性IgE、IL-4、IL-13的相关性分析采用Pearson相关性分析。结果AR组小鼠动物行为学评分均大于5分;病理组织HE染色显示AR组鼻黏膜纤毛大量脱落,组织间质水肿、小血管扩张、腺体增生、炎性细胞浸润;血清OVA特异性IgE、IL-4、IL-13表达增加。AR组小鼠鼻黏膜miR-150表达较NC组降低(P<0.05);IRAK2 mRNA表达水平明显高于对照组(P<0.05);且AR组鼻黏膜IRAK2蛋白表达水平也比NC组增加(P<0.05)。AR组与NC组miR-150相对表达量与IRAK2 mRNA表达量和OVA特异性IgE、IL-4、IL-13的含量呈负相关(r=-0.841、-0.869、-0.834、-0.857,P<0.05)。结论miR-150在变应性鼻炎模型小鼠中表达降低且与IRAK2、OVA特异性IgE、IL-4、IL-13含量存在负相关,二者的表达差异可能在变应性鼻炎的发生发展中发挥作用。     

Inhibitory effect of miR-125b on hepatitis C virus core protein-induced TLR2/My D88 signaling in THP-1 cells

AIM: To investigate the role of mi R-125 b in regulating monocyte immune responses induced by hepatitis C virus(HCV) core protein.METHODS: Monocytic THP-1 cells were treated with various concentrations of recombinant HCV core protein, and cytokines and mi R-125 b expression in these cells were analyzed. The requirement of Tolllike receptor 2(TLR2) or My D88 gene for HCV core protein-induced immune responses was determined by the transfection of THP-1 cells with gene knockdown vectors expressing either TLR2 si RNA or My D88 si RNA. The effect of mi R-125 b overexpression on TLR2/My D88 signaling was examined by transfecting THP-1 cells with mi R-125 b mimic RNA oligos.RESULTS: In response to HCV core protein stimulation, cytokine production was up-regulated and mi R-125 b expression was down-regulated in THP-1 cells. The modulatory effect of HCV core protein on cellular events was dose-dependent and required functional TLR2 or My D88 gene. Forced mi R-125 b expression abolished the HCV core protein-induced enhancement of tumor necrosis factor-α, interleukin(IL)-6, and IL-10 expression by 66%, 54%, and 66%, respectively(P 0.001), by inhibiting My D88-mediated signaling, including phosphorylation of NF-k Bp65, ERK, and P38.CONCLUSION: The inverse correlation between mi R-125 b and cytokine expression after HCV core challenge suggests that mi R-125 b may negatively regulate HCVinduced immune responses by targeting TLR2/My D88 signaling in monocytes.     

LncRNA CYTOR promotes pancreatic cancer cell proliferation and migration by sponging miR-205-5p

Background/aimsStudies have found that LncRNA CYTOR is an important regulator of cancer. However, the function of lncRNA CYTOR in pancreatic cancer (PC) is unclear. This study amid to explore the regulation of lncRNA CYTOR in PC.MethodsThe expression of CYTOR and miR-205-5p in PC was detected by RT-qPCR. CCK-8 assay, colony formation assay and scratch test were conducted to detect the effects of CYTOR and miR-205-5p on proliferation and migration of PC cells. Target gene prediction and screening and luciferase reporter assays were used to verify downstream target genes of CYTOR and miR-205-5p. The expression of Cyclin-dependent protein kinase 6 (CDK6) was detected by Western blotting. The tumor growth in mice was detected by in vivo experiments in nude mice.ResultsThe expression of LncRNA CYTOR was significantly elevated in PC. Knockdown of CYTOR significantly inhibited cell proliferation and migration of PC cells. In vivo animal studies showed that CYTOR promoted tumor growth. MiR-205-5p was a direct target of CYTOR, and the expression levels of miR-205-5p were significantly reduced in PC cell lines. Furthermore, co-transfection of shCYTOR with miR-205-5p inhibitor partially abolished the effect of shCYTOR on cell proliferation and migration. In addition, CYTOR was negatively correlated with the expression of miR-205-5p. CDK6 was a direct target of miR-205-5p, and miR-205-5p mimic and sh CYTOR significantly reduced the expression levels of CDK6.ConclusionCYTOR can promote PC progression by modulating the miR-205-5p/CDK6 axis, which may be a potential therapeutic target for PC.     

Serum Levels of IL-10 and IL-22 Cytokines in Patients with Psoriasis

Background: As a chronic inflammatory condition, psoriasis results from an interaction between genetic and immunologic factors in a predisposing environment. In spite of compelling evidence for the role of T cells and cytokines in psoriasis, interleukin (IL)- 10 and IL-22 have not been sufficiently investigated. Objective: To assess the serum levels of IL-10 and IL-22 in patients with psoriasis compared to healthy controls. Methods: A total of 28 patients with psoriasis were compared with 28 age and sexmatched healthy subjects. Psoriasis Area and Severity Index (PASI) criteria were used to measure the severity of the disease. Serum levels of IL-10 and IL-22 were measured in both groups and compared. Results: The mean serum level of IL-10 was 89.5±18.7 in patients compared to 117.2±23.4 pg/ml in the controls (p=0.36). Also, serum level of IL-22 was 284.1±49.7 in patients versus 425.4±82.8 pg/ml in control group (p=0.17). There was a significant direct correlation between levels of IL-10 and IL-22 in patients group (p=0.0005). The clinical severity of psoriasis was significantly correlated with high levels of IL-22 (p<0.0001).Conclusions: The decreased levels of IL-10 in psoriatic patients and direct correlation between higher levels of IL-22 and disease severity support the clinical implication of both cytokines in psoriasis.     

敲除miR-223促进血管炎症和动脉粥样硬化的发生

目的研究miR-223对血管炎症和动脉粥样硬化的影响,为临床动脉硬化性疾病提供新的诊疗方向。方法miR-223敲除鼠与ApoE敲除鼠(ApoE KO)繁殖制备miR-223/ApoE双敲鼠(miR-223/ApoE DKO);检测小鼠血浆脂质水平;处死取材后检测主动脉根部及血管全长的斑块含量;通过免疫组化检测斑块的炎症细胞浸润;转录组学测序分析血管中炎症相关基因的表达;结合microRNA靶基因数据库寻找并验证其可能的靶基因。结果miR-223/ApoE双敲鼠主动脉根部及血管全长斑块量显著增加(P<0.05)。免疫组化染色显示,主动脉根部炎症细胞浸润增加;血管转录组学测序发现炎症相关基因血管细胞黏附分子1(VCAM-1)、白细胞介素1α(IL-1α)等在双敲鼠中显著上调。通过靶基因数据库筛选,发现白细胞介素6(IL-6)是miR-223的靶基因并且在双敲鼠的血管中表达显著上调;使用miR-223模拟物刺激成纤维细胞,显著抑制了IL-6的表达。结论miR-223抑制靶基因IL-6的表达降低炎症反应,敲除miR-223显著升高血管炎症水平促进动脉粥样硬化的进展。     

lncRNA PACER促进脓毒症急性肺损伤炎症反应的实验研究

目的探讨lncRNA PACER对小鼠脓毒症急性肺损伤炎症反应的促进作用。 方法分离和培养急性肺损伤和健康非吸烟者的肺泡巨噬细胞,以及获取细菌脂多糖(LPS)诱导的急性肺损伤(ALI)小鼠肺组织,采用qRT-PCR方法检测lncRNA PACER的表达;过表达或敲低PACER后,ELISA方法检测THP-1和RAW264.7细胞炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的表达;对LPS所致ALI小鼠,尾静脉注射PACER siRNA慢病毒后,ELISA检测小鼠血清炎性因子TNF-α、IL-6的表达,HE染色观察肺组织病理学变化。 结果ALI患者肺泡巨噬细胞和ALI小鼠肺组织中lncRNA PACER表达均显著升高(P<0.01);细胞过表达PACER后,细胞炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的表达升高,而敲低PACER后,炎性因子TNF-α、IL-6的表达则降低(P<0.01);ALI小鼠敲低PACER后,小鼠肺组织和血清中炎性因子TNF-α、IL-6的表达均显著降低(P<0.01),肺组织损伤程度明显减弱。 结论lncRNA PACER可显著促进脓毒症急性肺损伤炎症反应,为明确lncRNA PACER作为ALI防治的靶标提供了依据。     

肺结核患者的Th1/Th2细胞因子失衡

目的　探讨肺结核患者是否存在Th1/Th2反应失衡 ,以及与病情严重程度和治疗反应的关系。方法　对 10 0名健康对照和 12 4例未经治疗的菌阳肺结核患者检测血清IgE、IL 4和IFN γ ,比较两组间差异并分析其与病情严重程度、治疗后痰菌阴转情况及初复治的关系。结果　肺结核组血清IgE(1.0 4 8± 0 .4 96 )、IL 4 (0 .4 39± 0 .16 0 )显著高于健康对照组 (分别为 0 .86 7± 0 .2 89和 0 .4 2 1±0 0 2 4 ) ,而血清IFN γ(0 .2 13± 0 .0 17)显著低于健康对照组 (0 .2 2 4± 0 .0 0 5 )。病灶范围大或有空洞形成的患者血清IL 4显著高于病灶范围小的患者 ,复治患者IL 4和IFN γ显著低于初治患者。抗结核治疗 2个月后痰菌未转阴组与痰菌阴转组相比 ,前者治疗前血清IL 4较高而IFN γ较低。结论　未经治疗的肺结核患者存在Th1反应减弱 ,Th2反应增强 ,其中病灶范围大或有空洞形成的患者更为显著。而且Th1反应较弱者治疗效果较差。