Serum miR-21 Research Articles

MiR-141 is negatively correlated with TLR4 in neonatal sepsis and regulates LPS-induced inflammatory responses in monocytes.

Neonatal sepsis is an inflammatory system syndrome and a main cause of neonatal mortality. However, there is a lack of ideal biomarkers for early neonatal sepsis diagnosis. The aim of this study was to evaluate the clinical significance of miR-141 in sepsis in neonates, and explore the regulatory effects of miR-141 on inflammation in monocytes. This study used qRT-PCR to calculate the expression of miR-141 in the serum of septic neonates. The diagnostic values of procalcitonin (PCT) and serum miR-141 were evaluated by receiver operating characteristic (ROC) curves. The relationship between miR-141 and TLR4 was determined using luciferase reporter assay. An inflammation model was established using monocytes with lipopolysaccharide (LPS) treatment. ELISA assay was used to analyze the levels of pro-inflammatory cytokines. The expression of miR-141 in neonatal sepsis was significantly lower than healthy controls. ROC curves showed that miR-141 had diagnostic accuracy. LPS stimulation in monocytes led to a decrease in the expression of miR-141. A luciferase reporter assay proved that miR-141 targeted TLR4, and a negative correlation of miR-141 with TLR4 was found in septic neonates. ELISA results demonstrated that the overexpression of miR-141 inhibited LPS-induced inflammation in monocytes. In conclusion, serum decreased miR-141 expression served as a candidate diagnostic biomarker of neonatal sepsis. TLR4 is a target gene of miR-141, which may mediate the inhibitory effects of miR-141 overexpression on LPS-induced inflammation in monocytes. Therefore, miR-141 is expected to be a potential diagnostic biomarker and a therapeutic target in neonatal sepsis.

MicroRNA-155 influences cell damage in ischemic stroke via TLR4/MYD88 signaling pathway

ABSTRACT Cerebral ischemic stroke (CIS) is extremely harmful, and its treatment should be underpinned by understanding its pathogenic mechanism. This study was designed to determine the involvement of miR-155 in CIS development via the TLR4/MyD88 signaling pathway. First, we quantified serum miR-155 in patients with CIS and healthy individuals, and found high expression of miR-155 in such patients and a decrease in it in the patients after therapy (P < 0.05). Serum miR-155 demonstrated a favorable function in predicting the development and prognosis of CIS (P < 0.001). We also conducted a mouse assay, and found that knocking out miR-155 can improve the neurological function of mice and suppress protein TLR4 and MyD88 (all P < 0.05). Finally, we carried out a cell assay, and found enhancement in the activity of SH-SY5Y cells, decrease in their apoptosis, and protein TLR4 and MyD88 in them after suppression of miR-155 (all P < 0.05). Furthermore, we also found complete reverse by TLR4/MyD88 pathway inhibitor on the influence of increasing miR-155 on cells (P > 0.05). Therefore, with an increase in cases with CIS, miR-155 takes a part in the development of cell damage by activating TLR4/MyD88, and it is probably the key to diagnosing and treating CIS.

MiR-124 Is Downregulated in Serum of Acute Cerebral Infarct Patients and Shows Diagnostic and Prognostic Value

Acute cerebral infarct (ACI) is a severe subtype of ischemic stroke. microRNAs (miRNAs) are implicated in the pathogenesis of ACI. This study investigated the expression pattern and clinical implication of miR-124 in ACI patients. Serum samples were collected from 108 healthy people and 108 ACI patients at 24 h, 48 h, and 72 h. Serum miR-124 expression was tested using qRT-PCR. The levels of interleukin (IL)-6, IL-8, and C-reactive protein (CRP) were detected using ELISA kits. The correlations between miR-124 expression and infarct classification, infarct size, risk factors, and inflammatory factors were analyzed. The diagnostic efficacy of miR-124 in ACI was analyzed by the ROC curve. ACI patients were assigned to the miR-124 high/low expression group and the incidence of poor prognosis was compared between the two groups. miR-124 expression was poorly expressed in the serum of ACI patients. The area under the ROC curve of miR-124 in the diagnosis of ACI was 0.9527, the specificity was 91.67%, and the sensitivity was 93.52%. miR-124 expression in ACI patients was not affected by infarct classification, infarct size, low-density lipoprotein level, and homocysteine level. miR-124 expression was negatively correlated with IL-6, IL-8, and CRP in ACI patients. Low expression of miR-124 was positively correlated with the poor prognosis of ACI. miR-124 was poorly expressed in the serum of ACI patients and served as a biomarker for the diagnosis and prognosis. This study shall confer a promising novel target for the diagnosis and treatment of ACI.

Sensitive detection of microRNA-21 in cancer cells and human serum with Au@Si nanocomposite and lateral flow assay

We report a highly sensitive approach for detecting microRNA-21 (miR-21) in cancer cells and human serum by using Au@Si nanocomposite labeled lateral flow assay. The Au@Si nanocomposite was prepared by coating numerous 3–5 nm gold nanoparticles (GNP) on a silica nanoparticle (SiNP) with a diameter of 150 nm and used as colored label on the lateral flow assay for signal amplification. TEM results show there are around 1000 GNPs coated on the SiNP surface. The principle of miR-21 detection is based on on-strip DNA-microRNA hybridization reactions to form DNA-miR-21-DNA-Au@Si complexes, which are captured on the test zone of the lateral flow test strip and produce a visible red band. A thiol-modified detecting DNA probe (Det-DNA) and a biotin-modified capturing DNA probe (Cap-DNA), which are complementary to miR-21, were used to prepare the lateral flow test strips. After systematic optimization, the method can detect a minimum concentration of 1.0 pM miR-21, which is 60 times lower than that of the GNP-based lateral flow assay (Gao et al. Biosens & Bioelectro, 2014, 54, 578–584). The method was applied to detect miR-21 in cancer cells and spiked human serum with satisfactory results.

At the border of inflammation and fibrosis - serum expression of miR 29 in patients with inflammatory bowel disease

Introduction: Recently, the expression of micro ribonucleic acids (microRNAs) in peripheral blood and tissues in patients with chronic inflammatory bowel disease (IBD) has been studied as a biomarker for diagnosis and monitoring of activity. Aim: The aim of this study is to evaluate the correlation between serum microribonucleic acid 29 (miR-29) expression and different clinical parameters: activity, severity and extent of disease in IBD patients. Materials and Methods: Serum miR-29 expression was tested in 70 patients with IBD (35 with ulcerative colitis (UC), 35 patients with Crohn's disease (CD)) and 30 healthy control subjects through reverse transcriptase and real-time quantitative PCR (RT-qPCR). Results: The group of patients showed a mean serum miR-29 expression of 1.69 for CD, 1.23 for UC and 1.07 for the control group with a significant difference in expression between the groups. Conclusion: There is a significant relation between increased serum miR-29 expression and disease activity, severity and extent in patients with UC.

The expression of miR-191 - a modern biomarker for distinguishing ulcerative colitis from Crohn's disease

Introduction: Literature data shows that miR-191 can be used as a potential diagnostic biomarker to differentiate between Crohn's disease (CD) and ulcerative colitis (UC). Aim: The aim of the present study is to evaluate the serum expression of miR-191 in patients with chronic inflammatory bowel disease and to determine the role of microRNAs in distinguishing CD from UC. Material s and Methods: Serum miR-191 expression was tested in 70 patients with inflammatory bowel disease (IBD) (35 with UC, 35 patients with CD) and 30 healthy control subjects through reverse transcriptase and real-time quantitative PCR (RT-qPCR). Results: The results show that miR-191 has increased expression in CD, active disease, small intestinal localization, and chronic persistent form of UC, and extraintestinal manifestations. The correlation analysis shows a positive correlation between the expression of miR-191 and faecal calprotectin and CDAI. Conclusion: The serum expression of miR-191 was significantly higher in patients with CD compared to patients with UC. It correlated with the localization and activity in CD, and in UC there was only a correlation to the form of the course and the extraintestinal manifestations. It can be used to distinguish between the two diseases.

Circulating microRNA-221 as a diagnostic biomarker for hepatitis C virus-related hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world and the third cause of cancer-related mortality as estimated by the World Health Organization (WHO). A major risk factor for HCC development; is Hepatitis C virus (HCV). According to WHO, approximately 55–85% of patients with HCV will develop chronic hepatitis, which lead to progressive fibrosis and cirrhosis. There are no biomarkers for early detection of HCC, and most patients are diagnosed at advanced stages, which is associated with poor prognosis. MicroRNAs have been reported as biomarkers for various pathologic conditions and recently HCC is one of them. Aim: Explore the potential usefulness of serum miR-221 as a diagnostic biomarker of HCV-related HCC. Methods: This study was conducted on 40 chronic HCV patients and 10 healthy control subjects. Patients were divided into two groups: HCC and cirrhosis. All patients and controls were subjected to clinical assessment and laboratory investigations in addition to the evaluation of the level of serum miRNA-221 expression by RT-PCR. Results: MiRNA- 221 showed significant fold increase in both HCC and cirrhosis (P = 0.005) when compared to normal controls. ROC curve analysis for miR-221 yielded 85% sensitivity and 40% specificity for differentiation of HCC patients from non-HCC. Conclusion: Serum miR-221 has a strong potential to serve as one of the biomarkers of early HCC development. This can modify liver carcinogenesis, and can be used to improve preventive, diagnostic, and therapeutic strategies in HCC.Key words: MiRNA; Hepatocellular carcinoma; Hepatitis C Virus

Associations of expressions of serum MiR-124 and MiR-182 with endothelial function and inflammation level in patients with cerebral arteriosclerosis.

Associations of expressions of serum MiR-124 and MiR-182 with endothelial function and inflammation level in patients with cerebral arteriosclerosis.

Evaluation of Serum miR-21 Expression Level in Patients Suspected of Mycobacterium tuberculosis

Evaluation of Serum miR-21 Expression Level in Patients Suspected of Mycobacterium tuberculosis

New risk factors in diabetic nephropathy: microRNA-196-3p and microRNA-203

Aim The aim of this study was to evaluate changes in the serum levels of miR-98, miR-184, miR-185, miR-203 and miR-196a-3p in type 2 diabetes mellitus (T2DM) patients with diabetic nephropathy (DN) and to associate the changes in microRNA levels with urinary albumin excretion (UAE). Methods The study included 35 healthy individuals aged between 18-65 years, 40 T2DM patients with normoalbuminuria, 40 T2DM patients with microalbuminuria, and 35 T2DM patients with macroalbuminuria. Metabolic laboratory parameters, microalbumin levels in 24 hour urine samples were measured in all groups. Serum levels of vascular endothelial growth factor-A (VEGF-A) and transforming growth factor-β (TGF-β) were measured quantitatively by enzyme-linked immunosorbent assay (ELISA). Circulating miRNA levels were determined by Real-Time quantitative PCR. Results Serum miR-196a-3p levels were reduced in the normoalbuminuria group compared to the healthy control group. In the macroalbuminuria and microalbuminuria groups, miR-196a-3p levels were higher compared to the normoalbuminuria group. In addition, increase in miR-196a-3p levels in the macroalbuminuria group was more prominent than the microalbuminuria group. Serum miR-203 levels were significantly higher in the macroalbuminuria group compared to healthy controls, microalbuminuria and normoalbuminuria groups; these levels were also higher in the microalbuminuria group compared to normoalbuminuria group. In logistic regression analysis, serum miR-196a-3p and miRNA-203 levels were independently correlated to UAE. Conclusions Increased serum levels of miR-203 and miR-196a-3p are independent risk factors of UAE which is a marker of DN.

Exosomal MiR-107 As Novel Biomarker and Tumor Suppressor By Targeting Ywhah in Diffuse Large B-Cell Lymphoma

Introduction : microRNAs (miRNAs) could be released into the extracellular microenvironment and mediate cellular communication through exosomes. Circulation exosomal miRNAs have recently emerged as complimentary biomarkers and novel approaches for the non-invasive tests of early diagnosis or follow-up of neoplasms. Accumulating evidences have indicated that miR-107 plays vital functions in suppressing tumorigenesis. However, the significance of exosomal miR-107 in DLBCL remains unclear. Thus, this study aimed to explore candidate biomarkers and investigate the role of miR-107 in DLBCL as well as the molecular mechanisms involved. Methods : Differentially expressed miRNAs (DEMs) in DLBCL were identified based on Gene Expression Omnibus (GEO) datasets and verified in a cohort of DLBCL patients. The functions and biological pathways of DEMs were enriched by DAVID. Serum-derived exosomes of 42 DLBCL patients and 31 healthy volunteers were isolated with informed consents by Exo Easy Maxi Kit, and further detected by western blot and transmission electron microscopy (TEM). Receiver operating characteristic curves (ROC) were performed to evaluate the diagnostic value of DEMs. The biological function of miR-107 in DLBCL were evaluated by miR-107 Agomir. Survival analyses were performed by the Kaplan-Meier method. Potential targets genes of miR-107 were predicted by miRDB, PicTar, Targetscan and miRTarBase. Western blotting and confocal staining assay were used to detect the expression of YWHAH. Interaction of miR-107 and YWHAH was confirmed by dual-luciferase reporter assay. Results : 14 DEMs were identified in DLBCL through evaluating the miRNA microarray profile of GSE117063 (Fig. 1A-A) and GSE29493 (Fig. 1A-B). The biological processes of these DEMs mainly enriched in regulatory of transcription, protein phosphorylation and cell proliferation (Fig.1B). In order to explore candidate exosomal biomarkers for DLBCL, we then isolated serum-derived exosomes from DLBCL patients. As depicted in Fig.1C, exosomes not only expressed exosomal biomarkers Tsg101 and CD9 but also shown cup-shape morphology. We further assessed the expression of DEMs on exosomes by qRT-PCR. Compared with normal samples, only two down-regulated miRNAs (miR-107, miR-375) and one up-regulated miRNAs (miR-485) were statistically dysregulated in DLBCL patients (Fig.2A). ROC curve analysis demonstrated that exosomal miR-107, miR-375, and miR-485 might be potential biomarkers for DLBCL patients, with the AUC of 0.854, 0.769, and 0.703, respectively (Fig.2B). In addition, survival analysis revealed that low-expression of miR-107 in DLBCL patients significantly associated with undesirable clinical outcomes (Fig.2C, p&amp;lt;0.05). These data suggested that miR-107 may serve as a valuable prognostic biomarker and participated in progression of DLBCL. Down-regulation of miR-107 was confirmed in DLBCL cell lines by qRT-PCR (Fig.3A). To validate our hypothesis, we up-regulated miR-107 expression by miRNA Agomir, which could significantly reduce cell proliferation and migration, and promote cell apoptosis (Fig.3B-D). The regulatory mechanisms of miR-107 in DLBCL was further explored. 28 target genes of miR-107 were identified and shown in Fig.4A. KEGG pathway analysis revealed that the targeted genes mainly enriched in the PI3K-Akt, Hippo, and AMPK signaling pathways (Fig.4B). We found out that the target gene YWHAH (14-3-3η) was significantly upregulated in the DLBCL cells (Fig.4C-D). Moreover, dual-luciferase assay revealed that miRNA-107 perform anti-tumor effect through targeting the 3'UTR region of YWHAH (Fig.4E-F). As previously reported, YWHAH associated with tumor metastasis, chemoresistance, apoptosis resistance and poor prognosis in DLBCL. Thus, we suggested that miR-107 antagonized DLBCL progression through downregulating YWHAH. Conclusion : In this study, we demonstrated for the first time that serum exosomal miR-107, miR-375, and miR-485 could be applied as potential non-invasive biomarkers in early diagnosis of DLBCL patients. Furthermore, miR-107 inhibited DLBCL progression by targeting YWHAH, which will provide a promising therapeutic approach for DLBCL. Disclosures No relevant conflicts of interest to declare.

Grafting Islets to a Dissected Peritoneal Pouch to Improve Transplant Survival and Function.

Although the liver is the primary site for clinical islet transplantation, it poses several restrictions, especially limited tissue volume due to portal vein pressure. We evaluated the preperitoneal space as an extrahepatic islet transplant site to deliver high tissue volumes and sustain long-term graft function. A peritoneal pouch was formed by dissecting the parietal peritoneum from the transversalis fascia of mice. Syngeneic C57BL/6 donor islets were transplanted into the peritoneal pouch of diabetic mouse recipients. Blood glucose was monitored for islet function, and miR-375 was analyzed for islet damage. Islet graft morphology and vascularization were evaluated by immunohistochemistry. [F] fluoro-D-glucose positron emission tomography/computed tomography was used to image islet grafts. Transplantation of 300 syngeneic islets into the peritoneal pouch of recipients reversed hyperglycemia for >60 days. Serum miR-375 was significantly lower in the peritoneal pouch group than in the peritoneal cavity group. Peritoneal pouch islet grafts showed high neovascularization and sustained insulin and glucagon expression up to 80 days posttransplantation. A peritoneal pouch graft with high tissue volume (1000 islets) could be visualized by positron emission tomography/computed tomography imaging. Human islets transplanted into the peritoneal pouch of diabetic nude mice also reversed hyperglycemia successfully. Islets transplanted into a dissected peritoneal pouch show high efficiency to reverse diabetes and sustain islet graft function. The preperitoneal site has the advantages of capacity for high tissue volume, enriched revascularization and minimal inflammatory damage. It can also serve as an extrahepatic site for transplanting large volume of islets necessitated in islet autotransplantation.

Serum miRNAs associated with tumor-promoting cytokines in non-small cell lung cancer.

Tumor-promoting cytokines are a cause of tumor progression; therefore, identifying key regulatory microRNAs (miRNAs) for controlling their production is important. The aim of this study is to identify promising miRNAs associated with tumor-promoting cytokines in non-small cell lung cancer (NSCLC). We identified circulating miRNAs from 16 published miRNA profiles. The selected miRNAs were validated in the serum of 32 NSCLC patients and compared with 33 patients with other lung diseases and 23 healthy persons using quantitative real-time PCR. The cytokine concentration was investigated using the enzyme-linked immunoassay in the same sample set, with clinical validation of the miRNAs. The correlation between miRNA expression and cytokine concentration was evaluated by Spearman's rank correlation. For consistent direction, one up-regulated miRNA (miR-145) was found in four studies, and seven miRNAs were reported in three studies. One miRNA (miR-20a) and four miRNAs (miR-25-3p, miR-223, let-7f, and miR-20b) were reported in six and five studies. However, their expression was inconsistent. In the clinical validation, serum miR-145 was significantly down-regulated, whereas serum miR-20a was significantly up-regulated in NSCLC, compared with controls. Regarding serum cytokine, all cytokines [vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), and transforming growth factor β (TGF-β)], except tumor necrosis factor-α (TNF-α), had a higher level in NSCLC patients than controls. In addition, we found a moderate correlation between the TGF-β concentration and miR-20a (r = -0.537, p = 0.002) and miR-223 (r = 0.428, p = 0.015) and a weak correlation between the VEGF concentration with miR-20a (r = 0.376, p = 0.037) and miR-223 (r = -0.355, p = 0.046). MiR-145 and miR-20a are potential biomarkers for NSCLC. In addition, the regulation of tumor-promoting cytokine, through miR-20a and miR-223, might be a new therapeutic approach for lung cancer.

Evaluation of MicroRNA-210(miR-210) as a Diagnostic and Prognostic Biomarker in Pre-eclampsia Pregnancies

Objective: This study aims to assess the level of miR-210 in Egyptian women with pre-eclampsia (PE) and to evaluate its role in diagnosis and prognosis of the disease. Subjects & Methods: The study was conducted on 30 pregnant women with PE divided into two groups: group (I): included 15 cases with mild PE and group (II): included 15 cases of severe PE and, 20 healthy pregnant women with matched age and sex were included as control group. All women included in the study were subjected to, history taking full clinical examination, laboratory investigations included (CBC), (PT), (ALT, AST), (urea, creatinine), detection of protein in urine, as well as miRNA-210 gene expression by RT-PCR. Results: Patients with PE showed a highly significantly increase in serum miR-210 (P value <0.001) compared to control as well as, it was higher in severe PE than in mild PE (P value <0.001). MiR 210 have highly significant positive correlation with (systolic, diastolic, MABP), (Proteinuria) and (PT) a significant positive correlation with (AST), (ALT),.However no significant correlation was found with (Hb%) ,platelet count, PTT and INR, serum urea and creatinine. PTT was statistically significant (P value 0.002).The best cutoff value of PE (2.03) with sensitivity and specificity of 90.0% and 85.0% respectively. Conclusion: Expression of mir-210 is upregulated in pre-eclampsia and was higher in severe than in mild. Hence, the serum miR-210 can be used as a diagnostic, prognostic biomarker in PE patients and understanding pathophysiology.

Circulating microRNA-145 as a diagnostic biomarker for non-small-cell lung cancer: A systemic review and meta-analysis.

MicroRNAs (miRNAs), a class of small non-coding, highly stable RNAs, have been reported to have diagnostic value for variety types of cancers. To assess the diagnostic value of circulating miR-145 for non-small cell lung cancer (NSCLC) by using systemic review and meta-analysis. A systematic literature search was conducted in five databases until 20 February 2020 to identify diagnostic trials of miR-145 in the diagnosis of NSCLC. The quality of included studies was assessed by the QUADAS-2 tool with Review Manager 5.3, and the summary receiver operating characteristic (SROC) curve was plotted by STATA 13.1 software. A total of 1394 patients from 11 data sets in trials (published in nine studies) were recruited. The area under the curve of the SROC was 0.83. According to the meta regression, the specimen selection was considered the source of heterogeneity, the SROC in serum (0.90 (95% CI 0.87, 0.92), the sensitivity was 0.84 (95% CI 0.79, 0.89), and the specificity was 0.80 (95% CI 0.71, 0.89)) was obviously higher than that in plasma (SROC=0.75). Serum miR-145 might be served as a potentially useful biomarker for NSCLC diagnosis. However, due to the existing limited-quality research, more large-scale and multicenter studies are required for further verification.

Targeting miR-124/Ferroportin signaling ameliorated neuronal cell death through inhibiting apoptosis and ferroptosis in aged intracerebral hemorrhage murine model.

Incidence of intracerebral hemorrhage (ICH) and brain iron accumulation increases with age. Excess iron accumulation in brain tissues post‐ICH induces oxidative stress and neuronal damage. However, the mechanisms underlying iron deregulation in ICH, especially in the aged ICH model have not been well elucidated. Ferroportin1 (Fpn) is the only identified nonheme iron exporter in mammals to date. In our study, we reported that Fpn was significantly upregulated in perihematomal brain tissues of both aged ICH patients and mouse model. Fpn deficiency induced by injecting an adeno‐associated virus (AAV) overexpressing cre recombinase into aged Fpn‐floxed mice significantly worsened the symptoms post‐ICH, including hematoma volume, cell apoptosis, iron accumulation, and neurologic dysfunction. Meanwhile, aged mice pretreated with a virus overexpressing Fpn showed significant improvement of these symptoms. Additionally, based on prediction of website tools, expression level of potential miRNAs in ICH tissues and results of luciferase reporter assays, miR‐124 was identified to regulate Fpn expression post‐ICH. Higher serum miR‐124 levels were correlated with poor neurologic scores of aged ICH patients. Administration of miR‐124 antagomir enhanced Fpn expression and attenuated iron accumulation in aged mice model. Both apoptosis and ferroptosis, but not necroptosis, were regulated by miR‐124/Fpn signaling manipulation. Our study demonstrated the critical role of miR‐124/Fpn signaling in iron metabolism and neuronal death post‐ICH in aged murine model. Thus, Fpn upregulation or miR‐124 inhibition might be promising therapeutic approachs for this disease.

Circulating microRNA-21 as Stable Blood-Based Markers for Patients With ISR After PCI.

It recently has been reported that the in-stent restenosis (ISR) of expanded area after percutaneous coronary intervention (PCI) within six months can become a serious postoperative complication. A real-time quantitative PCR was used to analyze the expression of serum miR-21 in 33 ISR and 37 non-ISR patients after PCI. Expression of miR-21 was significantly higher in the ISR group compared with that in the NISR group, and a similar trend also occurred in factor- (TNF-α) level, Interleukin -6 (IL-6) level, and plaque area (PLA). However, a contrary trend occurred in the external elastic membrane area (EEM) and minimal lumen area (MLA). This study suggests that the increased expression of serum miR-21 is related to ISR after PCI, and miR-21 can be a new predictor of ISR.

Bovine serum miR-21 expression affected by mastitis

The expression levels of circulating microRNAs (miRNAs) can be affected by disease. The miRNA released from cells within exosomes can act as a remote communication tool and can participate in inflammatory response regulation. Therefore, circulating miRNA has the potential to be an indicator of local disease. The objective of this study was to investigate the serum level of bovine mastitis-related miRNAs. We found that miR-16 expression in serum was affected by hemolysis. The expression levels of miR-21 in serum were increased significantly in cows with mastitis compared with unaffected controls; however, the expression levels of miR-146a, miR-155, miR-222 and miR-383 in cows with mastitis were unchanged. We further verified the upregulation of miR-21 in the serum of cows with mastitis using a digital PCR system. Although the sensitivity and specificity of miR-21 in the serum to detect bovine mastitis was inferior to miRNA biomarkers in the milk, the significant increase of miR-21 in serum may reflect the impact of local inflammation on the systemic reaction.

Serum a proliferation-inducing ligand and MicroRNA-223 are associated with rheumatoid arthritis: diagnostic and prognostic implications

BackgroundCurrent blood-based tests for rheumatoid arthritis (RA) have inherent limitations, necessitating the need for additional new biomarkers for its diagnosis and monitoring disease activity and responsiveness to therapy. MicroRNAs (miRNAs) and a proliferation-inducing ligand (APRIL) are deregulated in RA and were linked to its pathogenesis. This study investigated serum levels of APRIL, miR-223 and miR-155 in RA patients, their potential as diagnostic and prognostic biomarkers, and their correlation with disease activity and clinicopathological data.MethodsOne hundred and twenty Egyptian patients with RA and 130 healthy controls were included. Serum miRNAs and APRIL were assayed by RT-qPCR and ELISA, respectively.ResultsSerum APRIL and miR-223 were significantly upregulated, while miR-155 was unchanged in RA patients compared to controls. Serum miR-223 discriminated RA patients from controls with AUC = 0.85, whereas serum APRIL superiorly distinguished the two groups with AUC = 1 (sensitivity and specificity = 100% at cutoff> 4.19 ng/ml) by receiver-operating-characteristic analysis. Serum miR-223 was a significant predictor for RA diagnosis in multivariate logistic regression analysis. In RA group, serum APRIL was positively correlated with disease activity score (DAS28-CRP). Serum miR-223 expression was positively correlated with serum miR-155, APRIL levels and with the presence of subcutaneous nodules. Serum miR-155 levels were correlated with antinuclear antibody titer in reverse direction.ConclusionOur results suggest serum APRIL and miR-223 could serve as potential biomarkers of RA, with miR-223 as a predictor of RA risk and APRIL as an excellent biomarker of disease activity. Our data could be implicated for accurate and blood-based non-invasive diagnosis and prognosis of RA.

151 Poster - Role of miR-21 in chemotherapeutic drug resistance of oral squamous cell carcinoma

Background: Head and neck carcinoma spreads malignancies tumors arising from the epithelium or squamous covering the upper aerodigestive tract including oral that metastasize to other organs. The oral squamous cell carcinoma primarily addressing Head and neck carcinoma cancer that could affect the majority of older people with approximately 650,000 new cases per year. The consumption of alcohol and tobacco might be the prime cause of OSCC risk. But the recurrence or relapse of the malignancies is the major obstacle of the convention chemotherapeutic drug like paclitaxel or carboplatin. We know the circulating miRNAs is noted as the potential biomarkers of OSCC but their role in drug resistance is yet to reveal. In the present study, an attempt made to find the expression of miR-21 in serum/ whole blood of relapse OSCC (rOSCC) and newly identified OSCC (nOSCC) (N = 40). The expression of miR-21 was evaluated in relation to different demographical and clinicopathological features aiming to identify correlation with relapse and their cancer clinical stages (perineural invasion and N-stage).