The diagnostic value of circulating microRNAs for middle-aged (40–60-year-old) coronary artery disease patients

OBJECTIVE:Circulating microRNAs have been recognized as promising biomarkers for various diseases. The present study aimed to explore the potential roles of circulating miR-149, miR-424 and miR-765 as non-invasive biomarkers for the diagnosis of coronary artery disease in middle-aged (40–60-year-old) patients.METHODS:Sixty-five stable coronary artery disease patients (49–57 years old), 30 unstable coronary artery disease patients (49–58 years old), and 32 non-coronary artery disease patients (49–-57 years old) who were matched for age, sex, smoking habits, hypertension and diabetes were enrolled in this study. Total RNA was isolated from plasma with TRIzol reagent. Circulating miRNA levels were measured by quantitative real-time polymerase chain reaction.RESULTS:Circulating miR-149 levels were decreased 4.49-fold in stable coronary artery disease patients (1.18 ± 0.84) and 5.09-fold in unstable coronary artery disease patients (1.04 ± 0.65) compared with non-coronary artery disease patients (5.30 ± 2.57) (p<0.001). Circulating miR-424 levels were reduced 3.6-fold in stable coronary artery disease patients (1.18 ± 0.60) and 5-fold in unstable coronary artery disease patients (0.86 ± 0.54) compared with non-coronary artery disease patients (4.35 ± 2.20) (p<0.001). In contrast, circulating miR-765 levels were elevated 3.98-fold in stable coronary artery disease patients (6.09 ± 2.27) and 5.33-fold in unstable coronary artery disease patients (8.17 ± 2.77) compared with non-coronary artery disease patients (1.53 ± 0.99) (p<0.001). Receiver operating characteristic curve analysis revealed that the respective areas under the curve for circulating miR-149, miR-424 and miR-765 were 0.938, 0.919 and 0.968 in stable CAD patients and 0.951, 0.960 and 0.977 in unstable coronary artery disease patients compared with non-coronary artery disease patients.CONCLUSION:Our results suggest that circulating miR-149, miR-424 and miR-765 might be novel, non-invasive biomarkers for the diagnosis of coronary artery disease in middle-aged patients. However, future prospective trials in large patient cohorts are necessary before reaching a solid conclusion.     

Deletion of Mir155 Prevents Fas-Induced Liver Injury through Up-Regulation of Mcl-1

Fas-induced apoptosis is involved in diverse liver diseases. Herein, we investigated the effect of Mir155 deletion on Fas-induced liver injury. Wild-type (WT) mice and Mir155 knockout (KO) mice were i.p. administered with the anti-Fas antibody (Jo2) to determine animal survival and the extent of liver injury. After Jo2 injection, the Mir155 KO mice exhibited prolonged survival versus the WT mice (P < 0.01). The Mir155 KO mice showed lower alanine aminotransferase and aspartate aminotransferase levels, less liver tissue damage, fewer apoptotic hepatocytes, and lower liver tissue caspase 3/7, 8, and 9 activities compared with the WT mice, indicating that Mir155 deletion prevents Fas-induced hepatocyte apoptosis and liver injury. Hepatocytes isolated from Mir155 KO mice also showed resistance to Fas-induced apoptosis, in vitro. Higher protein level of myeloid cell leukemia-1 (Mcl-1) was also observed in Mir155 KO hepatocytes compared to WT hepatocytes. A miR-155 binding site was identified in the 3′-untranslated region of Mcl-1 mRNA; Mcl1 was identified as a direct target of miR-155 in hepatocytes. Consistently, pretreatment with a siRNA specific for Mcl1 reversed Mir155 deletion–mediated protection against Jo2-induced liver tissue damage. Finally, restoration of Mir155 expression in Mir155 KO mice abolished the protection against Fas-induced hepatocyte apoptosis. Taken together, these findings demonstrate that deletion of Mir155 prevents Fas-induced hepatocyte apoptosis and liver injury through the up-regulation of Mcl1.miRNAs are a class of small noncoding RNAs that regulate gene expression through binding to specific sequences of their targeted mRNAs.¹ miR-155 was initially identified as a specific miRNA for hematopoietic and immune cells and is among the first miRNAs linked to immunity and inflammation.^2–4 Indeed, miR-155 has been shown to modulate the production of cytokines in various immune cells; the expression of Mir155 is up-regulated in multiple immune cell lineages on stimulation with Toll-like receptor ligands, inflammatory cytokines, and specific antigens.^5–9 Subsequent studies have shown that miR-155 also mediates functions outside the hematopoietic and immune systems.^10,11 In the liver, miR-155 has been shown to play a role in hepatocarcinogenesis,^12–15 although its mechanism of action remains to be further defined. miR-155 has also been shown to contribute to alcohol-induced liver injury through induction of tumor necrosis factor α production in macrophages.¹⁶ Interestingly, the level of miR-155 is increased in serum and plasma in patients with alcoholic and inflammatory liver injuries.^17,18 These observations suggest a potential role of miR-155 in liver injury and liver diseases. However, at present, the biological functions and mechanisms of miR-155 in liver cells have not been delineated.The current study aimed to determine the effect and mechanism of miR-155 in Fas and lipopolysaccharide (LPS)/d-galactosamine (D-GalN)–mediated liver injury in mice. Our data show that deletion of Mir155 protects against Fas-induced hepatocyte apoptosis and liver injury but not LPS/D-GalN–induced liver injury. The role of miR-155 in hepatocytes was demonstrated by in vitro studies using hepatocytes isolated from Mir155 knockout (KO) mice. Myeloid cell leukemia-1 (Mcl-1) was identified as a direct target of miR-155 in hepatocytes. Our results reveal a novel role of miR-155 in hepatocytes for regulation of Mcl1 and protection against Fas-induced apoptosis.     

Predictive Role of Lung Injury Prediction Score in the Development of Acute Respiratory Distress Syndrome in Korea

PurposeEarly recognition and therapeutic intervention are important in patients at high risk of acute respiratory distress syndrome (ARDS). The lung injury prediction score (LIPS) has been used to predict ARDS development; however, it was developed based on the previous definition of ARDS. We investigated the predictive role of LIPS in ARDS development according to its Berlin definition in the Korean population.Materials and MethodsThis was a retrospective study that enrolled adult patients admitted to the intensive care unit (ICU) at a single university-affiliated hospital in Korea from September 1, 2018, to August 31, 2019. LIPS at the time of ICU admission and the development of ARDS were evaluated.ResultsOf the 548 enrolled patients, 33 (6.0%) fulfilled the Berlin ARDS definition. The LIPS for non-ARDS and ARDS groups were 4.96±3.05 and 8.53±2.45, respectively (p<0.001); it was significantly associated with ARDS development (odds ratio 1.48, 95% confidence interval, 1.29–1.69; p<0.001). LIPS >6 predicted the development of ARDS with a sensitivity of 84.8% and a specificity of 67.2% [area under the curve (AUC)=0.82]. A modified LIPS model adjusted for age and severity at ICU admission predicted ICU mortality in patients with ARDS (AUC=0.80), but not in those without ARDS (AUC=0.54).ConclusionLIPS predicted the development of ARDS as diagnosed by the Berlin definition in the Korean population. LIPS provides useful information for managing patients with ARDS.     

MiR-146a and miR-155 polymorphisms in Egyptian patients with Behcet’s disease

IntroductionThe current study was designed to analyze whether polymorphisms of miR-146a and miR-155 are related to Behçet’s disease (BD) in the Egyptian population.Material and methodsA total of 96 unrelated BD patients and 100 healthy subjects were genotyped for miR-146a (rs2910164) and miR-155 (rs767649) using real-time polymerase chain reaction.ResultsThe results showed significant elevation in the frequency of rs2910164 GG and CC genotypes in BD patients compared with controls (adjusted OR = 22.156, 95% CI: 4.728–103.818; p < 0.001 and adjusted OR = 40.358, 95% CI: 8.928–182.440; p < 0.001, respectively). Also, the rs2910164 G allele conferred a higher risk of developing BD (adjusted OR = 3.665, 95% CI: 2.013–6.671; p < 0.001). MiR-146a (rs2910164) polymorphism was a risk factor for susceptibility to BD in dominant, recessive and additive models of inheritance (all p < 0.001), while the miR-155 (rs767649) polymorphism was a risk factor in the recessive model only (p = 0.021). GG and CG genotypes of rs2910164 were associated with higher Behcet’s disease current activity index (BDCAI) and ocular involvement compared with CC genotype (p = 0.005 and p = 0.004, respectively). Genotype AT of rs767649 was related to higher BDCAI (p = 0.026) compared with TT and AA genotypes.ConclusionsmiR-146a (rs2910164) and miR-155 (rs767649) are likely to play an important role in the Egyptian population in development of BD and also influence disease severity.     

Combined identification of long non-coding RNA CCAT1 and HOTAIR in serum as an effective screening for colorectal carcinoma

Long non-coding RNAs (lncRNAs) CCAT1 and HOTAIR have been shown to play an important regulatory role in cancer biology, and CCAT1 and HOTAIR are upregulated in several cancers, however, its value in the diagnosis of colorectal cancer (CRC) is unclear. Therefore, the aim of this study is to evaluate the clinical significance of plasma CCAT1 and HOTAIR as a biomarker in the screening of CRC. In our study, we found that the levels of HOTAIR (P < 0.05) and CCAT1 (P < 0.05) were significantly higher in plasma of CRC patients than that of the healthy control. Moreover, the levels of lincRNA-p21 (P < 0.05) were obviously decreased in plasma of CRC patients as compared to those of healthy control. There was highly correlated for CCAT1 (R = 0.752, mean differences = -0.06 ± 1.20), HOTAIR (R = 0.739, mean differences = -0.26 ± 0.76) and lincRNA-p21 (R = 0.848, mean differences = -0.41 ± 0.89) in plasma and serum. By receiver operating characteristic curve (ROC) analysis, plasma CCAT1 provided the higher diagnostic performance for detection of CRC (the area under the ROC curve (AUC), 0.836; P < 0.001; sensitivity, 75.7%; specificity, 85.3%). Moreover, CCAT1 combining with HOTAIR could provide a more effective diagnosis performance (AUC, 0.954, P < 0.001, sensitivity, 84.3%; specificity, 80.2%). Most importantly, this combination was effective to detect CRC at an early stage (85%). In conclusion, our results demonstrated that increased plasma HOTAIR and CCAT1 could be used as a predictive biomarker for CRC screening, and that combination of HOTAIR and CCAT1 had a higher positive diagnostic rate of CRC than HOTAIR or CCAT1 alone.     

Twelve-month kinetics of circulating fibrosis-linked microRNAs (miR-21, miR-29, miR-30, and miR-133a) and the relationship with extracellular matrix fibrosis in dilated cardiomyopathy

IntroductionA single measurement of any biomarker may not reflect its full biological meaning. The kinetics of fibrosis-linked microRNAs and their relationship with extracellular matrix (ECM) fibrosis in dilated cardiomyopathy (DCM) have not been explored.Material and methodsWe evaluated 70 consecutive DCM patients (48 ±12.1 years, left ventricular ejection fraction 24.4 ±7.4%). All patients underwent right ventricular endomyocardial biopsy in order to quantify ECM fibrosis and measure collagen volume fraction (CVF). Circulating microRNAs (miR-21-5p, miR-29b, miR-30c-5p, and miR-133a-3p) were measured with quantitative polymerase chain reaction (PCR) at baseline and at 3 and 12 months.ResultsBased on the biopsy results, two groups of patients were identified: with (n = 24, 34.3%) and without (n = 46, 65.7%) ECM fibrosis. Except for a single measurement of miR-29b at 3 months (DCM with fibrosis: 6.03 ±0.72 vs. DCM without fibrosis: 6.4 ±0.75 ΔCq; p < 0.05), baseline, 3- and 12-month kinetics of microRNAs did not differ between the two groups. Moreover, 12-month microRNA kinetics did not differ in patients with new-onset DCM (duration < 6 months; n = 35) and chronic DCM (> 6 months; n = 35). Only miR-29 at 3 months correlated with CVF (r = –0.31; p < 0.05), whereas other microRNAs did not correlate with CVF either at 3 or at 12 months.ConclusionsRegardless of ECM fibrosis status or duration of the disease, 12-month patterns of circulating microRNAs are similar in DCM. Correlations between microRNAs, measured at 3 and 12 months, are lower than expected. In this study, regardless of the time point, circulating microRNAs were not able to differentiate between DCM patients with versus without fibrosis.     

Role of miR-155 in the Pathogenesis of Herpetic Stromal Keratitis

Ocular infection with herpes simplex virus 1 can result in a chronic immunoinflammatory stromal keratitis (SK) lesion that is a significant cause of human blindness. A key to controlling SK lesion severity is to identify cellular and molecular events responsible for tissue damage and to manipulate them therapeutically. Potential targets for therapy are miRNAs, but these are minimally explored especially in responses to infection. Here, we demonstrated that Mir155 expression was up-regulated after ocular herpes simplex virus 1 infection, with the increased Mir155 expression occurring mainly in macrophages and CD4⁺ T cells and to a lesser extent in neutrophils. In vivo studies indicated that Mir155 knockout mice were more resistant to herpes SK with marked suppression of T helper cells type 1 and 17 responses both in the ocular lesions and the lymphoid organs. The reduced SK lesion severity was reflected by increased phosphatidylinositol-3,4,5-trisphosphate 5-phosphatase 1 and interferon-γ receptor α-chain levels in activated CD4⁺ T cells in the lymph nodes. Finally, in vivo silencing of miR-155 by the provision of antagomir-155 nanoparticles to herpes simplex virus 1–infected mice led to diminished SK lesions and corneal vascularization. In conclusion, our results indicate that miR-155 contributes to the pathogenesis of SK and represents a promising target to control SK severity.Ocular infection with herpes simplex virus 1 (HSV-1) can result in a chronic tissue-damaging response in the stroma, which is considered to be largely the consequence of a host inflammatory response to the infection.¹ This concept is strongly supported by animal model studies in which lesions were shown to be mainly orchestrated by CD4⁺ T cells with neutrophils and macrophage largely responsible for the tissue damage.^2–5 Several effective control measures for stromal keratitis (SK) are suggested.⁶ These include approaches that influence cellular infiltration and activation of the proinflammatory mediators of SK.⁶ One potential means of modulating SK lesions that so far has received minimal attention is to manipulate the expression of miRNA species that affect either virus or host events during SK.A prime miRNA candidate for consideration is miR-155 because this miRNA can influence the expression of several immune events that contribute to tissue damage.^7–10 For example, animals unable to produce miR-155 because of gene knockout may develop milder lesions in some models of autoimmunity,^8,11–13 and suppressing miR-155 expression, as can be achieved by treatment with antagomirs, holds promise as a means of therapy for autoimmunity.¹³ However, the absence of miR-155 can result in higher susceptibility to some virus infections and some tumors in part because protective CD8⁺ T-cell responses are diminished.^14–17 In fact, overexpression of miRNA can result in enhanced CD8⁺ T-cell–mediated immune protection with some tumors.¹⁷Few studies have focused on the role of miR-155 in situations in which the immune response to an infectious agent may be a principal cause of tissue damage. This is the situation in ocular lesions of the cornea after HSV-1 infection. Here, we have compared the disease outcome after HSV-1 infection in miRNA-155 knockout (Mir155^−/−) mice and in mice in which miR-155 was suppressed by antagomir therapy with controls. We show that HSV-1 infection resulted in up-regulation of miR-155, which was mainly produced by the inflammatory cells and T cells in the cornea. Suppression of miR-155 production resulted in milder lesions that were associated with diminished responses of T helper cells type 1 (Th1) and type 17 (Th17) and reduced inflammatory cytokine production. We also demonstrated that miR-155 suppression resulted in increased phosphatidylinositol-3,4,5-trisphosphate 5-phosphatase 1 (Ship1) and IFN-γ receptor α-chain (IFN-γRα) levels, molecules known to be required for IFN-γ expression and Th1 differentiation.^17,18 These results indicate that miR-155 regulates differentiation and effector function of Th1 cells. Thus, our results suggest that miR-155 could be a promising therapeutic target to treat SK.     

The clinical implications of serum ferritin in Kawasaki disease: a helpful biomarker for evaluating therapeutic responsiveness,coronary artery involvement and the tendency of macrophage activation syndrome

IntroductionSerum ferritin (SF) is an acute-phase reactant in inflammatory diseases. Our aim was to analyze the clinical implications of SF in Kawasaki disease (KD).Methods244 KD children were divided into 6 subgroups. SF, inflammatory mediators and blood cell counts were detected.Results(1) SF dramatically increased in the acute phase of KD and maintained after IVIG therapy; (2) SF increased in IVIG-nonresponsive KD patients (AUC = 0.705; sensitivity: 57.10%; specificity: 82.90%); SF positively correlated with the internal diameter of the coronary artery (AUC = 0.603; sensitivity: 92.30%; specificity: 37.70%); (3) SF increased in 4 patients with the macrophage activation syndrome (MAS)/MAS tendency (979.03 ±474.19 μg/l).ConclusionsSF is implied to be a helpful biomarker for forecasting IVIG-nonresponsive KD, coronary artery abnormalities (CAAs) and MAS tendency.     

Interpreting Clinical Reaction Time Change and Recovery After Concussion: A Baseline Versus Norm-Based Cutoff Score Comparison

ContextPreseason testing can be time intensive and cost prohibitive. Therefore, using normative data for postconcussion interpretation in lieu of preseason testing is desirable.ObjectiveTo establish the recovery trajectory for clinical reaction time (RT_clin) and assess the usefulness of changes from baseline (comparison of postconcussion scores with individual baseline scores) and norm-based cutoff scores (comparison of postconcussion scores with a normative mean) for identifying impairments postconcussion.DesignCase-control study.SettingMultisite clinical setting.Patients or Other ParticipantsAn overlapping sample of 99 participants (age = 19.0 ± 1.1 years) evaluated within 6 hours postconcussion, 176 participants (age = 18.9 ± 1.1 years) evaluated at 24 to 48 hours postconcussion, and 214 participants (age = 18.9 ± 1.1 years) evaluated once they were cleared to begin a return-to-play progression were included. Participants with concussion were compared with 942 control participants (age = 19.0 ± 1.0 years) who did not sustain a concussion during the study period but completed preseason baseline testing at 2 points separated by 1 year (years 1 and 2).Main Outcome Measure(s)At each time point, follow-up RT_clin (ie, postconcussion or year 2) was compared with the individual year 1 preseason baseline RT_clin and normative baseline data (ie, sex and sport specific). Receiver operating characteristic curves were calculated to compare the sensitivity and specificity of RT_clin change from baseline and norm-based cutoff scores.ResultsClinical reaction time performance declined within 6 hours (18 milliseconds, 9.2% slower than baseline). The decline persisted at 24 to 48 hours (15 milliseconds, 7.6% slower than baseline), but performance recovered by the time of return-to-play initiation. Within 6 hours, a change from baseline of 16 milliseconds maximized combined sensitivity (52%) and specificity (79%, area under the curve [AUC] = 0.702), whereas a norm-based cutoff score of 19 milliseconds maximized combined sensitivity (46%) and specificity (86%, AUC = 0.700). At 24 to 48 hours, a change from baseline of 2 milliseconds maximized combined sensitivity (64%) and specificity (61%, AUC = 0.666), whereas a norm-based cutoff score of 0 milliseconds maximized combined sensitivity (63%) and specificity (62%, AUC = 0.647).ConclusionsNorm-based cutoff scores can be used for interpreting RT_clin scores postconcussion in collegiate athletes when individual baseline data are not available, although low sensitivity and specificity limit the use of RT_clin as a stand-alone test.     

Detection of miR-1246, miR-23a and miR-451 in sera of colorectal carcinoma patients: a case-control study in Cairo University hospital

BackgroundColorectal cancer (CRC) has high morbidity and mortality rates. Invasive techniques and other laboratory tests with variable sensitivity and specificity are currently used in diagnosis. Micro ribonucleic acids (miRNAs) have bio vital roles in cell proliferation and apoptosis. Dys-regulation of miRNAs is linked to tumour genesis. The objective of this study was to evaluate the specificity and sensitivity of serum non-invasive biomarkers (micro-RNAs), miR-1246, miR-23a, and miR-451in CRC patients.MethodsPeripheral expression of three miRNAs (miR-1246, miR-23a and miR-451) was investigated in sera of 37 CRC Egyptian patients and 30 healthy controls, using quantitative real-time polymerase chain reaction trying to reach the optimal non-invasive combination of miRNAs.ResultsSerum miR-1246 was up-regulated in sera of CRC patients compared to normal controls (fold change = 3.55; P<0.001) and showed 100% sensitivity and 80% specificity in diagnosis of CRC. Serum miR-451 was significantly down-regulated in CRC patients (fold change = -4.86; p= 0.014), whereas, miR-23a was down-regulated but this was not statistically significant.ConclusionUp-regulation of miR-1246 and down-regulation of miR-451 in the sera of primary CRC Egyptian patients were confirmed with high sensitivity and specificity. Large-scale studies on a wider spectrum of miRNAs in Egyptian CRC patients are needed.     

Acoustic radiation force impulse elastography in differentiating renal solid masses: a preliminary experience

New diagnostic methods are required to diagnose renal mass. Thus, we assessed virtual tissue quantification (VTQ) of acoustic radiation force impulse (ARFI) elastography in differentiation of renal solid masses. Forty-two patients with renal masses were assessed by VTQ in terms of measurement of the shear wave velocity (SWV). The masses were divided into three groups. They were clear cell carcinoma (CCC) angiomyolipoma (AML), and pseudotumor. The differences among the three groups in SWV, as well as between masses and its surrounding parenchyma, were investigated. Receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic performance. We found that the SWV among the three groups were significant different (F = 6.976, P = 0.003) and the SWV of pseudotumor (3.14 ± 0.75 m/s) was significantly higher than CCC (2.46 ± 0.45 m/s) and AML (2.49 ± 0.63 m/s) (P = 0.007 and 0.001 respectively). There were no significant difference between CCC and AML in SWV (P = 0.719). For each group, there was no significant difference between the mass and its surrounding parenchyma (P = 0.693, 0.892, and 0.714, respectively). Between pseudotumor and CCC, the optimal cut-off value of SWV for differential diagnoses was 3.07 m/s; and the area under the ROC curve (AUC) was 0.78 (95% CI: 0.560 to 0.924) (P = 0.004), the sensitivity and specificity were 100% and 58.3%, respectively. Between pseudotumor and AML, the optimal cut-off value of SWV for differential diagnoses was 3.03 m/s, thus AUC curve was 0.786 (95% CI: 0.591 to 0.918) (P = 0.002), the sensitivity and specificity were 100% and 58.3%, respectively. No significant difference was found between AML and CCC (P = 0.587) and the AUC was 0.562. To conclude, our results support that ARFI has potential value in differentiation between CCC and pseudotumor, or between AML and pseudotumor, however, it fails to make a distinction between CCC and AML.     

Efficacy of bisphosphonates in detection of early enamel caries using NIR fluorescence imaging and inhibition of caries progression

NIR fluorescence imaging using bisphosphonate-Indocyanine green has been indicated for early interproximal caries detection. This study assessed diagnostic accuracy of caries detection by NIR fluorescence imaging with OsteoSense 750^® (OS750) in vitro and ex vivo, and to analyze the therapeutic efficacy of a bisphosphonate (Etidronate) in inhibiting enamel caries progression in vitro.Methods: Four experiments were conducted using extracted human teeth; 1) to calculate the infiltration rate of OS750 into interproximal white spot lesions using fluorescence microscope, 2) to assess diagnostic accuracy of interproximal natural white spot lesions using desktop NIR fluorescence imaging device in vitro setting, 3) to assess diagnostic accuracy of artificially created deeper enamel carious lesion (0.5 mm~1.0 mm) using NIR fluorescence image through the head-mount display in ex vivo setting, 4) to compare the progression on the enamel caries lesions treated by Etidronate, NaF and distilled-water. Diagnostic accuracy was analyzed using sensitivity, specificity and receiver operating curves (ROC). The caries progression was calculated with micro-CT and was statistically analyzed using a two-way ANOVA and the Tukey HDS post-hoc test.Results: 1) The infiltration rate of OS750 was 101.83% ± 8.66 (Min: 90.10%, Max: 133.94%). 2) The average of sensitivity and specificity in vitro setting experiments were 86.7% ± 4.4% and 70% ± 11%, respectively. The average of area under the ROC curves (AUC) was 0.883 ± 0.059 indicating excellent performance. 3) The mean sensitivity and specificity in ex vivo setting was 82.97% ± 15% and 76.78% ± 13.27% respectively. 4) The carious lesion volume treated by Etidronate was significantly smaller at post treatment-1 (p<0.05) and treatment-2 (p<0.01) than the control. There was no significant difference in lesion volume in the Etidronate and NaF group at the time point of post treatment-1.Conclusion: This study suggests that bisphosphonates contribute to both early diagnosis of enamel caries and inhibition of caries progression.     

Expression of miRNAs and ZEB1 and ZEB2 correlates with histopathological grade in papillary urothelial tumors of the urinary bladder

Histopathological grading of papillary urothelial tumors (PUTs) of the urinary bladder is subjective and poorly reproducible. We investigated the relationship between the expression of frequently deregulated microRNAs (miRNAs) as well as their target genes (ZEB1/ZEB2) and bladder cancer histopathological grade in an attempt to find a miRNA that might allow more reliable grading of PUTs. We measured the expression levels of four miRNAs (miR-145, miR-205, miR-125b, and miR-200c) in 120 formalin-fixed, paraffin-embedded bladder tumor tissue samples using real-time PCR assays. ZEB1 and ZEB2 expression was assessed in the same bladder tissues by immunohistochemistry. MiR-205 distinguished low-grade papillary urothelial carcinoma (LG) from high-grade papillary urothelial carcinoma (HG), and miR-145 distinguished HG from infiltrating carcinoma (CA) with an area under the receiver operator characteristic curve (AUC) of 0.992 and 0.997, respectively (sensitivity/specificity of 95.8/96.7 % and 100/91.7 %, respectively; p?<?0.05). The expression level of miR-125b was significantly lower in LG than in PUNLMP, with an AUC value of 0.870 (93.3 % sensitivity and 84.2 % specificity; p?<?0.05). ZEB1 immunoreactivity was more frequently detected in HG than in LG (57 % vs 13 %, p?<?0.01) and in HG than in CA (57 % vs 17 %, p?<?0.01). ZEB2 immunoreactivity was more frequent in CA than in HG (83 % vs 54 %, p?<?0.05). ZEB1/ZEB2 and miRNAs expression seems to reliably distinguish between different grades of PUTs of the urinary bladder. They might well serve as useful complementary diagnostic biomarkers for grading of papillary urothelial tumors.     

Initial Abdominal CT and Laboratory Findings Prior to Diagnosis of Crohn’s Disease in Children

PurposeTo identify initial abdominal computed tomography (CT) and laboratory findings prior to a diagnosis of Crohn’s disease (CD) in children.Materials and MethodsIn this retrospective study, patients (≤18 year-old) who were diagnosed with CD from 2004 to 2019 and had abdominal CT just prior to being diagnosed with CD were included in the CD group. Patients (≤18 years old) who were diagnosed with infectious enterocolitis from 2018 to 2019 and had undergone CT prior to being diagnosed with enterocolitis were included as a control group. We assessed the diagnostic performances of initial CT and laboratory findings for the diagnosis of CD using logistic regression and the area under the curve (AUC).ResultsIn total, 107 patients (50 CD patients, 57 control patients) were included, without an age difference between groups (median 13 years old vs. 11 years old, p=0.119). On univariate logistic regression analysis, multisegmental bowel involvement, mesenteric vessel engorgement, higher portal vein/aorta diameter ratio, longer liver longitudinal diameter, lower hemoglobin (≤12.5 g/dL), lower albumin (≤4 g/dL), and higher platelet (>320×10³/µL) levels were significant factors for CD. On multivariate analysis, multisegmental bowel involvement [odds ratio (OR) 111.6, 95% confidence interval (CI) 4.778–2605.925] and lower albumin levels (OR 0.9, 95% CI 0.891–0.993) were significant factors. When these two features were combined, the AUC value was 0.985 with a sensitivity of 96% and specificity of 100% for differentiating CD.ConclusionMultisegmental bowel involvement on CT and decreased albumin levels can help differentiate CD from infectious enterocolitis in children prior to a definite diagnosis of CD.     

Associations of single nucleotide polymorphisms in precursor-microRNA (miR)-125a and the expression of mature miR-125a with the development and prognosis of autoimmune thyroid diseases

It is important to search the biomarker to predict the development and prognosis of autoimmune thyroid diseases (AITDs) such as Hashimoto''s disease (HD) and Graves'' disease (GD). MicroRNA (miR) bind directly to the 3′ untranslated region of specific target mRNAs to suppress the expression of proteins, promote the degradation of target mRNAs and regulate immune response. miR-125a is known to be a negative regulator of regulated upon activation normal T cell expressed and secreted (RANTES), interleukin (IL)-6 and transforming growth factor (TGF)-β; however, its association with AITDs remains unknown. To clarify the association between AITDs and miR-125a, we genotyped the rs12976445 C/T, rs10404453 A/G and rs12975333 G/T polymorphisms in the MIR125A gene, which encodes miR-125a, using direct sequencing and polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) methods in 155 patients with GD, 151 patients with HD and 118 healthy volunteers. We also examined the expression of miR-125a in peripheral blood mononuclear cells (PBMCs) from 55 patients with GD, 79 patients with HD and 38 healthy volunteers using quantitative real-time PCR methods. We determined that the CC genotype and C allele of the rs12976445 C/T polymorphism were significantly more frequent in patients with HD compared with control subjects (P < 0·05) and in intractable GD compared with GD in remission (P < 0·05). The expression of miR-125a was correlated negatively with age (P = 0·0010) and down-regulated in patients with GD compared with control subjects (P = 0.0249). In conclusion, miR-125a expression in PBMCs and the rs12976445 C/T polymorphism were associated with AITD development and prognosis.     

Diagnostic significance of miR-106a in gastric cancer

As one of the most common malignant tumors, gastric cancer still lacks tumor markers with enough specificity and sensitivity. Therefore the development of novel tumor markers is necessary for early diagnosis in clinics. MicroRNA (miR) has been known to be of unique expressional patterns in various tumors and may work as potential tumor markers for clinical use. This study thus explored the significance of plasma miR-106a in clinical diagnosis of gastric cancer and its effects on proliferation of cancer cells. Plasma miR-106a levels were quantified by real-time quantitative fluorescent PCR methods in 80 cases of gastric cancer patients and healthy individuals to analyze the correlation between miR-106a and clinical features. MiR-106 inhibitor was further transfected into human gastric carcinoma cells for further cell proliferation using CCK-8 approach. MiR-106a was significantly up-regulated in gastric cancer patient plasma samples compared to healthy individuals (P<0.01). The area under ROC curve was 0.895 (95% CI: 0.846~0.943). It has a specificity of 93.8% and a sensitivity of 77.5% in diagnosing gastric cancer. MiR-106a level was also correlated with cancer differentiation stage, lymph node metastasis, TNM stage and tumor size (P<0.05). The down-regulation of miR-106 in gastric carcinoma cells inhibited cell proliferation (P<0.05). MiR-106a was significantly up-regulated in gastric cancer patients and can facilitate the in vitro proliferation of tumor cells. It may work as a biological marker for gastric cancer.