scholarly journals Is MicroRNA-127 a Novel Biomarker for Acute Pancreatitis with Lung Injury?

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Na Shi ◽  
Lihui Deng ◽  
Weiwei Chen ◽  
Xiaoxin Zhang ◽  
Ruijie Luo ◽  
...  
Keyword(s):  
Acute Pancreatitis ◽  
Respiratory Failure ◽  
Lung Injury ◽  
Healthy Volunteers ◽  
Sodium Taurocholate ◽  
Control Group ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Potential Marker ◽  
Polymerase Chain

Background and Aims. The aim of this study was to determine the expression of microRNA-127 (miR-127) in both rat models and patients of acute pancreatitis (AP) with lung injury (LI). Methods. Rats were administrated with retrograde cholangiopancreatography injection of 0.5% or 3.5% sodium taurocholate to induce AP with mild or severe LI and were sacrificed at 6, 12, and 24 h. Rats from the control group received a laparotomy only. Plasma from a prospective cohort of AP patients was collected. The levels of miR-127 in the tissues and plasma were detected using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Results. The upregulation of miR-127 in the lungs of rats was detected in the groups of AP with severe LI at 6 h and 24 h, whereas it was scarcely detectable in plasma. In the pilot study that included 18 AP patients and 5 healthy volunteers, the plasma miR-127 level was significantly downregulated in AP patients with respiratory failure compared with the healthy volunteers (P=0.014) and those without respiratory failure (P=0.043). Conclusion. miR-127 might serve as a potential marker for the identification of AP with LI.

scholarly journals The Expression of miR-23a and miR-146a in the Saliva of Patients with Periodontitis and Its Clinical Significance

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Lanhua Kang ◽  
Ning Li ◽  
Lexiu Wang
Keyword(s):  
Area Under The Curve ◽  
Inflammatory Factors ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Interleukin 17 ◽  
Study Group ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Polymerase Chain ◽  
Basic Treatment

Background. This study is aimed at exploring the significance of the expression of miR-23a and miR-146a in patients with periodontitis and their correlations with inflammatory factors. Methods. A total of 120 patients with chronic periodontitis admitted to the department of stomatology in Yantai Yuhuangding Hospital from August 2017 to December 2018 were enrolled as a study group, and 80 healthy volunteers in physical examination during the same period were enrolled as a control group. The expression of miR-23a, miR-146a, interleukin-1β (IL-1β), interleukin-6 (IL-6), and interleukin-17 (IL-17) in the saliva of people in the two groups was determined using the quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). Results. The study group showed significantly higher relative expression of saliva miR-23a and miR-146a than the control group. The area under the curve (AUC) of saliva miR-23a and miR-146a for diagnosing periodontitis was 0.857 and 0.886, respectively. The expression of saliva miR-23a and miR-146a increased with the deterioration of periodontitis in the patients. After basic treatment, the study group showed significantly decreased expression of saliva miR-23a and miR-146a. Patients in the study group showed significantly higher levels of saliva IL-1β, IL-6, and IL-17 than those in the control group, and their saliva miR-23a and miR-146a were positively correlated with their saliva IL-1β, IL-6, and IL-17, respectively. Conclusion. Saliva miR-23a and miR-146a can be used as biomarkers for the diagnosis and assessment of periodontitis, and they may have regulatory relationships with IL-1β, IL-6, and IL-17.

scholarly journals Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Qrt Pcr ◽  
Highly Sensitive ◽  
One Step ◽  
Polymerase Chain

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.

Metformin Attenuates the Metabolic Disturbance and Depression-like Behaviors Induced by Corticosterone and Mediates the Glucose Metabolism Pathway

2021 ◽  
Author(s):  
Yong Hao ◽  
Yingpeng Tong ◽  
Yanhong Guo ◽  
Xiaoe Lang ◽  
Xinxin Huang ◽  
...  
Keyword(s):  
Glucose Metabolism ◽  
Tricarboxylic Acid Cycle ◽  
Antidepressant Activity ◽  
Tricarboxylic Acid ◽  
Serum Glucose ◽  
Metabolomic Analysis ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Metabolism Pathway ◽  
Polymerase Chain

Abstract Background Metabolism disturbances are common in patients with depression. The drug metformin has been reported to exhibit antidepressant activity. The purpose of this study was to investigate metabolism disturbances induced by corticosterone (CORT) and determine if metformin can reverse these effects and their accompanying depression-like behaviors. Methods Rats were exposed to corticosterone with or without metformin administration. Depression-like behaviors were tested. Gene expression was confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis. In addition, the metabolites were quantified by LC-MS/MS analysis. Results Metformin attenuated the depression-like behaviors induced by CORT. Furthermore, metformin reversed disturbances in body weight, serum glucose, and triglyceride levels, as well as hepatic TG levels induced by CORT. Metformin normalized the alterations in the expression of glucose metabolism-related genes (PGC-1α, G6pc, Pepck, Gck, PYGL, Gys2, PKLR, GLUT4) and insulin resistance-related genes (AdipoR1, AdipoR2) in the muscles and livers of rats induced by CORT. Metabolomic analysis showed that metformin reversed the effects of CORT on 11 metabolites involved in the pathways of the tricarboxylic acid cycle, glycolysis, and gluconeogenesis (3-phospho-D-glycerate, β-D-fructose 6-phosphate, D-glucose 6-phosphate, and pyruvate). Conclusion Our findings suggest that metformin can attenuate metabolism disturbances and depression-like behaviors induced by CORT mediating the glucose metabolism pathway.

Quantitative PCR Analysis for Bcl-2/IgH in a Phase III Study of Yttrium-90 Ibritumomab Tiuxetan As Consolidation of First Remission in Patients With Follicular Lymphoma

2009 ◽  
Vol 27 (36) ◽  
pp. 6094-6100 ◽  
Author(s):  
Lindsey Goff ◽  
Karin Summers ◽  
Sameena Iqbal ◽  
Jens Kuhlmann ◽  
Michael Kunz ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Follicular Lymphoma ◽  
Quantitative Polymerase Chain Reaction ◽  
Phase Iii ◽  
Pcr Analysis ◽  
Control Group ◽  
Ibritumomab Tiuxetan ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Yttrium 90

Purpose The randomized First-Line Indolent Trial (FIT) was conducted in patients with advanced follicular lymphoma (FL), to evaluate the safety and efficacy of yttrium-90 (90Y) ibritumomab tiuxetan given as consolidation of complete or partial remission. This study of minimal residual disease was undertaken in parallel, to determine the rate of conversion from bcl-2 polymerase chain reaction (PCR) –detectable to –undetectable status and the corresponding effect on progression-free survival (PFS). Patients and Methods Blood samples from 414 patients (90Y-ibritumomab, n = 208; control, n = 206) were evaluated using real-time quantitative polymerase chain reaction (RQ-PCR); 186 were found to have the bcl-2 rearrangement and were thus eligible for inclusion in the RQ-PCR analysis. Results Overall, 90% of treated patients converted from bcl-2 PCR–detectable to –undetectable disease status, compared with 36% in the control group. Treatment significantly prolonged median PFS in patients converting to bcl-2 PCR-undetectable status (40.8 v 24.0 months in the control group; P < .01, hazard ratio [HR], 0.399). In patients who had bcl-2 PCR-detectable disease at random assignment, treatment significantly prolonged median PFS (38.4 v 8.2 months in the control group; P < .01, HR, 0.293). Conclusion Eradication of PCR-detectable disease occurred more frequently after treatment with 90Y-ibritumomab tiuxetan and was associated with prolongation of PFS.

scholarly journals Pilot study of DNA methylation in the pathogenesis of chronic rhinosinusitis with nasal polyps

2015 ◽  
Vol 53 (4) ◽  
pp. 345-352
Author(s):  
Y.B. Zheng ◽  
Y. Zhao ◽  
L.Y. Yue ◽  
P. Lin ◽  
Y.F. Liu ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Polymerase Chain Reaction ◽  
Chronic Rhinosinusitis ◽  
Nasal Polyps ◽  
Cpg Islands ◽  
Inferior Turbinate ◽  
Control Group ◽  
Chain Reaction ◽  
Bisulphite Sequencing ◽  
Polymerase Chain

Background: DNA methylation has been implicated in the pathogenesis of allergy and atopy. This study aimed to identify whether DNA methylation also plays an important role in the pathogenesis of nasal polyps (NP). Methodology: NP tissues were obtained from 32 patients with chronic rhinosinusitis with bilateral NP. Biopsies of inferior turbinate mucosa (ITM) were taken from 18 patients who underwent rhinoseptoplasty (control group). The methylated genes, which were detected by DNA methylation microarray, were validated by methylation-specific polymerase chain reaction, bisulphite sequencing, real-time polymerase chain reaction and immunohistochemistry. Results: DNA methylation microarray identified 8,008 CpG islands in 2,848 genes. One hundred and ninety-eight genes were found to have a methylated signal in the promoter region in NP samples compared with ITM samples. The four top genes that changed, COL18A1, EP300, GNAS and SMURF1, were selected for further study. The methylation frequency of COL18A1 was significantly higher in NP samples than in ITM samples. Conclusions: DNA methylation might play an important role in the pathogenesis of NP. Promoter methylation of COL18A1 was found to be significantly increased in NP tissues, further studies are necessary to confirm the significance of these epigenetic factors in the mechanisms underlying the development or persistence of NP.

scholarly journals Long non-coding RNA H19 deficiency ameliorates bleomycin-induced pulmonary inflammation and fibrosis

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Xiaoyu Wan ◽  
Xinbei Tian ◽  
Jun Du ◽  
Ying Lu ◽  
Yongtao Xiao
Keyword(s):  
Pulmonary Fibrosis ◽  
Pulmonary Inflammation ◽  
Chain Reaction ◽  
Potential Therapeutic Target ◽  
Qrt Pcr ◽  
Stat3 Signaling ◽  
Non Coding Rna ◽  
Poor Understanding ◽  
Polymerase Chain ◽  
Long Non Coding Rna

Abstract Background The poor understanding of pathogenesis in idiopathic pulmonary fibrosis (IPF) impaired development of effective therapeutic strategies. The aim of the current study is to investigate the roles of long non-coding RNA H19 (lncRNA H19) in the pulmonary inflammation and fibrosis of IPF. Methods Bleomycin was used to induce pulmonary inflammation and fibrosis in mice. The mRNAs and proteins expression in lung tissues was determined by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. H19 knockout (H19−/−) mice were generated by CRISPR/Cas9. Results The expression of H19 mRNA was up-regulated in fibrotic lungs patients with IPF as well as in lungs tissues that obtained from bleomycin-treated mice. H19−/− mice suppressed bleomycin-mediated pulmonary inflammation and inhibited the Il6/Stat3 signaling. H19 deficiency ameliorated bleomycin-induced pulmonary fibrosis and repressed the activation of TGF-β/Smad and S1pr2/Sphk2 in the lungs of bleomycin-treated mice. Conclusions Our data suggests that H19 is a profibrotic lncRNA and a potential therapeutic target for IPF.

scholarly journals Quantitation of 35S Promoter in Maize DNA Extracts from Genetically Modified Organisms Using Real-Time Polymerase Chain Reaction, Part 2: Interlaboratory Study

2005 ◽  
Vol 88 (2) ◽  
pp. 558-573 ◽  
Author(s):  
Max Feinberg ◽  
Sophie Fernandez ◽  
Sylvanie Cassard ◽  
Chrystèle Charles-Delobel ◽  
Yves Bertheau ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Interlaboratory Study ◽  
Performance Criteria ◽  
Tolerance Interval ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Polymerase Chain

Abstract The European Committee for Standardization (CEN) and the European Network of GMO Working Laboratories have proposed development of a modular strategy for stepwise validation of complex analytical techniques. When applied to the quantitation of genetically modified organisms (GMOs) in food products, the instrumental quantitation step of the technique is separately validated from the DNA extraction step to better control the sources of uncertainty and facilitate the validation of GMO-specific polymerase chain reaction (PCR) tests. This paper presents the results of an interlaboratory study on the quantitation step of the method standardized by CEN for the detection of a regulatory element commonly inserted in GMO maize-based foods. This is focused on the quantitation of P35S promoter through using the quantitative real-time PCR (QRT-PCR). Fifteen French laboratories participated in the interlaboratory study of the P35S quantitation operating procedure on DNA extract samples using either the thermal cycler ABI Prism® 7700 (Applied Biosystems, Foster City, CA) or Light Cycler® (Roche Diagnostics, Indianapolis, IN). Attention was focused on DNA extract samples used to calibrate the method and unknown extract samples. Data were processed according to the recommendations of ISO 5725 standard. Performance criteria, obtained using the robust algorithm, were compared to the classic data processing after rejection of outliers by the Cochran and Grubbs tests. Two laboratories were detected as outliers by the Grubbs test. The robust precision criteria gave values between the classical values estimated before and after rejection of the outliers. Using the robust method, the relative expanded uncertainty by the quantitation method is about 20% for a 1% Bt176 content, whereas it can reach 40% for a 0.1% Bt176. The performances of the quantitation assay are relevant to the application of the European regulation, which has an accepted tolerance interval of about ±50%. These data were fitted to a power model (r2 = 0.96). Thanks to this model, it is possible to propose an estimation of uncertainty of the QRT-PCR quantitation step and an uncertainty budget depending on the analytical conditions.

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