Journal: Cells

Article Title: The Effect of Ursodeoxycholic Acid (UDCA) on Serum Expression of miR-34a and miR-506 in Patients with Chronic Cholestatic Liver Diseases

doi: 10.3390/cells14151137

Figure Lengend Snippet: Basal serum expression levels of miR-34a and miR-506 in patients with PBC and PSC before the introduction of UDCA treatment. miR-34a ( A ) expression was significantly elevated in PBC patients compared to age- and sex-matched healthy controls, while no significant difference was observed in PSC patients due to high intra-group variability. miR-506 ( B ) expression was markedly elevated—by several-dozen-fold—in PBC patients relative to controls. Comparisons between experimental groups were performed using the Mann–Whitney U test. Data are presented as mean ± SEM. Abbreviations: PBC, primary biliary cholangitis; PSC, primary sclerosing cholangitis; UDCA, ursodeoxycholic acid.

Article Snippet: The levels of miR-34a (478048_mir) and miR-506 (478958_mir), along with miR-16 (477860_mir), which was used as an endogenous control, were measured using TaqMan ® Advanced miRNA Assays (Applied Biosystems).

Techniques: Expressing, MANN-WHITNEY

Journal: Cells

Article Title: The Effect of Ursodeoxycholic Acid (UDCA) on Serum Expression of miR-34a and miR-506 in Patients with Chronic Cholestatic Liver Diseases

doi: 10.3390/cells14151137

Figure Lengend Snippet: The effect of UDCA treatment on the serum expression of miR-34a and miR-506. UDCA suppressed the expression of miR-34a ( A ) and miR-506 ( C ) in patients with PBC but not PSC ( B ). Each symbol represents one patient. A Student’s paired t -test was used to test statistical significance. The median duration of UDCA administration was 44 months (range: 22–56 months). PBC, primary biliary cholangitis; PSC, primary sclerosing cholangitis; UDCA naive, values before the introduction of ursodeoxycholic acid; UDCA, after treatment with UDCA.

Article Snippet: The levels of miR-34a (478048_mir) and miR-506 (478958_mir), along with miR-16 (477860_mir), which was used as an endogenous control, were measured using TaqMan ® Advanced miRNA Assays (Applied Biosystems).

Techniques: Expressing

Journal: Cells

Article Title: The Effect of Ursodeoxycholic Acid (UDCA) on Serum Expression of miR-34a and miR-506 in Patients with Chronic Cholestatic Liver Diseases

doi: 10.3390/cells14151137

Figure Lengend Snippet: Effects of UDCA on LPS-induced expression of TREM-2, miR-34a, and ADAM17 in NHC cell lines. qPCR analysis demonstrated that TREM-2 ( A ) was not induced by LPS stimulation alone; however, co-treatment with UDCA resulted in activation of this gene. The LPS-induced expression of miR-34a ( B ) and ADAM17 ( C ) mRNA was significantly suppressed by co-treatment with UDCA. Data are presented as mean ± SEM from four independent experiments. Statistical analysis was performed using two-way ANOVA followed by Fisher’s least significant difference (LSD) test. Abbreviations: ADAM17, a disintegrin and metalloprotease 17; LPS, lipopolysaccharide from Escherichia coli; NHC, normal human cholangiocytes; TREM-2, triggering receptor expressed on myeloid cells 2; UDCA, ursodeoxycholic acid.

Article Snippet: The levels of miR-34a (478048_mir) and miR-506 (478958_mir), along with miR-16 (477860_mir), which was used as an endogenous control, were measured using TaqMan ® Advanced miRNA Assays (Applied Biosystems).

Techniques: Expressing, Activation Assay

Journal: International Journal of Molecular Sciences

Article Title: Expression Analysis of Circulating miR-21, miR-34a and miR-122 and Redox Status Markers in Metabolic Dysfunction-Associated Steatotic Liver Disease Patients with and Without Type 2 Diabetes

doi: 10.3390/ijms26062392

Figure Lengend Snippet: Expressions of miR-21 ( A ), miR-34a ( B ) and miR-122 ( C ) in the studied population. miRNA expression levels are presented as relative expression values calculated using the 2 −ΔCt method, where ΔCt is determined as the difference between the Ct value of the miRNA of interest and the Ct value of the exogenous control for each sample. Expression levels of all three miRNAs are adjusted for age, sex, BMI, physical activity, antihypertensives and/or CVD therapy. Statistical comparisons between groups are indicated by p -values, with significant differences ( p < 0.05) highlighted in red.

Article Snippet: Reverse transcription (RT) and real-time PCR (qPCR) were performed using the 7500 Real-Time PCR System (Thermo Fischer Scientific, Waltham, MA, USA). cDNA synthesis and qPCR reactions were performed using the TaqMan Advanced miRNA cDNA Synthesis Kit (Thermo Fisher Scientific, Waltham, MA, USA), HOT FIREPol ® Probe qPCR Mix Plus (Solis Biodyne, Tartu, Estonia) and TaqMan Advanced miRNA Assays: 477975_mir for miR-21-5p, 478048_mir for miR-34a-5p, and 477855_mir for miR-122-5p (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s protocol. cDNA synthesis included a poly(A) tailing reaction (37 °C for 45 min, 65 °C for 10 min, pause at 4 °C), an adaptor ligation reaction (16 °C for 60 min, pause at 4 °C), a reverse transcription (RT) reaction (42 °C for 15 min, 85 °C for 5 min, pause at 4 °C), and miRNA amplification (95 °C for 5 min, 14 cycles at two temperatures—95 °C for 3 s and 60 °C for 30 s, 99 °C for 10 min, hold at 4 °C).

Techniques: Expressing, Control, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Expression Analysis of Circulating miR-21, miR-34a and miR-122 and Redox Status Markers in Metabolic Dysfunction-Associated Steatotic Liver Disease Patients with and Without Type 2 Diabetes

doi: 10.3390/ijms26062392

Figure Lengend Snippet: ROC analysis of miR-21 discriminatory power for T2D in MASLD. Abbreviations: AUC, area under the curve; CI, confidence interval; SE, standard error.

Article Snippet: Reverse transcription (RT) and real-time PCR (qPCR) were performed using the 7500 Real-Time PCR System (Thermo Fischer Scientific, Waltham, MA, USA). cDNA synthesis and qPCR reactions were performed using the TaqMan Advanced miRNA cDNA Synthesis Kit (Thermo Fisher Scientific, Waltham, MA, USA), HOT FIREPol ® Probe qPCR Mix Plus (Solis Biodyne, Tartu, Estonia) and TaqMan Advanced miRNA Assays: 477975_mir for miR-21-5p, 478048_mir for miR-34a-5p, and 477855_mir for miR-122-5p (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s protocol. cDNA synthesis included a poly(A) tailing reaction (37 °C for 45 min, 65 °C for 10 min, pause at 4 °C), an adaptor ligation reaction (16 °C for 60 min, pause at 4 °C), a reverse transcription (RT) reaction (42 °C for 15 min, 85 °C for 5 min, pause at 4 °C), and miRNA amplification (95 °C for 5 min, 14 cycles at two temperatures—95 °C for 3 s and 60 °C for 30 s, 99 °C for 10 min, hold at 4 °C).

Techniques: