Journal: Journal of Clinical Laboratory Analysis

Article Title: Candidate Biomarkers Associated With Circulating Tumor Cell Status in Metastatic Colorectal Cancer

doi: 10.1002/jcla.70013

Figure Lengend Snippet: The expression profiles of DEM groups with RNA‐seq data in COAD and READ datasets.

Article Snippet: Clinical information for the entire sample is provided in Table . cDNA synthesis was carried out using the miRCURY LNA miRNA RT Kit (QIAGEN GmbH, Hilden, Germany) on the SimpliAmp Thermal Cycle (Applied Biosystems, Foster City, CA), following the manufacturer's protocol.

Techniques: Expressing

Journal: Journal of Clinical Laboratory Analysis

Article Title: Candidate Biomarkers Associated With Circulating Tumor Cell Status in Metastatic Colorectal Cancer

doi: 10.1002/jcla.70013

Figure Lengend Snippet: Normalized qPCR expression of selected significantly downregulated miRNAs and their ROC curve analysis (A) hsa‐miR‐199a‐5p (B) hsa‐miR‐326 (C) hsa‐miR‐500b‐5p ( p < 0.05, AUC > 0.7).

Article Snippet: Clinical information for the entire sample is provided in Table . cDNA synthesis was carried out using the miRCURY LNA miRNA RT Kit (QIAGEN GmbH, Hilden, Germany) on the SimpliAmp Thermal Cycle (Applied Biosystems, Foster City, CA), following the manufacturer's protocol.

Techniques: Expressing

Journal: Journal of Clinical Laboratory Analysis

Article Title: Candidate Biomarkers Associated With Circulating Tumor Cell Status in Metastatic Colorectal Cancer

doi: 10.1002/jcla.70013

Figure Lengend Snippet: Comparison of microarray and qPCR results for selected three miRNAs in CTC(+)/CTC(−), CTC(−)/HIs, and mCRC/HIs groups.

Article Snippet: Clinical information for the entire sample is provided in Table . cDNA synthesis was carried out using the miRCURY LNA miRNA RT Kit (QIAGEN GmbH, Hilden, Germany) on the SimpliAmp Thermal Cycle (Applied Biosystems, Foster City, CA), following the manufacturer's protocol.

Techniques: Comparison, Microarray

Journal: Journal of Clinical Laboratory Analysis

Article Title: Candidate Biomarkers Associated With Circulating Tumor Cell Status in Metastatic Colorectal Cancer

doi: 10.1002/jcla.70013

Figure Lengend Snippet: Comparison of logFC values for the selected three miRNAs across different datasets: Microarray and qPCR analyses compare HIs and mCRC groups, while TCGA‐COAD and TCGA‐READ datasets compare HIs and CRC groups (incl. mCRC).

Article Snippet: Clinical information for the entire sample is provided in Table . cDNA synthesis was carried out using the miRCURY LNA miRNA RT Kit (QIAGEN GmbH, Hilden, Germany) on the SimpliAmp Thermal Cycle (Applied Biosystems, Foster City, CA), following the manufacturer's protocol.

Techniques: Comparison, Microarray

Journal: Frontiers in Immunology

Article Title: Hsa-miR-31 Governs T-Cell Homeostasis in HIV Protection via IFN-γ-Stat1-T-Bet Axis

doi: 10.3389/fimmu.2021.771279

Figure Lengend Snippet: miR-31 is correlated with disease progression during both acute and chronic HIV-1 infection. (A) Unsupervised clustering of the 251 miRNAs. After normalization and filtering of the microarray data, 251 miRNAs were retained for further analysis. Average linkage hierarchical clustering was performed using a centered correlation metric. Twenty-three samples from the FBD study were clustered into 2 groups: the left cluster was a mixture of elite controllers, viremic controllers and progressors; the right was mainly progressors with one exception. (B) Venn diagram showing the numbers of candidate miRNAs filtered with different criteria. The miRNAs in the lower left and right circles were generated by significance analysis of microarrays (SAM) of participants stratified by the CD4+ T cell count (<250 cells/μL vs. >450 cells/μL) and viral load (<2000 copies/mL vs. >10000 copies/mL), respectively. The identified 15 miRNA candidates were marked in red in (A) . (C) Correlation between expression levels of miR-31 and CD4+ T cell counts in HIV-1 infected individuals (FBD, former blood donor cohort). miR-31 expression was quantified by quantitative RT-PCR, and the relationship between relative level of miR-31 and CD4+ T cell count was examined by Spearman correlation (n = 50). Red dots represent patients that eventually reached the defined endpoints. (D) Kaplan-Meier survival curves of FDB patients stratified by median whole blood miR-31 level during the late phase of chronic infection. (E–G) Kaplan-Meier survival curves of another HIV patient cohort (an acute-phase prospective men who have sex with men (MSM) cohort) stratified by plasma miR-31 levels before and after infection. Absolute CD4+ T cell count below 350 cells/μL, initiation of long-term ART, progression to AIDS and death were defined as endpoints of the study. Patients were separated into two groups stratified by the median miR-31 level in plasma collected before infection (E) , during acute infection phase (F) , during early phase of chronic infection (G) .

Article Snippet: PAXgene Blood miRNA Kit (QIAGEN, Hilden, Germany) was used for extraction and purification of total RNA, including miRNA, from whole blood stabilized in PAXgene Blood RNA Tubes (PreAnalytix, BD, UK).

Techniques: Biomarker Discovery, Infection, Microarray, Generated, Cell Counting, Expressing, Quantitative RT-PCR, Clinical Proteomics

Journal: Frontiers in Immunology

Article Title: Hsa-miR-31 Governs T-Cell Homeostasis in HIV Protection via IFN-γ-Stat1-T-Bet Axis

doi: 10.3389/fimmu.2021.771279

Figure Lengend Snippet: Loss of miR-31 triggers CD4+ T cell activation. (A) miR-31 levels in different immune cell subtypes. Data were obtained from a miRNA RTqPCR data from Rossi et al ’s work (see the text for reference). (B) Comparison of absolute naïve CD4+ T cell counts in blood of HIV-1 infected individuals (FBD, n = 50) stratified by miR-31 expression. (C, D) Correlation between miR-31 levels and frequencies of CD38+ T cells (C) or HLA-DR+ T cells in blood of HIV infected individuals (FBD, n=44) (E) Gene set enrichment analysis (GSEA) of “naïve” signature in antagomiR-31- versus antagoNC- treated naïve CD4+ T cells. NES, normalized enrichment score. (F) Heatmap of representative genes associated with activation versus naïve state of T cells. Shown is log2 fold changes of gene expression in antagomiR-31-treated naïve CD4+ T cells relative to that in antagoNC-treated cells (n=3). (G) Effects of antagomiR-31 treatment on CD25 expression of naïve CD4+ T cells, assessed by frequency of CD25+ cells and median fluorescent intensity (MFI) of CD25. Average fold changes were 4.45 and 2.25, respectively (n = 8). Blue, antagomir-31 treated group; red, antagoNC-treated group. Representative FACS data for CD25 were shown in the left panel. (H) Schema of the in vitro assay used for examining the role of miR-31 in HIV-1 infection. CD4+ T cells were sorted, followed by transfection with antagomiR-31 or antagoNC. After 48 hours, cells were stimulated with a mix of anti-CD3 and anti-CD28 antibodies and infected with HIV-1 IIIB 5 days later. (I, J) Cells and supernatants were collected on days 5 and 11 post infection and respectively subjected to flow cytometry for determination of P24-expressing CD4+ T cells (I) and ELISA for quantification of released P24 proteins (J) (n = 3).

Article Snippet: PAXgene Blood miRNA Kit (QIAGEN, Hilden, Germany) was used for extraction and purification of total RNA, including miRNA, from whole blood stabilized in PAXgene Blood RNA Tubes (PreAnalytix, BD, UK).

Techniques: Activation Assay, Comparison, Infection, Expressing, Gene Expression, In Vitro, Transfection, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Toxicology Reports

Article Title: Placental microRNAs: Responders to environmental chemicals and mediators of pathophysiology of the human placenta

doi: 10.1016/j.toxrep.2020.08.002

Figure Lengend Snippet: miRNAs in the placenta that are responsive to exposure to environmental toxicants.

Article Snippet: Toxic Metals , Placenta, JEG-3 cells , N = 32 , 861 , miR-26a, miR-155 , AllPrep DNA/RNA/miRNA Univerisal Kit (Qiagen) , Human miRNA Oligo microarray (Agilent); qRT-PCR , miRNA U6 , (Brooks, Martin et al. 2016).

Techniques: Extraction, Microarray, Isolation

Journal: Oncology Letters

Article Title: Lectin-captured plasma microRNAs predict response and survival in patients with non-small cell lung cancer treated with nivolumab

doi: 10.3892/ol.2026.15669

Figure Lengend Snippet: Identification of circulating OAA1-captured miRNAs associated with lung cancer and resistance to nivolumab. (A) Lectin histochemistry using OAA1 on surgically resected lung cancer tissues (×200 magnification). The left panel shows OAA1 lectin staining in normal lung tissue, whereas the right panel shows OAA1 lectin staining in lung cancer tissue. (B) In the discovery phase, plasma samples from two patients with NSCLC with PD were collected before and after nivolumab treatment. After plasma purification using an OAA1 column, microarray analysis was performed to identify candidate miRNAs (miR-320a, miR-320b, and miR-3613-5p) associated with nivolumab resistance. In the validation phase, plasma samples from 48 patients with NSCLC (before nivolumab treatment) were analyzed with and without OAA1 column purification. The levels of the identified miRNAs were quantified using TaqMan qPCR, and statistical analyses were conducted to evaluate their predictive and prognostic values as biomarkers for nivolumab resistance. (C) Microarray analysis of OAA1-captured plasma miRNAs in patients with PD after nivolumab treatment reveals several upregulated candidate miRNAs, including miR-320a, miR-320b, and miR-3613-5p. (D) Relative levels of miR-320a, miR-320b, and miR-3613-5p in pre-treatment plasma from patients with NSCLC (n=48) and healthy controls (n=11) with and without OAA1 enrichment. Plasma miRNA levels were quantified by reverse transcription-qPCR. Relative miRNA levels were normalized to miR-16-5p and calculated using the 2 −ΔΔCq method, with the mean level of healthy volunteers serving as the calibrator. Statistical significance was determined using the Mann-Whitney U test. Data are presented as median with 95% confidence intervals. miRNA/miR, microRNA; NSCLC, non-small cell lung cancer; OAA1, Oscillatoria agardhii agglutinin 1; PD, progressive disease; qPCR, quantitative polymerase chain reaction.

Article Snippet: After thorough vortex mixing, the miRNA was purified using the NucleoSpin miRNA Plasma kit (Macherey-Nagel, Germany) with DNA digestion treatment, according to the manufacturer's protocol.

Techniques: Staining, Clinical Proteomics, Purification, Microarray, Biomarker Discovery, Reverse Transcription, MANN-WHITNEY, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Lectin-captured plasma microRNAs predict response and survival in patients with non-small cell lung cancer treated with nivolumab

doi: 10.3892/ol.2026.15669

Figure Lengend Snippet: Differential expression of miR-320a, miR-320b, and miR-3613-5p in pre-treatment plasma from patients with non-small cell lung cancer with PR or SD + PD. (A) Relative levels of miR-320a, miR-320b, and miR-3613-5p without OAA1 enrichment. (B) With OAA1 column enrichment. Plasma miRNA levels were quantified by reverse transcription-quantitative polymerase chain reaction and are shown as relative abundance using the 2 −ΔCq method normalized to miR-16-5p. Statistical significance was determined using the Mann-Whitney U test. Data are presented as median with 95% confidence intervals. miR, microRNA; OAA1, Oscillatoria agardhii agglutinin 1; SD, stable disease; PD, progressive disease; PR, partial response.

Article Snippet: After thorough vortex mixing, the miRNA was purified using the NucleoSpin miRNA Plasma kit (Macherey-Nagel, Germany) with DNA digestion treatment, according to the manufacturer's protocol.

Techniques: Quantitative Proteomics, Clinical Proteomics, Reverse Transcription, Real-time Polymerase Chain Reaction, MANN-WHITNEY

Journal: Oncology Letters

Article Title: Lectin-captured plasma microRNAs predict response and survival in patients with non-small cell lung cancer treated with nivolumab

doi: 10.3892/ol.2026.15669

Figure Lengend Snippet: ROC curve analysis of miRNAs for predicting non-response to immune checkpoint inhibitor therapy in patients with non-small cell lung cancer treated with nivolumab. (A) ROC curves for circulating miR-320a, miR-320b, and miR-3613-5p without OAA1 column enrichment. (B) With OAA1 enrichment. AUC, area under the curve; CI, confidence interval; miR, microRNA; OAA1, Oscillatoria agardhii agglutinin 1; ROC, receiver operating characteristic.

Article Snippet: After thorough vortex mixing, the miRNA was purified using the NucleoSpin miRNA Plasma kit (Macherey-Nagel, Germany) with DNA digestion treatment, according to the manufacturer's protocol.

Techniques:

Journal: Oncology Letters

Article Title: Lectin-captured plasma microRNAs predict response and survival in patients with non-small cell lung cancer treated with nivolumab

doi: 10.3892/ol.2026.15669

Figure Lengend Snippet: Kaplan-Meier analysis for OS based on pre-treatment plasma levels of miR-320a, miR-320b, and miR-3613-5p in patients with non-small cell lung cancer treated with nivolumab (n=48). (A) Kaplan-Meier curves comparing the OS between the high- and low-expression groups for each miRNA without OAA1 column enrichment. (B) Kaplan-Meier curves for the same miRNAs with the OAA1 enrichment. The number of patients in each group is indicated in the legends. Cutoff values for high/low expression were determined using receiver operating characteristic analysis. P-values are shown; *P<0.05. miR, microRNA; OAA1, Oscillatoria agardhii agglutinin 1; OS, overall survival.

Article Snippet: After thorough vortex mixing, the miRNA was purified using the NucleoSpin miRNA Plasma kit (Macherey-Nagel, Germany) with DNA digestion treatment, according to the manufacturer's protocol.

Techniques: Clinical Proteomics, Expressing

Journal: Biomedical reports

Article Title: Lipolysis by downregulating miR-92a activates the Wnt/β-catenin signaling pathway in hypoxic rats.

doi: 10.3892/br.2020.1340

Figure Lengend Snippet: Figure 1. Analysis of miR‑92a expression in the epididymal fat of hypoxic rats. (A) B3L, B6L and B9L were from normoxic rats. (B) D2L, D6L, and D9L were from hypoxic rats. (C) Relative expression of miRNAs in the normoxic and hypoxic rats. miR‑92a expression levels in the hypoxic rats were reduced compared with the normoxic rats based on the miRNA micro array analysis. miR, microRNA; Rno, Rattus norvegicus.

Article Snippet: The following reagents and instruments were used: miRcute miRNA Isolation kit (Tiangen Biotech Co., Ltd.), miRcute miRNA cDNA First Strand Synthesis kit (Tiangen Biotech Co., Ltd.), miRcute MiRNA Quantitative Fluorescence Detection kit (cat. no. FP401; Tiangen Biotech Co., Ltd.), SuperReal PreMix SYBR‐Green (cat. no. FP204; Tiangen Biotech Co., Ltd.), TIANScript II cDNA First Strand Synthesis kit (cat. no. KR107; Tiangen Biotech Co., Ltd.) and RT‐qPCR amplifier (BIOER FQD‐96A; Hangzhou Bioer Co., Ltd.).

Techniques: Expressing, Microarray

Journal: Biomedical reports

Article Title: Lipolysis by downregulating miR-92a activates the Wnt/β-catenin signaling pathway in hypoxic rats.

doi: 10.3892/br.2020.1340

Figure Lengend Snippet: Figure 3. Analysis of miR‑92a binding with Fzd10. (A) Wild‑type and mutant‑type binding sequences of miR‑92a with Fzd10. (B) Dual luciferase reporter assay. Wild‑type miR‑92a bound and degraded wild‑type Fzd10 mRNA in the Fzd10+miR92a group. Single Fzd10 or mutated Fzd10 mRNA did not result in altered fluorescence values of the empty carrier pRL‑TK, and miR‑92a was not significantly altered the fluorescence values of the mutated Fzd10 mRNA in the mFzd10+miR92a group. miR/miRNA, microRNA; Fzd10, frizzled 10; mFzd10, mutated Fzd10.

Article Snippet: The following reagents and instruments were used: miRcute miRNA Isolation kit (Tiangen Biotech Co., Ltd.), miRcute miRNA cDNA First Strand Synthesis kit (Tiangen Biotech Co., Ltd.), miRcute MiRNA Quantitative Fluorescence Detection kit (cat. no. FP401; Tiangen Biotech Co., Ltd.), SuperReal PreMix SYBR‐Green (cat. no. FP204; Tiangen Biotech Co., Ltd.), TIANScript II cDNA First Strand Synthesis kit (cat. no. KR107; Tiangen Biotech Co., Ltd.) and RT‐qPCR amplifier (BIOER FQD‐96A; Hangzhou Bioer Co., Ltd.).

Techniques: Binding Assay, Luciferase, Reporter Assay, Fluorescence