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This study aimed to identify and validate circulating microRNAs (miRNAs) as novel biomarkers for GDM.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>A two-phase cross-sectional study enrolled 55 GDM patients and 55 matched healthy pregnant controls. In the discovery phase, RNA sequencing (RNA-seq) of peripheral blood RNA from a randomly selected subset (5 GDM patients, 5 controls) identified differentially expressed miRNAs. The validation phase employed reverse transcription and quantitative PCR (qPCR) in the entire cohort to confirm identified miR-326 and miR-532-3p expression. Bioinformatics analyses (Gene Ontology [GO] and Kyoto Encyclopedia of Genes and Genomes [KEGG]) investigated their functional roles using a consensus-based approach across multiple databases.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      RNA-seq revealed significant upregulation of miR-326 (mean normalized counts\u2009=\u200914.59,\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.05; log\n                      <jats:sub>2<\/jats:sub>\n                      [Fold Change]\u2009=\u20090.93,\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.01) and miR-532-3p (mean normalized counts\u2009=\u200946.45,\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.05; log\n                      <jats:sub>2<\/jats:sub>\n                      [Fold Change]\u2009=\u20090.66,\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.01) in GDM patients, a finding corroborated by cross-referencing with the Gene Expression Omnibus (GEO) database. qPCR validation confirmed significantly higher expression for both miRNAs in GDM (\n                      <jats:italic>P<\/jats:italic>\n                      \u2009&lt;\u20090.001), with strong negative correlations to fasting, 1-hour and 2-hour postprandial glucose levels. Receiver operating characteristic (ROC) curve analysis revealed excellent diagnostic performance (area under the curve [AUC]: 0.95 [95% CI: 0.91\u20130.99] for miR-326 and 0.96 [95% CI: 0.93\u20130.99] for miR-532-3p), robustly confirmed by bootstrap resampling. Functional analyses linked these miRNAs to phosphoinositide 3-kinase (PI3K)\/Akt and Rap1 signaling pathways.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>This study provides compelling evidence for circulating miR-326 and miR-532-3p as a synergistic pair of biomarkers for GDM. Their high diagnostic accuracy and mechanistic insights into metabolic dysregulation position them as a promising complementary tool for early GDM risk assessment and for understanding GDM pathophysiology.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s40618-026-02829-z","type":"journal-article","created":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T08:48:49Z","timestamp":1770281329000},"page":"1635-1650","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Circulating miR-326 and miR-532-3p: novel biomarkers for early gestational diabetes mellitus detection with high diagnostic accuracy"],"prefix":"10.1007","volume":"49","author":[{"given":"Hui-Zhen","family":"Lin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhi-Kun","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun-Ju","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhi-Shen","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin-Lin","family":"Tan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheng","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shi-Han","family":"Ruan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bing-Yi","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ze-Fan","family":"Ruan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao-Sheng","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi-Guang","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2138-888X","authenticated-orcid":false,"given":"Long","family":"Xie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,2,5]]},"reference":[{"issue":"1","key":"2829_CR1","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1038\/s41572-019-0098-8","volume":"5","author":"HD McIntyre","year":"2019","unstructured":"McIntyre HD, Catalano P, Zhang C, Desoye G, Mathiesen ER, Damm P (2019) Gestational diabetes mellitus. 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