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MicroRNAs (miRNAs) play crucial roles in regulating various biological processes, including parasite development and host\u2013pathogen interactions, but their functions in schistosomes remain largely unexplored. This study aimed to investigate the differentially expressed miRNAs (DEMs) between mated male (MM) and single-sex male (SM) worms to gain insights into their regulatory roles in schistosome reproduction. Total RNA was extracted from 28-day-old MM and SM worms, followed by small RNA sequencing to identify DEMs. Bioinformatics analyses were used to predict the biological functions of DEM target genes. Comparative analysis with previously published miRNA datasets helped identify potentially significant miRNAs. Quantitative PCR (qPCR) validated the expression of selected miRNAs and mRNA levels of some target genes. A total of 20 DEMs were identified, with 9 upregulated in MM worms and 11 in SM worms. These DEMs may regulate processes such as intracellular transport, RNA processing, and cellular homeostasis. The study provides novel insights into the biological differences between SM and MM worms, suggesting that these miRNAs could be key regulators of parasite development and host adaptation, with potential diagnostic and therapeutic applications in schistosomiasis.<\/jats:p>\n          <jats:p>\n            <jats:bold>Graphical abstract<\/jats:bold>\n          <\/jats:p>","DOI":"10.1007\/s00436-025-08489-x","type":"journal-article","created":{"date-parts":[[2025,4,24]],"date-time":"2025-04-24T08:36:19Z","timestamp":1745483779000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Comparative microRNAs profile of Schistosoma japonicum male worms derived from single-sex and bisexual infections: Implications of the multifunctional role of microRNA"],"prefix":"10.1007","volume":"124","author":[{"given":"Haoran","family":"Zhong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danlin","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bowen","family":"Dong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luobin","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ke","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiqiang","family":"Fu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinming","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guiquan","family":"Guan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yamei","family":"Jin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,4,24]]},"reference":[{"issue":"7","key":"8489_CR1","doi-asserted-by":"publisher","first-page":"e0003965","DOI":"10.1371\/journal.pntd.0003965","volume":"9","author":"P Cai","year":"2015","unstructured":"Cai P, Gobert GN, You H, Duke M, McManus DP (2015) Circulating miRNAs: potential novel biomarkers for hepatopathology progression and diagnosis of schistosomiasis japonica in two murine models. 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