{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:17:41Z","timestamp":1785406661910,"version":"3.56.0"},"reference-count":46,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T00:00:00Z","timestamp":1781654400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32270545"],"award-info":[{"award-number":["32270545"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32070508"],"award-info":[{"award-number":["32070508"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["32270545"],"award-info":[{"award-number":["32270545"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"award":["32070508"],"award-info":[{"award-number":["32070508"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Open Project Funding of Shanghai Key Laboratory of Forensic Medicine","award":["KF202402"],"award-info":[{"award-number":["KF202402"]}]},{"DOI":"10.13039\/501100004772","name":"Natural Science Foundation of Ningxia","doi-asserted-by":"publisher","award":["2025AAC031004"],"award-info":[{"award-number":["2025AAC031004"]}],"id":[{"id":"10.13039\/501100004772","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Priority Academic Program Development of Jiangsu Higher Education"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>Phormia regina (Meigen, 1826) (Diptera: Calliphoridae), a necrophagous blow fly of significant forensic value, plays a crucial role in postmortem interval (PMI) estimation. The intra-puparial period of this species constitutes approximately half of its entire immature developmental duration. Therefore, precise aging of intra-puparial specimens is essential to improve the accuracy of PMI estimation. This study was performed at a constant 25 \u00b0C, examining intra-puparial morphology, histological sections, and body weight across different developmental stages. Additionally, full-length transcriptome profiling was conducted via a hybrid sequencing strategy combining PacBio single-molecule real-time (SMRT) sequencing and Illumina short-read next-generation sequencing (NGS). Morphological studies revealed the intra-puparial morphological characteristics, the reconstruction process of tissues and organs, and the continuous body weight loss during the intra-puparial period. Transcriptomic research identified a total of 425,349 full-length non-chimeric (FLNC) sequences. After redundancy removal, 84,852 transcript sequences were obtained, of which 46,325 transcripts were annotated. Using day-0 puparia (D0) as the control, differentially expressed gene (DEG) analysis was performed on samples from subsequent developmental stages (D1\u2013D5), identifying 4242, 7964, 9509, 10,526, and 10,011 DEGs, respectively. Results from GO enrichment and KEGG pathway analyses provided reasonable explanations for the behavioral traits of different developmental stages. Eight candidate genes for intra-puparial development were screened, most of which were highly expressed at different time points during the intra-puparial period. Quantitative real-time PCR (qRT-PCR) results showed consistent gene expression trends with the RNA-seq data, confirming the reliability of the RNA-seq results. This study provides key morphological and molecular markers for P. regina in forensic PMI estimation.<\/jats:p>","DOI":"10.3390\/insects17060642","type":"journal-article","created":{"date-parts":[[2026,6,18]],"date-time":"2026-06-18T00:49:42Z","timestamp":1781743782000},"page":"642","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrated Morphological and Transcriptomic Analyses of Gene Regulatory Mechanisms in Different Intra-Puparial Developmental Stages of Phormia regina"],"prefix":"10.3390","volume":"17","author":[{"given":"Jiani","family":"Yang","sequence":"first","affiliation":[{"name":"Department of Forensic Medicine, Soochow University, Ganjiang East Road, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruonan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Forensic Medicine, Soochow University, Ganjiang East Road, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui","family":"Zhu","sequence":"additional","affiliation":[{"name":"Criminal Police Branch, Shizuishan Public Security Bureau, Shengli East Road, Shizuishan 753299, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lan","family":"Gao","sequence":"additional","affiliation":[{"name":"Criminal Police Branch, Shizuishan Public Security Bureau, Shengli East Road, Shizuishan 753299, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenbin","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biodiversity and Biosafety, Nanjing Institute of Environmental Sciences, Ministry of Ecology and Environment, Jiangwangmiao Street, Nanjing 210042, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiya","family":"Gu","sequence":"additional","affiliation":[{"name":"Department of Forensic Medicine, Soochow University, Ganjiang East Road, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7097-6525","authenticated-orcid":false,"given":"Yu","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Forensic Medicine, Soochow University, Ganjiang East Road, Suzhou 215000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e25","DOI":"10.1016\/j.forsciint.2010.07.013","article-title":"Forensic entomology in Kuwait: The first case report","volume":"206","year":"2011","journal-title":"Forensic Sci. 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