Daniel Norman1 · Joey Lau1
Received: 22 March 2025 / Accepted: 18 January 2026 © The Author(s) 2026
Abstract
Background Laser capture microdissection (LCM) followed by RNA-sequencing is a powerful, widely applicable tool to analyze the transcriptome in regions of a tissue. Protocols for LCM of transplanted islets of Langerhans, particularly stem cell-derived islets (SC-islets) that have evaluated RNA quality, are lacking. This study demonstrates a robust protocol for LCM of SC-islets in multiple organ sites, generating high quality RNA.
Method SC-islets were transplanted to five organ sites in immunodeficient NOG-mice. Graft-containing organs were then sectioned, fixed in 75% ethanol, stained with the alcohol-based stain cresyl violet, and dehydrated before performing LCM. RNA was then extracted, and quality control was performed.
Results High RIN scores (RNA Integrity Number) were obtained from all organ sites, with the pancreas showing the most robust results, despite its known challenges due to high RNase content. Conversely, organs with small or dispersed grafts, such as the liver and omentum, exhibited lower RIN scores. This is likely due to the size of the dissected area correlating positively with RIN scores, potentially due to a more time-consuming LCM in these sites.
Conclusion Using this novel protocol, high-quality RNA from transplanted SC-islets can be obtained. Smaller and spread-out grafts pose a challenge in obtaining higher quality RNA, although possible.
Keywords Laser capture microdissection · Stem cell-derived islets · RNA quality · Transplantation · Type 1 diabetes
Honghong Zhang1 · Changlin Zhai1 · Huilin Hu1 · Gang Qian1 · Menghui Mao1
Received: 1 November 2025 / Accepted: 18 January 2026©The Author(s) 2026
Objective This study aimed to investigate gut microbiota composition and metabolic functions in patients with type 2 diabetes mellitus (DM) complicated by myocardial infarction (MI) and to explore potential mechanisms linking the gut microbiome to MI development.
Methods Sixty patients with DM complicated by MI and 52 patients with DM alone were initially recruited.After quality control, 29 DM +MI patients and 33 DM patients were included in the final analysis. Gut microbial profiles were characterized using shotgun metagenomic sequencing and bioinformatics analyses. Microbial diversity, composition, and gene functions were compared between groups based on KEGG, COG, and CAZy annotations.
Results Overall microbial diversity and metabolic profiles were comparable between the two groups; however, significant differences were observed in specific taxa and functional genes.Taxa enriched in the DM +MI group included Bacteroidales, Prevotellaceae, and Lachnospiraceae. In total, 510 KEGG orthology (KO) units and 21 pathways—including ABC transporters, quorum sensing, and general metabolic pathways—differed significantly between groups. Carbohydrate transport and metabolism, as well as glycoside hydrolase activity, represented the most enriched functional categories. Random forest models based on selected microbial species,KO units, and KEGG pathways achieved areas under the curve (AUCs) of0.868, 0.885, and 0.820, respectively.
Conclusion Patients with DM complicated by MI exhibit distinct gut microbial compositions and functional gene signatures compared with patients with DM alone. These microbiome-based markers may contribute to early risk stratification and provide potential targets for microbiota-focused interventions to mitigate MI risk in patients with diabetes.
Keywords Gut microbiome · Type 2 diabetes mellitus · Myocardial infarction · Metagenomics
This article is excerpted from the《Frontiers in Psychology》by Wound World
伤口世界平台生态圈,以“关爱人间所有伤口患者”为愿景,连接、整合和拓展线上和线下的管理慢性伤口的资源,倡导远程、就近和居家管理慢性伤口,解决伤口专家的碎片化时间的价值创造、诊疗经验的裂变复制、和患者的就近、居家和低成本管理慢性伤口的问题。
2019广东省医疗行业协会伤口管理分会年会
扫一扫了解详情:
任何关于疾病的建议都不能替代执业医师的面对面诊断。所有门诊时间仅供参考,最终以医院当日公布为准。
网友、医生言论仅代表其个人观点,不代表本站同意其说法,请谨慎参阅,本站不承担由此引起的法律责任。