Background: Papillary thyroid carcinoma (PTC) is the most common malignant endocrine tumour, and its incidence and prevalence are increasing considerably. Cellular heterogeneity in the tumour microenvironment is important for PTC prognosis. Spatial transcriptomics is a powerful technique for cellular heterogeneity study.Methods: In conjunction with a clinical pathologist identification method, spatial transcriptomics was employed to characterise the spatial location and RNA profiles of PTC‐associated cells within the tissue sections. The spatial RNA‐clinical signature genes for each cell type were extracted and applied to outlining the distribution regions of specific cells on the entire section. The cellular heterogeneity of each cell type was further revealed by ContourPlot analysis, monocle analysis, trajectory analysis, ligand–receptor analysis and Gene Ontology enrichment analysis.Results: The spatial distribution region of tumour cells, typical and atypical follicular cells (FCs and AFCs) and immune cells were accurately and comprehensively identified in all five PTC tissue sections. AFCs were identified as a transitional state between FCs and tumour cells, exhibiting a higher resemblance to the latter. Three tumour foci were shared among all patients out of the 13 observed. Notably, tumour foci No. 2 displayed elevated expression levels of genes associated with lower relapse‐free survival in PTC patients. We discovered key ligand–receptor interactions, including LAMB3–ITGA2, FN1–ITGA3 and FN1–SDC4, involved in the transition of PTC cells from FCs to AFCs and eventually to tumour cells. High expression of these patterns correlated with reduced relapse‐free survival. In the tumour immune microenvironment, reduced interaction between myeloid‐derived TGFB1 and TGFBR1 in tumour focus No. 2 contributed to tumourigenesis and increased heterogeneity. The spatial RNA‐clinical analysis method developed here revealed prognosis‐associated cellular heterogeneity in the PTC microenvironment.Conclusions: The occurrence of tumour foci No. 2 and three enhanced ligand–receptor interactions in the AFC area/tumour foci reduced the relapse‐free survival of PTC patients, potentially leading to improved prognostic strategies and targeted therapies for PTC patients.

Purpose:Prunella vulgaris (PV), a traditional Chinese medicine, has been used to treat patients with thyroid disease for centuries in China. The purpose of the present study was to investigate its bioactive ingredients and mechanisms against Hashimoto’s thyroiditis (HT) using network pharmacology and molecular docking technology to provide some basis for experimental research.Methods: Ingredients of the PV formula were retrieved from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Additionally, HT-related genes were retrieved from the UniProt and GeneCards databases. Cytoscape constructed networks for visualization. A protein–protein interaction (PPI) network analysis was constructed, and a PPI network was built using the Search Tool for the Retrieval of Interacting Genes (STRING) database. These key targets of PV were enriched and analyzed by molecular docking verification, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment.Results: The compound–target network included 11 compounds and 66 target genes. Key targets contained Jun proto-oncogene (JUN), hsp90aa1.1 (AKI), mitogen-activated protein kinase 1 (MAPK1), and tumor protein p53 (TP53). The main pathways included the AGE-RAGE signaling pathway, the TNF signaling pathway, the PI3K–Akt signaling pathway, and the mitogen-activated protein kinase signaling pathway. The molecular docking results revealed that the main compound identified in the Prunella vulgaris was luteolin, followed by kaempferol, which had a strong affinity for HT.Conclusion: Molecular docking studies indicated that luteolin and kaempferol were bioactive compounds of PV and might play an essential role in treating HT by regulating multiple signaling pathways.

Background: Thyroid tissue has a special immune microenvironment that is not well characterized. Whether immune cells have a prognostic value in the recurrence of papillary thyroid cancer (PTC) needs further investigation.Methods: Multinodular non-toxic goiter (MNG) was taken as normal tissue for the difficulty in obtaining completely normal thyroid tissue (normal thyroid function, no thyroiditis, and no nodules). We compared the composition of mononuclear cells (MNCs) in peripheral blood and thyroid tissues from MNG and PTC patients by high-dimensional flow cytometry profiling and verified the results by multiplex immunohistochemistry. The recurrence rates of PTC patients with different CD8⁺T cell subset signatures were compared using TCGA database.Results: We observed that the immune cell composition of MNG was different from that in peripheral blood. Thyroid tissue contains higher percentages of T cells and NK cells. Moreover, the percentages of memory T cells and Treg cells were higher in thyroid than in peripheral blood and increased in PTC tumors. We further focused on the antitumoral CD8⁺T cells and found that the expression patterns of PD-1, CD39, and CD103 on CD8⁺T cells were different between MNG and PTC. Importantly, we found higher percentages of PD-1⁺CD39⁺CD103⁺CD8⁺T and PD-1⁺CD39⁺CD103^-CD8⁺T cells in PTC tumor tissues from recurrent patients than non-recurrent patients. By analyzing PTC data from TCGA database, we found that the expression patterns of these molecules were associated with different pathologic types and genders among PTC patients. Moreover, patients with PD-1^hiCD39^loCD103^hiCD8^hi, PD-1^hiCD39^hiCD103^loCD8^hi, and PD-1^loCD39^hiCD103^hiCD8^hi expression patterns have a higher 10-year recurrence-free survival.Conclusion: The immune microenvironment in MNG tissue is distinct from that in peripheral blood and paratumor tissue. More memory CD8⁺T cells were detected in PTC, and expression patterns of PD-1, CD39, and CD103 on CD8⁺T cells were significantly different in physiology and gender and associated with the recurrence rate of PTC. These observations indicate that CD8⁺T cell signatures may be useful prognostic markers for PTC recurrence.

Introduction: Hashimoto’s thyroiditis (HT) is one of the most common immune-mediated diseases. It makes thyroid surgery more complicated and difficult because there may be adhesions between the thyroid gland and surrounding structures. However, it is still controversial whether HT patients carry a high risk for postoperative complications of thyroid surgery. The purpose of this study was to investigate the significance of HT for the postoperative complications of thyroid surgery. Methods: A search for studies assessing the postoperative complication risks of HT patients compared with that of patients with benign nodules (BNs) was performed in PubMed, EMBASE and Web of Science. Nine studies (20,118 cases, 1,582 cases of HT and 18,536 cases of BN) were identified, and the data from the relevant outcomes were extracted and analysed. Results: There were no significant differences between the HT group and BN group in recurrent laryngeal nerve palsy (RLNP) and permanent hypoparathyroidism (PHP). The rate of transient hypocalcaemia (THC) was significantly higher in the HT group (16.85%) than in the BN group (13.20%). Conclusions: The meta-analysis showed that HT only increased the risk of the postoperative complication THC compared to BN. Understanding the significance of HT in postoperative hypoparathyroidism after thyroid surgery would help clinicians perform sufficient preoperative (and postoperative) assessments and to optimise surgical planning.