ABSTRACT
Objective: To analyze the classification and functions of cell subsets in laryngeal carcinoma and metastatic lymph nodes, and to explore the evolution trajectory of epithelial cells to tumor cells. Methods: Single-cell RNA sequencing was performed on 5 cases of laryngeal cancer, matched metastatic lymph nodes and 3 normal tissues. Patients were admitted to Ningbo Medical Center Lihuili Hospital from October 22, 2019 to December 16, all patients were male, aged 53-70 years old. Cell subsets of the above-mentioned tissues were analyzed by the Seurat, and the biological functions of cell subpopulation were investigated by functional enrichment analysis. Malignant epithelial cells were identified using copy number variation (CNV). The evolutionary trajectory of epithelial cells to cancer cells was analyzed by cell trajectory analysis, and cancerous transitional cells were identified. The highly expressed genes in transitional cells were analyzed by the FindAllMarker of the Seurat and verified by immunohistochemistry. Results: A total of 66 969 high-quality cells were obtained in 9 major clusters: epithelial cells, T cells, B cells, fibroblasts, endothelial cells, myeloid cells, mast cells, plasmacytoid dendritic cells and nerve cells. The first 5 cell clusters were divided into 8, 6, 4, 3 and 2 subgroups, respectively. Four epithelial cell subsets (C0, C1, C2 and C5) were derived from tumor tissues and metastatic lymph nodes, and had high levels of CNV and tumor cell content. Cell trajectory analysis showed that the evolution trajectory of epithelial cells was from normal epithelial subpopulation C4 to early cancerous cell population C0, which differentiated into three major malignant cell subsets C1, C3, and C5. Epithelial cell C0 may represent the transitional cell population of carcinogenesis, and were enriched in biological processes such as epithelial-mesenchymal transformation and angiogenesis. C0 highly expressed sulforaphane (SFN) which may be related to the occurrence and development of cancer. Immunohistochemistry confirmed that SFN was highly expressed in tumor tissues and metastatic lymph nodes compared with paracancerous tissues. Conclusion: Single-cell sequencing may be used to elucidate the diversity of cells and functions in laryngeal carcinoma tissues and metastatic lymph nodes, and cell population C0 plays a key role in the evolution of cells.
Subject(s)
Aged , Humans , Male , Middle Aged , Carcinoma, Squamous Cell/pathology , DNA Copy Number Variations , Endothelial Cells/pathology , Laryngeal Neoplasms/pathology , Lymph Nodes/pathologyABSTRACT
Objective To understand the acceptability and relevant impact factors of provider initiated testing counseling (abbreviated as PITC) in Liangshan Yi autonomous prefecture,so as to promote strategies for the sustainable development of PITC in AIDS epidemic areas.Methods Four medical institutions were selected with rates of acceptance counted.Multi-factors logistic regression was used to analyze the impact factors from the completed questionnaires.Results Among the 413respondents,197 patients accepted the PITC,with an acceptance rate as 47.7％ (95％ CI:42.9％-52.3％).Factors as being female (OR=7.283,95％CI:3.933-13.465),acceptance of publicity (OR=1.855,95％ CI:1.013-3.395),worried about being infected of HIV/AIDS (OR=2.699,95％ CI:1.616-4.506) etc.were protect factors of PITC,while those who previously received HIV testing (OR=0.226,95％CI:0.124-0.409),afraid of taking blood test (OR=0.052,95％CI:0.024-0.106),knowing more AIDS related knowledge (OR=0.446,95％ CI:0.258-0.773) would significantly reduce the rate of acceptance.Conclusion Nearly half of the outpatients from medical institutions were willing to accept PITC in Liangshan Yi prefecture,thus provided important chance for HIV screening.Ways as strengthening related counseling before and after HIV test,launching effective HIV/AIDS-related publicity and advocating correct knowledge on blood testing,etc.need to be promoted,in order to improve the acceptability of PITC.