The regulatory processes in developmental biology research are significantly influenced by long non-coding RNAs (lncRNAs). However, the dynamics of lncRNA expression during human tooth development remain poorly understood. In this research, we examined the lncRNAs present in the dental epithelium (DE) and dental mesenchyme (DM) at the late bud, cap, and early bell stages of human fetal tooth development through bulk RNA sequencing. Developmental regulators co-expressed with neighboring lncRNAs were significantly enriched in odontogenesis. Specific lncRNAs expressed in the DE and DM, such as PANCR, MIR205HG, DLX6-AS1, and DNM3OS, were identified through a combination of bulk RNA sequencing and single-cell analysis. Further subcluster analysis revealed lncRNAs specifically expressed in important regions of the tooth germ, such as the inner enamel epithelium and coronal dental papilla (CDP). Functionally, we demonstrated that CDP-specific DLX6-AS1 enhanced odontoblastic differentiation in human tooth germ mesenchymal cells and dental pulp stem cells. These findings suggest that lncRNAs could serve as valuable cell markers for tooth development and potential therapeutic targets for tooth regeneration.
Humans ; RNA, Long Noncoding/metabolism* ; Odontogenesis/genetics* ; Tooth Germ/embryology* ; Cell Differentiation ; Gene Expression Regulation, Developmental ; Mesoderm/metabolism* ; Tooth/embryology* ; Gene Expression Profiling ; Sequence Analysis, RNA ; Dental Pulp/cytology*

Objective:To explore the distribution pattern of TCM syndromes in patients with pulmonary tuberculosis combined with coronary heart disease in Kunming area.Methods:A survey was conducted by research of questionnaires to the general information and TCM four diagnostic information of pulmonary tuberculosis patients (116 cases) with coronary heart disease admitted to our hospital from April 2019 to December 2020, and principal component analysis and clustering analysis were conducted. Frequency analysis and severity analysis methods were used for data processing.Results:The TCM symptoms of pulmonary tuberculosis patients with coronary heart disease were mainly cough [89.66%(104/116)], expectoration [73.28% (85/116)], dry mouth [70.69% (82/116)], chest tightness [64.66% (75/116)], fatigue [56.03%(65/116)], hot flashes [52.59% (61/116)], shortness of breath [50.00% (58/116)]; tongue color was more common with light red [43.10% (50/116)] and red [47.41% (55/116)]; the tongue shape was more common with cracked tongue [37.07% (43/116)], punctured tongue [27.59% (32/116)] and old tongue [23.28%(27/116)]; tongue coating with little or no coating [48.28%(56/116)], yellow and greasy coating [21.55% (25/116)] was more common; the pulse was usually thin (number) [54.31% (63/116)] and slippery (number) [25.86% (30/116)]. The principal component analysis method used a load coefficient >0.40 as the threshold to screen out the four diagnostic information of 10 principal components; the clustering analysis results were divided into three categories of TCM types: deficiency syndrome, excess syndrome, and mixed deficiency and excess syndrome. There were significant differences in TCM syndrome types among patients of different genders, ages, disease courses, and educational levels ( P<0.05). Deficiency syndrome was more common in male patients [41.18% (21/51)], and mixed syndrome of deficiency and excess was more common in female patients [63.08% (41/65)]; 43-59 years old patients were more likely to have excess syndrome [36.36%(24/66)], and ≥60 years old patients were more likely to have mixed syndrome [70.00% (35/50)]; patients with a course of disease <3 months had more excess syndrome [41.30% (19/46)], patients with a course of 3-12 months had more deficiency syndrome [57.14% (20/35)], and patients with a course of more than 1 year had more mixed syndrome of deficiency and excess [74.29% (26/35)]. male patients [54.17%(26/48)] were more serious in excess syndrome, and female patients [53.33% (24/45)]; were more serious in deficiency syndrome; deficiency syndrome [43-59 years old was 54.17% (42/84), ≥60 years old was 54.17% (12/24)] more serious in patients of different age groups; patients with a course of disease <3 months [56.86% (29/51)] were more serious in excess syndrome, and patients with a course of disease ≥3 months were more serious in mixed syndromes [3~12 months 52.38%(22/42), >1 year 53.33% (24/45)]. Conclusion:The pathogenesis of TCM in patients with pulmonary tuberculosis complicated with coronary heart disease in Kunming is deficiency in root and excess in superficiality; the syndrome is based on yin deficiency and qi deficiency, with blood stasis, phlegm heat, phlegm stasis and phlegm turbidity as the symptoms.