Role of METTL3-mediated m6A modification in osteogenic differentiation of periodontal ligament stem cells extracted from adult periodontal ligaments ex-vivo.

Periodontal ligament stem cells (PDLSCs) are identified as candidate cells for the regeneration of periodontal and alveolar bone tissues. This research was to analyze the effect of methyltransferase-like 3 (METTL3)-mediated m6A modification on the osteogenic differentiation of PDLSCs extracted from adult periodontal ligaments (PDLs) ex-vivo. From June 2022 to October 2022, 27 patients undergoing orthodontic treatment in our hospital were selected as the research population, with 31 teeth extracted in total. PDLSCs were isolated from PDLs by tissue block culture, and the results were analyzed. Then PDLSCs were induced to differentiate into osteoblasts, and changes in METTL3 and m6A levels during differentiation were observed. Additionally, abnormal METTL3 expression vectors were constructed and transfected into PDLSCs to observe the influence of METTL3 on the biological behavior and osteogenic differentiation of PDLSCs. PDLSCs isolated from ex-vivo PDLs were predominantly spindle-shaped, with high CD73, CD90 and CD105 levels and low CD11b, CD34 and CD45 levels, showing the characteristics of stem cells. Spearman correlation coefficients identified a positive connection between Runx2, Sp7, Alp, Bglap, METTL3 and m6A levels and osteogenic differentiation incubation time (P<0.05). As METTL3 expression was increased, the proliferation capacity and osteogenic differentiation ability of PDLSCs were enhanced (P<0.05), and the content of m6A was increased (P<0.05). However, the activity and osteogenic differentiation ability of PDLSCs was decreased after silencing METTL3 (P<0.05). In conclusion, METTL3-mediated m6A modification promoted the osteogenic differentiation of PDLSCs extracted from adult PDLs ex vivo. This study offered a novel understanding of the mechanisms underlying osteogenic differentiation, and implied a possible method for accelerating bone formation.

lncRNA-FMR6 directly binds SAV1 to increase apoptosis of granulosa cells in premature ovarian failure.

BACKGROUND: A regulatory mechanism of lncRNA binding to protein has been detected in premature ovarian failure (POF). Therefore, this study was expected to illustrate the mechanism of lncRNA-FMR6 and SAV1 regulating POF. METHODS: Follicular fluid and ovarian granulosa cells (OGCs) from POF patients and healthy volunteers were collected. Using RT-qPCR and western blotting, lncRNA-FMR6 and SAV1 expression were detected. KGN cells were cultured, and the subcellular localization analysis of lncRNA-FMR6 was carried out. In addition, KGN cells were treated with lncRNA-FMR6 knockdown/overexpression or SAV1 knockdown. Then, cell optical density (proliferation), apoptosis rate, Bax and Bcl-2 mRNA expression were explored by CCK-8, caspase-3 activity, flow cytometry and RT-qPCR analysis. By performing RIP and RNA pull-down experiments, the interactions among lncRNA-FMR6 and SAV1 was investigated. RESULTS: Up-regulation of lncRNA-FMR6 was shown in follicular fluid and OGCs of POF patients, and ectopic overexpression of lncRNA-FMR6 promoted KGN cells apoptosis and inhibited proliferation. lncRNA-FMR6 was localized in the cytoplasm of KGN cells. SAV1 bounding to lncRNA-FMR6 was negatively regulated by lncRNA-FMR6, and was down-regulated in POF. SAV1 knockdown promoted KGN cells proliferation and inhibited apoptosis, and partially eliminated the effect of lncRNA-FMR6 low expression on KGN cells. CONCLUSION: Overall, lncRNA-FMR6 accelerates POF progression by binding to SAV1.

SNHG1/miR-145-5p/KLF5 Axis Participates in Regulating the Proliferation and Migration of Oral Squamous Cell Cancer.

We aimed to clarify the molecular mechanism of lncRNA SNHG1 in regulating the OSCC process. Clinical samples of OSCC were collected for detecting the differential level of SNHG1 by qRT-PCR. Pathological indexes of OSCC patients were analyzed for uncovering the prognostic value of SNHG1. The interaction between SNHG1 and miR-145-5p was assessed through the bioinformatics method and dual-luciferase reporter assay. Their coregulation on proliferative and migratory functions of Tca8113 and CAL-27 cells was explored by the CCK-8, EdU, and Transwell assay. Finally, the regulatory effect of miR-145-5p on its downstream gene KLF5 was evaluated. SNHG1 was abnormally upregulated in OSCC samples and linked to a poor prognosis of OSCC patients. Serving as an oncogene, SNHG1 strengthened proliferative and migratory functions of Tca8113 and CAL-27 cells. miR-145-5p was a key downstream target inducing the oncogenic role of SNHG1 in the OSCC process with KLF5 as its downstream gene. SNHG1/miR-145-5p/KLF1 axis is responsible for driving the malignant process of OSCC.

Inhibitory effect of baicalin on orthodontically induced inflammatory root resorption in rats.

OBJECTIVE: This study investigated the inhibitory effect of baicalin on orthodontically induced inflammatory root resorption in rats. METHODS: Forty-five male Wistar rats were randomly divided into three groups of 15 rats each. Fifty grams of force was used to establish an orthodontic tooth movement model. Baicalin (40 mg/kg) was locally injected into rats in the baicalin group at 3-day intervals; concurrently, normal saline was injected into rats in the negative control group. On the 21st day after orthodontic treatment, the tooth movement distance and root resorption area ratio were measured. Histomorphology changes were observed by hematoxylin and eosin staining and immunohistochemistry. RESULTS: There was no significant difference in tooth movement distance between groups. The root resorption area ratio was significantly lower in the baicalin group than in the negative control group. Runx-2 expression was significantly higher in the baicalin group than in the negative control group, while tumor necrosis factor (TNF)-α expression was significantly lower in the baicalin group than in the negative control group. CONCLUSIONS: Baicalin inhibits orthodontically induced inflammatory root resorption by enhancing the expression of Runx-2 and reducing the expression of TNF-α, but does not affect tooth movement distance.

Prospective ECG-gated 320-row CT angiography of the whole aorta and coronary arteries.

OBJECTIVES: To investigate the feasibility of using a prospective ECG-gated wide-volume protocol in CT angiography (CTA) of the whole aorta and coronary arteries (CA). METHODS: A total of 61 consecutive patients with suspected acute aortic diseases underwent CTA of the whole aorta using a prospective ECG-gated wide-volume CT protocol without heart rate (HR) control. The exposure window was set at 40-50 % of R-R interval (HR ≥70 bpm) or 70-80 % of R-R interval (HR <70 bpm) in a single heartbeat. The image quality of the ascending aorta, aortic valve and CA was evaluated for motion artefacts. The mean attenuation was measured at different levels of the aorta. The radiation dose and contrast medium volume were recorded. RESULTS: All of the examinations were performed successfully. The image quality was acceptable in the ascending aorta, aortic valve (100 %) and CA (94.4 %). The mean radiation dose was 18.42 ± 5.02 mSv. Of 61 patients, 14 were diagnosed with aortic aneurysm and 35 were diagnosed with aortic dissection or intramural haematoma. Coronary artery stenosis was detected in 12 patients. CONCLUSION: For patients with aortic diseases, CTA of the whole aorta using a prospective ECG-gated wide-volume protocol has the potential to provide additional information about the CA and aortic valve with lower radiation exposure. KEY POINTS : • Computed tomography is increasingly used to assess the coronary arteries and the aorta. • This novel study investigates prospective ECG-gated wide-volume 320-row CT angiography. • Heart rate variation did not influence the image quality of coronary arteries. • Abnormalities in the coronary arteries and aorta can be simultaneously assessed noninvasively.