Background: Melanoma is typically regarded as the most dangerous form of skin cancer. Although surgical removal of in situ lesions can be used to effectively treat metastatic disease, this condition is still difficult to cure. Melanoma cells are removed in great part due to the action of natural killer (NK) and T cells on the immune system. Still, not much is known about how the activity of NK cell-related pathways changes in melanoma tissue. Thus, we performed a single-cell multi-omics analysis on human melanoma cells in this study to explore the modulation of NK cell activity.Materials and methods: Cells in which mitochondrial genes comprised > 20% of the total number of expressed genes were removed. Gene ontology (GO), gene set enrichment analysis (GSEA), gene set variation analysis (GSVA), and AUCcell analysis of differentially expressed genes (DEGs) in melanoma subtypes were performed. The CellChat package was used to predict cell–cell contact between NK cell and melanoma cell subtypes. Monocle program analyzed the pseudotime trajectories of melanoma cells. In addition, CytoTRACE was used to determine the recommended time order of melanoma cells. InferCNV was utilized to calculate the CNV level of melanoma cell subtypes. Python package pySCENIC was used to assess the enrichment of transcription factors and the activity of regulons in melanoma cell subtypes. Furthermore, the cell function experiment was used to confirm the function of TBX21 in both A375 and WM-115 melanoma cell lines.Results: Following batch effect correction, 26,161 cells were separated into 28 clusters and designated as melanoma cells, neural cells, fibroblasts, endothelial cells, NK cells, CD4+ T cells, CD8+ T cells, B cells, plasma cells, monocytes and macrophages, and dendritic cells. A total of 10137 melanoma cells were further grouped into seven subtypes, i.e., C0 Melanoma BIRC7, C1 Melanoma CDH19, C2 Melanoma EDNRB, C3 Melanoma BIRC5, C4 Melanoma CORO1A, C5 Melanoma MAGEA4, and C6 Melanoma GJB2. The results of AUCell, GSEA, and GSVA suggested that C4 Melanoma CORO1A may be more sensitive to NK and T cells through positive regulation of NK and T cell-mediated immunity, while other subtypes of melanoma may be more resistant to NK cells. This suggests that the intratumor heterogeneity (ITH) of melanoma-induced activity and the difference in NK cell-mediated cytotoxicity may have caused NK cell defects. Transcription factor enrichment analysis indicated that TBX21 was the most important TF in C4 Melanoma CORO1A and was also associated with M1 modules. In vitro experiments further showed that TBX21 knockdown dramatically decreases melanoma cells’ proliferation, invasion, and migration.Conclusion: The differences in NK and T cell-mediated immunity and cytotoxicity between C4 Melanoma CORO1A and other melanoma cell subtypes may offer a new perspective on the ITH of melanoma-induced metastatic activity. In addition, the protective factors of skin melanoma, STAT1, IRF1, and FLI1, may modulate melanoma cell responses to NK or T cells.

Background: Melanoma, a malignant neoplasm originating from melanocytes, is a form of skin cancer with rapidly increasing global incidence, often exacerbated by UV radiation[1]. Particularly, acral melanoma, characterized by its swift metastasis and poor prognosis, underscores the significance of further research into its heterogeneity. Single-cell sequencing has been widely utilized in the study of tumor heterogeneity; however, research related to melanoma remains to be further refined.

Aims: LY3023414 is a novel oral phosphatidylinositol 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) dual inhibitor designed for advanced cancers, for which a phase II clinical study was completed in March 2020; however, little is known about its effect on bone modelling/remodelling. In this study, we aimed to explore the function of LY3023414 in bone modelling/remodelling. Methods: The function of LY3023414 was explored in the context of osteogenesis (bone formation by osteoblasts) and osteoclastogenesis (osteoclast formation and bone resorption). Murine preosteoblast MC3T3-E1 cell line and murine bone marrow-derived macrophage cells (BMMs) were subjected to different treatments. An MTS cell proliferation assay was used to examine the cytotoxicity. Thereafter, different induction conditions were applied, such as MCSF and RANKL for osteoclastogenesis and osteogenic media for osteogenesis. Specific staining, a bone resorption assay, and quantitative real-time polymerase chain reaction (qRT-PCR) were subsequently used to evaluate the effect of LY3023414. Moreover, small interfering RNA (siRNA) was applied to knockdown Akt1 or Akt2 for further validation. Lastly, western blot was used to examine the exact mechanism of action. Results: LY3023414 attenuated PI3K/protein kinase B (Akt)/GSK3-dependent activation of β-catenin and nuclear factor-activated T cell 1 (NFATc1) during osteogenesis and osteoclastogenesis, respectively. LY3023414 mainly inhibited osteoclast formation instead of mature osteoclast function. Moreover, it suppressed osteogenesis both in the early stage of differentiation and late stage of calcification. Similarly, gene knockdown of Akt isoforms by siRNA downregulated osteogenic and osteoclastogenic processes, indicating that Akt1 and Akt2 acted synergistically. Conclusion: LY3023414 can suppress osteogenesis and osteoclastogenesis through inhibition of the PI3K/Akt/GSK3 signalling pathway, which highlights the potential benefits and side effects of LY3023414 for future clinical applications. Cite this article: Bone Joint Res 2021;10(4):237–249.

Genioplasty is the main way to treat diseases such as chin asymmetry, dysplasia and overdevelopment, which involve the three-dimensional direction abnormalities of the chin. Since this kind of surgery mainly uses intraoral incisions, the narrow surgical field of intraoral incisions and the surrounding important neurovascular tissues make it easy for complications, to occur during the osteotomy process, which results in greater surgical risks. The first craniofacial-plastic surgical robot (CPSR-I) system is developed to complete the precise positioning and improve the surgeon's force perception ability. The Kalman filtering method is adopted to reduce the interference of sensor signal noise. An adaptive fuzzy control system, which has strong robustness and adaptability to the environment, is designed to improve the stability of robot-assisted surgical operations. To solve the problem of the depth perception, we propose an automatic bone drilling control strategy that combines position and force conditions to ensure that the robot can automatically stop when the bone is penetrated. On the basis of model surgery and animal experiments, preliminary experiments were carried out clinically. Based on the early results of 6 patients, the robot-assisted approach appears to be a safe and effective strategy for genioplasty.

Background: To explore the potential of electromagnetic (EM) navigation technology in the field of robot-assisted surgery, we set up a maxillofacial surgical robotic system (MSRS) guided by an EM navigation tool. Mandibular angle osteotomy was used to analyze the feasibility in confined surgical areas. Methods: Model and animal experiments were implemented to validate the system precision. Before the experiment, a customized dental splint was made and then fixed with a standard navigation part. An accurate 3D surgical plan was designed based on the preoperative CT scan. During the experiment, the splint was rigidly mounted on teeth for navigation registration, so the robot could position a specially designed template to guide the accurate osteotomy according to the preoperative plan. For the model experiment, a Coordinate Measuring Machine was used to measure the template’s position and angle. For the animal experiment, surgeons completed the surgery by moving a saw along the template, while a postoperative CT scan was carried out to calculate the precision. Results: All procedures were successfully completed, with no complications in any of the experimental animals. In the model experiment, the accuracy of the navigation position and angle was 0.44±0.19 mm and 3.5°±2.1°, respectively. In the animal experiment, the lateral osteotomy line error was 0.83±0.62 mm, the interior error was 1.06±1.03 mm, and the angle between the actual cutting plane and preoperative planning plane was 5.9°±4.7°. Conclusions: Robot-assisted surgery with EM navigation resulted feasible in the real operating environment. Moreover, this system’s precision could meet clinical needs, while the proposed procedure was safe and easy on animals. Consequently, this approach has the potential to be applied to clinical craniomaxillofacial practice in the near future.