Necroptosis-related LncRNAs in skin cutaneous melanoma: evaluating prognosis, predicting immunity, and guiding therapy.

BACKGROUND: An increasing amount of research has speculated that necroptosis could be a therapeutic strategy for treating cancer. However, understanding the prognostic value of the necroptosis-related long non-coding RNAs (NRLs) in skin cutaneous melanoma (SKCM, hereafter referred to as melanoma) remains poor and needs to be developed. Our research aims to construct a model based on NRLs for the prognosis of patients with melanoma. METHODS: We obtained the RNA-seq and clinical data from The Cancer Genome Atlas (TCGA) database and retrieved 86 necroptosis-related genes from the GeneCards database. The lncRNAs associated with necroptosis were identified via the Pearson correlation coefficient, and the prognostic model of melanoma was constructed using LASSO regression. Next, we employed multiple approaches to verify the accuracy of the model. Melanoma patients were categorized into two groups (high-risk and low-risk) according to the results of LASSO regression. The relationships between the risk score and survival status, clinicopathological correlation, functional enrichment, immune infiltration, somatic mutation, and drug sensitivity were further investigated. Finally, the functions of AL162457.2 on melanoma proliferation, invasion, and migration were validated by in vitro experiments. RESULTS: The prognostic model consists of seven NRLs (EBLN3P, AC093010.2, LINC01871, IRF2-DT, AL162457.2, AC242842.1, HLA-DQB1-AS1) and shows high diagnostic efficiency. Overall survival in the high-risk group was significantly lower than in the low-risk group, and risk scores could be used to predict melanoma survival outcomes independently. Significant differences were evident between risk groups regarding the expression of immune checkpoint genes, immune infiltration, immunotherapeutic response and drug sensitivity analysis. A series of functional cell assays indicated that silencing AL162457.2 significantly inhibited cell proliferation, invasion, and migration in A375 cells. CONCLUSION: Our prognostic model can independently predict the survival of melanoma patients while providing a basis for the subsequent investigation of necroptosis in melanoma and a new perspective on the clinical diagnosis and treatment of melanoma.

Liquid-liquid phase separation throws novel insights into treatment strategies for skin cutaneous melanoma.

BACKGROUND: In recent years, there has been growing evidence indicating a relationship between liquid-liquid phase separation (LLPS) and cancer development. However, to date, the clinical significance of LLPS in skin cutaneous melanoma (SKCM, hereafter referred to as melanoma) remains to be elucidated. In the current study, the impact of LLPS-related genes on melanoma prognosis has been explored. METHODS: LLPS-related genes were retrieved from the DrLLPS database. The prognostic feature for LLPS in melanoma was developed in The Cancer Genome Atlas (TCGA) dataset and verified in the GSE65904 cohort. Based on risk scores, melanoma patients were categorized into high- and low-risk groups. Thereafter, the differences in clinicopathological correlation, functional enrichment, immune landscape, tumor mutational burden, and impact of immunotherapy between the two groups were investigated. Finally, the role of key gene TROAP in melanoma was validated by in vitro and in vivo experiments. RESULTS: The LLPS-related gene signature was developed based on MLKL, PARVA, PKP1, PSME1, RNF114, and TROAP. The risk score was a crucial independent prognostic factor for melanoma and patients with high-risk scores were related to a worse prognosis. Approximately, all immune-relevant characteristics, such as immune cell infiltration and immune scores, were extremely evident in patients with low-risk scores. The findings from the in vitro and in vivo experiments indicated that the viability, proliferation, and invasion ability of melanoma cells were drastically decreased after the knockdown of TROAP. CONCLUSION: Our gene signature can independently predict the survival of melanoma patients. It provides a basis for the exploration of the relationship between LLPS and melanoma and can offer a fresh perspective on the clinical diagnosis and treatment of the disease.

A smartphone-based online platform for clinical skills training and assessment with standardized patients: platform development and pilot study outcomes.

There are limitations and difficulties in the management of traditional in-person standardized patient (SP) practice. The latest developments in online communication tools and the COVID-19 pandemic have promoted the needs for online clinical skills training objectively. However, existing commercial online platforms may not meet the requests for SP-based medical simulation. This paper described the methodology applied to develop a smartphone-based online platform for the management of clinical skills training and assessment with remote SPs, and aimed to determine whether this new platform is acceptable or useful through a pilot run in September 2020. The post-run survey including questionnaire inspired by technological acceptance model and determinants of the perceived ease of use was used to assess the acceptability and usefulness of the platform. Twenty four-year students of clinical medicine participated in the pilot study with twenty SPs and ten faculties. Data from the post-run survey showed that there was a general recognition that the platform is easy to use among all the users. Two questions regarding the usefulness of the platform showed significant differences between the SPs/faculties and the students. More SPs found the platform useful as a training method than the students did. The faculties showed more attempts than the students to use this platform for clinical skills training in the future. This smartphone-based online platform was widely accepted among the tested students, SPs and faculties, which meets the requests and challenges of the new era. It provides an effective approach for clinical skills training and assessment with remote SPs.

Immune Landscape and an RBM38-Associated Immune Prognostic Model with Laboratory Verification in Malignant Melanoma.

BACKGROUND: Current studies have revealed that RNA-binding protein RBM38 is closely related to tumor development, while its role in malignant melanoma remains unclear. Therefore, this research aimed to investigate the function of RBM38 in melanoma and the prognosis of the disease. METHODS: Functional experiments (CCK-8 assay, cell colony formation, transwell cell migration/invasion experiment, wound healing assay, nude mouse tumor formation, and immunohistochemical analysis) were applied to evaluate the role of RBM38 in malignant melanoma. Immune-associated differentially expressed genes (DEGs) on RBM38 related immune pathways were comprehensively analyzed based on RNA sequencing results. RESULTS: We found that high expression of RBM38 promoted melanoma cell proliferation, invasion, and migration, and RBM38 was associated with immune infiltration. Then, a five-gene (A2M, NAMPT, LIF, EBI3, and ERAP1) model of RBM38-associated immune DEGs was constructed and validated. Our signature showed superior prognosis capacity compared with other melanoma prognostic signatures. Moreover, the risk score of our signature was connected with the infiltration of immune cells, immune-regulatory proteins, and immunophenoscore in melanoma. CONCLUSIONS: We constructed an immune prognosis model using RBM38-related immune DEGs that may help evaluate melanoma patient prognosis and immunotherapy modalities.

Application of an indocyanine green surgical fluorescence imaging system in sentinel lymph node biopsy of acral malignant melanoma.

BACKGROUND: Regional lymph node status is an independent influencing factor for the prognosis of acral malignant melanoma, and the accuracy of sentinel lymph node biopsy (SLNB) is directly related to the judgment of regional lymph node status. This study aimed to explore the application value of indocyanine green (ICG) surgical fluorescence imaging system in the SLNB of acral malignant melanoma. METHODS: A total of 34 patients with acral malignant melanoma were admitted to the Department of Burn and Plastic Surgery in Jiangsu Provincial People's Hospital from January 2020 to March 2020. Among these patients, 22 required SLNB. ICG and methylene blue (MB) were combined to intraoperatively trace the sentinel lymph nodes (SLNs). The total number of SLNs detected during the operation was counted. We compared the number, detection rate, as well as the detection rate and false negative rate of positive SLNs of SLNs detected by ICG, MB, and ICG combined with MB. RESULTS: A total of 56 SLNs were detected in the 22 patients, among which 55 were detected by ICG (98%), 41 were detected by MB (71%), and 56 (100%) were detected by ICG combined with MB, and the average number of SLNs were 2.5, 1.64, and 2.55, respectively. A total of nine SLNs were detected, of which nine were detected by ICG (100%), seven by MB (78%), and nine by ICG combined with MB (100%). Patients with negative SLNs had no recurrence at the 6-month follow-up. CONCLUSIONS: Compared with MB, the ICG fluorescent imaging system can improve the detection rate of SLNs in patients with acral malignant melanoma. Also, ICG combined with MB was superior to ICG alone.

Application of dermal regenerative template in reconstructing skin defects after plantar malignant melanoma excision.

PURPOSE: The excision of plantar malignant melanoma frequently leads to wide skin defects on the plantar surface. This study aimed to investigate the advantages and feasibility of dermal regenerative template reconstructing plantar blemishes caused by malignant melanoma. METHODS: 28 patients identified with plantar malignant melanoma were included in this retrospective article. Eighteen patients received immediate skin grafts after wide excision skin graft (SG) group), whereas the remaining 10 patients were treated with dermal regenerative template (DRT) (Lando ®, Shenzhen TsingCare Medical Co. Ltd) 14 days before skin grafts (DRT group) and the postoperative survival rate in the two groups was analyzed. During the 6-month follow-up, we compared the scar index, plantar pain, and recurrent skin graft ulcer incidence on the skin grafts area. RESULTS: Postoperative survival rate in the DRT group (91.75% ± 7.64%) was higher than in the SG group (80.51% ± 7.17%). The DRT group showed less scar formation on Vancouver scar scale (VSS index): 3.40 ± 1.07 than the SG group (VSS index: 6.33 ± 0.68). The dermal regenerative template alleviated plantar pain and decreased the incidence of ulcer on the skin grafts area. CONCLUSIONS: The dermal regenerative template not only improves the survival rate of skin grafts but also alleviates scar condition, plantar pain and recurrent skin graft ulcer. This study provides a new reconstructive strategy in plantar skin defects after the excision of malignant melanoma.

Diagnostic Key Points and Surgical Management of Necrotizing Fasciitis: A Retrospective Study.

Necrotizing fasciitis (NF) is a fatal disease with a high mortality rate that can be easily misdiagnosed. The aim of this study was to improve the diagnostic rate of NF and overall survival. We conducted a single-center, retrospective, noncontrolled study involving 36 patients who were admitted to our department between December 2017 and October 2019, and summarized the diagnostic key points and timing of surgical treatment. All patients were diagnosed at our department and underwent multiple courses of treatment. The records included information regarding underlying diseases, bacterial culture results, laboratory risk indicator for necrotizing fasciitis (LRINEC) score, number of procedures, and type of antibiotics. All 36 cases of NF were cured and showed good patient condition on follow-up; the mean number of surgeries was three, and the mean duration of hospitalization was 37 days (range, 21-83 days). The LRINEC scores of 16 patients were ≥8 points. Seventeen patients with underlying diabetic disease had higher inflammatory index scores than those without diabetes. The LRINEC scores of patients with (n = 17) and without (n = 19) DM were 7.40 ± 2.99 and 3.80 ± 2.39, respectively (P < .01). Cases of NF that were treated with early incision and surgical abscess drainage required fewer surgeries and a shorter length of hospitalization. Thus, surgeons should be more aware of NF and aim to make an early and accurate diagnosis using various approaches. Complete surgical debridement plays an essential role in NF treatment, and diabetes mellitus is a significant adverse factor that exacerbates the severity of NF. Negative-pressure techniques are useful in cases involving nonanaerobic infections and cause minimal complications.

Circular RNA circRNA_0082835 promotes progression and lymphatic metastasis of primary melanoma by sponging microRNA miRNA-429.

To identify how circular RNA circRNA_0082835 impacts melanoma cells and lymphatic metastasis to observe whether it exerts effects through its action mechanism of sponging microRNA miR-429. Clinical baseline information was collected, and clinical samples were used for detection on circRNA_0082835 and EZH2. The expression of circRNA_0082835, EZH2, and miR-429 was detected by quantitative real-time PCR (RT-qPCR). Cell proliferation was tested with cell counting kit-8 (CCK-8). Flow cytometry was applied to examination of cell cycle levels. Cell invasion and migration were observed by transwell and wound healing. The expression of Wnt/ß-catenin pathway, cell cycle and epithelial-mesenchymal transition (EMT) marker proteins was analyzed by western blot. Dual-luciferase determined the binding of miR-429 and circ_0082835. As a result, the expression of circRNA_0082835 was increased and that of miR-429 was decreased with the increase in lymphatic metastasis level. CircRNA_0082835 expression was downregulated by circ_0082835 interference, upregulated by EZH2 interference and also downregulated after transfection of both shRNA-circ_0082835 and shRNA-EZH2. Inhibiting circ_0082835 and EZH2 suppressed the proliferation, invasion and migration, regulated the cell cycle levels, inhibited Wnt/ß-catenin and attenuated EMT in melanoma cells. Inhibition of circ_0082835 and/or EZH2 elevated miR-429 expression. The binding among miR-429 and circ_0082835 was verified. MiR-429 inhibitor reversed the effect of circ_0082835 interference while having no significant impact on EZH2. In conclusion, circRNA_0082835 sponges miR-429 to affect the anti-tumor effect of miR-429 in primary melanoma and lymphatic metastasis.