Identification of diagnostic molecules and potential therapeutic agents for atopic dermatitis by single-cell RNA sequencing combined with a systematic computing framework that integrates network pharmacology.

Atopic dermatitis (AD) is composed of highly flexible cellular participants. To better understand its pathobiology and molecular regulation mechanisms, it is necessary to combine single-cell RNA sequencing (scRNA-seq) with new computing frameworks or specific technologies, which may contribute to the development of better treatments for AD. The scRNA-seq data of GSE180885 and bulk RNA-seq data of GSE193309 were obtained from Gene Expression Omnibus (GEO) database, and the scRNA-seq data was analyzed by Seurat package to identify the cell types in AD. The genes related to the activity of AD topical drugs were obtained from the ChEMBL database, which provided a variety of bioactivity data such as multiple drugs and targets. AD-related genes were obtained from DisGeNET and CTD databases synthesizing human disease-related genes; the intersection of AD-related genes from these three sources with differentially expressed genes (DEGs) between non-diseased AD and normal human skin (NHS) samples and differential cell type marker genes was taken. The proximity analysis of drug gene network was performed based on the gene with the largest area of receiver operating characteristic (ROC) curve. Ten distinct cell types of AD and NHS were identified, except for phagocytes cells. Three hub genes, F10 and CALCRL and CTSB, were obtained. The area under the curve of ROC based on CTSB expression was the largest, which was 60.15%. By binding drug CTSB-related gene interaction network, we identified 145 potential drugs. Among them, the score of DB07045 and CTSB docking was the lowest, and molecular docking and molecular dynamics (MD) simulation confirmed the close and stable binding of DB07045 and cathepsin B. This work identified diagnostic molecules and potential therapeutic drugs of AD by scRNA-seq combined with a systematic computing framework of network pharmacology, which may provide valuable clues for drug design.

Exploring a 7-gene prognostic model based on ferroptosis for efficiently guiding immunotherapy in melanoma patients.

PURPOSE: Although skin cutaneous melanoma (SKCM) is a relatively immunotherapy-sensitive tumor type, there is still a certain fraction that benefits less from treatment. Ferroptosis has been demonstrated to modulate tumor progression in many cancer types. This study focused on ferroptosis-related genes to construct a prognostic model for SKCM patients. MATERIALS AND METHODS: Gene expression profiles of SKCM samples were obtained from public databases. Unsupervised consensus clustering was used to determine molecular subtypes related to ferroptosis. Least absolute shrinkage and selection operator (LASSO) and stepwise Akaike information criterion (stepAIC) were applied to construct a prognostic model based on differentially expressed genes between two molecular subtypes. RESULTS: C1 and C2 subtypes were identified with differential prognosis and immune infiltration. A 7-gene prognostic model was constructed to classify samples into high-FPRS and low-FPRS groups. Low-FPRS group with favorable prognosis had higher immune infiltration and more enriched immune-related pathways than the high-FPRS group. The two groups showed distinct sensitivity to immunotherapy, with the low-FPRS group predicted to have more positive response to immunotherapy than the high-FPRS group. A nomogram based on the FPRS score and clinical features was built for more convenient use. CONCLUSIONS: The critical role of ferroptosis involved in SKCM development was further validated in this study. The prognostic model was efficient and stable to be applied in clinical conditions to support clinicians in determining personalized therapy for SKCM patients especially those with metastasis.

Study on the intervention effect and mechanism of bacillus Calmette-Guerin polysaccharide and nucleic acid injection on atopic dermatitis by targeting the transient receptor potential vanilloid subtype 1 pathway.

Background: This study aimed to explore the mechanism of Bacillus Calmette-Guerin polysaccharide and nucleic acid injection (BCG-PSN) targeting the transient receptor potential vanilloid subtype 1 (TRPV1) pathway for atopic dermatitis (AD) in mice. Methods: Experiment 1: a total of 30 Kunming (KM) mice were randomized into blank control, model, BCG-PSN low-dose (25 g/kg), BCG-PSN medium-dose (75 g/kg), BCG-PSN high-dose (225 g/kg), and positive drug (hydrocortisone 25 mg/kg) control groups. The AD model mice were established by induction with 2,4-Dinitrochlorobenzene (DNCB). After treatment in groups, the symptom score and scratching frequency in skin lesions were observed. The levels of immunoglobulin E (IgE), interleukin (IL)-4, IL-31, and IL-13 in serum were detected, as well as the levels of tumor necrosis factor-α (TNF-α), TRPV1, and nuclear factor (NF)-κB p65 in skin lesions in each group. Experiment 2: the optimal dose of BCG-PSN in Experiment 1 was adopted. A total of 20 KM mice were randomized into blank control, model, BCG-PSN, and BCG-PSN + PAC (PAC-14028) groups. The symptom score and scratching frequency in skin lesions were observed. The levels of IgE, IL-4, IL-31, and IL-13 in serum were detected, as well as the levels of TNF-α and TRPV1 in skin lesions in each group. Results: In Experiment 1, compared with the blank control group, the ear tissues of mice in model groups developed AD, with increased symptom score, scratching frequency, levels of IgE, IL-4, IL-31, and IL-13 in serum and levels of TNF-α, TRPV1, and NF-κB p65 in skin lesions. Compared with the model group, BCG-PSN low-dose, BCG-PSN medium-dose, BCG-PSN high-dose, and positive drug control groups had reduced AD symptoms, decreased symptom score, and decreased scratching frequency, with declined expression of each inflammatory substance, including the greatest decrease in the medium-dose group. In Experiment 2, after BCG-PSN was combined with PAC, the inflammation indexes decreased compared with those in the model group, and increased compared with those in the BCG-PSN group. Conclusions: Intramuscular BCG-PSN can target the TRPV1 pathway, inhibit inflammation, and improve the symptoms of AD mice.

Analysis on the pathogenic fungi in patients with superficial mycosis in the Northeastern China during 10 years.

This study analyzed the infection of superficial mycosis and the relationship between the distribution characteristics of pathogenic fungi and age, time and sex in Northeast China in the past 10 years. We would like to provide a theoretical basis for the diagnosis and treatment of related diseases. From December 2008 to December 2018, 5,374 superficial mycoses from Northeast China were selected. The fungal species were identified by fungal microscopy, fungal culture, and species identification. Besides, the relationship between sex, age, time and the distribution of superficial mycosis and pathogenic fungi was analyzed. Among the 5,374 patients, the top three infections were tinea pedis (n=1,538, 28.62%), tinea cruris (n=1,018, 18.94%) and tinea corporis (n=938, 17.45%). The top three pathogens were Trichophyton rubrum (n=2,849, 48.65%), Trichophyton mentagrophytes (n=947, 16.14%) and Candida spp. (n=804, 13.70%). The main pathogenic fungi were dermatophytes. The age group with the highest incidence of tinea capitis was children (n=372, 6.92%). The highest incidence rate of tinea pedis was in 31-69-year adults (n=905, 16.84%); Malassezia mainly affects young people aged 15-30. Yeast and mold mostly invade the elderly patients >60 years old. The incidence of tinea cruris, tinea pedis and tinea corporis in male patients was higher than that in female patients. The incidence of onychomycosis in female patients was higher than that in male patients (P<0.05). The isolation rate of Candida, Mold, Microsporum canis, Malassezia and Sporothrix increased year by year, while that of Trichophyton rubrum, Trichophyton mentagrophyte, Trichophyton schoenleinii and Epidermophyton floccosum decreased. From December 2008 to December 2018, dermatophytes were the main pathogens of superficial mycosis in Northeast China. The distribution of disease species and pathogenic fungi varied in different gender, age and time.

ZNF23 Suppresses Cutaneous Melanoma Cell Malignancy via Mitochondria-Dependent Pathway.

BACKGROUND/AIMS: Cutaneous melanoma is one of the leading causes of cancer deaths with an increasing incidence worldwide. A KRAB-containing zinc finger protein member, zinc finger 23 (ZNF23), was reduced in some types of tumors and inhibited cell growth by inducing cell cycle arrest. However, the role of ZNF23 expression is still poorly understood in melanoma. METHODS: The level of ZNF23 expression was detected in cutaneous melanoma, adjacent normal skin tissues and cutaneous melanoma cell lines using immunohistochemistry and western blotting. The correlations between ZNF23 expression and other clinicopathologic parameters were analyzed in melanoma patients. Ectopic expression of ZNF23 plasmid was transfected into melanoma cells, SK-MEL-1 and SK-MEL-28. MTT, flow cytometry and transwell assay were used to measure cell proliferation, apoptosis, invasion and migration abilities, respectively. Mitochondrial functions and structures were detected by mitochondrial membrane potential assay and Transmission electron microscopy (TEM) method in melanoma cells transfected with overexpressing ZNF23 plasmid or empty vector. Western blotting was performed to detect the levels of ZNF23, p53, p27, Bcl-2 and cleaved caspase-3 after overexpressing of ZNF23 in melanoma cells. RESULTS: ZNF23 was elevated in adjacent normal skin tissues compared with melanoma tissues. Patients with low level of ZNF23 expression exhibited higher incidence of lymphoid metastasis, thicker size of tumors and worse outcome. By using Cox's regression analysis, ZNF23 expression, tumor thickness and lymph node metastasis were the independent prognostic factors for overall survival (p < 0.05). Results from cellular experiments indicated that ectopic expression of ZNF23 induced cell apoptosis by activation of caspase-3, p27, p53 expression and down-regulation of Bcl-2 through mitochondria-dependent pathway. CONCLUSIONS: Decreased ZNF23 was contributed to melanoma progression and poor survival with mitochondria-dependent pathway. It indicated that ZNF23 could be a promising therapeutic biomarker candidate for cutaneous melanoma.

Reversal effect of melanoma differentiation associated gene-7/interleukin-24 on multidrug resistance in human hepatocellular carcinoma cells.

Multidrug resistance is the main cause for failure of chemotherapy. Melanoma differentiation associated gene-7/interleukin-24 (mda-7/IL-24) has been implicated in the inhibition of human tumor cell proliferation. However, the reversing effect of mda-7/IL-24 on multidrug resistance of human hepatocellular carcinoma (HCC) is not fully clear. In this study, we investigated the effects of overexpression of the mda-7/IL-24 gene in human HCC. We established mda-7/IL-24 overexpressing BEL-7402/5-fluorouracil (5-FU) cell lines and their drug sensitivity to 5-FU and doxorubicin (DOX) which were investigated by MTT. Furthermore, we investigated the apoptotic rate and the intracellular accumulation of Rhodamine-123 and DOX by flow cytometry. We also studied the expression of multidrug resistance gene 1 (MDR1), lung resistance-related protein (LRP), and multidrug resistance-related protein 1 (MRP1) by real-time polymerase chain reaction and Western blotting. Transcriptional activation of AP-1 and NF-κB was determined by luciferase reporter assay. The drug sensitivity of 5-FU or DOX, the apoptotic rate, and the intracellular accumulation of Rhodamine-123 and DOX were increased, while the mRNA and protein expression levels of MDR1, LRP, and MRP1 were reduced. The transcriptional activation of AP-1 and NF-κB was suppressed in mda-7/IL-24 overexpressing BEL-7402/5-FU cells. Our results demonstrated that mda-7/IL-24 could restore the drug sensitivity through the downregulation of MDR1, MRP1, and LRP expression, as well as the transcriptional activation of AP-1 and NF-κB and effectively reverse MDR.