ABSTRACT
BACKGROUND: Tryptophan metabolism dysregulation has been observed in vitiligo. However, drawing a mechanistic linkage between this metabolic disturbance and vitiligo pathogenesis remains challenging. OBJECTIVE: Aim to reveal the characterization of tryptophan metabolism in vitiligo and investigate the role of tryptophan metabolites in vitiligo pathophysiology. METHODS: LC-MS/MS, dual-luciferase reporter assay, ELISA, qRT-PCR, small interfering RNA, western blotting, and immunohistochemistry were employed. RESULTS: Kynurenine pathway activation and KYAT enzyme-associated deviation to kynurenic acid (KYNA) in the plasma of stable non-segmental vitiligo were determined. Using a public microarray dataset, we next validated the activation of kynurenine pathway was related with inflammatory-related genes expression in skin of vitiligo patients. Furthermore, we found that KYNA induced CXCL10 upregulation in keratinocytes via AhR activation. Moreover, the total activity of AhR agonist was increased while the AhR concentration per se was decreased in the plasma of vitiligo patients. Finally, higher KYAT, CXCL10, CYP1A1 and lower AhR expression in vitiligo lesional skin were observed by immunohistochemistry staining. CONCLUSION: This study depicts the metabolic and genetic characterizations of tryptophan metabolism in vitiligo and proposes that KYNA, a tryptophan-derived AhR ligand, can enhance CXCL10 expression in keratinocytes.
Subject(s)
Chemokine CXCL10 , Keratinocytes , Kynurenic Acid , Receptors, Aryl Hydrocarbon , Skin , Tryptophan , Up-Regulation , Vitiligo , Humans , Vitiligo/metabolism , Vitiligo/genetics , Vitiligo/blood , Chemokine CXCL10/metabolism , Chemokine CXCL10/genetics , Receptors, Aryl Hydrocarbon/metabolism , Receptors, Aryl Hydrocarbon/genetics , Tryptophan/metabolism , Tryptophan/blood , Kynurenic Acid/blood , Kynurenic Acid/metabolism , Male , Keratinocytes/metabolism , Skin/metabolism , Skin/pathology , Adult , Female , Basic Helix-Loop-Helix Transcription Factors/metabolism , Basic Helix-Loop-Helix Transcription Factors/genetics , Kynurenine/metabolism , Kynurenine/blood , Cytochrome P-450 CYP1A1/genetics , Cytochrome P-450 CYP1A1/metabolism , Middle Aged , Case-Control Studies , Signal Transduction , Young AdultABSTRACT
Vitiligo is an acquired autoimmune dermatosis characterized by patchy skin depigmentation, causing significant psychological distress to the patients. Genetic susceptibility, environmental triggers, oxidative stress, and autoimmunity contribute to melanocyte destruction in vitiligo. Due to the diversity and complexity of pathogenesis, the combination of inhibiting melanocyte destruction and stimulating melanogenesis gives the best results in treating vitiligo. The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that can regulate the expression of various downstream genes and play roles in cell differentiation, immune response, and physiological homeostasis maintenance. Recent studies suggested that AhR signaling pathway was downregulated in vitiligo. Activation of AhR pathway helps to activate antioxidant pathways, inhibit abnormal immunity response, and upregulate the melanogenesis gene, thereby protecting melanocytes from oxidative stress damage, controlling disease progression, and promoting lesion repigmentation. Here, we review the relevant literature and summarize the possible roles of the AhR signaling pathway in vitiligo pathogenesis and treatment, to further understand the links between the AhR and vitiligo, and provide new potential therapeutic strategies.
Subject(s)
Receptors, Aryl Hydrocarbon , Vitiligo , Humans , Antioxidants/metabolism , Melanocytes , Receptors, Aryl Hydrocarbon/metabolism , Skin/pathology , Vitiligo/metabolismABSTRACT
Vitiligo is an autoimmune disease characterized by depigmentation. Kaempferol is a flavonoid compound with broad anti-inflammatory and antioxidant properties. The purpose of this study was to investigate the effect of kaempferol on melanogenesis in PIG1 normal human skin melanocytes and its response to oxidative stress. The effect of kaempferol on melanin synthesis in PIG1 normal human skin melanocytes was explored by measuring tyrosinase activity, melanin content, mRNA and protein expression of key enzymes and expression of related pathway proteins. The effects of kaempferol pretreatment on cell viability, apoptosis, ROS level and HO-1 protein level under H2 O2 stimulation were explored. When treated with kaempferol, the tyrosinase activity and melanin content of PIG1 cells increased, the mRNA and protein expressions of TYR, TRP1, TRP2 and MITF increased, and the phosphorylation level of ERK1/2 increased. Upon the stimulation of H2 O2 , kaempferol reduced the production of ROS, decreased apoptosis and increased the protein expression of HO-1 in PIG1 cells. In addition, kaempferol inhibited oxidative stress-induced melanin reduction and promoted melanin synthesis in PIG1 cells and protected against H2 O2 -induced oxidative stress damage.
Subject(s)
Melanins , Monophenol Monooxygenase , Humans , Reactive Oxygen Species/metabolism , Monophenol Monooxygenase/genetics , Monophenol Monooxygenase/metabolism , Kaempferols/pharmacology , Melanocytes/metabolism , Oxidative Stress , RNA, Messenger/metabolismABSTRACT
The prognosis of skin cutaneous melanoma (SKCM) remains poor, and patients with SKCM show a poor response to immunotherapy. Thus, we aimed to identify necroptosis-related biomarkers, which can help predict the prognosis of SKCM and improve the effectiveness of precision medicine. Data of SKCM were obtained from The Cancer Genome Atlas (TCGA) and GEO databases. TCGA samples were classified into two clusters by consensus clustering of necroptosis-related genes. Univariate Cox and least absolute shrinkage and selection operator regression analyses led to the identification of 11 genes, which were used to construct a prognostic model. GSE65904 was used as the test set. Principal component, t-distributed stochastic neighbor embedding, and Kaplan-Meier survival analyses indicated that samples in the train and test sets could be divided into two groups, with the high-risk group showing a worse prognosis. Univariate and multivariate Cox regression analyses were performed, and a nomogram, calibration curve, and time-dependent receiver operating characteristic curve were constructed to verify the efficacy of our model. The 1-, 3-, and 5-year areas under the receiver operating characteristic curves for the train set were 0.702, 0.663, and 0.701 and for the test set were 0.613, 0.627, and 0.637, respectively. Moreover, we performed Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses between the high- and low-risk groups. Single sample gene set enrichment analysis, immune cell infiltration analysis, tumor microenvironment scores, immune checkpoint analysis, and half-maximal inhibitory concentration prediction indicated that the high-risk group showed weaker antitumor immunity; further, the response to immune checkpoint inhibitors was worse, and the high-risk group was sensitive to fewer antitumor drugs. Tumor mutational burden analysis, Kaplan-Meier survival analysis, and correlation analysis between risk score and RNA stemness score revealed that the high-risk group with low tumor mutational burden and high RNA stemness score was potentially associated with poor prognosis. To conclude, our model, which was based on 11 necroptosis-related genes, could predict the prognosis of SKCM; in addition, it has guiding significance for the selection of clinical treatment and provides new research directions to enhance necroptosis against SKCM.
Subject(s)
Melanoma , Skin Neoplasms , Humans , Melanoma/genetics , Melanoma/pathology , Skin Neoplasms/genetics , Skin Neoplasms/pathology , Necroptosis/genetics , Prognosis , Biomarkers, Tumor/genetics , RNA , Tumor Microenvironment/genetics , Melanoma, Cutaneous MalignantABSTRACT
Background: A large number of autologous melanocytes are required for surgical treatment of depigmentation diseases such as vitiligo. The purpose of this experiment is to explore the application of melanocytes induced by mesenchymal stem cells to clinical treatment. Therefore, we have induced mouse bone marrow mesenchymal stem cells (BMMSCs) into melanocytes (miMels) in the previous experiment. This experiment continues the previous experiment to further study the biological functions of miMels and their application in tissue engineering. Methods: We examined whether miMels can produce active tyrosinase, melanin, and response to α-MSH. The ability of miMels to produce melanin to keratinocytes was tested by co-culture. By applying miMels to tissue-engineered skin, the survival and function of miMels on the surface of nude mice were verified. Results: MiMels can produce active tyrosinase and melanin, and can pass melanin to the co-cultured keratinocytes. Under the stimulation of α-MSH, the active tyrosinase and melanin content of miMels increased. We tried to apply it to the establishment of tissue-engineered skin and obtained tissue-engineered skin containing miMels. Then we tried to transplant tissue-engineered skin on the back skin of nude mice and succeeded. The transplanted miMels survived in local tissues, synthesized active tyrosinase and melanin, and expressed the marker protein of melanocytes. Conclusion: In short, miMels can be used as a cell source for tissue engineering skin. MiMels not only have a typical melanocyte morphology but also have the same biological functions as normal melanocytes. What's more important is its successful application in mouse tissue-engineered experiments.
ABSTRACT
BACKGROUND: Alopecia areata is a common non-scarring alopecia, mainly manifested as sudden localized patchy alopecia. It is currently believed to be related to autoimmune, genetic, emotional stress, and endocrine factors. OBJECTIVE: The aim of this study was to evaluate the efficacy and safety of the mixed preparation of piperine, capsaicin, and curcumin on alopecia areata treatment. METHODS: Sixty patients were enrolled in this study and divided into 2 groups randomly: topical treated with the mixed preparation (case) twice daily and 5%minoxidil (control) once daily for 3 months. The degree of hair loss was assessed by SALT and dermoscopy. RESULTS: On the completion of the study, compared with baseline, statistically significant regrowth occurred in both groups (p < 0.05). The mean SALT scores and hair follicle status under trichoscopy at baseline and at the end of 12 weeks in the mixed preparation group and in the minoxidil group were comparable, respectively. The effective rate of mixed preparation group was 63.33% and minoxidil group was 70%. Adverse symptoms were temporary and no serious adverse event occurred. CONCLUSION: Based on our findings, the mixed preparation of piperine, capsaicin, and curcumin is effective in treating alopecia areata, but it has not been shown to be superior to minoxidil in short-term therapy.
Subject(s)
Alopecia Areata , Curcumin , Humans , Alopecia Areata/diagnosis , Minoxidil , Capsaicin/adverse effects , Curcumin/adverse effects , Administration, Topical , Alopecia/drug therapy , Alopecia/diagnosisABSTRACT
Ferroptosis is a non-apoptotic regulated cell death process, and much research has indicated that ferroptosis can induce the non-apoptotic death of tumor cells. Ferroptosis-related genes are expected to become a biological target for cancer treatment. However, the regulation of ferroptosis-related genes in skin cutaneous melanoma (SKCM) has not been well studied. In the present study, we conducted a systematic analysis of SKCM based on RNA sequencing data and clinical data obtained from The Cancer Genome Atlas (TCGA) database and the FerrD database. SKCM patients from the GSE78220 and MSKCC cohorts were used for external validation. Applying consensus clustering on RNA sequencing data from TCGA the generated ferroptosis subclasses of SKCM, which were analyzed based on the set of differentially expressed ferroptosis-related genes. Then, a least absolute shrinkage and selection operator (LASSO)-Cox regression was used to construct an eight gene survival-related linear signature. The median cut-off risk score was used to divide patients into high- and low-risk groups. The time-dependent receiver operating characteristic curve was used to examine the predictive power of the model. The areas under the curve of the signature at 1, 3, and 5 years were 0.673, 0.716, and 0.746, respectively. Kaplan-Meier survival analysis showed that the prognosis of high-risk patients was worse than that of low-risk patients. Univariate and multivariate Cox regression analyses showed that the risk signature was a robust independent prognostic indicator. By incorporating risk scores with tumor staging, a nomogram was constructed to predict prognostic outcomes for SKCM patients. In addition, the immunological analysis showed different immune cell infiltration patterns. Programmed-death-1 (PD-1) immunotherapy showed more significant benefits in the low-risk group than in the high-risk group. In summary, a model based on ferroptosis-related genes can predict the prognosis of SKCM and could have a potential role in guiding targeted therapy of SKCM.
ABSTRACT
Previous studies have shown the participation of the cytokines interleukin (IL) 17A and IL22 in the development of vitiligo. The aryl hydrocarbon receptor (AhR) functions in the pathogenesis of vitiligo and can modulate cytokine production. The aim of the present study was to determine the relationship between AhR activation and the secretion of IL17A and IL22 in CD4+ T cells in vitiligo. A total of 20 newly diagnosed patients with progressive, unstable vitiligo and 20 healthy controls were recruited. CD4+ T cells and skin samples were collected. Immunohistochemistry, ELISA, reverse transcription-quantitative PCR, western blotting and RNA interference experiments were performed. The expression of AhR was significantly lower in the CD4+ T cells and skin, both lesional and nonlesional, of patients with vitiligo compared with healthy subjects. AhR expression was markedly lower in nonlesional compared with lesional skin of patients with vitiligo. The expression levels of IL17A and IL22 were significantly higher in patients with vitiligo compared with healthy subjects. Knockdown of AhR significantly increased the production of IL17A and markedly decreased IL22 levels in the CD4+ T cells of patients with vitiligo. Ginkgo biloba extract EGb 761 activated AhR, inhibited IL17A secretion and enhanced IL22 release in the CD4+ T cells of patients with vitiligo. In conclusion, reduced AhR expression is associated with progressive, unstable vitiligo. Activation of AhR with G. biloba extract EGb 761 may have therapeutic potential for decreasing IL17A levels and increasing IL22 levels in patients with vitiligo.
ABSTRACT
BACKGROUND: Sweet, spicy or greasy food, staying up late, and using electronic products for a long time are common bad habits nowadays. Their role in skin diseases has been paid much attention. OBJECTIVE: The aim of this study was to investigate whether unhealthy lifestyles would affect the skin sebum content, SC hydration, and pH and how do they affect. METHODS: A total of 300 volunteers were enrolled, and a multifunctional skin physiology monitor measured the three skin biophysical properties on the forehead and dorsal hand. Lifestyle factors were evaluated by a self-administered questionnaire. RESULTS: Eating oily, sweet, spicy food, and staying up late increased the sebum content of the forehead significantly. Dorsal hand SC hydration was higher in people eating more sweet food and oily food, and forehead SC hydration was higher in people eating more sweet food and go to bed earlier. Eating sweet food could increase pH in both forehead and dorsal hand. The forehead pH decreased in using electronic products over 6 hours a day or staying up late. There are significant differences in sebum, hydration, and pH value among different age groups. In males, the pH was lower than females, but the sebum was higher. CONCLUSION: Sebum content, SC hydration, and pH are affected by unhealthy lifestyles, age, and gender.
Subject(s)
Skin Physiological Phenomena , Skin , Female , Humans , Hydrogen-Ion Concentration , Life Style , Male , SebumABSTRACT
Vitiligo is a refractory disfiguring skin disease. However, the aetiology and pathogenesis of vitiligo have not been fully defined. Previous studies have shown that exosomes from normal human keratinocytes improve melanogenesis by up-regulating the expression of melanogenesis-related proteins. Several microRNAs (miRNAs) have been demonstrated to be effective in modulating melanogenesis via exosomes. In the present study, it was found that the effect of exosomes derived from keratinocytes in vitiligo lesions in regulating melanin synthesis is weakened. Furthermore, miR-200c was detected to be significantly down-regulated in exosomes from keratinocytes in vitiligo lesions. In addition, miR-200c enhanced the expression of melanogenesis-related genes via suppressing SOX1 to activate ß-catenin. In conclusion, our study revealed that the effect of exosomes secreted by keratinocytes in vitiligo lesions exhibited a weaker capacity in promoting melanogenesis of melanocytes. Moreover, the expression of miR-200c, which mediates melanogenesis in exosomes secreted by keratinocytes in vitiligo lesions, is down-regulated, which may be one of the pathogenesis in vitiligo. Therefore, keratinocyte-derived exosomal miR-200c may be a potential target for the treatment of vitiligo.
Subject(s)
Down-Regulation/genetics , Exosomes/genetics , Keratinocytes/metabolism , Melanins/biosynthesis , MicroRNAs/genetics , Vitiligo/genetics , Vitiligo/pathology , Base Sequence , Epidermis/pathology , Exosomes/ultrastructure , Humans , Melanocytes/metabolism , MicroRNAs/metabolism , Models, Biological , Pigmentation/genetics , SOXB1 Transcription Factors/metabolism , beta Catenin/metabolismABSTRACT
BACKGROUND: Leucoderma, a depigmentation of the skin, is a common disease in humans, and has been observed in cattle, horses and buffalo as well. OBJECTIVES: To analyze the correlation between melanin stem cells and the differentiation and proliferation of melanocytes (MCs). MATERIAL AND METHODS: Magnetic cell separation was used to separate melan-A+ cells and PAX3+ cells, which were cultured in vitro. The L-DOPA staining was used to observe cell morphology; Cell Counting Kit-8 (CCK8) was used to determine the cell proliferation rate; and flow cytometry (FCM) was used to determine cell cycle changes. The relative mRNA levels of melanocyte-inducing transcription factor (MITF), dopachrome tautomerase (DCT) and melan-A in cells were determined with reverse-transcription polymerase chain reaction (RT-PCR). RESULTS: The number of MC dendrites increased and extended continually during in vitro culture following magnetic cell separation. The proportion of positive L-DOPA staining cells increased from a baseline 40.70% to 82.03%, and the cell proliferation rate increased from 335.0% at D3 to 1577.4% at D20. The results of FCM showed that the cell proportion at the G1 stage in the D20 group was significantly lower than the D3 group; the cell proportion at the G2/M stage also decreased significantly. The expression of MITF and melan-A increased as the culture time increased, while the expression of DCT decreased. CONCLUSIONS: The number of MC stem cells decreased and mature MCs increased gradually, indicating that MC stem cells can gradually differentiate into mature MCs during in vitro culture following magnetic cell separation.
Subject(s)
Melanocytes , Animals , Cattle , Cell Differentiation , Cell Separation , Cells, Cultured , Horses , Magnetic Phenomena , Melanins , TechnologyABSTRACT
BACKGROUND: Increased human endogenous retroviruses E clone 4-1 (HERV-E clone 4-1) mRNA expression is observed in systemic lupus erythematosus (SLE) patients and associates with the disease activity. In this study, we want to further investigate the mechanism of HERV-E clone 4-1 mRNA upregulation and its roles in SLE progression. METHODS: CD4+ T cells were isolated from venous blood of SLE patients or healthy controls and qRT-PCR was used to detect HERV-E clone 4-1 mRNA expression. We then investigated the regulation of Nuclear factor of activated T cells 1 (NFAT1) and Estrogen receptor-α (ER-α) on HERV-E clone 4-1 transcription and the functions of HERV-E clone 4-1 3' long terminal repeat (LTR) on DNA hypomethylation and IL-17 release. RESULTS: We found HERV-E clone 4-1 mRNA expression was upregulated in CD4+ T cells from SLE patients and positively correlated with SLE disease activity. This is associated with the activation of Ca2+/calcineurin (CaN)/NFAT1 and E2/ER-α signaling pathway and DNA hypomethylation of HERV-E clone 4-1 5'LTR. HERV-E clone 4-1 also takes part in disease pathogenesis of SLE through miR-302d/Methyl-CpG binding domain protein 2 (MBD2)/DNA hypomethylation and IL-17 signaling via its 3'LTR. CONCLUSIONS: HERV-E clone 4-1 mRNA upregulation is due to the abnormal inflammation/immune/methylation status of SLE and it could act as a potential biomarker for diagnosis of SLE. HERV-E clone 4-1 also takes part in disease pathogenesis of SLE via its 3'LTR and the signaling pathways it involved in may be potential therapeutic targets of SLE.
Subject(s)
CD4-Positive T-Lymphocytes/immunology , DNA-Binding Proteins/immunology , Endogenous Retroviruses/genetics , Interleukin-17/immunology , Lupus Erythematosus, Systemic/immunology , MicroRNAs/immunology , Adult , Cells, Cultured , DNA Methylation/genetics , DNA Methylation/immunology , Endogenous Retroviruses/immunology , Female , Humans , Lupus Erythematosus, Systemic/diagnosis , Lupus Erythematosus, Systemic/pathology , Male , RNA, Messenger/genetics , RNA, Messenger/immunology , Signal Transduction/immunologyABSTRACT
BACKGROUND: This study aims to screen the key genes and possible signaling pathways involved in the differentiation and proliferation of human melanocytes (MCs) by in vitro culture of mixed skin cells. This will be helpful to further study the mechanisms and treatment strategies of pigment-related diseases such as vitiligo. METHODS: Mixed skin cells were obtained by digesting and separating normal human foreskin tissues. Ribonucleic acid (RNA) was extracted from sorting cells and high-throughput transcriptome sequencing was performed at different culture time points. Differentially expressed genes (DEGs) were obtained by comparing the expression abundance of genes at different culture time points. Then the key genes and signaling pathways involved in the differentiation and proliferation of MCs were screened and verified by real-time quantitative polymerase chain reaction (qPCR) test. RESULTS: Twenty one DEGs were finally screened for further qPCR validation, mainly involved in 4 signaling pathways. The expressions of Wnt5A, Wnt5B, FZD2 and FZD3 in Wnt pathway were continuously up-regulated, and that of Wnt4 gene was continuously down-regulated, however, all the above hadn't been verified by qPCR. The expressions of COL5A2, COL6A3, ITGB1, ITGA4, ITGAV, AKT3, PIK3CD, PIK3R1 and PIK3R2 in phosphoinositide 3-kinase (PI3K) pathway were continuously up-regulated, of which PIK3CD, PIK3R2, COL5A2, ITGA4, ITGAV and AKT3 were verified by qPCR. PDGFB and GRB2 gene expressions were down-regulated in platelet-derived growth factor (PDGF) pathway, while PDGFRB was continuously up-regulated, of which PDGFB and PDGFRB were verified. The DHRS3, DHRS9, RDH10 and SDR16C5 genes in retinol metabolic pathway were continuously down-regulated and verified by qPCR. CONCLUSION: We suggested that Wnt5A gene in Wnt/ß-catenin classical pathway, integrin combining with extracellular matrix through PI3K signaling pathway, retinoic acid catabolism-related genes could promote the differentiation and proliferation of MCs; however, PDGFB gene might have a negative regulatory effect on the growth of MCs.
Subject(s)
Biomarkers/metabolism , Hair Follicle/cytology , Stem Cells/metabolism , Antigens, CD34/metabolism , Cell Differentiation/physiology , Humans , Intramolecular Oxidoreductases/metabolism , Keratinocytes/metabolism , Melanins/metabolism , Melanocytes/metabolism , PAX3 Transcription Factor/metabolismSubject(s)
Foot Diseases/pathology , Melanoma, Amelanotic/pathology , Skin Neoplasms/pathology , Female , Humans , Middle AgedABSTRACT
To clarify the characteristic growth of melanocytes (MCs) and Keratinocytes (KCs) in vitro and discuss the mechanism of culturing autologous melanocytes in the treatment of vitiligo. Epidermis cells derived from normal skin tissues were isolated and cultured in vitro. Melanocytes in DOPA staining were observed. The expression level of markers in MCs was detected by qRT-PCR and the percentage of MCs and KCs were detected by flow cytometry. Cells derived from normal skin tissues mainly included KCs, MCs, and fibroblasts. There were significant differences between the percentage of KC, MC, fibroblasts (P < 0.05), and the expression of Microphthalmia-associated transcription factor (P < 0.05) and Tyrosinase-related protein-2 (P < 0.05) in the second, 10th, 20th, and 30th day. Significant differences were also found between the average numbers of MC stained by DOPA (P < 0.05) and the average percentage of MCs in the 10th, 20th, and 30th Day (P < 0.05). But there were no significant differences between the average percentage of KCs in the 10th, 20th, and 30th Day (P > 0.05) detected by flow cytometry. The number of MCs co-cultured with KCs in vitro reached the maximum in the 20th Day and this co-cultured model may contribute to the growth of MCs which could be used in the treatment of vitiligo.
Subject(s)
Keratinocytes/cytology , Melanocytes/cytology , Skin/cytology , Vitiligo/pathology , Adult , Case-Control Studies , Cell Differentiation , Cell Proliferation , Cells, Cultured , Coculture Techniques , Female , Humans , In Vitro Techniques , Keratinocytes/metabolism , Male , Melanocytes/metabolism , Skin/metabolism , Vitiligo/metabolismABSTRACT
To describe the prevalence of human papillomavirus (HPV) and its genotype distribution among females in the suburb of Shanghai. A total of 33 562 participants were enrolled in this study from January to December 2016. HPV GenoArray test kit was used to perform HPV genotyping and was also used in DNA amplification and HybriBio's proprietary flow-through hybridization technique. The overall prevalence of HPV was 18.98% and the top ten genotypes of HPV infection were HPV 16 (3.36%), HPV 58 (2.65%), HPV 52 (2.48%), HPV 51 (1.58%), HPV 54 (1.40%), HPV 68 (1.32%), HPV 18 (1.23%), HPV 6 (1.15%), HPV 56 (1.10%), and HPV 33 (1.07%). Single infection (4749, 14.15%) was the most common types among all the infected cases. Significant differences were found among age groups and month groups in terms of simple and multiple infection (P < 0.05), pure HR, LR and mixed HPV infection (P < 0.05). The prevalence of HR and LR HPV infection among females in the suburb of Shanghai is high, prevalence of single and multiple infection, pure HR, LR and mixed infection is correlated with the age and month.
Subject(s)
Papillomaviridae/genetics , Papillomavirus Infections/epidemiology , Papillomavirus Infections/virology , Adolescent , Adult , Aged , Aged, 80 and over , Cervix Uteri/cytology , Cervix Uteri/virology , Child , China/epidemiology , Coinfection/epidemiology , DNA, Viral/genetics , Female , Genotype , Genotyping Techniques , Human papillomavirus 16/genetics , Human papillomavirus 16/isolation & purification , Human papillomavirus 18/genetics , Human papillomavirus 18/isolation & purification , Humans , Middle Aged , Nucleic Acid Hybridization , Papillomaviridae/classification , Papillomaviridae/isolation & purification , Prevalence , Young AdultABSTRACT
BACKGROUND: Previous studies have reported that ultraviolet B (UVB) inhibits DNA methyltransferase1 (DNMT1) activity in CD4+ T cells from systemic lupus erythematosus (SLE) patients. Silent mating type information regulation 2 homolog 1 (SIRT1) is a type of Class III histone deacetylases (HDACs), and has been reported to play roles in the pathogenesis of different autoimmune diseases and can modulate DNMT1 activity. Moreover, aryl hydrocarbon receptor (AhR) has been reported to link UVB with SLE. However, the exact mechanisms by which DNMT1 activity is inhibited by UVB in lupus CD4+ T cells remain largely unknown. OBJECTIVE: To elucidate the exact mechanisms by which DNMT1 activity is inhibited by UVB in lupus CD4+ T cells. METHODS: Twenty-two newly diagnosed active SLE patients and 30 healthy controls were enrolled in the study. CD4+ T cells were isolated, cultured and treated. DNMT1 activity assay, quantitative real-time PCR (qRT-PCR), Western blotting, RNA interference using small interfering RNA and Chromatin Immunoprecipitation (ChIP) assay were employed. RESULTS: DNMT1 activity was inhibited in si-SIRT1-transfected CD4+ T cells, and increased by the established SIRT1 activator, SRT1720. Moreover, the mRNA and protein expression of SIRT1 were suppressed by UVB exposure in lupus CD4+ T cells. UVB-inhibited DNMT1 activity was reversed by SRT1720 in si-control-transfected lupus CD4+ T cells, but not in si-SIRT1-transfected lupus CD4 + T cells. Furthermore, AhR activation by VAF347 reduced the mRNA and protein expression of SIRT1. ChIP using an antibody against AhR in normal CD4+ T cells revealed a 16-fold stronger signal at the site about 1.6kb upstream from the translation start site of the SIRT1 promoter. Finally, UVB could activate AhR and inhibit the mRNA and protein expression of SIRT1. AhR knockdown abrogated the inhibition of UVB-mediated SIRT1 mRNA and protein expression and DNMT1 activity in lupus CD4+ T cells. CONCLUSION: UVB suppressed SIRT1 expression via activating AhR, and subsequently inhibited DNMT1 activity in CD4+ T cells from SLE patients.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/radiation effects , CD4-Positive T-Lymphocytes/radiation effects , DNA (Cytosine-5-)-Methyltransferase 1/metabolism , Lupus Erythematosus, Systemic/enzymology , Receptors, Aryl Hydrocarbon/radiation effects , Sirtuin 1/metabolism , Ultraviolet Rays , Adult , Basic Helix-Loop-Helix Transcription Factors/genetics , Basic Helix-Loop-Helix Transcription Factors/metabolism , CD4-Positive T-Lymphocytes/enzymology , CD4-Positive T-Lymphocytes/immunology , Case-Control Studies , Cells, Cultured , Gene Expression Regulation, Enzymologic , Humans , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/genetics , Lupus Erythematosus, Systemic/immunology , Promoter Regions, Genetic , RNA Interference , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptors, Aryl Hydrocarbon/genetics , Receptors, Aryl Hydrocarbon/metabolism , Signal Transduction/radiation effects , Sirtuin 1/genetics , TransfectionABSTRACT
Large numbers of autogenous melanocytes (Mcs) are required when conducting studies on tissue engineering of skin and performing surgical treatment of depigmentation diseases. This study was conducted to explore the possibility of inducing differentiation of bone marrow mesenchymal stem cells (MSCs) into Mcs as a means of providing autogenous Mcs for purposes of tissue engineering and clinical treatment. MSCs were harvested from the bone marrows of black mice; and after six passages, hydrocortisone, insulin, transferrin and fibroblast growth factor were applied to induce their differentiation into Mcs. The morphological and ultrastructural characteristics of the newly differentiated cells were observed. The transcription and expression of multiple markers were examined using qRT-PCR, Western blot, and immunofluorescence analysis. Cell cycle phases and yields of Mcs were analyzed by flow cytometry. Following 120-180 days induction, differentiated cells were morphologically similar to Mcs, and mature melanosomes were observed. Multiple markers of Mcs, but not melanoma cells, were expressed by the differentiated cells. Most induced Mcs were in phase G1 or S, and yield of target cells was â¼80%. Mcs induced from bone marrow MSCs for periods of 120-180 days represent a potential source of autogenous Mcs.
Subject(s)
Cell Differentiation , Melanocytes/cytology , Mesenchymal Stem Cells/cytology , Animals , Cell Culture Techniques/methods , Cell Cycle , Cells, Cultured , Male , Mice , Mice, Inbred C57BL , Tissue Engineering/methodsABSTRACT
BACKGROUND: Previous studies have reported that the DNA of T cells from systemic lupus erythematosus (SLE) patients contains global hypomethylation that may contribute to the development of SLE. Human endogenous retroviruses (HERVs) are encoded within the genomes of all higher eukaryotes and are of special interest where autoimmune disorders are concerned. Until recently, minimal effort has been made to identify specific HERVs associated with SLE and to explore their precise mechanism of expression. OBJECTIVE: To examine the expression of HERVs associated with SLE and elucidate the effect of ultraviolet B (UVB) exposure on SLE-associated HERV expression in CD4+ T cells from patients with SLE. METHODS: Fifteen patients with SLE and 10 healthy controls were enrolled in the study. The mRNA expression of selected HERVs and the methylation status of the long terminal repeats (LTRs) in SLE-related HERVs in CD4+ T cells were investigated. Furthermore, CD4+ T cells treated with 5-aza-deoxycytidine (5-aza C) and UVB were analyzed. Reverse-transcription PCR (RT-PCR), quantitative real-time PCR (qRT-PCR) and bisulfite sequencing analysis were employed. RESULTS: HERV-E mRNA expression was higher in lupus CD4+ T cells than in cells from healthy controls, whereas the mRNA expression levels of HERV-K, HERV-K10 and HERV-W were comparable in SLE patients and healthy controls. Additionally, the HERV-E mRNA expression level was positively correlated with SLE disease activity. Furthermore, the HERV-E LTR methylation level was decreased and was negatively correlated to the HERV-E mRNA expression level in lupus CD4+ T cells. Finally, lupus CD4+ T cells showed markedly decreased HERV-E LTR2C methylation levels and increased HERV-E mRNA expression after treatment with 5-aza C or UVB. CONCLUSION: HERV-E is involved in the development of SLE. HERV-E transcription may be activated via inhibition of LTR methylation in lupus CD4+ T cells.