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Objective To compare and analyze the differences in the transcriptomics between mycelium and early yeast phases of Sporothrix schenckii (S.schenckii),and to realize the changes in transcriptome expression profiles during mycelium-to-yeast transformation.Methods A standard strain of S.schenckii (ATCC 10268) was subjected to 96-hour culture with Sabouraud medium at 25 ℃ or 36-hour culture with brain-heart infusion medium at 37 ℃ to obtain the mycelium and yeast form of S.schenckii,and then,their transcriptomes were sequenced.Functional annotation was performed for screened unigenes by comparison using several databases (such as NR,Swiss-Prot,KEGG,COG,KOG,GO and Pfam),coding sequence prediction,and gene expression analysis in each sample.Finally,the differentially expressed genes were subjected to pattern clustering,functional annotation and enrichment analysis.Results A total of 14.76 Gb valid data (clean data) were obtained,and functional annotation results were acquired in 28 094 of 43 863 assembled unigene clusters.Compared with S.schenckii in mycelium phase,there were 10 969 up-regulated genes and 199 down-regulated genes in S.schenckii in yeast phase.These differentially expressed genes were involved in protein phosphorylation,intracellular protein transport,cellular protein modification,small guanosine triphosphate-mediated signal transduction,vesicle-mediated transport,translation,intracellular signal transduction,microtubule formation,adenosine triphosphate synthesis,coupled proton transport and so on.Sixteen genes in the mitogen-activated protein kinase (MAPK) pathway and two-component signaling pathway,which were two important signal transduction pathways involved in fungal morphogenesis,and 16 genes involved in chitin synthesis and metabolism were all confirmed to be up-regulated in S.schenckii in yeast phase.Conclusions Compared with S.schenckii in mycelium phase,great changes in gene expression profiles were observed in S.schenckii in yeast phase.These differentially expressed genes are involved in many functions,suggesting that the dimorphic transition of S.schenckii is regulated by a multi-gene network system.
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BACKGROUND:The development of keloid is a progress of fibrosis in wound healing, and involves various fibrosis-related cytokines. Bone morphogenetic protein 7 (BMP7), Gremlin and high mobility group box-1 (HMGB1) play an important role in fibrosis of many organs, but their role in keloid tissue has rarely been reported. OBJECTIVE:To investigate the role of BMP7, Gremlin, vascular endothelial growth factor (VEGF) and HMGB1 in the development of keloid. METHODS:The protein levels and distribution of BMP7, Gremlin, VEGF and HMGB1 in 20 cases of keloid and 20 cases of normal skin were detected by immunohistochemistry and western blot analysis, respectively. And the correlations among expression levels of BMP7, Gremlin, VEGF and HMGB1 in keloid were analyzed. RESULTS AND CONCLUSION:In keloid tissue, the expression levels of Gremlin, VEGF and HMGB1 were significantly higher than that in normal skin (P<0.01), while the expression levels of BMP7 were significantly lower (P<0.01). The levels of Gremlin were negatively correlated with the levels of BMP7 (r=-0.539, P<0.05). And the levels of VEGF were positively correlated with the levels of HMGB1 (r=0.56, P<0.05). The overexpression of Gremlin and decreased expression of BMP7, as wel as the increased expression of HMGB1 and VEGF, may contribute to the pathogenesis of fibrosis in the development of keloid.
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BACKGROUND:YY1 is mainly expressed in the undifferentiated epidermic cells in mouse basal lamina, and the expression level is gradual y down-regulated as the differentiation towards suprabasal lamina. The differential expression indicates that, YY1 is one of the regulators in the process of epidermic cells differentiation. OBJECTIVE:To observe the effects of YY1 over-expression on the differentiation of HaCaT cells infected with lentivirus. METHODS:Lentivirus-YY1 was transferred into the HaCaT cells by using Lipofectamine 2000. After selection of the puromycin, monoclonal celllines were established, and the control group were lentivirus-infected HaCaT cells and uninfected HaCaT cells. The expression of YY1 was detected by using western blot analysis. cells in Lentivirus-YY1-HaCaT group and HaCaT-YY1 group were further divided into two subgroups according to the calcium concentration in culture medium, cells were either cultured in low-calcium medium (0.12 mmol/L) for 48 hours, or cultured in low-calcium medium (0.12 mmol/L) for 24 hours and in high-calcium medium (0.35 mmol/L) for additional 24 hours. Keratin K1, K10, K14, and involucrin, filaggrin and loricrin after over-expression of YY1 were detected with western blot analysis. RESULTS AND CONCLUSION:The HaCaT cells were successful y infected with lentivirus-YY1, and we obtained over-expression of YY1 protein in monoclonal celllines under high-calcium concentrations, the over-expressed YY1 could decrease the expression of K1, involucrin and loricrin, thereby preventing the process of epidermal keratinocytes and maintaining the cells in an undifferentiated state. Lentivirus can efficiently infect human immortalized epidermal cellHaCaT, and YY1 may the important factor of inhibiting the differentiation of basal epidermal cells and maintaining the undifferentiated proliferation status.
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Objective To investigate the incidence of insomnia in patients with nasopharyngeal carcinoma during radiotherapy and analyze the influencing factors.Method A sleep quality questionnaire for patients with nasopharyngeal carcinoma during radiotherapy was designed and used for the investigation of the incidence of insomnia as well as the influencing factors related to insomnia.Results The incidence of insomnia was 42.2%.Univariate analysis revealed that radioactive xerostomia,nausea,hospital environmental noise,radioactive oral mucositis,worry about job,worry about prognosis,burp,vomiting,worry about family influence were related to insomnia in nasopharyngeal carcinoma patients during radiotherapy;multivariate analysis showed that radioactive oral mucositis,environmental noise and job stress were the independent factors leading to insomnia(All P<0.05).Conclusions There is a high incidence of insomnia in patients with nasopharyngeal carcinoma during radiotherapy.Radioactive oral mucositis,environmental noise and job stress might be the independent factors relative with insomnia during radiotherapy.