ABSTRACTPurpose:RNA activation (RNAa) is a mechanism of gene activation triggered by promoter-targeted small double stranded RNAs (dsRNAs), also known as small activating RNAs (saRNAs). Myogenic regulatory factor MyoD is regarded as the master activator of myogenic differentiation cascade by binding to enhancer of muscle specific genes. Stress urinary incontinence (SUI) is a condition primarily resulted from urethral sphincter deficiency. It is thus expected that by promoting differentiation of adipose-derived stem cells (ADSCs) into myoblasts by activating MyoD gene through RNAa may offer benefits to SUI.Materials and Methods:Rats ADSCs were isolated, proliferated in vitro, and identified by flow cytometry. Purified ADSCs were then transfected with a MyoD saRNA or control transfected. Real-time polymerase chain reaction (RT-PCR) and western blotting were used to detect MyoD mRNA and protein expression, respectively. Immunocytochemical staining was applied to determine the expression of desmin protein in transfected cells. Cell viability was measured by using CellTiter 96® AQueous One Solution Cell Proliferation Assay kit.Results:Transfection of a MyoD saRNA (dsMyoD) into ADSCs significantly induced the expression of MyoD at both the mRNA and protein levels, and inhibited cell proliferation. Desmin protein expression was detected in dsMyoD treated ADSCs 2 weeks later.Conclusion:Our findings show that RNAa mediated overexpression of MyoD can promote transdifferentiation of ADSCs into myoblasts and may help treat stress urinary incontinence (SUI)–a condition primarily resulted from urethral sphincter deficiency.

Cordyceps is a fastidious pathogenic fungus infecting insects, and recent years have witnessed rapid progress in its medical properties. In this study, a wild isolate, C. cicadae MP12, was characterized through in vitro cultivation and its nuclear small-subunit (SSU) ribosomal DNA (rDNA) data. In vitro culture of C. cicadae MP12 was established by growing its fruiting bodies in a solid matrix. C. cicadae MP12 was inoculated into Cryptotympana atrata cicada pupae for in vivo culture, where the fungi developed its fruiting body as well. The contents of adenosine and cordycepin in dried fruiting bodies after culture were 1421.45µg/g and 1398.12 µg/g, respectively. Therefore, the established cultures from this study could be used for the production of various medically important metabolic substances.