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Objectives: Nerve growth factor (NGF) induces neuron transdifferentiation of adrenal medulla chromaffin cells (AMCCs) and consequently downregulates the secretion of epinephrine (EPI), which may be involved in the pathogenesis of bronchial asthma. Mammalian achaete scute-homologous 1 (MASH1), a key regulator of neurogenesis in the nervous system, has been proved to be elevated in AMCCs with neuron transdifferentiation in vivo. This study aims to explore the role of MASH1 in the process of neuron transdifferentiation of AMCCs and the mechanisms., Methods: Rat AMCCs were isolated and cultured. AMCCs were transfected with siMASH1 or MASH1 overexpression plasmid, then were stimulated with NGF and/or dexamethasone, PD98059 (a MAPK kinase-1 inhibitor) for 48 hours. Morphological changes were observed using light and electron microscope. Phenylethanolamine-N-methyltransferase (PNMT, the key enzyme for epinephrine synthesis) and tyrosine hydroxylase were detected by immunofluorescence. Western blotting was used to test the protein levels of PNMT, MASH1, peripherin (neuronal markers), extracellular regulated protein kinases (ERK), phosphorylated extracellular regulated protein kinases (pERK), and JMJD3. Real-time RT-PCR was applied to analyze the mRNA levels of MASH1 and JMJD3 . EPI levels in the cellular supernatant were measured using ELISA., Results: Cells with both tyrosine hydroxylase and PNMT positive by immunofluorescence were proved to be AMCCs. Exposure to NGF, AMCCs exhibited neurite-like processes concomitant with increases in pERK/ERK, peripherin, and MASH1 levels (all P <0.05). Additionally, impairment of endocrine phenotype was proved by a signifcant decrease in the PNMT level and the secretion of EPI from AMCCs (all P <0.01). MASH1 interference reversed the effect of NGF, causing increases in the levels of PNMT and EPI, conversely reduced the peripherin level and cell processes (all P <0.01). MASH1 overexpression significantly increased the number of cell processes and peripherin level, while decreased the levels of PNMT and EPI (all P <0.01). Compared with the NGF group, the levels of MASH1, JMJD3 protein and mRNA in AMCCs in the NGF+PD98059 group were decreased (all P <0.05). After treatment with PD98059 and dexamethasone, the effect of NGF on promoting the transdifferentiation of AMCCs was inhibited, and the number of cell processes and EPI levels were decreased (both P <0.05). In addition, the activity of the pERK/MASH1 pathway activated by NGF was also inhibited., Conclusions: MASH1 is the key factor in neuron transdifferentiation of AMCCs. NGF-induced neuron transdifferentiation is probably mediated via pERK/MASH1 signaling.

Objectives: At present, there are few studies on the sexual activity of young and middle-aged men in China. This study aims to explore the factors that affect the frequency of intercourse among young and middle-aged men in China., Methods: Data for 923 men aged 20-60 years were collected in the Health Management Center, Third Xiangya Hospital, Central South University from January 2019 to March 2019, and a questionnaire survey (including basic conditions and sexual function-related scales) was carried out in the subjects. Finally, the data were analyzed by multivariate logistic regression analysis., Results: The age ( P <0.01), number of children ( P <0.01), total cholesterol ( P <0.05), low-density lipoprotein ( P <0.05), erectile function ( P <0.01) and premature ejaculation ( P <0.05) was significantly correlated with the frequency of intercourse in the young and middle-aged men in China., Conclusions: The sexual frequency in the middle-young men in China is closely related to factors such as age, number of children, total cholesterol, low-density lipoprotein, and sex function.