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【Objective】 To investigate the effects of miR-29b-3p on congenital heart disease and its mechanism. 【Methods】 The expression level of miR-29b-3p in serum from CHD patients and normal individuals， and in cells was detected by RT-qPCR. Dimethylsulfoxide （DMSO） was used to induce P19 cells to differentiate into cardiomyocytes. Western blot was used to measure the expression levels of cardiogenesis-associated genes， and phosphatase and tensin homolog （PTEN） level in cells. The proliferation and migration of cardiomyocytes were measured by CCK-8 and Transwell assay， respectively. Dual-luciferase gene reporter assay was used to verify the targeted relationship between miR-29b-3p and PTEN. 【Results】 Compared with that of normal individuals， the expression of miR-29b-3p in CHD patients was decreased. During differentiation， miR-29b-3p level was higher at late stage than that at early stage. Downregulated miR-29b-3p inhibited the differentiation of P19 cells into cardiomyocytes， and inhibited cell proliferation and migration. miR-29b-3p targeted PTEN. The increased PTEN level induced by miR-29b-3p knockdown inhibited the differentiation of P19 cells， and proliferation and migration of cardiomyocytes. 【Conclusion】 miR-29b-3p was downregulated in the serum of CHD patients. The downregulation of miR-29b-3p inhibited the differentiation of P19 cells， proliferation and migration of cardiomyocytes by targeting and regulating PTEN.
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Objective Based on the observation of the changes of symptoms, histopathology, visceral sensitivity, mast cell activation, autophagy, and Beclin-1 and Claudin-2 expression in rats, we established and evaluated a new rat model of diarrhea-predominant irritable bowel syndrome (D-IBS) induced by restraint-stress combined with capsaicin (CAP) administration.Methods Forty healthy 5-week old male Sprague Dawley (SD) rats were randomly divided into normal group, model group I, model group II and model group III, with 10 rats in each group.The D-IBS model was established by restraint-stress combined with intragastric administration of CAP (2 mL/100 g body weight, 0.125% in group I, 0.250% in group II, 0.500% in group III), tail clipping and forelimb restriction for 30 minutes every day for 2 weeks.The rats in the control group were treated with saline for 2 weeks.The number of contraction of abdominal wall and arched back were measured by Power Lab instrument.The mast cell activation was detected using aldehyde-magenta-orange G staining.Light and electron microscopic examinations were performed to detect the morphology and autophagy of colonic tissues.The expressions of Beclin-1 and Claudin-2 in the colonic mucosa were detected by streptavidin-biotin complex (SABC) immunohistochemical staining.Results All rats in the model group III died during the experiment.Compared with the control group and model group I, the stool frequency was increased and the visceral sensitivity threshold decreased in the model group II, and there were statistically significant differences between the model group II and the control and model groups I (P < 0.05).The colonic mucosa, mucosal epithelium and glands in each group showed normal morphology and there was no submucosal vasodilatation and diffuse inflammatory cell infiltration.Except for the control group, round purple-reddish staining spots were observed in the rat mucosal stroma or submucosa in the model groups I and II, indicating an increased expression of mast cells.The autophagy, expressions of Beclin-1 and Claudin-2 in the colonic epithelium were significantly increased in the model group II compared with control group and model group I (P< 0.05).Conclusions The model of D-IBS induced by restraint-stress combined with capsaicin is characterized by increased diarrhea, visceral hypersensitivity, increased mast cell expression and autophagy of intestinal epithelial cells, and disruption of the intestinal mucosal barrier.This model is simple to set up and shows similar symptoms of human irritable bowel syndrome.Therefore, it is worthy of popularization and application.
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Objective To investigate the effect of hyperthermia on the expression of E6 and E7 genes of human papillomavirus (HPV) type 6 and 11 in HPV-infected human skin. Methods Tissue samples were obtained from the lesions of condyloma accuminatum (CA) in 6 patients after informed consent. Each sample was divided into 4 parts: one was embedded and directly stored at -80 ℃; the other 3 parts were placed in culture medium and the surface of the samples was irradiated for 30 minutes with a thermotherapy apparatus at 37℃, 42 ℃, 45 ℃, respectively, then the samples were taken out and stored at -80 ℃. RNA was extracted from the specimens, real time quantitative PCR (qPCR) was performed to detect the expression of E6 and E7 genes of HPV-6 and -11. Results Of the 6 patients, 2 were infected with HPV-6 and -11 respectively, 4 with both HPV-6 and HPV-11. The expression of E6 and E7 mRNA decreased with the increase in irradiation temperature. The relative mRNA expression levels at 37 ℃, 42 ℃ and 45 ℃ were 1.00 ± 0.00, 0.61 ± 0.17, 0.27 ± 0.15, respectively, for HPV-6 E6 gene, 1.00 ± 0.00, 0.56 ± 0.21, 0.16 ± 0.11 respectively, for HPV-6 E7 gene, 1.00 ± 0.00, 0.60 ± 0.22, 0.16 ± 0.08, respectively, for HPV-I1 E6 gene, 1.00 ± 0.00, 0.55 ± 0.15, 0.24 ± 0.06, respectively, for HPV-11 E7 gene; statistical difference was noted among them between the specimens irradiated at different temperature (all P < 0.01). Conclusion Hyperthermia can remarkably suppress the expression of HPV-6/I 1 E6 and E7 genes, which may be a possible mechanism under the regression of warts induced by local hyperthermia.