Objective To investigate the effects of low-to-moderate intensity treadmill exercise on pain and anxiety-like behaviors in rats with chronic constriction injury of the sciatic nerve(CCI),and to explore the neural mechanism of the exercise-related brain-derived neurotrophic factor(BDNF)/tropomyosin receptor kinase B(TrkB)-cAMP-response element binding protein(CREB)pathway in relieving pain and anxiety behaviors in CCI rats.Methods Thirty-two D rats were divided randomly into four groups:sham group,CCI group,sham+exercise(Sham+Exe)group,and CCI+exercise(CCI+Exe)group.Rats in the exercise groups underwent treadmill training for 4 weeks.The paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)were measured before and at different time points after the operation.The elevated plus maze(EPM)and open field test(OFT)were used to evaluate anxiety-like behaviors in the rats.mRNA and protein expression levels of BDNF,TrkB,and CREB in the hippocampus were detected by real-time quantitative reverse transcription PCR and Western Blot,respectively.Results(1)The PWT and PWL on the operative side of the rats were significantly lower in the CCI compared with the sham group at 7,14,21,28,and 35 days after the operation(P＜0.001).The PWT on the ipsilateral side was significantly increased in the CCI+Exe group after 21 days compared with the CCI group(P＜0.05),and the PWL on the ipsilateral side increased significantly after 14 days(P＜0.05).(2)The EPM result showed that rats in the CCI group spent a significantly lower proportion of time in the open arms(P＜0.001)and significantly more time in the closed arms compared with the sham group(P＜0.01).Rats in the CCI+Exe group spent significantly more time in the open arms than the CCI group(P＜0.05).(3)The OFT result showed that rats in the CCI group spent a significantly lower proportion of time in the central area of the open field compared with the sham group(P＜0.001),while the percentage of time was significantly increased in the CCI+Exe group compared with the CCI group(P＜0.05).(4)BDNF,TrkB,and CREB mRNA and protein levels in the hippocampus were significantly lower in the CCI group compared with the sham group(P＜0.05,P＜0.01).Four-week treadmill exercise increased the mRNA and protein expression levels of BDNF,TrkB,and CREB in the hippocampus of CCI rats(P＜0.05).Conclusions Four weeks of treadmill exercise alleviates mechanical and thermal hyperalgesia and anxiety induced by chronic pain in CCI rats.Up-regulation of the BDNF/TrkB-CREB pathway may be one of the mechanisms by which exercise relieves chronic pain and improves anxiety.

Objective:To investigate the expression of microRNA (miR) -26a in human skin fibroblasts during photoaging induced by ultraviolet A (UVA) , and to evaluate the effect of up-or down-regulation of miR-26a expression on the methylation level of the whole genome, the target gene enhancer of zeste homolog 2 (EZH2) and cell aging.Methods:Some human skin fibroblasts were irradiated with 10 J/cm 2 UVA once a day for 7 consecutive days, RNA was extracted on days 0, 3 and 7, and real-time quantitative reverse PCR (RT-PCR) was performed to determine the expression of miR-26a; miR-26a mimics and inhibitors were transfected into fibroblasts to up-or down-regulate the expression of miR-26a respectively, and fluorescence microscopy and RT-PCR were performed to determine the expression of miR-26a and evaluate the transfection efficiency. Some human skin fibroblasts were divided into 6 groups: blank control group receiving no treatment, UVA group treated with UVA irradiation according to the above method, miR-26a mimic group transfected with miR-26a-mimics, UVA+miR-26a mimic group transfected with miR-26a-mimics followed by UVA irradiation, miR-26a inhibitor group transfected with miR-26a inhibitors, UVA+miR-26a inhibitor group transfected with miR-26a inhibitors followed by UVA irradiation. On day 7, cells in each group were collected after the end of UVA irradiation. Then, flow cytometry was performed to detect cell cycle, DNA methylation quantitative detection kit was used to detect the methylation level of whole genome, RT-PCR was conducted to determine the mRNA expression of EZH2 (a histone-lysine N-methyltransferase enzyme) , DNA methyltransferase 1 (DNMT1) and miR-26a, and Western blot analysis was performed to determine the protein expression of EZH2 and DNMT1. Statistical analysis was carried out by using one-way analysis of variance and least significant difference- t test. Results:Compared with the unirradiated control group, the expression of miR-26a gradually increased in the UVA irradiation group over time during the culture, and there was a significant difference in the expression of miR-26a between the two groups after 7 days of UVA irradiation ( t=5.295, P < 0.05) . Strong fluorescence signals were observed in the miR-26a mimic-or miR-26a inhibitor-transfected fibroblasts, suggesting a high transfection efficiency. Flow cytometry showed that the proportion of cells at G1 phase significantly differed among the blank control group, UVA group, miR-26a mimic group, UVA+miR-26a mimic group, miR-26a inhibitor group, and UVA+miR-26a inhibitor group (52.82% ± 2.56%, 78.56% ± 4.34%, 53.63% ± 3.13%, 89.52% ± 4.17%, 54.39% ± 3.86%, 65.34% ± 4.78%, respectively; F=46.728, P < 0.01) , and significantly higher in the UVA group than in the blank control group ( t=8.848, P < 0.01) , higher in the UVA+miR-26a mimic group than in the miR-26a mimic group and UVA group ( t=11.922, 3.154, P < 0.01, < 0.05, respectively) , and higher in the UVA+miR-26a inhibitor group than in the miR-26a-inhibitor group ( t=3.087, P < 0.05) , but significantly lower in the UVA+miR-26a inhibitor group than in the UVA group ( t=3.547, P < 0.05) . Detection of the genome-wide methylation level showed that the methylation level ( A450 value) significantly differed among the above groups (0.676 ± 0.024, 0.323 ± 0.043, 0.506 ± 0.035, 0.169 ± 0.024, 0.602 ± 0.036, 0.422 ± 0.029, respectively, F=97.402, P < 0.01) , and significantly lower in the UVA group than in the blank control group ( P < 0.01) , lower in the UVA+miR-26a mimic group than in the miR-26a mimic group and UVA group (both P < 0.01) , and lower in the UVA+miR-26a inhibitor group than in the miR-26a inhibitor group ( P < 0.01) , but significantly higher in the UVA+miR-26a inhibitor group than in the UVA group ( P < 0.05) . RT-PCR and Western blot analysis showed significant differences in the mRNA and protein expression of EZH2 and DNMT1 respectively among the 6 groups (both P < 0.05) , which were significantly lower in the UVA group than in the blank control group ( P < 0.05) , lower in the UVA+miR-26a mimic group than in the miR-26a mimic group and UVA group (both P < 0.05) , and lower in the UVA+miR-26a inhibitor group than in the miR-26a inhibitor group ( P < 0.05) , but significantly higher in the UVA+miR-26a inhibitor group than in the UVA group ( P < 0.05) . Conclusion:In the UVA irradiation-induced photoaging of skin fibroblasts, miR-26a expression was up-regulated, cellular proliferative activity and genome-wide methylation level decreased; up-regulation of miR-26a expression could down-regulate the expression of its target gene EZH2 and methylation-related gene DNM1, and promote cell photoaging, while down-regulation of miR-26a expression could up-regulate the expression of EZH2 and DNMT1, and inhibit cell photoaging.

There are few clinical reports on the diagnosis and treatment of upper urinary tract stones secondary to urinary diversion. This study included 30 patients with upper urinary tract stones secondary to urinary diversion, and all of which were successfully managed. The individualized treatment with the ureteroscopy and/or percutaneous nephrolithotripsy with antegrade, retrograde, or a combination of antegrade and retrograde is safe and feasible.