Oleanolic acid (OA) has a wide variety of bioactivities such as hepatoprotective, anti-inflammatory and anti-cancer activity and is used for medicinal purposes in many Asian countries. In the present study, the effect of OA on induction of autophagy in human hepatocellular carcinoma HepG2 and SMC7721 cells and the related mechanisms were investigated. MTT assay showed that OA significantly inhibited HepG2 and SMC7721 cells growth. OA treatment enhanced formation of autophagic vacuoles as revealed by monodansylcadaverine (MDC) staining. At the same time, increasing punctuate distribution of microtubule-associated protein 1 light chain 3 (LC3) and an increasing ratio of LC3-II to LC3-I were also triggered by OA incubation. In addition, OA-induced cell death was signifi cantly inhibited by autophagy inhibitors 3-methyladenine (3-MA) and chloroquine (CQ) pretreatment. And we found out that OA can suppress the PI3K/Akt1/mTOR signaling pathway. Furthermore, our data suggested that OA-triggered autophagy was ROS-dependent as demonstrated by elevated cellular ROS levels by OA treatment. When ROS was cleared by N-acetylcysteine (NAC), OA-induced LC3-II convertsion and cell death were all reversed. Taken together, our results suggest that OA exerts anticancer eff ect via autophagic cell death in hepatocellular carcinoma.
Acetylcysteine ; Asian Continental Ancestry Group ; Autophagy* ; Carcinoma, Hepatocellular* ; Cell Death ; Chloroquine ; Humans ; Microtubule-Associated Proteins ; Oleanolic Acid* ; Vacuoles

Objective:To investigate the application effect of the compound flap transfer in tubularized incised plate (TIP) urethroplasty.Methods:Children with hypospadias who received TIP in the Department of Pediatric Urology, the Affiliated Xuzhou Children’s Hospital of Xuzhou Medical University from January 2018 to March 2022 were prospectively divided into compound flap transfer group (group A) and traditional TIP group (group B) by random number method. Group A was treated with TIP modified by compound flap transfer, and group B was treated with traditional TIP covered with fascia.The improvement of the compound flap transfer for TIP operation is mainly reflected in the multi-layer cover of the new urethra and the free skin embedded in the incision of the penile head. According to the intention-to-treat (ITT) principle, the final full analysis set (FAS) includes both group A and group B. Some patients were excluded from the analysis of the primary endpoint events due to reasons such as loss to follow-up or treatment group switch. Additionally, the final per-protocol set (PPS) consisting of group A and group B, which adheres to the study protocol, is subjected to statistical analysis.Independent sample t-test or Wilcoxon rank sum test was used for average age, the width of the penile head, degree of chordee, length of neourethra, and operative time. The classification of hypospadias, proportion of dorsal tunica albuginea plication, and incidence of postoperative complications were compared using the Chi-square test or Fisher’s exact test, P < 0.05 was considered statistically significant. Results:According to ITT principles, 50 children were included in FAS group A, ranging in age from 8 months to 15 years and 2 months, with an average age of 4 years. Group B included 50 children, ranging in age from 10 months to 14 years and 9 months, with an average age of 4 years and 1 month. Thirty-seven children in PPS group A were included, ranging in age from 1 year 2 months to 12 years 1 month, with an average age of 4 years. Group B consisted of 41 children, ranging in age from 1 year 2 months to 11 years 9 months, with an average age of 4 years 2 months. Statistical analysis showed that no matter FAS set or PPS, there were no statistically significant differences in the mean age, hypospadias type, average penile head width, average penile subcurvature number, number of dorsal tunica albuginea plication, and the average length of plastic neourethra between group A and group B ( P > 0.05). The average operative time of group A was higher than that of group B. The difference was statistically significant ( P < 0.05). The postoperative follow-up was 3 to 48 months, with an average follow-up of 2 years and 3 months. The complication rate of group A was lower than that of group B [10.81% (4/37) vs 29.27% (12/41)], and the difference was statistically significant ( P< 0.05). Urethral fistula occurred in 3 cases (8.11%), respectively and 6 cases (14.63%) in the two groups, the difference was not statistically significant ( P>0.05); urethral stricture occurred in 1 case (2.70%) and 5 cases (12.20%) respectively, the difference was not statistically significant ( P>0.05), There were 0 case and 1 case (2.44%) of urethral orifice descending or urethral dehiscence respectively, and the difference was not statistically significant ( P> 0.05). Conclusion:TIP with an improved compound flap transfer can reduce the overall postoperative complication rate and is worthy of promotion.

Objective:To investigate the application effect of the compound flap transfer in tubularized incised plate (TIP) urethroplasty.Methods:Children with hypospadias who received TIP in the Department of Pediatric Urology, the Affiliated Xuzhou Children’s Hospital of Xuzhou Medical University from January 2018 to March 2022 were prospectively divided into compound flap transfer group (group A) and traditional TIP group (group B) by random number method. Group A was treated with TIP modified by compound flap transfer, and group B was treated with traditional TIP covered with fascia.The improvement of the compound flap transfer for TIP operation is mainly reflected in the multi-layer cover of the new urethra and the free skin embedded in the incision of the penile head. According to the intention-to-treat (ITT) principle, the final full analysis set (FAS) includes both group A and group B. Some patients were excluded from the analysis of the primary endpoint events due to reasons such as loss to follow-up or treatment group switch. Additionally, the final per-protocol set (PPS) consisting of group A and group B, which adheres to the study protocol, is subjected to statistical analysis.Independent sample t-test or Wilcoxon rank sum test was used for average age, the width of the penile head, degree of chordee, length of neourethra, and operative time. The classification of hypospadias, proportion of dorsal tunica albuginea plication, and incidence of postoperative complications were compared using the Chi-square test or Fisher’s exact test, P < 0.05 was considered statistically significant. Results:According to ITT principles, 50 children were included in FAS group A, ranging in age from 8 months to 15 years and 2 months, with an average age of 4 years. Group B included 50 children, ranging in age from 10 months to 14 years and 9 months, with an average age of 4 years and 1 month. Thirty-seven children in PPS group A were included, ranging in age from 1 year 2 months to 12 years 1 month, with an average age of 4 years. Group B consisted of 41 children, ranging in age from 1 year 2 months to 11 years 9 months, with an average age of 4 years 2 months. Statistical analysis showed that no matter FAS set or PPS, there were no statistically significant differences in the mean age, hypospadias type, average penile head width, average penile subcurvature number, number of dorsal tunica albuginea plication, and the average length of plastic neourethra between group A and group B ( P > 0.05). The average operative time of group A was higher than that of group B. The difference was statistically significant ( P < 0.05). The postoperative follow-up was 3 to 48 months, with an average follow-up of 2 years and 3 months. The complication rate of group A was lower than that of group B [10.81% (4/37) vs 29.27% (12/41)], and the difference was statistically significant ( P< 0.05). Urethral fistula occurred in 3 cases (8.11%), respectively and 6 cases (14.63%) in the two groups, the difference was not statistically significant ( P>0.05); urethral stricture occurred in 1 case (2.70%) and 5 cases (12.20%) respectively, the difference was not statistically significant ( P>0.05), There were 0 case and 1 case (2.44%) of urethral orifice descending or urethral dehiscence respectively, and the difference was not statistically significant ( P> 0.05). Conclusion:TIP with an improved compound flap transfer can reduce the overall postoperative complication rate and is worthy of promotion.