Transcription Factor FOXO3a Overexpression Inhibits the Progression of Neuroblastoma by Regulating the miR-21/SPRY2/ERK Axis.

OBJECTIVE: Neuroblastoma is one of the most common extracranial solid tumors in children. The forkhead transcription factor FOXO3a has been implicated in the progression of a variety of human diseases. Here, we aim to identify the effects of FOXO3a on the malignancy of neuroblastoma. METHODS: Bioinformatics analysis was employed to identify differentially expressed genes related to neuroblastoma and the downstream regulator of FOXO3a. FOXO3a expression was examined in SH-SY5Y neuroblastoma cells. Interactions between FOXO3a and microRNA-21 (miR-21) were then identified using bioinformatics analysis and dual-luciferase reporter assay. After ectopic expression and depletion experiments in SH-SY5Y cells, cell malignant phenotypes were assessed by cell counting kit-8 and Transwell assays. FOXO3a-overexpressing neuroblastoma cells were xenografted into nude mice to validate the role of FOXO3a in tumor growth. RESULTS: Downregulated expression of FOXO3a was observed in neuroblastoma cells, with a negative correlation between FOXO3a and miR-21 expression. FOXO3a bound to the promoter region of miR-21 to downregulate its expression, resulting in inhibition of SH-SY5Y cell malignant phenotypes. Additionally, miR-21 targeted SPRY2 by binding to the 3'UTR of the mRNA encoding SPRY2, activating the extracellular signal-regulated kinase (ERK) pathway. FOXO3a disrupted the binding of miR-21 to SPRY2 and inactivated ERK to suppress the malignant phenotypes of SH-SY5Y cells as well as tumor growth in vivo. CONCLUSIONS: In conclusion, FOXO3a may inhibit the progression of neuroblastoma by suppressing the miR-21 expression and facilitating SPRY2-dependent ERK pathway inactivation.

Inguinal hernia repair by Bianchi incision in boys: a retrospective study.

OBJECTIVE: Open inguinal hernia repair by Bianchi incision is a potential alternative technique for the treatment of IH. This study aims to retrospectively analyze boys with IH, who underwent open IH repair by Bianchi incision. METHODS: A total of 3300 boys (1-144 months) with IH from April 2007 to September 2015 were enrolled into this study. An open high scrotal incision (Bianchi incision) to ligate the processus proximal to the internal inguinal ring was performed in patients for IH repair. Then, all patients were followed up after 7 days, 1 month, and 1 year. Operation time, hernia recurrence, hydrocele, testicular atrophy, cosmetic results, and the satisfaction of parents were evaluated. RESULTS: Among these 3300 boys, 1662 (50.36%) and 1349 (40.88%) boys with IH were operated on the right and left side, respectively, while 289 (8.76%) patients underwent bilateral surgery. The average operation time was 13.0 ± 2.3 min for unilateral cases and 25.2 ± 4.2 min for bilateral cases. Furthermore, among these 3300 boys, 309 boys (9.36%) were lost to follow-up, and the remaining 2991 boys underwent a total of 3245 IH repairs. The complications included 20 recurrences who were repaired with the same technique, one wound rupture, and one acquired undescended testis. No infection, obvious scrotal hematoma, testicular atrophy, and vas deferens injury were found during the follow-up. In most instances, the scars were invisible, obtaining an excellent cosmetic effect. CONCLUSION: Inguinal hernia repair by Bianchi incision is a safe, easy and effective technique with cosmetic benefits, which could be a reliable alternative for the treatment of pediatric inguinal hernia.

MicroRNA-149 is associated with clinical outcome in human neuroblastoma and modulates cancer cell proliferation through Rap1 independent of MYCN amplification.

PURPOSE: We evaluated the clinical relevance of microRNA-149 (miR-149) in neuroblastoma (NB) and its functional roles in regulating NB proliferation in vitro. METHODS: QRT-PCR was used to evaluate miR-149 expression in NB cell lines and primary NB tumors. Association between endogenous miR-149 expression in primary NB tumors and their host patients' clinicopathological factors and overall survival (OS) were statistically evaluated. In SH-SY5Y, an MYCN-non-amplified, and LAN5, an MYCN-amplified NB cell lines, miR-149 was either upregulated or downregulated by lentiviral transduction, to evaluate its effect on NB proliferation in vitro. Possible downstream target of miR-149, Ras-related protein 1 (Rap1), was evaluated by qRT-PCR and western blot in lentiviral-transduced NB cells. Moreover, Rap1 was either upregulated or downregulated in lentiviral-transduced NB cells to further evaluate its effect on miR-149-mediated NB proliferation in vitro. RESULTS: MiR-149 is markedly downregulated in both in vitro NB cell lines and in vivo NB primary tumors. Low miR-149 expression is predominantly associated with Stage 3 or 4 primary NB tumors, and poor OS among NB patients. In SH-SY5Y and LAN5 cells, lentivirus-induced miR-149 upregulation inhibited, whereas miR-149 downregulation promoted NB proliferation in vitro, despite MYCN-amplification status. Rap1 expression, at both mRNA and protein levels, was inversely associated with miR-149 in NB. In addition, Rap1 upregulation or downregulation reversely regulated miR-149-mediated NB proliferation in vitro. CONCLUSION: MiR-149 is downregulated in NB and closely associated with NB patients' clinical outcome. MiR-149 also functionally modulates NB cell proliferation in vitro, possibly through inverse-regulation on Rap1.

[Modified Snodgrass technique for hypospadias].

OBJECTIVE: To investigate the clinical effect of modified Snodgrass surgical technique in the treatment of hypospadias. METHODS: We retrospectively analyzed the clinical data about 212 cases of hypospadias treated by urethroplasty from January 2008 to October 2016, 94 with the modified Snodgrass technique, namely with a silk line in addition to the urethral suture to make easier postoperative removal of the suture (group A), and the other 118 with the conventional Snodgrass technique (group B). The urethral suture was removed at 10 days after surgery for the patients in group A. We compared the success rate of surgery and incidence of postoperative complications between the two groups. RESULTS: Compared with group B, group A showed a significantly higher success rate of surgery (81.36% vs 91.49%, P <0.05) but lower incidence rates of postoperative incisional infection (12.71% vs 4.26%, P <0.05) and urinary fistula (16.10% vs 6.38%, P <0.05). No statistically significant difference was found in the incidence of urethral stenosis between the two groups (2.54% vs 2.13%, P >0.05). CONCLUSIONS: The modified Snodgrass technique can improve the success rate of surgery and reduce the incidence rates of incisional infection and urinary fistula, which deserves wide clinical application.

[Effect of epidermal growth factor and testosterone on androgen receptor activation in urethral plate fibroblasts in hypospadias].

OBJECTIVE: To investigate androgen receptor (AR) expression and the effect of epidermal growth factor (EGF) and testosterone on AR expression level.
 METHODS: EGF or different concentrations of testosterone were incubated with the primary urethral plate fibroblasts from patients with hypospadias. The levels of AR expression in the fibroblasts were detected by immunocytochemical assays and graphical analysis.
 RESULTS: There was no significant difference in AR activation under physiological concentrations (3×10(-8) mol/L) of testosterone between the control and the distal hypospadias group (P>0.05). However, there was a significant decrease in AR activation in the proximal hypospadias group compared to that in the control group (P<0.001). Under the concentration of 3×10(-6) mol/L, the effects of testosterone on AR activation were dramatically different in the three groups (control group>distal hypospadias group>proximal hypospadias group, P<0.001). AR activation level in the group of proximal hypospadias was improved most obviously when EGF and physiological concentration of testosterone were employed in the urethral plate fibroblasts from hypospadias patients (P<0.001), and it was improved more in the distal hypospadias group than that in the control group (P=0.02).
 CONCLUSION: AR expression and activation in the urethral plate fibroblasts from hypospadias patients are abnormal. EGF can be used to improve AR activation in fibroblasts from different types of hypospadias, especially in the proximal type.

[EFFECTIVENESS OF ADVANCED SKIN FLAP AND V-SHAPED VENTRAL INCISION ALONG THE ROOT OF PENILE SHAFT FOR CONCEALED PENIS].

OBJECTIVE: To investigate effectiveness of advanced skin flap and V-shaped ventral incision along the root of penile shaft for concealed penis in children. METHODS: Between July 2007 and January 2015, 121 boys with concealed penis were treated with advanced skin flap and V-shaped ventral incision along the root of penile shaft. The age varied from 18 months to 13 years (mean, 7.2 years). Repair was based on a vertical incision in median raphe, complete degloving of penis and tacking its base to the dermis of the skin. Advanced skin flap and a V-shaped ventral incision along the root of penile shaft were used to cover the penile shaft. RESULTS: The operation time ranged from 60 to 100 minutes (mean, 75 minutes). Disruption of wound occurred in 1 case, and was cured after dressing change; and primary healing of incision was obtained in the others. The follow-up period ranged from 3 months to 7 years (median, 24 months). All patients achieved good to excellent cosmetic results with a low incidence of complications. The results were satisfactory in exposure of penis and prepuce appearance. No obvious scar was observed. The penis had similar appearance to that after prepuce circumcision. CONCLUSION: A combination of advanced skin flap and V-shaped ventral incision along the root of penile shaft is a simple, safe, and effective procedure for concealed penis with a similar appearance result to the prepuce circumcision.