The Efficacy of Active Fistulation in Duckett Procedure for Proximal Hypospadias in Children.

PURPOSE: To explore the efficacy of active fistulation in the treatment of proximal hypospadias in children by comparing one-stage and two-stage Duckett  procedure. MATERIALS AND METHODS:  A total of sixty-seven children who were diagnosed with proximal hypospadias and underwent Duckett operation at our hospital between January 2013 and January 2021 were selected for this study. These subjects were divided into two groups: the research group (n=36), using two-stage Duckett procedure with active fistulation, and the control group (n=31), using one-stage Duckett procedure. The incidence of postoperative complications and the score of pediatric penile perception Scale were compared between the two groups. RESULTS:  The research group exhibits significantly lower incidence rate of urethral fistula (8.3% Vs 16.1%) and urethral stricture (5.6% Vs 12.9%) in comparison to the control group (P<0.01). Furthermore, the analysis of Pediatric Penile Perception Scale scores indicates that the research group achieves significantly higher scores in terms of urethral shape, penile skin shape, and overall appearance than the control group (P<0.05).  Conclusion: In the treatment of proximal hypospadias in children, The active fistulation within the two-stage Duckett procedure significantly reduces the rate of stage 1 postoperative complications and improves parental satisfaction. The active fistulation may offer a more promising option for the treatment of proximal hypospadias in children.

Adult hair follicle stem cells differentiate into neuronal cells in explanted rat intestinal tissue.

Hair follicle stem cells (HFSCs) are adult stem cells located in the outer root sheath of the follicle bulge with high neural plasticity, which promise a potential for the stem cell therapy for neurological diseases. Hirschsprung's disease (HD) is characterized by the absence of ganglia in the distant bowel. In this study, we elucidated the capacity of HFSCs to differentiate into neuronal cells in the aganglionic colon from embryonic rat. HFSCs were isolated from adult Sprague-Dawley (SD) rats and formed spheres that could be passaged. The cultured HFSCs expressed neural crest stem cells (NCSCs) markers such as SOX10, CD34, and nestin, which indicated their neural crest lineage. Subsequent differentiation assays demonstrated that these cells could give rise to neural progeny that expressed neuronal or glial markers. The aganglionic colon from the embryonic intestine was applied as in vitro explant to test the capacity of proliferation and differentiation of HFSCs. The HFSCs expressing GFP or RFP integrated in intestinal explants and maintained proliferative capacity. Moreover, the HFSCs differentiated into Tuj1- or S100ß-positive cells in the cultured intestinal explants. The results proposed that the HFSCs might be an alternative source of neural stem cells for the HD therapy.

Analysis of The Curative Effect of Tubularized Incised Plate Urethroplasty for Distal Hypospadias With The Dysplastic Corpus Spongiosum Covering Technique.

PURPOSE: To investigate the use of tubularized incised plate (TIP) urethroplasty for distal second- and third-degree hypospadias to free the dysplastic forked corpus spongiosum and Buck's fascia, which are used as a covering material for the new urethra, thereby reducing the incidence of urinary fistula and other complications in the coronal sulcus. MATERIALS AND METHODS: Clinical data of 113 patients with distal hypospadias treated with TIP urethroplasty from January 2017 to December 2020 were retrospectively analyzed. The study group comprised 58 patients (use of dysplastic corpus spongiosum and Buck's fascia to cover the new urethra), and the control group comprised 55 patients (use of dorsal Dartos fascia to cover the new urethra). RESULTS: All children were followed up for more than 12 months. In the study group, 4 patients developed urinary fistulas, 4 developed a urethral stricture, and no case developed glans fissure. In the control group, 11 patients developed urinary fistulas, 2 developed a urethral stricture, 3 developed a glans cracking. CONCLUSION: Using the dysplastic corpus spongiosum to cover the new urethra increases the amount of tissue in the coronal sulcus and reduces the incidence of urethral fistula, but it may increase the incidence of urethral stricture.

PAX3 Promotes Cell Migration and CXCR4 Gene Expression in Neural Crest Cells.

Neural crest (NC) cells are a multipotent cell population with powerful migration ability during development. C-X-C chemokine receptor type 4 (CXCR4) is a chemokine receptor implicated to mediate NC migration in various species, whereas the underlying mechanism is not well documented yet. PAX3 is a critical transcription factor for the formation of neural crest and the migration and differentiation of NCs. In this study, we retrieved a potential PAX3 binding element in the promoter of the CXCR4 gene, and we further found that PAX3 could promote the expression of CXCR4 and facilitate the migration of NCs. We finally demonstrated that PAX3 could bind the promoter region of CXCR4 and increase CXCR4 transcription by luciferase assay and electrophoretic mobility shift assay (EMSA). These findings suggested that PAX3 is a pivotal modulator of NC migration via regulating CXCR4 expression.

Prospective identification and culture of rat enteric neural stem cells (ENSCs).

Hirschprung's disease (HD), a very common congenital abnormality in children, occurs mainly due to the congenital developmental defect of the enteric nervous system. The absence of enteric ganglia from the distal gut due to deletion in gut colonization by neural crest progenitor cells may lead to HD. The capacity to identify and isolate the enteric neuronal precursor cells from developing and mature tissues would enable the development of cell replacement therapies for HD. However, a mature method to culture these cells is a challenge. The present study aimed to propose a method to culture enteric neural stem cells (ENSCs) from the DsRed transgenic fetal rat gut. The culture medium used contained 15 % chicken embryo extract, basic fibroblast growth factor, and epidermal growth factor. ENSCs were cultured from embryonic day 18 in DsRed transgenic rat. Under inverted microscope and fluorescence staining, ENSCs proliferated to form small cell clusters on the second day of culture. The neurospheres-like structure were suspended in the medium, and there were some filaments between the adherent cells from day 3 to day 6 of the culture. The neurospheres were formed by ENSCs on day 8 of the culture. Network-like connections were formed between the adherent cells and differentiated cells after adding 10 % FBS. The differentiated cells were positive for neurofilament and glial fibrillary acidic protein antibodies. The present study established a method to isolate and culture ENSCs from E18 DsRed transgenic rats in the terminal stage of embryonic development. This study would offer a way to obtain plenty of cells for the future research on the transplantation of HD.

Wogonin Induced Mitochondrial Dysfunction and Endoplasmic Reticulum Stress in Human Malignant Neuroblastoma Cells Via IRE1α-Dependent Pathway.

Wogonin, a flavonoid isolated from Scutellaria baicalensis Georgi, has been reported to exhibit a variety of biological effects including anti-cancer effects. It has a pro-apoptotic role in many cancer types. However, the molecular mechanisms of wogonin in treating neuroblastoma remain elusive. In the present study, two malignant neuroblastoma cell lines (SK-N-BE2 and IMR-32 cells) were treated with different doses of wogonin (0-150 µM). Wogonin showed significant cytotoxic effects in SK-N-BE2 and IMR-32 cells in a dose- and time-dependent manner. Treatment of SK-N-BE2 and IMR-32 cells with 75 µΜ wogonin for 48 h significantly promoted apoptosis, the release of cytochrome c, altered the expression of certain members of Bcl-2 family (Bcl-2, Bax and Bid), and increased the activation of caspase-3, caspase-8, caspase-9, and PARP-1, which demonstrated that the cytotoxic effect of wogonin in SK-N-BE2 and IMR-32 cells is mediated by mitochondrial dysfunction. Moreover, wogonin induced the expression of endoplasmic reticulum (ER) stress-related proteins (GRP78/Bip and GRP94/gp96) and activation of caspase-12 and caspase-4 in SK-N-BE2 and IMR-32 cells. In addition, wogonin increase the expression of IRE1α and TRAF2, and phosphorylation of ASK1 and JNK in SK-N-BE2 and IMR-32 cells. Knockdown of IRE1α by siRNA not only markedly inhibited wogonin-induced up-regulation of IRE1α and TRAF2, and phosphorylation of ASK1 and JNK but also reduced wogonin-induced cytotoxic effects and mitochondrial dysfunction in SK-N-BE2 and IMR-32 cells. These results indicated that wogonin could induce apoptosis, mitochondrial dysfunction, and ER stress in SK-N-BE2 and IMR-32 cells by modulating IRE1α-dependent pathway.

[Expression change and significance of homebox gene-A10 in rat cryptorchidism].

OBJECTIVE: To explore the effects of HOXA10 gene expression in undescended and descended testis. METHODS: Twenty-four male Sprague Dawley rats were surgically prepared for unilateral cryptorchidism model. Their undescended and descended testes were removed at days 7 (infancy period, group B7) and 60 (sexual maturity period, group B60) post-operation. And contralateral descended testis served as controls (group A7, group A60). The expression of HOXA10 with histologic changes in experimental cryptorchidism were detected with one-step real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. RESULTS: Compared with the control group, the levels of HOXA10 gene for groups B7 and B60 decreased markedly to 0.78 ± 0.11 (P < 0.01) and 0.56 ± 0.03 (P < 0.01) respectively.However the levels of HOXA10 gene and protein in group B60 were significantly lower than those in group B7 (P < 0.01). CONCLUSIONS: There is a lower expression of HOXA10 in undescended testes than in normally developed counterparts. And the expression of HOXA10 decreases with the elapsing time of cryptorchidism.

Growth-promoting effect of environmental endocrine disruptors on human neuroblastoma SK-N-SH cells.

Bisphenol A (BPA), a monomer component of polycarbonate plastics and epoxy resins, is widely used in many consumer products. Zearalenone (ZEA), a non-steroidal estrogenic mycotoxin, is present in high concentrations in dairy products and cereals. Numerous researches describe a possible correlation between environmental endocrine disruptors and human tumors, but only a few papers concerned solid tumors in childhood. We investigated the effects of BPA and ZEA on the proliferation in the human neuroblastoma SK-N-SH cell line. Cell counting kit-8 and flow cytometric analysis were used to determine whether BPA and ZEA promote cell proliferation. The results indicated that BPA and ZEA-mediated increase in cell proliferation is significant (p<0.05). To explore the possible underlying mechanism, additive effect of the estrogen receptor antagonist ICI182780 or insulin-like growth factor I (IGF-I) was observed. ICI182780 could inhibit these proliferative effects of BPA and ZEA. However, no synergistic or additive growth-promoting effect was noted when IGF-1 was added. These results suggested that BPA and ZEA can promote the proliferation of SK-N-SH cells, and the estrogen receptor pathway may be involved in this effect.