Identification and analysis of MSC-Exo-derived LncRNAs related to the regulation of EMT in hypospadias.

OBJECTIVE: This study aims to screen the differentially expressed long non-coding RNAs (DELncRNAs) related to the regulation of epithelial-mesenchymal transition (EMT) in hypospadias in mesenchymal stem cell-derived exosomes (MSC-Exons) and explore the potential mechanism of these lncRNAs for the EMT in hypospadias. METHODS: In this study, the microarray data related to MSC-Exos and hypospadias were downloaded from Gene Expression Omnibus (GEO). Besides, the lncRNAs highly expressed in MSC-Exos and the differentially expressed mRNAs and lncRNAs in children with hypospadias were screened, respectively. In addition, the lncRNAs enriched in MSC-Exos and differentially expressed lncRNAs in hypospadias were intersected to obtain the final DElncRNAs. Moreover, the co-expression interaction pairs of differentially expressed lncRNAs and mRNAs were analyzed to construct a Competing Endogenous RNA (ceRNA) network. Finally, the candidate lncRNAs in exosomes were subjected to in vitro cell function verification. RESULTS: In this study, a total of 4 lncRNAs were obtained from the microarray data analysis. Further, a ceRNA regulatory network of MSC-Exo-derived lncRNAs related to the regulation of EMT in hypospadias was constructed, including 4 lncRNAs, 2 mRNAs, and 6 miRNAs. The cell function verification results indicated that the exosomes secreted by MSCs may transport HLA complex group 18 (HCG18) into target cells, which promoted the proliferation, migration, and EMT of these cells. CONCLUSION: MSC-Exo-derived lncRNA HCG18 can enter target cells, and it may be involved in the regulation of EMT in hypospadias through the ceRNA network.

The construction of a nomogram to predict the prognosis and recurrence risks of UPJO.

Objective: This study was conducted to explore the risk factors for the prognosis and recurrence of ureteropelvic junction obstruction (UPJO). Methods: The correlation of these variables with the prognosis and recurrence risks was analyzed by binary and multivariate logistic regression. Besides, a nomogram was constructed based on the multivariate logistic regression calculation. After the model was verified by the C-statistic, the ROC curve was plotted to evaluate the sensitivity of the model. Finally, the decision curve analysis (DCA) was conducted to estimate the clinical benefits and losses of intervention measures under a series of risk thresholds. Results: Preoperative automated peritoneal dialysis (APD), preoperative urinary tract infection (UTI), preoperative renal parenchymal thickness (RPT), Mayo adhesive probability (MAP) score, and surgeon proficiency were the high-risk factors for the prognosis and recurrence of UPJO. In addition, a nomogram was constructed based on the above 5 variables. The area under the curve (AUC) was 0.8831 after self cross-validation, which validated that the specificity of the model was favorable. Conclusion: The column chart constructed by five factors has good predictive ability for the prognosis and recurrence of UPJO, which may provide more reasonable guidance for the clinical diagnosis and treatment of this disease.

Nephroblastoma-specific dysregulated gene SNHG15 with prognostic significance: scRNA-Seq with bulk RNA-Seq data and experimental validation.

Wilms tumor (WT) is the most common malignancy of the genitourinary system in children. Currently, the Integration of single-cell RNA sequencing (scRNA-Seq) and Bulk RNA sequencing (RNA-Seq) analysis of heterogeneity between different cell types in pediatric WT tissues could more accurately find prognostic markers, but this is lacking. RNA-Seq and clinical data related to WT were downloaded from the Therapeutically Applicable Research to Generate Effective Treatments (TARGET) database. Small nucleolar RNA host gene 15 (SNHG15) was identified as a risk signature from the TARGET dataset by using weighted gene co-expression network analysis, differentially expressed analysis and univariate Cox analysis. After that, the functional mechanisms, immunological and molecular characterization of SNHG15 were investigated at the scRNA-seq, pan-cancer, and RNA-seq levels using Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), ESTIMATE, and CIBERSORT. Based on scRNA-seq data, we identified 20 clusters in WT and annotated 10 cell types. Integration of single-cell and spatial data mapped ligand-receptor networks to specific cell types, revealing M2 macrophages as hubs for intercellular communication. In addition, in vitro cellular experiments showed that siRNAs interfering with SNHG15 significantly inhibited the proliferation and migration of G401 cells and promoted the apoptosis of G401 cells compared with the control group. The effect of siRNAs interfering with SNHG15 on EMT-related protein expression was verified by Western blotting assay. Thus, our findings will improve our current understanding of the pathogenesis of WT, and they are potentially valuable in providing novel prognosis markers for the treatment of WT.

Identification of the expression patterns and potential prognostic role of m6A-RNA methylation regulators in Wilms Tumor.

BACKGROUND: To explore the potential role of m6A methylation modification in Wilms Tumor (WT) by m6A-RNA Methylation (m6A) regulators. METHODOLOGY: The association of m6A modification patterns with immune and prognostic characteristics of tumors was systematically evaluated using 19 m6A regulators extracted from Wilms Tumor's samples in public databases. A comprehensive model of "m6Ascore" was constructed using principal component analysis, and its prognostic value was evaluated. RESULTS: Almost all m6A regulators were differentially expressed between WT and normal tissues. Unsupervised clustering identified three distinct m6A clusters that differed in both immune cell infiltration and biological pathways. The m6Ascore was constructed to quantify m6A modifications in individual patients. Our analysis suggests that m6Ascore is an independent prognostic factor for WT and can be used as a novel predictor of WT prognosis. CONCLUSIONS: This study comprehensively explored and systematically characterized m6A modifications in WT. m6A modification patterns play a critical role in the tumor immune microenvironment (TIME) and WT prognosis. m6Ascore provides a more comprehensive understanding of m6A modifications in WT and offers a practical tool for predicting WT prognosis. This study will help clinicians to identify valid indicators of WT to improve the poor prognosis of this disease. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at https://www.aliyundrive.com/drive/folder/64be739cd6956a741fb24670baeea53422be6024 .

An exploration into the application of specialty-orientated CBL pedagogy in undergraduate teaching in pediatric surgery.

Objective: This study aims to identify whether the specialty-oriented case-based learning (CBL) pedagogy contributes to the teaching of basic theory and practical operation in undergraduate clinical teaching in pediatric surgery, and to assess the satisfaction of undergraduates. Methods: A total of 72 undergraduates in Grade 2016 who interned at Qilu Hospital of Shandong University were enrolled in this study. All these undergraduates voluntarily participated in this experimental study. They were randomly divided into the experimental group (the CBL group, n = 36) and the control group [the traditional lecture-based learning (LBL) group, n = 36] with the assistance of random number tables. In the control group, a traditional pedagogy was adopted and the knowledge in the textbook was explained according to the syllabus. In the experimental group, a specialty-oriented CBL pedagogy was adopted under the guidance of clinical instructors. After the teaching, a comparison was drawn between both groups in respect of the theoretical exam and practical exam scores. In addition, the teaching results were evaluated by a questionnaire survey. Results: The average theoretical exam scores and comprehensive scores of undergraduates in the CBL group were higher than those in the LBL group (P < 0.05). There was no significant difference in the practical exam scores between the CBL group and the LBL group (P > 0.05). However, those undergraduates in the CBL group attained higher scores in doctor-patient communication and perioperative diagnosis and treatment (P < 0.05). According to the questionnaire survey, the undergraduates in the CBL group had higher satisfaction than those in the LBL group. Besides, this specialty-oriented CBL pedagogy had higher performance in improving their ability to solve problems independently and cultivating and expanding their knowledge compared with the traditional pedagogy. Meanwhile, this specialty-oriented CBL pedagogy can cultivate the critical thinking of undergraduates, which could increase their learning efficiency and improve their interest in learning. Conclusion: This specialty-oriented CBL pedagogy could improve the mastery of professional knowledge, course satisfaction, doctor-patient communication ability in clinical practice, and perioperative diagnosis and treatment ability of these undergraduates. Therefore, it is worthwhile to recommend and popularize this pedagogy in undergraduate clinical teaching in pediatric surgery.

Downregulated TICAM1 is a prognostic biomarker and associated with immune tolerance of Wilms tumor patients.

BACKGROUND: TIR domain containing adaptor molecule 1 (TICAM1) is a coding gene participating in immune and inflammation responses to malignant cells. However, the role of TICAM1 in Wilms tumor (WT) is rarely known. MATERIALS AND METHODS: The expression level of TICAM1 was calculated in the WT TARGET cohort and validated using the GSE66405 cohort. The Kaplan-Meier method was employed to investigate the potential clinical value of TICAM1 and the association between its expression level and clinical features. The influence of TICAM1 on immune infiltration was examined by ESTIMATE, CIBERSORT and MCPcounter algorithms. IC50 of chemotherapeutic drugs was calculated by "pRRophetic" R package. RESULTS: TICAM1 was downregulated in WT patients with worse prognosis and a more advanced clinical stage. Moreover, a low expression level of TICAM1 contributed to less immune cell infiltration, few protective immune cells and more antitumor immune cells. CONCLUSIONS: TICAM1 exerts a significant impact on the prognosis, progression and immune infiltration condition of WT.

Association of a GATA Binding Protein 4 Polymorphism with the Risk of Hypospadias in the Chinese Children.

PURPOSE: GATA binding protein 4 (GATA4) has been implicated in the etiology of congenital malformation of the urogenital system. The present study investigated the influence of GATA4 polymorphisms on susceptibility to hypospadias. METHODS: We genotyped 4 potentially functional polymorphisms (rs12458, rs12825, rs884662, and rs904018) in GATA4 in the hospital-based case-control study including 410 child patients and 520 nonmalformed individuals by the TaqMan MGB method. Risk associations were assessed using unconditional logistic regression, adjusted for potential confounding factors. RESULTS: A significant association was found between rs12458 (3'-UTR of GATA4) and susceptibility to hypospadias (p = 0.008). Compared with rs12458 AA genotype individuals, those harboring the variant allele (rs12458 AT/TT) were correlated with significantly higher risk of hypospadias (AT/TT vs. AA: OR = 1.42, 95% CI = 1.17-2.35, p = 0.036). Furthermore, the rs12458T allele showed significantly decreased activity in a luciferase reporter assay, indicating a possible role of rs12458 variant in regulating the combination of microRNAs with the GATA4 mRNA. CONCLUSIONS: The present results indicate that the functional GATA4 rs12458 variant confers individuals' susceptibility to hypospadias, possibly through regulating the GATA4 expression level.

Pediatric laparoscopic partial cystectomy for the treatment of benign bladder tumors and urachal cysts.

OBJECTIVE: To investigate the feasibility and efficacy of carrying out pediatric laparoscopic partial cystectomies (LPC) when treating benign bladder tumors and urachal cysts. METHODS: Retrospectivey analyzing 4 clinical cases involving children with bladder tumors, which were collected from October 2017 to December 2018. In these clinical cases, there were 3 male children and 1 female child, aged from 4.5 to 9.4 years old, with an average age of 6.5 years. An intraperitoneal laparoscopic partial cystectomy was performed in the treatment of 3 of these patients with benign bladder tumors and in 1 patient with an urachal cyst. The surgical procedures included a partial cystectomy and a complete intracavitary bladder suture. RESULTS: All 4 cases were successful and no operation was transferred to opensurgery. The operation time was 100-120 min, with an average time of 108 min. The intraoperative blood loss was 10-20 ml, with an average loss of 15 ml. 6 h after the operation, the patients still maintained a fluid diet and 1 case of hematuria had occurred, with the catheter removed 12 days after the operation. No postoperative urine leakage, intestinal adhesion or intestinal obstruction occurred, and the average postoperative hospitalization time was 14 days. CONCLUSION: Laparoscopic partial cystectomy is a safe and feasible method to be used for the treatment of benign bladder tumors and urachal cysts. It presents the advantages of being minimally invasive, and having a quick recovery and short hospitalization time. It is an alternative surgical method for the treatment of pediatric benign bladder tumors.

Experience Summary of Laparoscopic Treatment for Pediatric Ureteral Polyps.

Objective: To investigate surgical techniques and challenges of laparoscopic in treating pediatric ureteral polyps under laparoscopy. Methods: The clinical data of 7 of pediatric ureteral polyps patients who were admitted to the hospital from July 2015 to January 2020 were analyzed retrospectively. There were 6 males and 1 female from 7.7 to 13.9 years old at the mean age of 10.4. Before surgery, all children performed urinary B ultrasound, magnetic resonance urography (MRU), and renal radionuclide scanning. Six cases were observed on the left lateral and 1 on the right. The lesions of 5 cases were located at the ureteropelvic junction, 1 in the upper ureter and 1 in the middle ureter. The polyps were treated intraoperatively by the resecting of the lesion segment and simple polypectomy to retain the attached part of the original diseased segment of the ureter. All surgeries were performed under laparoscopy and B-ultrasound was performed during follow up after surgery. Results: All 7 surgeries were performed successfully under the laparoscope. The surgery time was 80-110 min, and the average surgery time was 97.5 min. The intraoperative bleeding was 10-25 ml and the average postoperative hospital stay was 6 d. Postoperative hematuria occurred in 1 case. Neither urinary leakage nor urinary tract infection was reported post surgery. Preoperative affected pyelectasis of all patients was 2.0-3.7 cm. Three months postoperatively, the affected pyelectasis was measured at 1.2-3.0 cm. No recurrence of polyps was reported after surgery. During the follow-up to April 2020, there was no significant change in the kidney size of all patients, and hydronephrosis was alleviated compared with that before surgery. Conclusions: Laparoscopy is a safe, effective and minimally invasive surgical technique for pediatric multiple ureteral polyps. The surgery plan was designed according to the location and size of polyps, including segmental ureterectomy of polyps + pyeloureterostomy, segmental ureterectomy of polyps + ureter - ureteral anastomosis.

The expression and clinical significance of different forms of LILRA3 in systemic lupus erythematosus.

OBJECTIVE: Our previous study has shown that functional leukocyte immunoglobulin-like receptors A3 (LILRA3) contributes to susceptibility and subphenotypes of systemic lupus erythematosus (SLE). However, the mechanism remains unclear. We aimed to evaluate the role of LILRA3 in SLE. METHODS: One hundred twenty-six SLE patients and 48 healthy controls were recruited in this study. Functional studies were performed using intracellular flow cytometry and ELISA. RESULTS: Both LILRA3 levels in serum and CD14+ monocytes were significantly elevated in SLE patients compared with healthy controls. Elevated LILRA3 level was found positively correlated with SLEDAI. Furthermore, more elevated LILRA3 levels were found in patients with higher SLEDAI, presence of lupus nephritis, and thrombocytopenia. CONCLUSIONS: Both LILRA3 levels in serum and CD14+ monocytes significantly increased in SLE and positively correlated with disease activity and severity. The upregulation of LILRA3 expression may serve as a biomarker of disease activity and severity of SLE. KEY POINTS: • LILRA3 contributes to susceptibility and subphenotypes of SLE; LILRA3 is elevated in SLE patients. • Increased LILRA3 correlated with disease activity and severity. • LILRA3 may serve as a biomarker of disease activity and severity of SLE.

Expression of phosphatase of regenerating liver-3 is associated with prognosis of Wilms' tumor.

OBJECTIVE: The current study was undertaken to explore the clinical and prognostic value of phosphatase of regenerating liver-3 (PRL-3) expression in Wilms' tumor. METHODS: Seventy-six patients with Wilms' tumor in Qilu Hospital from January 2003 to July 2009 were enrolled in the study. Protein expression level of PRL-3 was examined by immunohistochemical staining, and the correlation between PRL-3 expression and histopathological parameters, clinical variables, and outcome of patients with Wilms' tumor were analyzed. RESULTS: We found that 19% of patients with unfavorable histology had tumor recurrence and 16% of patients died following the operation. PRL-3 was expressed in 15 out of 76 tumors (19%) and expressed highly in unfavorable histology Wilms' tumor (P=0.04). PRL-3 protein expression level was correlated to 2.5-fold increase in recurrence rate of Wilms' tumor (P=0.06) without any statistically significant difference. However, in favorable histology Wilms' tumor, PRL-3 expression was correlated to an increase of 3.4-fold in recurrence rate (P=0.03). CONCLUSION: The expression of PRL-3 protein was correlated with an increased recurrence rate of favorable histology Wilms' tumor. PRL-3 may serve as a promising biomarker for predicting patients with high risk of Wilms' tumor. Further investigations are warranted to investigate the clinical function of PRL-3 in Wilms' tumor.

Carboplatin-based Nanomedicine to Enhance the Anticancer Effect in SK-NEP-1 WilmsꞌTumor Cells.

Wilms tumor (WT) is the most common pediatric malignant primary renal tumor. Carboplatin (CRB), a platinum compound is widely used in the treatment of multiple cancers including ovarian, lung, head and neck, and wilm's tumor. However, lower uptake of CRB in cancer cells and toxicity concerns in healthy cells often limited its clinical outcome. The aim of this study was to investigate the antitumor effect of CRB on SK-NEP-1 wilm's cancer cells. Earlier, CRB was formulated in nanoparticulate formulations and characterized its biophysical parameters. SK-NEP-1 cell growth in vitro was assessed by MTT. Then, apoptosis potential was investigated by TUNEL, Hoechst, and colony formation assay. CRB treatment resulted in inhibition of cell proliferation of SK-NEP-1cells in a dose-dependent manner. TUNEL, Hoechst, and colony formation assay demonstrated that CRB was more effective in killing wilm's cancer cell when encapsulated in nanoparticle formulations. Overall, the present study demonstrates that CRB treatment resulted in marked inhibition of cell proliferation and cell apoptosis. These results may pave way for the effective treatment of wilm's tumor in clinical models.

Therapeutic effect of oridonin on mice with prostate cancer.

OBJECTIVE: To investigate the therapeutic effect and the related mechanism of oridonin on mice with prostate cancer. METHODS: Sixty BALB/C male nude mice were selected. A model of RM-1 cell transplantation tumor of prostate cancer was built by the subcutaneous inoculation of RM-1 cells. After that, those 60 experimental mice were randomly divided into groups A, B and C. Each group had 20 mice. Mice in group A were treated with 0.2 mL of normal saline (0.9%) by intraperitoneal injection once a day; mice in group B received intraperitoneal injection of 1.875 mg/mL of oridonin once a day; and mice in group C received intraperitoneal injection of 7.5 mg/mL of oridonin once a day. Mice in the three groups were treated uninterruptedly for 5 weeks and were all killed. Then, tumors were excised and weighed to calculate their growth inhibitory rate, volume increment and anti-tumor rate. Thymus and spleen of mice in the three groups were collected to calculate the thymus and spleen index. Immunohistochemical staining was applied to observe the expression of caspase-3 in prostate cancer tissue of mice of the three groups. RESULTS: The qualities and volume increment of tumors in groups B and C were significantly lower than those of group A (P < 0.05); the qualities and volume increment of tumors in groups C were evidently lower than those of group B (P < 0.05); the tumor volume increment and anti-tumor rate in group C were obviously higher than those of group B (P < 0.05); the thymus and spleen indexes of groups B and C were distinctly higher than those of group A (P < 0.05); comparison of the thymus and spleen indexes between group B and group C showed no statistical differences (P > 0.05). Immumohistochemical staining revealed that the caspase-3 protein in prostate cancer tissue of mice of group A expressed negatively with colorless or light-colored karyon; while the caspase-3 protein in prostate cancer tissue of mice of group B expressed positively with dark-colored karyon, centralized distribution and granular sensation; and the caspase-3 in prostate cancer tissue of mice of group C showed strong positive expression with big and darker colored karyon and dense distribution. CONCLUSIONS: Oridonin can inhibit the growth of RM-1 prostate cancer cells effectively and have great therapeutic effects on RM-1 cell transplantation tumor of prostate cancer.

Effect of the artificial somato-autonomic neuroanastomosis on defecation after spinal cord injury and its underlying mechanisms.

A new artificial somatic-autonomic neuroanastomosis has been established in male rats with spinal cord injury (SCI). Anorectal manometry and neural retrograde tracing were conducted in this animal model to analyze the mechanisms and the effects on recovery of anorectal function. The left L4 ventral root (L4VR) was intradurally micro-anastomosed to the L6 ventral root (L6VR) to establish the new regenerated neural pathway. Three months later the spinal cord was completely transected at the T9-10 level. Eight weeks later the model rats were randomly divided into two groups. The rats in the group 1 (n=8) were applied for anorectal manometry, and those in the group 2 (n=4) were used for neural retrograde tracing study with fluorogold (FG) and dextran tetramethylrhodamine (TMR). The results of anorectal manometry showed the new reflex pathway could induce rectum to contract and simultaneously electric activity of external anal sphincter (EAS) to become weak or disappearing (indicating synergetic relaxation of EAS). FG and TMR dual labeled neurons with round and elliptical shape were mainly observed in L4 angulus anterior of model rats. The regenerated neural pathways were effective to improve the rectum external sphincter synergetic status and restore the anorectal function.