Non-coding RNAs Expression Profiles Study Of Fetal Rats With Anorectal Malformations And Clinical Evaluation Of One-Stage Laparoscopically Surgery For Newborns With Anorectal Malformations

PART 1 Non-coding RNAs Expression Profiles Study of Fetal Rats with Anorectal MalformationsBackground Anorectal malformations(ARMs)are the most common congenital malformation of the gut,and regulated by multiple signal transduction pathways.The microRNA(miRNA)and IncRNA expression profiles and their biologial functions in anorectal malformations(ARMs)remain unclear.The aim of our study was to evaluate miRNA,IncRNA,and mRNA expression profiles in the ARM fetal rats.Methods and Materials ARM was induced with ethylenethiourea(ETU)on gestational day 10.Cesarean deliveries were performed to harvest the embryos on gestional gay 20.For the extraction of total RNA,1 cm termianl hindgut samples were obtained from three fetal rats that had similar weights.The microarrays and quantitative RT-PCR analysis were conducted to evaluate the miRNA,lncRNA,and mRNA expression profiles in normal fetal rats(n=6)and ARM fetal rats(n=6).Results In total,33 miRNAs,164 lncRNA,and 772 mRNAs were significantly and differentially expressed in termianl hindgut tissues of ARM group versus control group.Functional annotation was performed to understand the functions and pathways of differentially expressed mRNAs.And we constructed a miRNA-target gene regulatory network including 25 differentially expressed miRNAs and 76 mRNAs.Furthermore,the credibility of the microarray-based results were validated by using qRT-PCR.Conclusion The miRNA,lncRNA,and mRNA expression in terminal hindgut tissue in ARM fetal rat might provide a basis for further research on the pathogenesis of ARMs.PART 2 A Comparative Study of One-stage and Three-stage Laparoscopic-assisted Anorectoplasty in Anorectal Malformations with Rectoprostatic Fistula and Rectobulbar FistulaAbstract The aim of this study was to compare the clinical outcomes of 1-stage and 3-stage surgical procedures to treat anorectal malformations(ARMs)with rectoprostatic and rectobulbar fistula using laparoscopic-assisted anorectoplasty(LAARP).Methods A total of 56 patients with ARMs and rectoprostatic and rectobulbar fistula who underwent LAARP from January 2011 to May 2014 in our institution were included in the study.They were divided into 2 groups according to the stage of procedure.The patients’ data and postoperative complications were compared between the 2 groups.The Krickenbeck classification was used for assessing the bowel functions.Results About 20 ARM newborns(rectoprostatic fistula/8,rectobulbar fistula/12)successfully underwent an one-stage LAARP,and about 36 ARM children(rectoprostatic fistula/20,rectobulbar fistula/16)underwent a three-stage LAARP(colostomy,LAARP,and closure of colostomy).The average age at the LAARP procedure in one-stage group was significantly lower than that in three-stage group(43.1 ±8.1 hours vs 4.9± 1.2 months;P=.00).The average operative time during the definitive procedure was 132.2± 15.9 minutes in the one-stage group and 120.5± 12.7 minutes in the three-stage group(P=.13).There was only 5 to 10 mL of blood loss during the LAARP procedure both the groups(P=.75).There were no significant differences between the 2 groups in postoperative hospital stay during the definitive procedure(10.2±2.3 days vs 8.5±2.2 days;P=.22).The rate of surgical site infection and dehiscence was 5%(1/20)in the one-stage group and 5.6%(2/36)in three-stage group(P=1.00).During the period of follow-up,the rate of voluntary bowel movement was 90%(18/20)in one-stage group and 94.4%(34/36)in three-stage group(P=.94).Free from soiling or grade I soiling was 80%(16/20)in one-stage group and 83.3%(30/36)in three-stage group(P=1.00);grade II soiling was found in 1(5%)patients in one-stage group and 5.6%in three-stage group(P=0.42).Two patients(10%)in one-stage group and 3 patients(8.3%)in three-stage group suffered from grade I constipation(P=0.83);while 2(10%)patients in one-stage group and 2 patients(5.6%)in three-stage group had grade Ⅱ constipation(P=0.42).No patients in the two groups suffered from grade Ⅲsoiling and grade Ⅲ constipation.Conclusion The one-stage LAARP procedure for neonate with rectoprostatic and rectobulbar fistula can achieve comparable midterm outcomes as the conventional three-stage LAARP procedure.It provides an alternative method to rectify the ARMs with rectoprostatic fistula and rectobulbar fistula without colostomy.