Dendritic targeted mRNA expression via a cis-acting RNA UTR element.

Local translation in neurites is considered as an important mechanism to modulate synaptic plasticity of neurons. However, it is hard to specifically express a protein-coding gene in neurites. Recently, the 5'-UTR of Tick-borne encephalitis virus (TBEV) is reported to be able to drive its RNA to the dendrites of infected neurons, as a cis-acting RNA element. To construct a neurite specific gene expression system, present study tested the ability of 5'-UTR of TBEV to bring a mRNA (mCherry CDS) to the neurites for targeted expression. We showed that both the 5'-UTR of TBEV and the 3'-UTR of Actb gene could bring the protein coding mRNA to neurites, and the TBEV 5'-UTR bearing mRNA was more robust targeted into neurites. About the safety of the TBEV 5'-UTR, there was no obvious cytotoxicity to the neurons when adding either cis-acting RNA element to the protein-expressing plasmid vectors. Given the short length and high efficiency of the TBEV 5'-UTR, the 5'-UTR of TBEV were assemble into an AAV plasmid to produce virus particles for expressing protein-coding gene in vivo. After two weeks infection, the TBEV 5'-UTR infected neurons expressed more mCherry protein in their neurites. In conclusion, as a short while high efficient cis-acting RNA element, TBEV 5'-UTR could be useful in neural system research and locally express synaptic proteins more precisely.

Gonadal tumor risk in pediatric and adolescent phenotypic females with disorders of sex development and Y chromosomal constitution with different genetic etiologies.

Objectives: This retrospective study sought to investigate the risk and proportion of gonadal neoplasms in phenotypic female pediatric patients with DSD and the presence of the Y chromosome and different genetic backgrounds in a single Chinese center. Materials and Methods: From January 2012 to December 2020, pediatric and adolescent patients with DSD and the presence of the Y chromosome who had unambiguous female genitalia and underwent bilateral gonadectomy or gonadal biopsy were included in this study. Patients' demographics, karyotype, laboratory test results, gross pathology, and histology of gonadal tissue were all collected. The patients were divided into three groups based on their different genetic backgrounds, and the percentage of gonadal tumors was calculated to assess the risk of gonadal tumor and malignancy by etiology. Results: A total of 22 patients with DSD and an unambiguous female phenotype with a Y chromosome were recruited. The mean age was 10.91 ± 4.99 years (9 months to 19 years). Gonadal neoplasia was confirmed in six (27.3%) cases by pathological examination of surgical gonadal tissue samples. Among 44 gonadal samples from these 22 patients, the following were identified: five gonadoblastomas, three dysgerminomas, and two Leydig cell tumors. The youngest patient with a tumor was a 2-year-old girl with 46,XY complete gonadal dysgenesis (46,XY CGD or Swyer syndrome) and bilateral gonadoblastoma. Patients with 46,XY complete gonadal dysgenesis (4/6; 66.7%) had the highest tumor occurrence rate. Among 10 patients with Turner syndrome with the presence of the Y chromosome, only one patient was diagnosed with a gonadal tumor. Leydig cell tumor was diagnosed in only one of six patients with 46,XY androgen synthesis/action disorders. Conclusion: Pediatric patients with 46,XY complete gonadal dysgenesis had a significantly increased risk of developing gonadal tumors and underwent prophylactic gonadectomy as soon as the diagnosis was confirmed, whereas those with Turner syndrome with Y chromosome and 46,XY androgen synthesis/action disorders had a relatively low risk. In view of the limited number of patients, a large multicenter study with close follow-ups is needed to support these conclusions.

SIDT1-dependent absorption in the stomach mediates host uptake of dietary and orally administered microRNAs.

Dietary microRNAs have been shown to be absorbed by mammals and regulate host gene expression, but the absorption mechanism remains unknown. Here, we show that SIDT1 expressed on gastric pit cells in the stomach is required for the absorption of dietary microRNAs. SIDT1-deficient mice show reduced basal levels and impaired dynamic absorption of dietary microRNAs. Notably, we identified the stomach as the primary site for dietary microRNA absorption, which is dramatically attenuated in the stomachs of SIDT1-deficient mice. Mechanistic analyses revealed that the uptake of exogenous microRNAs by gastric pit cells is SIDT1 and low-pH dependent. Furthermore, oral administration of plant-derived miR2911 retards liver fibrosis, and this protective effect was abolished in SIDT1-deficient mice. Our findings reveal a major mechanism underlying the absorption of dietary microRNAs, uncover an unexpected role of the stomach and shed light on developing small RNA therapeutics by oral delivery.

[Diagnosis and treatment of ovotesticular disorders of sex development in children].

OBJECTIVE: To investigate the diagnosis and treatment of ovotesticular disorders of sex development (DSD) in children. METHODS: We reviewed the clinical data of 9 cases of ovotesticular DSD admitted in our department from 1988 to 2007. RESULTS: The patients ranged in age from 9 months to 9 years, 7 raised as males and 2 as females. As for the karyotype, 4 cases were 46,XX, 2 were 46,XX/46,XY, 1 was 46,XY, and the other 2 had no karyotype data. All of them presented with obscure external genitalia: perineal or penoscrotal hypospadias with or without cryptorchidism in males and hypertrophy of the clitoris in females. They were diagnosed with ovotesticular DSD by gonad biopsy and underwent genitoplasty. CONCLUSION: The gender assignment of the ovotesticular DSD patient was chiefly based on the development of external genitalia, dominant gonad, karyotype and the parent's will. Laparoscopic technology is recommended in gonad biopsy and orchiopexy during the treatment of ovotesticular DSD.

Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents.

Erectile dysfunction (ED) is defined as the inability to attain or keep an erection of the penis, and this has become a prevalent male sexual disorder. Rodents are employed by many studies to research the physiology/pathology of erectile function. Erectile function in rodents can be evaluated by measuring the intracavernosal pressure (ICP). In practice, ICP can be monitored following electrical stimulation of the cavernous nerves (CNs). The arterial pressure of the carotid artery (the mean arterial pressure) is used as the reference for ICP. Using ICP recording protocols, many key parameters of erectile function can be measured from the ICP response curve. The ICP measurement provides more information than the apomorphine-induced penile erection test, and is cheaper than telemetric monitoring of the corpus spongiosum penis, making this method the most popular one to evaluate erectile function. However, compared to the easily-performed APO-induced erectile function test, successful ICP recordings require attention to detail, practice, and adherence to the operation method. In this work, an introduction to ICP recording in rats is provided to complement the procedure efficiently.

Laparoscopic Resection and End-to-End Ureteroureterostomy for Midureteral Obstruction in Children.

PURPOSE: Midureteral obstruction is an extremely rare condition. This retrospective study assessed the outcomes of laparoscopic ureteroureterostomy (UU) in 13 children with midureteral obstructions. METHODS: Records of patients with midureteral obstruction who underwent laparoscopic end-to-end UU between July 2011 and August 2015 were reviewed. The medical records comprised patient demographic data, intraoperative details, postoperative outcomes, and related complications. Renal ultrasound, magnetic resonance urography, and radioisotope renography were used for pre- and postoperative assessment. Success was defined as removal of symptomatic obstruction or improvement of hydronephrosis. RESULTS: Thirteen patients from 3.0 months to 12 years old were identified: 6 patients presented with asymptomatic hydronephrosis (according to the renal ultrasonography), 3 with intermittent abdominal pain, and 2 with gross hematuria. The remaining 2 patients presented with urinary extravasation after inguinal hernia repair and the Soave procedure, respectively. All patients underwent laparoscopic end-to-end UU. Procedures were performed using either the laparoscopic transperitoneal approach or laparoscopic retroperitoneal approach. All surgeries were performed successfully except for one case that required open surgery because of difficult anastomosis. There were no intraoperative or immediate postoperative complications. The etiology of the midureteral obstruction included congenital midureteral stricture, ureteral polyp, retrocaval ureter, and iatrogenic injury. The mean surgical time was 148.4 min, and postoperative hospital stay was 4.0-9.0 d. Postoperative outcomes were successful in all 13 patients (100%); median follow-up was 19.1 months. CONCLUSION: Laparoscopic UU was found to be feasible and safe for the management of midureteral obstruction in children.

Epidemiologic study of 230 cases of testicular/paratesticular tumors or masses: 15-year experience of a single center.

PURPOSE: This study retrospectively investigated the cases of testicular tumors/masses treated in our center from 2002 to 2017 and analyzed their epidemiologic features. METHODS: Data were collected by searching our center's database using "testicular tumor" or "testicular mass" as keywords. Patients not operated in our hospital were excluded. Preoperative serum alpha-fetoprotein (AFP) levels were reviewed in germ cell tumor (GCT) cases and analyzed to predict malignancy in various age groups. RESULTS: In total, 230 cases were identified: 151 were benign (78 in the left, 72 in the right, and 1 bilateral) with 3.63years mean age during the operation, and 79 were malignant (42 in the left, 36 in the right, and 1 bilateral) with 2.21years mean age during the operation. Main pathological diagnoses were mature teratoma (92, 40.00%), yolk sac tumor (53, 23.04%), dermoid cyst (23, 10.00%), embryonic carcinoma (15, 6.53%), immature teratoma (14, 6.09%), benign cyst (8, 3.48%), Leydig cell tumor (6, 2.61%), and paratesticular rhabdomyosarcoma (5, 2.17%). All GCT cases with AFP >1000ng/ml, >100ng/ml, >20ng/ml were malignant in <7-, 7-9-, and ≥10-month-old groups, respectively. CONCLUSIONS: Radical inguinal orchiectomy without biopsy is suggested in 7-9- and ≥10-month-old cases with AFP >100ng/ml and >20ng/ml, respectively. TYPE OF STUDY: Retrospective Study. LEVEL OF EVIDENCE: Level III-IV.

Efficacy and safety of percutaneous administration of dihydrotestosterone in children of different genetic backgrounds with micropenis.

BACKGROUND: Exogenous androgen supplement is an optional treatment for micropenis; however, its use in childhood is controversial due to potential side effects. METHODS: Twenty-three children (mean age: 4.07±3.4 years) with micropenis of unknown causes harboring the 46,XY karyotype were recruited in an open prospective study. Androgen receptor (AR), steroid 5α-reductase-2 (SRD5A2), and SRY genes were sequenced; 2.5% dihydrotestosterone (DHT) transdermal gel (0.1-0.3 mg/kg/day) was applied and titrated within the normal DHT serum reference ranges. Stretched penile length (SPL) was measured before therapy, and after 1, 3 and 6 months of DHT gel treatment, respectively. RESULTS: Two patients were found with AR gene mutations and five patients with SRD5A2 gene mutations. Average stretched penile lengths (SPLs) were 1.68±0.6 cm at baseline and 2.2±0.66 cm, 2.6±0.59 cm and 2.9±0.55 cm (mean ± 1 SD) after 1, 3 and 6 months of treatment, respectively. Fourteen cases (61%) reached standard penile length ranges (>-2.5 SD) and medication was discontinued; six cases (26%) were satisfied with the improved penile lengths despite failing to reach the aged matched standards. Three infants (13%) discontinued the medication after 3 months due to anxiety about the potential side effects. No significant side effects were found except the elevated DHT serum levels after therapy. CONCLUSIONS: Short term and local application of DHT at low doses in patients with micropenis could accelerate penile growth effectively without evident side effects; however, precautions still need be taken due to the paucity of long term study and the lack of ideal DHT dosage.

Using conventional 3- and 5-mm straight instruments in laparoendoscopic single-site pyeloplasty in children.

BACKGROUND: Laparoendoscopic single-site surgery (LESS) remains limited to a few relatively simple procedures in the field of pediatric surgery. We performed LESS pyeloplasty in children using conventional straight instruments and ports, and reviewed the perioperative data to evaluate the feasibility of this technique in pediatric patients. METHODS: Twenty-two consecutive patients with unilateral pelvic ureteral junction obstruction underwent LESS pyeloplasty. Their ages ranged from 2 to 134 months (mean: 56.9 months). Two 3-mm and one 5-mm conventional ports were placed in the single incision inside the umbilicus. Conventional 3-mm laparoscopic instruments and a 30-degree 5-mm camera were used for Anderson-Hynes pyeloplasty. A F4.7 or F3 pigtail stent was introduced through one of the instrument ports. A drainage was placed next to the anastomosis, which exited via the umbilical ring incision. The stent was removed 6 weeks postoperatively. RESULTS: All cases were successfully completed with no need for extra-umbilical incisions. The mean operative time was 198 minutes (range: 150-270 minutes). All patients could tolerate oral food intake on postoperative day 1. One case (4.5%) developed symptoms of anastomosis obstruction and required open renal stoma. No other significant complications occurred. CONCLUSION: LESS pyeloplasty is technically feasible in children. However, further experience and longer follow-up are necessary to appropriately evaluate the benefits and limitations of this technique.