Exosomal miR-34s panel as potential novel diagnostic and prognostic biomarker in patients with hepatoblastoma.

PURPOSE: The aim of this study is to identify the diagnostic values of serum exosomal miRNA-34s of patients with HB in a large Asian group and explore the prognostic value of the exosomal miRNA-34s panel compared with other risk factors. METHODS: We retrospectively reviewed 89 children with HB. Among these patients, 63 patients were included as training group to build the diagnostic model for HB. 26 patients were defined as the validation group. The expressions of miRNA-34s were detected by real-time PCR. The comparison of diagnostic and prognostic performance of serum exosomal miRNA-34s was measured using the area under ROC curve (AUC). RESULTS: For patients in the training group, expression of miRNA-34a, miRNA-34b and miRNA-34c was significantly lower in patients with HB compared with control group in serum exosomes. Between HB training group and the control group, exosomal miRNA-34a, miRNA-34b and miRNA-34c had no significant differences compared with the AFP level in diagnosing HB. The performance of the exosomal miRNA-34s panel in differentiating the HB training group from the control group was superior to the AFP level. The value of the exosomal miRNA-34s panel in predicting prognosis of patients with HB was superior to other risk factors in both training group and validation group. CONCLUSIONS: In this study, we found that the expression of exosomal miRNA-34a, miRNA-34b and miRNA-34c was significantly lower in patients with HB compared with the control group, and we confirmed the exosomal miRNA-34s panel could be defined as a diagnostic and prognostic biomarker for patients with HB. LEVEL OF EVIDENCE: Level II. TYPE OF STUDY: Retrospective Study.

Hedgehog signaling pathway regulates liver regeneration in the Fah-/- knockout mice model xenografted by human hepatocytes.

BACKGROUND: The aim of this study is to evaluates the hypothesis that Hh pathway activation occurs in the Fah-/- Nod/Scid mice model and plays a role in regulating liver regeneration after functional human hepatocytes xenograft. METHODS: Fah-/- Nod/Scid mice were established in Shandong Cancer Hospital affiliated to Shandong University. Xeno-regeneration of Fah-/- Nod/Scid mice livers was then transplanted by human hepatocytes. Hh signaling pathway associated factors were detected by RT-PCR and western blot. All statistical calculations were carried out using the SPSS.16.0 software. RESULTS: Fah-/- mice were healthy and reproduced normally while treated with NTBC. NTBC-OFF Fah-/- mice with HHT gradually gained weight by six weeks after human hepatocyte transplantation. NTBC-OFF Fah-/- mice causes a progressive increase in ALT, AST, GGT and AP, which were used to monitor hepatocyte inflammation. However, liver function in NTBC-OFF Fah Fah-/- mice with HHT returned to normal. Hedgehog pathway activation and Hh-target genes were enhanced follows human hepatocytes transplantation which indicated liver regeneration associated closely with Hh signaling pathway. Moreover, Hh signaling was inhibited by cyclopamine after HHT. CONCLUSIONS: The current study provides novel evidence that Hedgehog signaling pathway regulation is required for optimal regeneration of NTBC OFF Fah-/- mice with HHT.

HBV inhibits LPS-induced NLRP3 inflammasome activation and IL-1ß production via suppressing the NF-κB pathway and ROS production.

BACKGROUND & AIMS: Hepatitis B virus (HBV) has developed strategies to evade immune responses. However, the mechanisms involved remain unclear. The NLRP3 inflammasome plays crucial roles in antiviral host defense and its downstream factor IL-1ß has been shown to inhibit HBV infection in vivo. This study aims to assess whether HBV can affect the NLRP3 inflammasome signaling pathways and shed light on the underlying mechanisms HBV utilizes to evade host innate immune responses. METHODS: HBV inhibition of the lipopolysaccharide (LPS)-induced NLRP3 inflammasome activation was evaluated by Western blot, quantitative RT-PCR, flow cytometry and immunofluorescence. RESULTS: Kupffer cells expressed significantly more NLRP3 and IL-1ß after LPS stimulation; whereas, chronic HBV infection suppressed LPS-induced NLRP3 and pro-IL-1ß expression as well as IL-1ß maturation. This inhibitory activity is mediated by HBeAg, and is involved in the inhibition of NF-κB signal pathway and reactive oxygen species (ROS) production. The inhibitory effect of HBeAg was confirmed in patients with chronic hepatitis B (CHB) and hepatocellular carcinoma by comparing the levels of IL-1ß and NLRP3-related proteins in para-carcinoma tissues from HBeAg-positive or negative patients. Moreover, chronic HBV infection increases the susceptibility of mice to S. typhimurium infection, possibly via inhibiting the NLRP3 inflammasome activation and IL-1ß production. CONCLUSIONS: HBeAg inhibits LPS-induced NLRP3 inflammasome activation and IL-1ß production via suppressing NF-κB pathway and ROS production. This finding provides a novel mechanism for HBV-mediated suppression of innate immune responses, and identifies new therapeutic targets for chronic HBV infection and related diseases. LAY SUMMARY: HBeAg suppresses LPS-induced NLRP3 inflammasome activation and IL-1ß production in two ways, one is to repress NLRP3 and pro-IL-1ß expression via inhibiting NF-κB phosphorylation, and the other is to repress caspase-1 activation and IL-1ß maturation via inhibiting ROS production. This effect contributes to the HBV persistence and immune tolerance.

Combined low miR-34s are associated with unfavorable prognosis in children with hepatoblastoma: A Chinese population-based study.

PURPOSE: The aim of this study is to identify the association between miR-34's family and the prognosis of HB in a large Asian cohort and to explore the interaction of miR-34 with other independent risk factors in the process of affecting prognosis of HB. METHODS: We retrospectively reviewed 78 children with HB (36 female, 42 male) managed in our institutions between 2007 and 2014. The expression of miR-34 was detected by real-time PCR. Prognostic factors were evaluated using Kaplan-Meier curves and Cox proportional hazards models. RESULTS: For the entire cohort of 76 patients, The normalized real-time PCR results showed that all three miRNAs were deregulated in tumor tissues as compared with corresponding noncancerous tissue samples. Descriptive survival statistics and Kaplan-Meier curves suggested that AFP levels, metastases, vascular invasion, PRETEXT stage and miR-34 had prognostic significance in this relatively selected cohort. After that we made miR-34 into different combinations. The results demonstrated that combined low miR-34a and miR-34b (HR:2.212, P=0.016), combined low miR-34a and miR-34c (HR:1.984, P=0.025) and combined low miR-34a, miR-34b and miR-34c (HR:3.569, P=0.001) were independent prognostic factors of HB. We further conduct stratified analysis of the impact of other identified risk factors on the combined low of three miR-34. CONCLUSIONS: In this study, we found that miR-34s were deregulated in tumor tissues compared with corresponding noncancerous tissue samples. We also confirmed that combined low miR-34 is an independent prognostic factor related with HB.

Long-term outcome of penile appearance and sexual function after hypospadias repairs: situation and relation.

OBJECTIVE: The aim of this study was to assess the long-term cosmetic and sexual outcomes of hypospadias surgery performed in childhood and to analyze the relation between them. METHODS: A long-term follow-up was made to 174 patients who were operated for hypospadias in this institute between 1984 and 1992. Their records were analyzed retrospectively, and a detailed questionnaire was mailed to them. Responses from returned questionnaires were pooled and analyzed. RESULTS: Forty-three (24.7%) of 174 patients finished the questionnaire. The mean patient age of the 43 patients was 21.6 years. Twenty-three (53.5%) of 43 patients were dissatisfied with penile appearance. The main reason for dissatisfaction was smaller penile size and curvature. Thirty-six (83.7%) of 43 patients were satisfied with overall sexual function. Of these 43 patients, 76.7% reported good quality of erection, but 23.3% reported the existence of problems during erection. The main complaint was smaller penile size and curvature. There were 17 patients (39.5%) with ejaculation problems, mainly including impotent ejaculation. Sixteen (88.9%) of the 18 patients who had experienced sexual intercourse reported no problems during intercourse. Depending on the severity of hypospadias, these patients with proximal hypospadias were more dissatisfied with penile appearance than those with distal hypospadias (76.5 vs. 38.5%, P < 0.025). And they complained more problems during erection (41.2 vs. 11.5%, P < 0.05), ejaculation (100 vs. 0%), and sexual intercourse (50 vs. 0%, P < 0.05), and were less likely to experience sexual intercourse (23.5 vs. 53.8%, P < 0.05), compared to those with distal hypospadias. Depending on their self-evaluation on penile appearance, these patients who were satisfied with penile appearance had less problems in achieving erection (5.0 vs. 39.1%, P < 0.025) and ejaculation (20 vs. 56.5%, P < 0.025) and were more likely to experience sexual intercourse (60 vs. 26.1%, P < 0.05), compared to those who were dissatisfied. The self-rated satisfaction scale correlates positively with the penile appearance and sexual function (r = 0.469, P < 0.01). CONCLUSIONS: Patients who had been operated for hypospadias have a significantly careful concern on penile appearance. Penile size can obviously impact satisfaction with penile appearance and sexual function. Some patients have erection, ejaculation, and intercourse problems in adulthood, but majority have a rather normal sexual function. The more severe the hypospadias, the more dissatisfactory the long-term outcome. Better cosmetic outcome is related to better sexual outcome. Although remaining challenging, it is necessary to follow up hypospadias patients after surgeries into adulthood.