Gong Chen

Targeting long non-coding RNA-TUG1 inhibits tumor growth and angiogenesis in hepatoblastoma

Hepatoblastoma is the most common liver tumor of early childhood, which is usually characterized by unusual hypervascularity. Recently, long non-coding RNAs (lncRNA) have emerged as gene regulators and prognostic markers in several cancers, including hepatoblastoma. We previously reveal that lnRNA-TUG1 is upregulated in hepatoblastoma specimens by microarray analysis. In this study, we aim to elucidate the biological and clinical significance of TUG1 upregulation in hepatoblastoma. We show that TUG1 is significantly upregulated in human hepatoblastoma specimens and metastatic hepatoblastoma cell lines. TUG1 knockdown inhibits tumor growth and angiogenesis in vivo, and decreases hepatoblastoma cell viability, proliferation, migration, and invasion in vitro. TUG1, miR-34a-5p, and VEGFA constitutes to a regulatory network, and participates in regulating hepatoblastoma cell function, tumor progression, and tumor angiogenesis. Overall, our findings indicate that TUG1 upregulation contributes to unusual hypervascularity of hepatoblastoma. TUG1 is a promising therapeutic target for aggressive, recurrent, or metastatic hepatoblastoma.

microRNA-222 modulates liver fibrosis in a murine model of biliary atresia.

microRNA-222 (miR-222) has been shown to initiate the activation of hepatic stellate cells, which plays an important role in the pathogenesis of liver fibrosis. The aim of our study was to evaluate the role of miR-22 in a mouse model of biliary atresia (BA) induced by Rhesus Rotavirus (RRV) infection. New-born Balb/c mice were randomized into control and RRV infected groups. The extrahepatic bile ducts were evaluated. The experimental group was divided into BA group and negative group based on histology. The expression of miR-222, protein phosphatase 2 regulatory subunit B alpha (PPP2R2A), proliferating cell nuclear antigen (PCNA) and phospho-Akt were detected. We found that the experimental group showed signs of cholestasis, retardation and extrahepatic biliary atresia. No abnormalities were found in the control group. In the BA group, miR-222, PCNA and Akt were highly expressed, and PPP2R2A expression was significantly inhibited. Our findings suggest that miR-222 profoundly modulated the process of fibrosis in the murine BA model, which might represent a potential target for improving BA prognosis.

Serum microRNA microarray analysis identifies miR-4429 and miR-4689 are potential diagnostic biomarkers for biliary atresia

This study aimed to investigate pathogenesis and novel diagnostic biomarkers of biliary atresia (BA). Serum samples from infants with BA and non-BA neonatal cholestasis (NC) were collected for miRNA microarray analysis, and then differentially expressed miRNAs were screened. Differentially expressed miRNAs were validated by qRT-PCR using an independent serum samples from infants with BA and NC. Diagnostic utility of validated miRNAs was further analyzed using serum samples by receiver-operating characteristic curve analysis. Totally, 13 differentially expressed miRNAs were identified including 11 down-regulated and 2 up-regulated ones. Target genes of hsa-miR-4429 and hsa-miR-4689 were significantly involved in FoxO signaling pathway. Eight differentially expressed miRNAs were chosen for validation by qRT-PCR analysis, and four miRNAs (hsa-miR-150-3p, hsa-miR-4429, hsa-miR-4689 and hsa-miR-92a-3p) were differentially expressed. The area under the curve of hsa-miR-4429 and hsa-miR-4689 was 0.789 (sensitivityu2009=u200983.33%, specificityu2009=u200980.00%) and 0.722 (sensitivityu2009=u200966.67%, specificityu2009=u200980.00%), respectively. Differentially expressed miRNAs including hsa-miR-4429 and hsa-miR-4689 might play critical roles in BA by regulating their target genes, and these two miRNAs may have the potential to become diagnostic biomarkers.

Interleukin-33 overexpression is associated with gamma-glutamyl transferase in biliary atresia

Interleukin-33 (IL-33) plays a crucial role in inflammation. However, it is not clear whether IL-33 levels are of clinical significance for patients with biliary atresia (BA). The purpose of this study was to determine correlations between serum IL-33 levels and the clinicopathologic features of BA. Serum samples were collected from 18 BA infants, 12 nonicteric choledochal cyst (CC) infants with normal liver function, and 10 healthy controls (HCs). Serum IL-33 levels were measured with an enzyme-linked immunosorbent assay (ELISA). Routine liver function tests were performed on the serum samples. qRT-PCR and Western blot analysis were used to detect IL-33 expression in BA liver biopsy tissues. Hepatic lobule localization of IL-33 expression in the hepatic lobule was conducted by immunohistochemistry (IHC). IL-33 levels in serum collected from BA infants were significantly elevated in comparison with CC and HC patients. Furthermore, the elevated serum levels of IL-33 in BA infants were correlated with gamma-glutamyl transferase (GGT) levels. The expression of IL-33 mRNA and protein levels were up-regulated in BA liver biopsy tissues in comparison with CC patients. IHC analysis revealed increased positive immunostaining for IL-33 in BA liver tissues as compared to that in CC tissues. These results suggest that IL-33 may play an important role in the pathogenesis of BA. In addition, the correlation of serum IL-33 levels with GGT levels may provide a novel marker for the diagnosis of BA.

Outcomes of thoracoscopy versus thoracotomy for esophageal atresia with tracheoesophageal fistula repair: A PRISMA-compliant systematic review and meta-analysis.

Background:A thoracoscopic approach for repair of esophageal atresia (EA) with tracheoesophageal fistula (TEF) has become a standard procedure in many pediatric surgical centers. However, whether thoracotomy or thoracoscopy offer advantages in terms of surgical outcomes is not known. Methods:To evaluate the efficacy and safety of thoracoscopic repair (TR) versus conventional open repair (COR) for EA with TEF.PubMed, Cochrane Library, and EMBASE were searched to identify relevant literature until 2016.Studies comparing surgical outcomes of patients undergoing TR versus COR for EA with TEF were reviewed.The quality of each included study was assessed using the Newcastle–Ottawa scale score. A fixed or random-effect model was applied depending on heterogeneity tests. Results:Eight observational clinical studies involving 452 patients were included in this meta-analysis. The meta-analysis of 2 major postoperative complications (leaks and strictures) did not show significant differences between TR and COR. Overall estimates of the odds ratio (OR) of TR versus COR for leaks and strictures were: 1.57 (95% confidence interval [CI], 0.77–3.20; Pu200a=u200a0.22) and 0.90 (95% CI, 0.27–2.97; Pu200a=u200a0.86), respectively. However, meta-analysis of operation time (ORu200a=u200a19.59, 95% CIu200a=u200a0.77–38.40, Pu200a=u200a0.04), timing of extubation (ORu200a=u200a−2.50, 95% CIu200a=u200a−3.39 to −1.62, Pu200a<u200a0.001), time to 1st oral feeding (ORu200a=u200a−2.58, 95% CIu200a=u200a−3.79 to −1.36, Pu200a<u200a0.001), and duration of hospital stay (ORu200a=u200a−10.76, 95% CIu200a=u200a−16.39 to −5.12, Pu200a<u200a0.001) showed significant differences.No randomized controlled trial was included, and most studies had small sample sizes and were based on retrospective analysis. Conclusion:TR and COR show a similar complication rates of leaks and strictures for EA/TEF repair. Although associated with a longer operative time, TR has the advantages of an earlier time to extubation and 1st oral feeding, and shorter hospital stay.

Improved Outcome of Biliary Atresia with Postoperative High-Dose Steroid

Objective. The dosage, duration, and the benefits of high-dose steroid treatment and outcome in biliary atresia (BA) remain controversial. In this study, we evaluated the impact of high-dose steroid therapy on the outcome of BA after the Kasai procedure. Methods. Intravenous prednisolone administration was started 1 week after surgery, followed by 8 to 12 weeks of oral prednisolone. Total bilirubin (TB) levels (3, 6, and 12 months after surgery), early onset of cholangitis, and two-year native liver survival were evaluated. Results. 53.4%, 56.9%, and 58.1% of the patients in the high-dose steroid group were jaundice-free 3, 6, and 12 months after surgery, respectively; these values were significantly higher than the 38.7%, 39.4%, and 43.3% of the low-dose steroid group. One year after surgery, the incidence of cholangitis in the high-dose group (32.0%) was lower than that in the low-dose group (48.0%). Infants with native liver in the high-dose group had a better two-year survival compared to those in the low-dose steroid group (53.7% versus 42.6%). Conclusions. The high-dose steroid protocol can reduce the incidence of cholangitis, increase the jaundice-free rate, and improve two-year survival with native liver after the Kasai operation.

Forkhead box A3 attenuated the progression of fibrosis in a rat model of biliary atresia

Biliary atresia is a rare, devastating disease of infants where a fibroinflammatory process destroys the bile ducts, leading to fibrosis and biliary cirrhosis, and death if untreated. The cause and pathogenesis remain largely unknown. We tried to investigate factors involved in biliary atresia, especially forkhead box A3 (Foxa3), which might exert a role in the treatment of liver disease. We used RNA sequencing to sequence the whole transcriptomes of livers from six biliary atresia and six choledochal cysts patients. Then, we employed a rat disease model by bile duct ligation (BDL) and adenovirus transduction to address the function of Foxa3 in biliary atresia. We found that tight junction, adherence junction, cell cycle, apoptosis, chemokine singling, VEGF and MAPK signaling pathways were enriched in biliary atresia livers. We showed that Foxa3 expression was notably decreased in liver samples from biliary atresia patients. More importantly, we found that its lower expression predicted a poorer overall survival of biliary atresia patients. Rats that received BDL surgery and Foxa3 expression adenovirus resulted in a significant decrease in the deposition of collagen, and expression of profibrotic cytokines (transforming growth factor-β and connective tissue growth factor) and fibrosis markers (α-smooth muscle actin, collagen I and collagen III), as compared with rats that received BDL surgery and control adenovirus. Our data suggested a protection role for Foxa3 during the progression of liver fibrosis in biliary atresia, and thereby supported increasing Foxa3 as a targeted treatment strategy.

Saturation of stool color in HSV color model is a promising objective parameter for screening biliary atresia

PURPOSEnWe aimed to study whether saturation in HSV color model could be a parameter for acholic stool and utilized for designing a mobile application for screening biliary atresia (BA).nnnMETHODSnSaturations of the colors in the three validated stool color cards (Taiwan, Japan, Britain) were read using PHOTOSHOP. Stools from 40 BA patients and 40 in-hospital neonates with pneumonia were photographed and analyzed with color-analyzing mobile applications.nnnRESULTSnSaturations of normal colors in the published stool cards were all >50% (67%~99%, median 85%) and were all <50% (7~47%, median 25%) for abnormal colors. With saturation<60% as a cutoff line, acholic stools could be identified with a sensitivity of 100% and a specificity of 85%.nnnCONCLUSIONnSaturation of stool color in HSV model is a promising objective parameter for acholic stool and could be utilized in designing mobile APPs for screening BA.nnnLEVEL OF EVIDENCEnStudy of diagnostic test, level II.

The Appropriate Timing and Dose of Obeticholic Acid in Patients With Primary Biliary Cirrhosis

Dear Editor: We read with great interest the article by Hirschfield et al, who evaluated the efficacy and safety of obeticholic acid (OCA) in a randomized, controlled trial of patients with primary biliary cirrhosis (PBC) who had exhibited an inadequate response to ursodeoxycholic acid (UDCA) therapy. They concluded that daily doses of OCA, ranging from 10 to 50 mg, significantly reduced the serum concentrations of alkaline phosphatase (ALP), g-glutamyl transpeptidase (g-GT) and alanine aminotransferase in this population. These are impressive findings that raise 2 important issues. First, Hirschfield et al enrolled patients with PBCwho had been on a stable dose of UDCA for 6 months before screening, and documented biochemical responses to OCA that were maintained during a 12-month, open-label extension trial. This is a very convincing result. The question remains, when should patients with an inadequate response to UDCA start taking OCA? Should it be when the inadequate response to UDCA becomes evident, or immediately after the diagnosis of PBC? Two additional randomized phase II trials have been undertaken in patients with PBC treated with OCA for 3 months. The first compared UDCA in combination with 1 of 3 doses of OCA (10, 25, or 50mg) with UDCA and placebo in patients who presented with persistently increased serum ALP concentration >1.5 times the upper limit of normal while on a stable dose of UDCA. Clinically significant reductions in ALP concentration were observed with all 3 doses of OCA compared with placebo. The second study evaluated OCA as monotherapy, comparing the effects of 10 or 50 mg OCA daily with placebo. Compared with placebo, significant relative and absolute decreases in ALP concentration were reported, as well as significant improvements in serum g-GT, conjugated bilirubin, C-reactive protein, immunoglobulin M, and tumor necrosis factor-a concentrations. These 2 studies suggest that OCA is effective immediately after the diagnosis of PBC with or without UDCA. Establishing the appropriate time to start OCA would make this conclusion even more convincing. Second, at what dose should patients start to take OCA? Hirschfield et al tested 3 doses of OCA (10, 25, and 50 mg), and concluded that 10 mg once daily was the most effective, with the incidence and severity of pruritus least among those who received this dose. Pruritus was reported by a high proportion of the participants, likely a consequence of the higher doses of OCA. A 12-month extension trial (at doses of 25 mg) showed a consistent biochemical response, and a long-term phase III trial of OCA in UDCA-treated patients, based on an up-titration assessment of OCA in 5-mg increments every 3 months up to 25 mg, is currently under way (EudraCT Number: 2011 –004728–36). Another phase III study is examining the safety and efficacy of once-daily OCA in PBC in patients incompletely responsive to or unable to tolerate UDCA. These patients were randomized to one of 3 groups: placebo, 10 mg OCA, or 5 mg OCA for 6 months titrated to 10 mg OCA based on clinical response (NCT01473524). These studies may help to illuminate the appropriate starting and maintenance doses of OCA. Therefore, the long-term efficacy of OCA, including the optimum timing of initiation, dose, and its clinical benefit in patients with advanced disease, still needs to be confirmed in longer prospective studies.

Surgical outcome and etiologic heterogeneity of infants with biliary atresia who received Kasai operation less than 60 days after birth: A retrospective study

Abstract This study aimed to analyze the impact of etiologic heterogeneity and operation age on prognosis of infants with biliary atresia (BA) who received Kasai operation prior to 60 days of age. From 2004 to 2010, 158 infants received Kasai operation before turning 60 days old. According to Davenport 2012 classifications, 4 groups of BA were defined: cystic BA, syndrome BA, and associated malformation, cytomegalovirus (CMV)-associated BA, and isolated BA. Native (autologous) liver survival rates and incidence of cholangitis 2 years after operation, as well as jaundice clearance rates 3 months after operation, were recorded. Although infants who received the operation between 51 and 60 days of age had a better jaundice clearance 3 months after operation and lower incidence of cholangitis as compared with those under 40 or between 41 and 50 days of age, there was no significant difference in survival rates. Among types of BA, infants with cystic BA had the best prognosis. In the syndrome BA and associated malformations group, as well as CMV-associated group, infants who received the operation early (<40 days of age) had a worse outcome as compared with those who received the operation between 41 and 50 days or 51 and 60 days of age. Both clinical etiologic heterogeneity and operation age may influence BA prognosis.