Molecular characterization of exons 6, 8 and 9 of ABCB4 gene in children with Progressive Familial Intrahepatic Cholestasis type 3

Aim of work: To estimate the frequency of mutations involving exons 6, 8 and 9 of Adenosine triphosphate-binding cassette, subfamily B, member 4 (ABCB4) gene among children with progressive intrahepatic cholestasis with high γ-GT activity (PFIC3).

Subjects and methods: Cross sectional study was conducted on 30 children with PFIC3. Genotyping was performed by sequencing analysis of exons 6, 8 and exon 9 of ABCB4 gene.

Results: Heterozygous synonymous polymorphic variant was detected in exon 6 (rs 1202283) and in exon 8 (rs 2109505). No mutations in studied exons were detected.

Conclusion: Exons 6, 8 and 9 mutations of ABCB4 gene are not common among Egyptian children with PFIC3.     

妊娠期肝内胆汁淤积症合并妊娠期糖尿病的母儿结局分析

目的:探讨妊娠期肝内胆汁淤积症(ICP)合并妊娠期糖尿病(GDM)对母儿结局的影响。方法:选取2012年1月至2013年2月在我院住院分娩的13例ICP合并GDM孕妇为ICP+GDM组,将同期住院分娩的69例单纯ICP孕妇归为ICP组,对两组孕妇的母儿结局进行回顾性比较分析。结果:两组孕妇的子痫前期、胎膜早破、剖宫产、产后出血发生率比较,无明显差异(P0.05);ICP+GDM组孕妇围产儿Apgar小于7分、新生儿肺炎、早产发生率明显高于ICP组,差异有统计学意义(P均0.05);ICP+GDM组孕妇围产儿平均出生体重低于ICP组,差异有统计学意义(P0.05)。结论:妊娠期肝内胆汁淤积症合并妊娠期糖尿病将进一步加重围产儿不良结局,对于此类孕妇,应加强监护和管理,适时终止妊娠,以改善围产儿结局。     

熊去氧胆酸联用思美泰对妊娠期肝内胆汁淤积症重症患者瘙痒程度、妊娠结局及肝功能的影响

目的:探讨熊去氧胆酸联用思美泰对妊娠期肝内胆汁淤积症(ICP)重症患者瘙痒程度、妊娠结局及肝功能的影响。方法:选取2016年6月到2017年12月期间在我院接受治疗的ICP重症患者76例,根据随机数字表法将患者均分为研究组(n=38)和对照组(n=38)。对照组采用熊去氧胆酸进行治疗,研究组采用熊去氧胆酸联用思美泰进行治疗。比较两组患者治疗前、治疗7d后的Ribaha皮肤瘙痒程度评分、天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、总胆汁酸(TBA)、血清总胆红素(TB)的水平,比较两组患者的妊娠结局。结果:治疗7d后两组患者的瘙痒程度评分均明显降低,且研究组低于对照组(P0.05)。治疗7d后两组患者的AST、ALT、TBA、TB水平均明显降低,且研究组患者的AST、ALT、TBA、TB水平低于对照组,差异有统计学意义(P0.05)。两组患者的新生儿窒息发生率和新生儿体质量比较无统计学差异(P0.05),研究组患者早产、胎儿窘迫、羊水胎粪污染的发生率低于对照组,差异有统计学意义(P0.05)。结论:熊去氧胆酸联用思美泰可有效缓解ICP重症患者的瘙痒程度,改善肝功能和妊娠结局,临床疗效显著,具有积极的临床意义。     

FIC1, a P-type ATPase linked to cholestatic liver disease,has homologues (ATP8B2 and ATP8B3) expressed throughout the body

ATP8B1/FIC1 is a member of the Type IV P-type ATPase family, which function as ATP dependent aminophospholipid translocases (APLT). We identified two familial intrahepatic cholestasis type 1 (FIC1) homologues, ATP8B2 and ATP8B3, with 53% and 45% amino acid identity, respectively. The expression profile for each gene was determined using a 73-tissue human RNA expression array. The subfamily of FIC1-like proteins is expressed in a wide range of tissues. Given that mutations in FIC1 result in liver disease, these proteins may have important roles in other organs in which they are candidates for genetic and acquired diseases.     

Epigenetic regulation of miR-124 by Hepatitis C Virus core protein promotes migration and invasion of intrahepatic cholangiocarcinoma cells by targeting SMYD3

Hepatitis C Virus core protein (HCVc) plays important roles in the development of intrahepatic cholangiocarcinoma (ICC). MicroRNAs (miRNAs) contribute to tumor progression by interacting with downstream target genes. However, the regulation and role of miRNAs in HCV-related intrahepatic cholangiocarcinoma (HCV-ICC) is poorly understood. In this study, we found that miR-124 was down-regulated in HCV-ICC and the induction of DNMT1 by HCVc mediated the suppression of miR-124. Over-expression of miR-124 suppressed cell migration and invasion in vitro, and reduced the protein levels of SMYD3 and downstream target genes (c-Myc and MMP9). Knockdown of SMYD3 inhibited cell migration and invasion resembling that of miR-124 over-expression. In conclusion, our studies indicate that low miR-124 levels mediated by HCVc via DNMT1 promote ICC cell migration and invasion by targeting SMYD3.     

人肝内胆管癌细胞PRL-3 的表达与侵袭转移研究

目的:探讨PRL-3在人肝内胆管癌侵袭转移中的作用.方法:利用小RNA技术干扰肝内胆管癌细胞株PRL-3表达,并采用细胞划痕实验和Transwell体外侵袭实验评价PRL-3对肝内胆管癌细胞侵袭转移能力的影响.结果:RT-PCR和Western blot结果均显示转染PRL-3特异性siRNA-2组PRL-3表达明显降低(P＜0.05).PRL-3 siRNA-2组在划痕培养24h后划痕区域宽度占初始划痕区域宽度的百分比为(62.12±6.28)％,阴性对照组为(23.88±2.55)％,空白对照HCCC-9810组为(21.20±6.07)％.PRL-3siRNA-2组细胞的划痕两端距离相比明显较宽,分别与阴性对照组细胞和空白对照HCCC-9810组细胞相比均有统计学意义(P＜0.05),后两者无显著性差异(P＞0.05).Transwell体外侵袭实验结果显示,PRL-3 SiRNA-2组细胞侵袭能力明显减弱,穿膜细胞数为(19.40±2.30)个/HP,明显少于阴性对照组(64.00±2.73)个/HP和正常HCCC-9810组(67.20±3.l1)个/HP,差异有显著性(P＜0.05);正常HCCC-9810组和阴性对照组无明显差异(P＞0.05).结论:PRL-3特异性siRNA能够抑制肝内胆管癌细胞HC-CC-9810中内源性PRL-3的表达,并可以明显抑制肝内胆管癌细胞的迁移侵袭能力.     

Expression profiles of MUC mucins and trefoil factor family (TFF) peptides in the intrahepatic biliary system: physiological distribution and pathological significance

Mucin secreted by mucosal epithelial cells plays a role in the protection of the mucosal surface and also is involved in pathological processes. So far, MUC1-4, 5AC, 5B, 6-8, 11-13 and 15-17 genes coding the backbone mucin core protein have been identified in humans. Their diverse physiological distribution and pathological alterations have been reported. Trefoil factor family (TFF) peptides are mucin-associated molecules co-expressed with MUC mucins and involved in the maintenance of mucosal barrier and the biological behavior of epithelial and carcinoma cells. Intrahepatic biliary system is a route linking the bile canaliculi and the extrahepatic bile duct for the excretion of bile synthesized by hepatocytes. Biliary epithelial cells line in the intrahepatic biliary system, secreting mucin and other molecules involved in the maintenance and regulation of the system. In this review, the latest information regarding properties, expression profiles and regulation of MUC mucins and TFF peptides in the intrahepatic biliary system is summarized. In particular, we focus on the expression profiles and their significance of MUC mucins in developmental and normal livers, various hepatobiliary diseases and intrahepatic cholangiocarcinoma.     

Blocking intrahepatic deletion of activated CD8+ T cells by an altered peptide ligand

BACKGROUND: Activated CD8(+) T cells are retained by the healthy liver where the majority undergo apoptosis. The intrahepatic apoptosis of activated CD8(+) T cells is enhanced by the presence of SIINFEKL peptide. It is of great interest to identify strategies for maintaining intrahepatic T cell number and function in the presence of SIINFEKL peptides. AIM: Our aim was to test if low affinity peptides can block SIINFEKL peptide induced T cell deletion. METHODS: We used an in vivo model of intrahepatic CD8(+) T cell deletion with peptides of different affinities. RESULTS AND DISCUSSION: We show that the intrahepatic deletion of CD8(+) T cells by SIINFEKL peptide results in loss of in vivo cytotoxic T lymphocyte function. In contrast we show that a low affinity peptide (G4) does not result in intrahepatic deletion of CD8(+) T cells. High concentrations G4 peptide can however block intrahepatic deletion of activated CD8(+) T cells, and prevent loss of in vivo cytotoxicity due to SIINFEKL peptide. This is the first demonstration of blocking of SIINFEKL peptide induced CD8(+) T cell deletion in the liver, with enhancement of in vivo cytotoxicity.