Fenofibrate in a patient with primary biliary cholangitis with an inadequate response to bezafibrate

To the Editor,A 48-year-old male patient presented to our hospital with abnormal liver function.The results of his liver function tests were as follows:alanine transaminase,118 U/L(normal range,5-35 U/L);aspartate transaminase,86 U/L(8-40 U/L);alkaline phosphatase(ALP),246 U/L(40-150 U/L);andγglutamyl transferase(γ-GT),792 U/L(17-53 U/L).His immunoglobulin(Ig)G and IgM levels were 13.0 g/L(7.51-15.60 g/L)and 2.44 g/L(0.460-3.040 g/L),respectively,and his autoimmune antibody tests were positive for antinuclear antibodies and M2 subtype antimitochondrial antibodies.

Retrorsine Cooperates with Gut Microbiota to Promote Hepatic Sinusoidal Obstruction Syndrome by Disrupting the Gut Barrier

Background and Aims:Hepatic sinusoidal obstruction syndrome(HSOS)is a life-threatening syndrome,and a cause is exposure to pyrrolizidine alkaloid(PA)-containing products.It is well-established that retrorsine(RTS),a rep-resentative Pas,insults hepatic sinusoidal endothelial cells and ensues congestion of hepatic sinusoids.However,little known about the impact of Pas on gut microbiota and intesti-nal barrier and inflammation in HSOS.Methods:Mice were gavaged with or without nonabsorbable antibiotics(ABX),followed by a single dose of RTS.The gut microbiota was examined by 16S rDNA sequencing.Results:ABX pretreat-ment significantly reversed RTS-induced liver damage.RTS altered gut microbiota composition,increasing Gram-nega-tive bacteria and resulting in a sharp elevation of circulating lipopolysaccharides(LPS)in HSOS mice.Gut decontamina-tion with ABX alleviated RTS-induced intestine inflamma-tion,protected against disruption of the intestinal epithelial barrier and gut vascular barrier(GVB),and suppressed he-patic LPS-NF-κB pathway activation in RTS-induced HSOS.Importantly,the LPS level was positively correlated with MELD score in patients with HSOS.Elevated LPS in patients with HSOS confirmed that Gram-negative bacteria were in-volved in the pathogenesis of HSOS.Conclusions:RTS,a PA,cooperated with gut dysbiosis to cause intestinal inflam-mation and gut barrier compromise that increased transport of gut-derived LPS into the liver through the portal vein,which contributed to the pathology of HSOS.Modulating the gut microbiota,protecting the intestinal barrier,and sup-pressing intestinal inflammation with prebiotics or antibiot-ics might be a useful pharmacologic intervention in HSOS.

Predictive Model of Ursodeoxycholic Acid Treatment Response in Primary Biliary Cholangitis

Background and Aims:Although ursodeoxycholic acid(UDCA)treatment in primary biliary cholangitis is effective in many patients,there are still many people who respond poorly to it.Identifying and intervening these patients early is important.Therefore,exploring the risk factors and proposing a predictor index to predict the UDCA treatment nonresponse earlier among primary biliary cholangitis patients were the aims of this research.Methods:A total of 135 primary biliary cholangitis patients treated with UDCA(13–15 mg/kg/d)were enrolled in this retrospective study.The response to treatment was evaluated based on Paris I criteria.The univariate and logistic multivariate regression analyses were adopted to determine the independent risk factors and propose a predictor index.Receiver operating characteristic curve was used to evaluate the predictive ability of the predictor index.Results:Total bilirubin,albumin,globulin,immunoglobin M,and aspartate aminotransferase-to-platelet ratio index were the five independent risk factors associating with early biochemical nonresponse to UDCA treatment.Based on these factors,we established a predictor index with the predictive value being 0.886(sensitivity:82.80%,specificity:84.40%).Conclusions:We developed a predictor index that had an accurate prediction of the early biochemical nonresponse to UDCA treatment,which is expected to provide valuable information for the high-risk group before treatment begins.

Construction and expression of hepatitis B virus vector encoding TC-tagged core protein

Virus tagged with greenfluorescent protein(GFP)contributes to the visualization and study of the virus in living cells.However,the hepatitis B virus(HBV)particle,which is a compact virion with limited internal space,cannot be incorporated with GFP tag as a large fragment.It was recently reported that protein genetically inserted with a smaller size tetracysteine(TC)tag could be specially labeled by a biarsenicalfluorescent dye in living cells.In this study,we constructed a recombinant HBV vector encoding TC-tagged core protein for biarsenical labeling of HBV virion.TC tag was genetically inserted near the immunodominant c/e1 site of HBV core protein by mutagenesis.Western blot and enzyme-linked immu-nosorbent assay(ELISA)analysis showed that the TC-tagged core protein,hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)could be expressed in cells transfected with the recombinant HBV vector,which is similar to the cells transfected with wild-type HBV vector.Reverse transcription-polymerase chain reaction(RT-PCR)and Southern blot analysis showed that HBV virion formation was affected by the genetic insertion of TC tag into core protein in some degree,but cells transfected with the HBV vector could still produce HBV virions incorporated with TC-tagged core proteins.Taken together,the recombinant HBV vector can serve as a useful tool to produce HBV virions incorporated with TC-tagged core proteins to befluorescently labeled by biarsencial dye for visualizing and studying HBV in living cells.

Cloning of human XAF1 gene promoter and assay of its transcription activity in a variety of cell lines

To investigate the regulation of tumor sup-pressor XAF1 gene expression in digestive system cancers,we studied XAF1 gene promoter transcription activity and mRNA level in digestive system cancer cell lines(human hepatoma cell line HepG2,human colon cancer cell line LoVo,and human gastric cancer cell line AGS)and nontumor cell lines(human embryonic liver cell line L02(L02 cells)and human embryonic kidney 293 cells[HEK293 cells])as controls.1395-bp-promoter fragment of XAF1 gene was amplified by polymerase chain reaction(PCR)and cloned into pGL3-basic vector and pEGFP-1 vector to assay its promoter transcription activity.The plasmids were transfected into a variety of cell lines by lipofectamine 2000.The promoter transcription activity was determined by dual-luciferase report assay,and enhanced greenfluorescent protein(EGFP)-positive cells were detected byfluorescence microscope.The expression of XAF1 mRNA in HEK293 and L02 were significantly higher than that in any of the three digestive system cancer cell lines.The dual-luciferase reporter assay showed that the promoter transcription activity in digestive system tumor cell lines transfected with pGL3-XAF1p promoter was apparently lower than that of both HEK293 and L02 cells.Expression of greenfluorescent protein(GFP)under the control of XAF1 promoter in the three digestive system cancer cell lines was lower than that of both HEK293 and L02 cells.The activities of pGL3-XAF1p in the three digestive system cancer cell lines after treatment with heat stress were significantly lower than those in the unstressed cells.The results suggested that remarkably down-regulated XAF1 mRNA expression in digestive system cancer cell lines may be due to loss of transcription activity of XAF1 promoter.