Thus, whilst useful in terms of generating hypotheses, the results of this more recent review are also inconclusive with respect to confirming the relative immunogenicity between plasma-derived find more and recombinant FVIII products.

Early in 2013, a study by the European Pediatric Network for Hemophilia Management (PedNet) and Research of Determinants of Inhibitor Development (RODIN) was published [14]. This multicentre, observational (mainly prospective, partially retrospective) cohort study evaluated 574 consecutive patients with severe haemophilia A (FVIII activity < 0.01 U mL−1) born between 1 January 2000 and 1 January 2010, making it the largest study to date in PUPs at high risk of developing inhibitors. Over a total of 29 679 exposure days, inhibitory antibodies developed in 177 of the children for a cumulative

incidence of 32.4%. Interestingly, and in contrast to the systematic reviews discussed above [7, 13], no difference was found in the risk of inhibitor development between plasma-derived and recombinant products (adjusted hazard ratio 0.96 [95% CI 0.62–1.49]). Although the study provided useful new evidence in relation to the use of recombinant products, it also has some important limitations. Patients were not assigned randomly to FVIII products and there was no apparent use of scores to account for differences in the propensity of various check details products to be associated during with inhibitor development [15]. The number of children treated with pdFVIII was small, involving only 4018 exposure days compared with 25 661 exposure days for patients treated

with rFVIII. Many patients treated with plasmatic FVIII received monoclonal products that contain only traces of VWF, but results were combined with those of patients receiving VWF-rich products. Finally, as with all studies, there is risk of a type 2 error, i.e. finding no difference when a difference does in fact exist. As is the case with the systematic reviews, the results of this study cannot be considered conclusive and the ‘jury is still out’ with regard to the relative risk of inhibitor development in PUPs treated with plasma-derived or recombinant FVIII concentrates. Switching among FVIII products has also been suggested as a risk factor for inhibitor development. In their cohort of PUPs, the PedNet/RODIN investigators found no increased risk of inhibitor development in patients who switched vs. those who did not switch FVIII products (adjusted hazard ratio 0.99 [95% CI 0.63–1.56]) [14]. The haemophiliac population also contains a small group of previously treated patients (PTPs) who initially respond to FVIII but develop inhibitors upon further exposure. Inhibitor development in PTPs treated with any type of FVIII concentrate was investigated in a systematic review of 33 independent cohorts of PTPs [16].

Thus, whilst useful in terms of generating hypotheses, the results of this more recent review are also inconclusive with respect to confirming the relative immunogenicity between plasma-derived RG7204 ic50 and recombinant FVIII products.

Early in 2013, a study by the European Pediatric Network for Hemophilia Management (PedNet) and Research of Determinants of Inhibitor Development (RODIN) was published [14]. This multicentre, observational (mainly prospective, partially retrospective) cohort study evaluated 574 consecutive patients with severe haemophilia A (FVIII activity < 0.01 U mL−1) born between 1 January 2000 and 1 January 2010, making it the largest study to date in PUPs at high risk of developing inhibitors. Over a total of 29 679 exposure days, inhibitory antibodies developed in 177 of the children for a cumulative

incidence of 32.4%. Interestingly, and in contrast to the systematic reviews discussed above [7, 13], no difference was found in the risk of inhibitor development between plasma-derived and recombinant products (adjusted hazard ratio 0.96 [95% CI 0.62–1.49]). Although the study provided useful new evidence in relation to the use of recombinant products, it also has some important limitations. Patients were not assigned randomly to FVIII products and there was no apparent use of scores to account for differences in the propensity of various Birinapant chemical structure products to be associated Farnesyltransferase with inhibitor development [15]. The number of children treated with pdFVIII was small, involving only 4018 exposure days compared with 25 661 exposure days for patients treated

with rFVIII. Many patients treated with plasmatic FVIII received monoclonal products that contain only traces of VWF, but results were combined with those of patients receiving VWF-rich products. Finally, as with all studies, there is risk of a type 2 error, i.e. finding no difference when a difference does in fact exist. As is the case with the systematic reviews, the results of this study cannot be considered conclusive and the ‘jury is still out’ with regard to the relative risk of inhibitor development in PUPs treated with plasma-derived or recombinant FVIII concentrates. Switching among FVIII products has also been suggested as a risk factor for inhibitor development. In their cohort of PUPs, the PedNet/RODIN investigators found no increased risk of inhibitor development in patients who switched vs. those who did not switch FVIII products (adjusted hazard ratio 0.99 [95% CI 0.63–1.56]) [14]. The haemophiliac population also contains a small group of previously treated patients (PTPs) who initially respond to FVIII but develop inhibitors upon further exposure. Inhibitor development in PTPs treated with any type of FVIII concentrate was investigated in a systematic review of 33 independent cohorts of PTPs [16].

[237, 238] 52. Referral for LT evaluation should be considered for

CNI patients before the development of brain damage, ideally at the time of diagnosis when the option of LT can be discussed. PLX-4720 mouse (1A) Autoimmune hepatitis (AIH) is a progressive inflammatory liver disorder characterized by increased aminotransferases, high serum levels of immunoglobulin G (IgG), and the presence of autoantibodies: antinuclear antibody (ANA), antismooth muscle antibody (ASMA), antiliver-kidney microsomal antibody (anti-LKM), with a potentially more aggressive course in children.[239] Type 1, characterized by positive ANA and/or ASMA, is more common,[240] although Type 2, characterized by a positive anti-LKM, is more frequently associated with fulminant liver failure.[240] In a study of 55 consecutive children with clinical and biochemical evidence of AIH, 27/55 (50%) had cholangiographic findings consistent with autoimmune sclerosing cholangitis (ASC).[240] ASC subsequently developed in a patient with AIH and ulcerative colitis. Conventional treatment includes prednisone with or without azathioprine for both AIH and AIH/ASC; ursodeoxycholic PLX3397 acid may be helpful for those with AIH/ASC.[241] LT is required in 10%-20% of children with AIH.[239] Despite a greater degree of immunosuppression required in the posttransplant period, outcomes are similar to the overall transplanted population

in terms of infectious or metabolic complications. The risk of late rejection is higher for those who receive LT for AIH, but this does not result in increased chronic rejection, steroid resistant rejection, or the need for retransplantation,[242] which differs from adults.[243] Pediatric patients transplanted for AIH may be at greater risk of developing ulcerative colitis after LT than adult patients.[244] The risk of relapse of AIH posttransplant is estimated to be 10%-35%[19, 245, 246]; however, criteria for recurrent AIH remain controversial. 53. LT is considered in patients with autoimmune hepatitis (AIH) who present Thalidomide with acute liver failure associated with encephalopathy and those who

develop complications of endstage liver disease not salvageable with medical therapy (2-B). 54. Children with AIH and families being evaluated for LT should be informed they may require more immunosuppression than children transplanted for other indications and remain at risk for recurrence of AIH. (2-B) Primary sclerosing cholangitis (PSC) is characterized by chronic inflammation and obliterative fibrosis of the intra- and/or extrahepatic biliary tree, leading to bile stasis and cirrhosis.[240, 241, 247] Children with biliary features consistent with PSC can have isolated biliary tract disease or have histologic characteristics may present prior to, coincident with, or subsequent to histological and biochemical features of autoimmune hepatitis (AIH) type 1.

Twenty-five healthy mountaineers were studied. Blood samples and duodenal biopsies were taken at baseline of 446 m as well as on day 2 (MG2) and 4 (MG4) after rapid ascent to 4559 m. Divalent metal-ion transporter 1 (DMT-1), ferroportin 1 (FP-1) messenger RNA (mRNA), and protein expression were analyzed in biopsy specimens by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. Serum hepcidin levels were analyzed by mass spectrometry. Serum iron,

Selleck Neratinib ferritin, transferrin, interleukin (IL)−6, and C-reactive protein (CRP) were quantified by standard techniques. Serum erythropoietin and growth differentiation factor 15 (GDF15) levels were measured by enzyme-linked immunosorbent assay (ELISA). Under hypoxia, erythropoietin peaked at MG2 (P < 0.001) paralleled by increased GDF15 on MG2 (P < 0.001). Serum iron and ferritin levels declined rapidly on MG2 and MG4 (P < 0.001). Duodenal DMT-1 and FP-1 mRNA expression increased up to 10-fold from baseline on MG2 and

MG4 (P < 0.001). Akt inhibitor Plasma CRP increased on MG2 and MG4, while IL-6 only increased on MG2 (P < 0.001). Serum hepcidin levels decreased at high altitude on MG2 and MG4 (P < 0.001). Conclusion: This study in healthy volunteers showed that under hypoxemic conditions hepcidin is repressed and duodenal iron transport is rapidly up-regulated. These changes may increase dietary iron uptake and allow release of stored iron to ensure a sufficient

iron supply for hypoxia-induced compensatory erythropoiesis. (Hepatology 2013; 58:2153–2162) Iron is an essential trace element required as a component of various molecules that sense, transport, and store oxygen.[1] Availability of sufficient Montelukast Sodium amounts of iron is critically important for normal and stress-induced erythropoiesis. Circulating iron levels are affected by intestinal absorption from the diet, iron transport capacity of the blood, iron losses via bleeding and cellular desquamation, and the release of iron from cells such as macrophages and hepatocytes.[2] Inorganic iron is absorbed at the brush border of duodenal enterocytes by the divalent metal-ion transporter 1 (DMT-1; SLC11A2) following reduction by a membrane-associated ferrireductase. Cytosolic iron can be exported by the basolateral iron transporter ferroportin (FP-1; SLC40A1)[3, 4] and subsequently undergoes oxidation by the multicopperoxidase hephaestin before being incorporated into circulating transferrin. Systemic iron content is tightly regulated,[1, 4, 5] because accumulation of intracellular iron causes cell and tissue damage, presumably by iron-catalyzed generation of reactive oxygen species.[5, 6] Hepcidin, a liver-derived 25 amino acid peptide hormone, has been identified as the key regulator of iron homeostasis[7, 8] (reviewed[6, 9]).

In contrast to controls, showing collecting DAPT ducts with continuous basement membrane (Fig. 4A), kidneys of 3-day CBDL mice showed severely altered collecting ducts, with ulceration of the epithelium and exfoliation of epithelial cells coalescing to cell casts within the lumens of collecting ducts, and frequent loss of basement membrane continuity corresponding to these areas (Fig.

4B). The functional relevance of these findings was demonstrated by leakage of portal-vein–injected and urinary-excreted UDCA/NBD/lysine in CBDL mice from the lumens of collecting ducts (Fig. 5), whereas sham-operated controls stained almost negative (not shown). In line and principally reflecting BA leakage in a well-established experimental condition, hepatic bile infarcts of the same CBDL mice stained positive with www.selleckchem.com/products/pexidartinib-plx3397.html fluorescent UDC/NBD/lysine (Supporting Fig. 3). Taken together, these findings demonstrate that collecting ducts represent early targets in cholemic nephropathy in CBDL mice and suggest that a potential tubulotoxic agent with the presumably highest local concentration or toxicity at the level of collecting ducts may lead to epithelial cell injury and basement membrane damage already as early as 3 days after CBDL. Because inflammation is a well-known trigger for renal fibrosis and subgroups of peripheral monocytes may also significantly contribute

to kidney fibrosis, we next tested renal VCAM-1 and macrophage/dendritic cell marker F4/80

expression. Renal VCAM-1 and F4/80 protein expression was induced over time. Because sham-operated controls did not Dichloromethane dehalogenase significantly differ over the various time points studied, only 8-week sham-operated animals are shown. VCAM-1 expression was primarily induced in tubular epithelial cells and, to a lesser degree, in endothelial cells as well as the interstitium increasing over time (Supporting Fig. 4A), accompanied by significant increased levels of VCAM-1 messenger RNA (mRNA) and protein expression (Supporting Fig. 4B,C). In contrast, F4/80 expression was primarily induced in cells of the renal interstitium and within glomeruli (Fig. 6A). This renal inflammatory response in CBDL mice was accompanied by pronounced overexpression of F4/80 and monocyte chemoattractant protein 1 (Mcp-1) mRNA already at week 3 after CBDL (Figs. 6B,C). To closely follow up the development of kidney fibrosis in CBDL mice, we compared renal collagen α1(I)) and transforming growth factor beta 1 (tgf-β1) mRNA, hydroxyproline levels, and Sirius Red–stained kidney sections at several time points after CBDL to controls. Significant renal fibrosis was already developed 3 weeks after CBDL and increased later (Fig. 7A). This was accompanied by induction of collagen α1(I) and tgf-β1 mRNA expression (Fig. 7B) and significantly elevated renal hydroxyproline levels (Fig. 7C). PAS staining after 3-day CBDL revealed tubular epithelial injury in both genotypes (Fig. 8A).

In contrast to controls, showing collecting Rapamycin mw ducts with continuous basement membrane (Fig. 4A), kidneys of 3-day CBDL mice showed severely altered collecting ducts, with ulceration of the epithelium and exfoliation of epithelial cells coalescing to cell casts within the lumens of collecting ducts, and frequent loss of basement membrane continuity corresponding to these areas (Fig.

4B). The functional relevance of these findings was demonstrated by leakage of portal-vein–injected and urinary-excreted UDCA/NBD/lysine in CBDL mice from the lumens of collecting ducts (Fig. 5), whereas sham-operated controls stained almost negative (not shown). In line and principally reflecting BA leakage in a well-established experimental condition, hepatic bile infarcts of the same CBDL mice stained positive with BGJ398 nmr fluorescent UDC/NBD/lysine (Supporting Fig. 3). Taken together, these findings demonstrate that collecting ducts represent early targets in cholemic nephropathy in CBDL mice and suggest that a potential tubulotoxic agent with the presumably highest local concentration or toxicity at the level of collecting ducts may lead to epithelial cell injury and basement membrane damage already as early as 3 days after CBDL. Because inflammation is a well-known trigger for renal fibrosis and subgroups of peripheral monocytes may also significantly contribute

to kidney fibrosis, we next tested renal VCAM-1 and macrophage/dendritic cell marker F4/80

expression. Renal VCAM-1 and F4/80 protein expression was induced over time. Because sham-operated controls did not ADAM7 significantly differ over the various time points studied, only 8-week sham-operated animals are shown. VCAM-1 expression was primarily induced in tubular epithelial cells and, to a lesser degree, in endothelial cells as well as the interstitium increasing over time (Supporting Fig. 4A), accompanied by significant increased levels of VCAM-1 messenger RNA (mRNA) and protein expression (Supporting Fig. 4B,C). In contrast, F4/80 expression was primarily induced in cells of the renal interstitium and within glomeruli (Fig. 6A). This renal inflammatory response in CBDL mice was accompanied by pronounced overexpression of F4/80 and monocyte chemoattractant protein 1 (Mcp-1) mRNA already at week 3 after CBDL (Figs. 6B,C). To closely follow up the development of kidney fibrosis in CBDL mice, we compared renal collagen α1(I)) and transforming growth factor beta 1 (tgf-β1) mRNA, hydroxyproline levels, and Sirius Red–stained kidney sections at several time points after CBDL to controls. Significant renal fibrosis was already developed 3 weeks after CBDL and increased later (Fig. 7A). This was accompanied by induction of collagen α1(I) and tgf-β1 mRNA expression (Fig. 7B) and significantly elevated renal hydroxyproline levels (Fig. 7C). PAS staining after 3-day CBDL revealed tubular epithelial injury in both genotypes (Fig. 8A).

In contrast to controls, showing collecting BMS-907351 chemical structure ducts with continuous basement membrane (Fig. 4A), kidneys of 3-day CBDL mice showed severely altered collecting ducts, with ulceration of the epithelium and exfoliation of epithelial cells coalescing to cell casts within the lumens of collecting ducts, and frequent loss of basement membrane continuity corresponding to these areas (Fig.

4B). The functional relevance of these findings was demonstrated by leakage of portal-vein–injected and urinary-excreted UDCA/NBD/lysine in CBDL mice from the lumens of collecting ducts (Fig. 5), whereas sham-operated controls stained almost negative (not shown). In line and principally reflecting BA leakage in a well-established experimental condition, hepatic bile infarcts of the same CBDL mice stained positive with Trichostatin A clinical trial fluorescent UDC/NBD/lysine (Supporting Fig. 3). Taken together, these findings demonstrate that collecting ducts represent early targets in cholemic nephropathy in CBDL mice and suggest that a potential tubulotoxic agent with the presumably highest local concentration or toxicity at the level of collecting ducts may lead to epithelial cell injury and basement membrane damage already as early as 3 days after CBDL. Because inflammation is a well-known trigger for renal fibrosis and subgroups of peripheral monocytes may also significantly contribute

to kidney fibrosis, we next tested renal VCAM-1 and macrophage/dendritic cell marker F4/80

expression. Renal VCAM-1 and F4/80 protein expression was induced over time. Because sham-operated controls did not Mannose-binding protein-associated serine protease significantly differ over the various time points studied, only 8-week sham-operated animals are shown. VCAM-1 expression was primarily induced in tubular epithelial cells and, to a lesser degree, in endothelial cells as well as the interstitium increasing over time (Supporting Fig. 4A), accompanied by significant increased levels of VCAM-1 messenger RNA (mRNA) and protein expression (Supporting Fig. 4B,C). In contrast, F4/80 expression was primarily induced in cells of the renal interstitium and within glomeruli (Fig. 6A). This renal inflammatory response in CBDL mice was accompanied by pronounced overexpression of F4/80 and monocyte chemoattractant protein 1 (Mcp-1) mRNA already at week 3 after CBDL (Figs. 6B,C). To closely follow up the development of kidney fibrosis in CBDL mice, we compared renal collagen α1(I)) and transforming growth factor beta 1 (tgf-β1) mRNA, hydroxyproline levels, and Sirius Red–stained kidney sections at several time points after CBDL to controls. Significant renal fibrosis was already developed 3 weeks after CBDL and increased later (Fig. 7A). This was accompanied by induction of collagen α1(I) and tgf-β1 mRNA expression (Fig. 7B) and significantly elevated renal hydroxyproline levels (Fig. 7C). PAS staining after 3-day CBDL revealed tubular epithelial injury in both genotypes (Fig. 8A).

Compared with IFX, ADA tended to have longer time to reinduction (median 22 vs 37 months respectively, p = 0.07) and longer time from reinduction to objective reinduction failure (median 11 vs 21, p = 0.60), albeit the latter was non-significant. Post-reinduction, factors associated at least 12 months continuation on same anti-TNF included, for both ADA and IFX, no current IBD extra-intestinal

manifestation(s) OR 10.5, 95% CI [1.5,71.4](p = 0.01), no psychological comorbidity OR 6.7 [1.05, 42.4] (each p < 0.05), female sex OR 3.9 [0.8, 21.3](p = 0.1). Again, female sex was associated with continuation at 12 months post-induction for ADA (Fisher's exact, p < 0.04) but not for IFX (p = 0.65). Conversely, concurrent thiopurine was associated with continuation (at 12 m) for IFX (p < 0.05) but not for ADA (p = 0.3) YAP-TEAD Inhibitor 1 clinical trial and there was a trend for prior bowel resection to be associated with continuation for IFX but no ADA (p = 0.06). Conclusions: Overall reinduction was very effective in regaining response to anti-TNF in many patients, and should be considered in patients as the next step where concomitant immunomodulators have already been added. Smoking rates are high in this cohort, implying its important role

in secondary LOR. Pharmacokinetic AZD0530 mw differences between ADA and IFX, with the potentially greater immunogenic potential of IFX, may explain why concurrent PLEK2 thiopurines were linked with more durable outcomes in IFX post-reinduction (but not ADA), and the differential response post-reinduction for ADA

between males and females. Further larger studies are needed in this area to best utilize anti-TNF reinduction to achieve optimal outcomes in the Australian PBS setting in CD. M OOI, J PANETTA, M ZHU, C CORTE, RWL LEONG Gastroenterology and Liver Services, Concord Hospital, Sydney Australia Background: Traditional forward-viewing colonoscopy (FVC) is impeded by a narrow field of view of <170 degrees with adenoma “miss-rates” of 24–42%. This limitation impairs dysplasia surveillance in chronic colitis. Chromoendoscopy improves visualization of dysplasia but has never been used with FUSE, which provides 330 degree visualization. This study compares FVC and FUSE with and without chromoendoscopy for the identification of dysplasia in IBD. Methods: This was a prospective, randomized-order, crossover tandem colonoscopy trial at an academic IBD center. Inclusion criteria were left sided or extensive colitis for >10 years disease duration, primary sclerosing cholangitis (PSC) or previous dysplasia. Patients underwent same-day, back-to-back tandem colonoscopy with FVC (PCF/CF 180/190, Olympus, Japan) and the FUSE colonoscope (EndoChoice, USA) under propofol sedation. Order randomization was computer-generated. Patients and endoscopist were masked to group allocation until immediately prior to colonoscopy.

miRNA profiles of these samples brought two important insights to the understanding of hepatocarcinogenesis. First, we found that

miRNA deregulation is an early event in which discernible difference was observed between miRNA profiles of primary HCCs and nontumorous liver through clustering analysis. However, no major change was observed between the miRNA profiles of primary HCCs and venous metastases. Second, unlike other cancers, no global miRNA down-regulation was detected Y27632 in primary HCCs. Instead, marked global miRNA down-regulation was detected in venous metastases, and this global miRNA down-regulation could exacerbate the preexisting miRNA deregulation in primary HCCs and facilitate metastasis formation. HBV, hepatitis B virus; HCC, hepatocellular carcinoma; miRNA, LY2157299 price microRNA; PCR, polymerase chain reaction; snoRNA, small nucleolar RNA; TLDA, TaqMan Low-Density Array. Twenty advanced HCC cases with intrahepatic metastasis as diagnosed with the presence of venous thrombi were selected from a large cohort of approximately 400 HCC patients who underwent curative surgical resection at Queen Mary Hospital, Hong Kong, between 1999 and 2009. Formalin-fixed and paraffin-embedded sections were retrieved from these cases, and the presence of venous thrombi was reviewed by an experienced liver pathologist (IOLN). In our

center, HCCs with gross tumor thrombosis in the portal vein are often inoperable. We therefore selected only those cases in which the venous thrombi were large selleck products enough for microdissection. Twenty HCC cases with medium-sized (3-10 mm) HCC thrombi were selected for this study. All patients were of Chinese origin, with a mean age of 51.5 years; 19 were male and one was female. Eighteen patients had chronic hepatitis B virus (HBV) infection, as shown by the positive serum HBV surface antigen status, and the remaining two patients had chronic hepatitis C viral (HCV) infection, as shown by the positive serum anti-HCV status. There were no patients with both HBV and HCV infection. Liver cirrhosis

was present in 13 patients. The demographic data and clinicopathological features of the patients are described in Supporting Table 1. Use of human samples was approved by the Institutional Review Board of the University of Hong Kong/Hospital Authority Hong Kong West Cluster. Paraffin sections were cut at a thickness of 4 μm, dewaxed, rehydrated, and lightly stained with hematoxylin. Nontumorous livers and their corresponding primary HCC tissues and venous thrombi were examined and microdissected with a 25-gauge needle under a dissecting microscope as described. 11, 12 Four to eight consecutive tissue sections were cut in each case to obtain enough microdissection material for further evaluation. miRNA extraction was performed with an miRNeasy FFPE kit (Qiagen, Valencia, CA).

miRNA profiles of these samples brought two important insights to the understanding of hepatocarcinogenesis. First, we found that

miRNA deregulation is an early event in which discernible difference was observed between miRNA profiles of primary HCCs and nontumorous liver through clustering analysis. However, no major change was observed between the miRNA profiles of primary HCCs and venous metastases. Second, unlike other cancers, no global miRNA down-regulation was detected click here in primary HCCs. Instead, marked global miRNA down-regulation was detected in venous metastases, and this global miRNA down-regulation could exacerbate the preexisting miRNA deregulation in primary HCCs and facilitate metastasis formation. HBV, hepatitis B virus; HCC, hepatocellular carcinoma; miRNA, Pexidartinib microRNA; PCR, polymerase chain reaction; snoRNA, small nucleolar RNA; TLDA, TaqMan Low-Density Array. Twenty advanced HCC cases with intrahepatic metastasis as diagnosed with the presence of venous thrombi were selected from a large cohort of approximately 400 HCC patients who underwent curative surgical resection at Queen Mary Hospital, Hong Kong, between 1999 and 2009. Formalin-fixed and paraffin-embedded sections were retrieved from these cases, and the presence of venous thrombi was reviewed by an experienced liver pathologist (IOLN). In our

center, HCCs with gross tumor thrombosis in the portal vein are often inoperable. We therefore selected only those cases in which the venous thrombi were large Staurosporine solubility dmso enough for microdissection. Twenty HCC cases with medium-sized (3-10 mm) HCC thrombi were selected for this study. All patients were of Chinese origin, with a mean age of 51.5 years; 19 were male and one was female. Eighteen patients had chronic hepatitis B virus (HBV) infection, as shown by the positive serum HBV surface antigen status, and the remaining two patients had chronic hepatitis C viral (HCV) infection, as shown by the positive serum anti-HCV status. There were no patients with both HBV and HCV infection. Liver cirrhosis

was present in 13 patients. The demographic data and clinicopathological features of the patients are described in Supporting Table 1. Use of human samples was approved by the Institutional Review Board of the University of Hong Kong/Hospital Authority Hong Kong West Cluster. Paraffin sections were cut at a thickness of 4 μm, dewaxed, rehydrated, and lightly stained with hematoxylin. Nontumorous livers and their corresponding primary HCC tissues and venous thrombi were examined and microdissected with a 25-gauge needle under a dissecting microscope as described. 11, 12 Four to eight consecutive tissue sections were cut in each case to obtain enough microdissection material for further evaluation. miRNA extraction was performed with an miRNeasy FFPE kit (Qiagen, Valencia, CA).