Methods: We investigate 72 patients who were diagnosed with recur

Methods: We investigate 72 patients who were diagnosed with recurred or metastatic gastric adenocarcinoma in a single center (Chungnam National University Hospital) during March, 2008 to July, 2012. The patient received either FOLFOX or FOLFIRI chemotherapy. Results: There were no significant difference between FOLFOX (42 patients) and FOLFIRI (30 patients) group in sex, age, prior surgery, histology and ECOG performance (P > 0.05). FOLFOX group showed response rate 52.4% (CR 21.4% and PR 31%) and

FOLFIRI group showed response rate 46.6% (CR 3.3%, PR 43.3%), but response rate showed no significant difference (P = 0.171). The Time to progression was longer among patients treated with FOLFOX (median, 8.58 month) than among those with FOLFIRI (median, 5.0 month), and this difference statistically

significant (P = 0.032). The overall survival showed no significant difference (P = 0.094), with the oxaliplatin group (28.96 month) Pictilisib in vitro being slightly longer than the irinotecan group (16.48 month). Grade 3/4 Hematologic toxicity (Neutropenia, Anemia and Thrombocytopenia) occurred similarly in both groups. Conclusion: Both combination therapy can be used effectively in recurred or metastatic gastric adenocarcinoma and there were no significant difference between FOLFOX and FOLFIRI in response rate and overall survival. Key Word(s): 1. stomach neoplasm; 2. chemotherapy Presenting Author: KUAN SIANG TAN Additional Authors: TAUFIQUE AHMED Corresponding Author: KUAN SIANG TAN Affiliations: Khoo Teck Puat Hospital Objective: To explore predictive factors associated with diagnosis of lesions, defined as ulcers and carcinomas on endoscopy. Methods: Clinicopathological data of 133 inpatients that underwent endoscopy for investigation of anaemia between October 2013 and Janurary 2014 were analyzed retrospectively. Patients were separated into two groups; patients who had endoscopic and /or histological findings of ulcers and carcinomas constitute the group with lesions and patients without CYTH4 lesions

constitute a control group. Patients were scored for each of the associated factors of anaemia including mean corpuscular volume (MCV), ferritin, iron saturation, vitamin B12 levels (B12), folate, presence of end stage renal failure (ESRF) and faecal occult blood (FOB) and a total score was computed for each patient. Univariate analysis was performed to analyze the above individual factors and the total scores for each group of patients. Results: There were 35 patients with lesions and 98 patients without lesions. Univaried analysis showed a high total score is suggestive of a lesion (OR = 1.218, P = 0.024), with low MCV (OR = 2.428, P = 0.021) and positive FOB (OR = 1.826, P = 0.027) individual predictors of a lesion.

Both receptors were absent in hepatocytes but were expressed by b

Both receptors were absent in hepatocytes but were expressed by bile ducts and ductules when present (e.g., ductules in FNH and I-HCA). The most obvious expression of VEGFR-1 and VEGFR-2 was present in sinusoidal macrophages, SECs, and VECs. Stromal cells and macrophages in fibrous scars and septa of FNH also expressed both receptors (Fig. 4). The patterns observed in the different tissues are summarized in Table 2. FNH and HCA are two nodular hepatic lesions of different etiological backgrounds. HCA is a benign, neoplastic lesion of several mutational backgrounds, whereas FNH is thought H 89 datasheet to represent a hyperplastic reaction following a vascular injury.3, 5 FNH and HCA contain various

morphological vascular abnormalities, the pathogenesis of which is not clear. Some vascular features are shared by the two lesions, AZD2014 concentration whereas some are lesionally restricted. Studies in transgenic mice have shown that overexpression of the angiogenic growth factor

Ang-1 results in hepatic vascular anomalies and generates hepatocellular nodules similar to FNH.14, 15 We hypothesized that the various abnormal vascular features prominent in human FNH and HCA are related to increased vascular remodeling with a central role for the angiopoietin system. To test this hypothesis, we investigated the gene and protein expression pattern of growth factors belonging to the angiopoietin system: Ang-1, Ang-2, and their cognate receptor Tie-2. VEGF-A and its receptors VEGFR-1 and VEGFR-2 were included in the analysis as these factors are known to act in concert with the angiopoietins.18 We observed a significant increase of Ang-1 in FNH and to a lesser extent in HCA in comparison with histologically normal livers, with a concurrent

increase in the Ang-1/Ang-2 ratio. In FNH, this increase existed next to a significant increase in Tie-2 expression. In contrast, changes in VEGF-A and VEGFR expression were not prominent in either type of lesion. Our results support the concept, schematically CYTH4 depicted in Fig. 5, that in FNH and HCA, the Ang-1/Tie-2 system may have a regulatory role in the development of the characteristic vascular features of these lesions without signs of robust involvement of the VEGF system. Studies addressing the molecular background of vascular remodeling in FNH and HCA are rare. Paradis et al.6 investigated 209 genes in FNH and were the first to report that Ang-1 gene expression was enhanced in FNH, whereas Ang-2 was decreased, but without a concurrent increase in Tie-2 as we observed. The same group also studied telangiectatic FNH and postulated that this FNH subtype resembles HCA more than it resembles FNH on the basis of the expression patterns of Ang-1 and Ang-2.7 In a similar pursuit to classify telangiectatic FNH, Bioulac-Sage et al.19 confirmed these results. In these two studies, the telangiectatic FNHs were monoclonal lesions, and this supports the concept that they represent an HCA subtype.

8A) This could be related to the cell cycle alterations observed

8A). This could be related to the cell cycle alterations observed upon p38α down-regulation. The mitotic delay could be reflected in an increase in polyploid nuclei (nonmitotic nuclei division) or binucleated cells (nonmitotic cytoplasm division). By identifying nuclei with 4,6-diamidino-2-phenylindole (DAPI) and cellular membranes with anti-β-catenin immunostaining, we found that p38α KO mice exhibited a higher rate of binucleated hepatocytes than WT mice both before and after Selleck AZD1152 HQPA BDL, whereas no differences were observed between BDL and sham groups in p38α KO animals (Fig. 8). In contrast, the percentage of binucleated

cells in WT sham livers was markedly reduced when they start proliferating after induction of cholestasis.

The two major groups of proteins that are regulated by p38 MAPK-mediated phosphorylation are protein kinases, such as MK2, and transcription factors, such as p53.1 p38α may negatively regulate AKT activity independently of PI3K, by regulating its interaction with PP2A or through the activation of MK2 (Fig. 2A). Indeed, MK2 mediates HSP27-dependent selleck activation of AKT by way of phosphorylation on Ser473.13 Accordingly, phosphorylation of MK2 on Thr334 and of AKT on Ser473 were markedly reduced in liver of p38α-deficient mice upon chronic cholestasis (Figs. 2B, 3A). Activation of AKT triggers a key antiapoptotic Alanine-glyoxylate transaminase signaling pathway in the liver. However, in our model of chronic cholestasis the lack of AKT activation did not lead to increased apoptosis (Supporting Fig. S5).

PDK1 and AKT are required for normal cell growth and liver regeneration after partial hepatectomy.17 Taking into account the absence of significant PDK1-mediated AKT phosphorylation on Thr308 upon chronic cholestasis (Fig. 3A), it seems that p38α/MK2-dependent AKT activation is essential for liver regeneration and hepatomegaly in this chronic disorder. In vitro in rat hepatoma cells, AKT activation increased cell size through mTOR-dependent and mTOR-independent pathways and the latter also involved inhibition of protein degradation.18 Accordingly, p38α deficiency may reduce hepatocyte growth during chronic cholestasis through down-regulation of both AKT and mTOR (Fig. 3A). Similar to yeast, in mammals two distinct protein kinase mTOR complexes have been characterized. mTORC1 is rapamycin-sensitive and controls protein synthesis, whereas mTORC2 is rapamycin-insensitive and controls the actin cytoskeleton.19 Hence, down-regulation of mTOR may contribute to reduce albumin levels in liver of p38α-deficient mice. The Akt/mTOR pathway may lead to activation of the p70 S6 kinase/S6 pathway. Our findings suggest that blockade of this pathway seems to be involved in the lack of hepatocyte proliferation and growth that occurs upon p38α deficiency during chronic cholestasis.

1A), suggesting that HBV+ mice were systemically immunotolerant t

1A), suggesting that HBV+ mice were systemically immunotolerant to HBV. Similar to infected human hepatocytes and liver tissues, IFN-α/β mRNA levels were lower in HBV+ than in HBV− hepatocytes (Fig. 1B), while immunosuppressive Tanespimycin cost cytokines significantly increased (Fig. 1C). These results collectively indicate that HBV infection induces hepatocyte-intrinsic innate immunotolerance. Evaluating adaptive immunity generated in HBV+ mice, we found that the percentage and absolute number of hepatic CD8+ T cell (Fig. 1D) was reduced, and moreover, inhibitory PD-1 expression on hepatic CD8+ T cells was almost 3-fold higher than in HBV− mice (Fig. 1E). To observe recall responses and to determine if HBV

persistence was established in HBV+ mice, pAAV/HBV1.2 plasmid was readministered. Two weeks later, HBV− mice eliminated HBV, but HBV+ mice remained HBV persistent (data not shown). Importantly, the percentage and absolute number of hepatic HBc-specific CD8+ T cells (detected by HBcAg93-100 pentamer staining) (Fig. 1F) as well as the percentage of hepatic IFN-γ+ CD8+ T cells (Fig. 1G) decreased significantly LBH589 research buy in HBV+ mice, indicating that HBV persistence impaired CD8+ T-cell responses. We also detected the specific response to LCMV infection by LCMV gp33 administration. Our data showed that the percentages of LCMV gp33+ CD8+ T cells were increased in both HBV− and HBV+ mice with no significant

differences (Fig. 1H). These results suggest that HBV-induced systemic immunotolerance is HBV-specific. All the results raised the possibility that impairing HBV-induced hepatocyte-intrinsic immune responses leads to systemic adaptive immunotolerance. To test whether intrinsic innate immunotolerance can be reversed in vivo, we constructed a dually functional vector containing an immunostimulatory ssRNA and an HBx-gene-silencing shRNA. We designed four different sequences encoding ssRNAs and HBx-shRNA, and inserted Smoothened them

into the shRNA pSIREN expression vector. Transfection with ssRNA1- and ssRNA4-containing vectors significantly enhanced IFN-α production in supernatants, while all four shRNA vectors effectively silenced HBx expression at both the messenger RNA (mRNA) and protein levels (Supporting Fig. 3A,B). We selected ssRNA4 and HBx-shRNA3 to construct the dual-function vector (Supporting Fig. 3C). The dual-function (dual), single immunostimulatory RNA (ssRNA), single HBx-shRNA (shRNA), or pSIREN (empty control) vectors were separately transfected into HBV-persistent HepG2.2.15 cells. Although shRNA and dual vectors significantly reduced HBx expression at both the mRNA and protein levels, the dual vector more effectively reduced HBV DNA replication and HBsAg/HBeAg production (Supporting Fig. 4A). Furthermore, the dual vector induced higher IFN-α, IFN-β, ISG15, and MxA production (Supporting Fig. 4B-D) as well as lower TGF-β and IL-10 (Supporting Fig. 4B).

Paraffin-fixed liver sections (5 μm thick) were deparaffinized an

Paraffin-fixed liver sections (5 μm thick) were deparaffinized and stained with hematoxylin and eosin. Pancytokeratin (56- and 64-kDa keratins; DAKO, Carpinteria, CA [1:300]) or K19 (polyclonal rat anti-K19 Troma III; Hybridoma Bank, University of Iowa, Iowa City, IA [1:200]) antibodies were used JQ1 to identify the biliary cysts.7, 8, 18 To detect the antigen of interest, serial liver tissue sections were immunostained as described.7, 8, 18 For all immunoreactions, negative controls were also included and showed no staining. The two main liver lobes were embedded in paraffin and serial 5-μm sections, cut and mounted on 0.1% poly-L-lysine–coated glass slides. Each sample

was immunostained with a pancytokeratin or K19 antibody to allow a correct discrimination

of the biliary cyst structures from the vessels. We used two different approaches: (1) samples labeled with pancytokeratin were used to calculate the relative area covered by the biliary cysts. For each main liver lobe, five random nonoverlapping fields were recorded by a digital camera (magnification ×10) for a total number of 10 fields per mouse. The cystic areas per field were then manually measured by two investigators blinded to Dabrafenib cost the treatment code using ImageJ software (National Institutes of Health, Bethesda, MD).19 The same samples, labeled with K19, underwent computer-assisted morphometric analysis using a motorized stage system to scan the whole liver lobes at magnification ×4 and the Metamorph software (Molecular Devices, Downington, PA). Data are expressed as the percentage of the whole liver lobe area occupied by K19-positive cells. The setup consisted in

a Nikon Eclipse TE2000U microscope (Nikon, Bloomfield, CT), a motorized stage system (Rockland, MA) and a photometric cool snap HQ Staurosporine digital camera (Roper Scientific, Tucson, AZ). Liver sections from treated and untreated animals were immunostained with phosphorylated-ERK (pERK) (Cell Signaling Technology, Danvers, MA) for Ki67 (Abcam, Cambridge, MA), and cleaved caspase-3 (CC3) (R&D Systems, Minneapolis, MN) antibodies to assess the percent of proliferating cystic cholangiocytes and to detect cells undergoing apoptosis. For this analysis, five random nonoverlapping fields taken at magnification ×40 per slide were recorded by a digital camera by two different observers blinded to the treatment code. Data are expressed as the percentage of the K19-positive cell area. Western blots on cell lysates were performed as described.7, 8 (See Supporting Information for additional details.) Cells were plated into 96-multiwell plates (5,000 cells/well) and serum-starved. After 24 hours, cells were treated with an increasing concentration of sorafenib (0.001, 0.01, 0.1, 1, and 10 μM) as described in Results.

1; Supporting Information Fig S1) RNA transcripts expressed fro

1; Supporting Information Fig. S1). RNA transcripts expressed from these clones are infectious on transfection and virus from transfections can be used to determine directly the susceptibility of different genotypes to PIs. In the current study, this in vitro phenotypic assay was used to investigate susceptibilities of genotypes 1-6 to the two structurally distinct PIs in advanced clinical trials (telaprevir and danoprevir). We have additionally genetically and phenotypically characterized a large number of mutations that developed on in vitro passage of different genotypes in subinhibitory concentrations of each PI and determined their effect on viral replication

fitness. This study provides the first evidence-based assessment of the applicability FK506 of PIs to nontype 1 genotypes using the full-length viral replication

cycle, and may contribute to the more effective use of antiviral therapy in future HCV management. HCV, hepatitis C virus; IFN, interferon; PIs, protease inhibitors; RBV, ribavirin. Construction and the successful expression of replication-competent virus from the intra- and intergenotypic recombinants J1b1b, J2a2a-T1066S, J3a3a, J4a4a-19, J5a5a-Q1247L, and J6a6a-V1040L with Jc117, 18 has been described.16 For reverse genetic studies, mutations were introduced using mutated primers with the Quick BYL719 Change Site-Directed Mutagenesis Kit (Stratagene). Modified fragments were verified by sequencing. Procedures for cell culturing, RNA synthesis, transfection, and immunostaining were described.16 Briefly, linearized and blunted DNA templates were cleaned by phenol/chloroform

extraction, followed by ethanol precipitation and RNA synthesized using T7 RNA Polymerase (Promega). RNA was transfected into Huh7.5 cells by electroporation and viral spread assessed by NS5A immunostaining with polyclonal sheep anti-NS5A serum. RNA transcripts from the replication-deficient JFH1-based genome containing the GND mutation in the NS5B polymerase served as a negative and that of Jc1 as positive control, respectively. The HCV-specific PI BILN 2061 (a gift from GlaxoSmithKline) was resuspended at 5 mM in dimethylsulphoxide, telaprevir, and danoprevir (both purchased from Acme Bioscience, Palo Alto, CA) at 20 mM dimethylsulphoxide. The effect of the different PIs on the replication of the intra- and intergenotypic recombinants was assessed as described.16 Briefly, Inositol oxygenase RNA was transfected into Huh7.5 cells and cultures incubated with or without PIs. Alternatively, naïve cells were infected with infectious supernatant, washed, and incubated with or without PIs. Antiviral efficacy was measured as relative inhibition of RNA replication by staining for NS5A and determining the percentage of HCV-positive cells or counting the number of foci forming units/mL. Each concentration was assayed in triplicate. Intra- and intergenotypic recombinants were passaged for 2 to 3 weeks under subinhibitory concentrations of PIs as described.

Patients enrolled did not have any current evidence of HE at incl

Patients enrolled did not have any current evidence of HE at inclusion. The main study outcomes were the occurrence of clear episodes of HE: a more objective endpoint than the amelioration of HE symptoms in patients already affected by HE at inclusion. Rifaximin is a semisynthetic, gut-selective antibiotic whose mechanism of antimicrobial action depends on inhibition of RNA synthesis.8, 9 There are several lines of evidence that rifaximin has poor solubility and is poorly absorbed, which results in a gut-specific action. In healthy individuals, as much as 97% of radiolabeled rifaximin is recovered; 96% in the stool and

only 0.32% in the urine.10 The systemic exposure to rifaximin in patients with Child A, B, and C cirrhosis compared to rifampin and neomycin is shown in Fig. 1.10 Dosing of rifaximin

can be approached in two broad ways: cyclical or continuous. In Italy, cyclical dosing is preferred and several clinical trials have shown benefit with PD-0332991 solubility dmso rifaximin treatment 2 weeks per month.8 The alternative is daily therapy with rifaximin that is currently being used in the United States. There are possible advantages and disadvantages to each approach; cyclical therapy reduces cost and exposure to antibiotic but adherence to the schedules may be difficult in patients who already have HE. Continuous therapy is more expensive and could have the potential to increase resistance to rifaximin. The daily dose most studied, regardless of cyclical or continuous, is 1200 mg whereas the most recent trial used 1100 mg/day.6 Thymidine kinase In some European countries, including Italy and Spain, rifaximin is available in packages containing only 12 tablets, which is the dosage needed for just 2 days of therapy, and this also may limit the availability of the drug for long term treatment. For example, in Italy, in order

to obtain the drug by means of the public health care system, the patient should consult the prescribing practitioner every 4 days! Being a gut-selective antibiotic, the majority of the action of rifaximin is concentrated on the gut microflora. The antibacterial properties of rifaximin include bactericidal activity at rifaximin concentrations greater than or equal to the minimal inhibitory concentration (MIC); at sub-MIC concentrations, rifaximin can change functioning of epithelial cells as well as virulence of the gut bacteria.10 Several randomized, placebo-controlled trials have been performed with rifaximin, most of them in Europe.8, 10 These trials have studied short-term management of the acute episode, long-term therapy and prevention of recurrence.9 A summary of the trials with their key findings is shown in Table 1. Rifaximin has been studied in the context of intestinal pathogens and the concentration needed to inhibit 50% (MIC50) and 90% of microorganism growth (MIC90) have been established for 1607 pathogens; the highest MIC reached was 1024 μg/mL.

Pain Ranking – The severity of headache was recorded immediately

Pain Ranking – The severity of headache was recorded immediately prior to the first dose and 30 minutes, 1 hour, 2 hours, and 4 hours postdose on a 4-point scale (0 = no pain; 1 = mild pain, ability to perform normal daily activities; 2 = moderate pain, disturbing normal activities; 3 = severe pain, disabling activities, requiring bed rest). In addition, the patients documented the presence

of associated symptoms (nausea, buy Ibrutinib vomiting, photophobia, phonophobia, and osmophobia) at 2 hours postdose. Headache free was defined as conversion from moderate or severe pain to no pain (score of 2 or 3 reducing to 0) at 2 hours without taking rescue or a second dose. Headache improvement was defined as an improvement learn more from severe or moderate (grade 2 or 3) at baseline to mild or none (grade 0 or 1) after dosing. Headache recurrence was defined as a return to moderate or severe pain within 48 hours of primary treatment following initial improvement to mild or no pain at 2 hours. Safety and tolerability were assessed by comparing the incidence of AEs. The AEs were documented on a symptom checklist (including somnolence, dizziness, malaise, EPS [extrapyramidal symptoms], paresthesia, dry mouth, nausea, chest discomfort, abdominal

pain) at 4 hours after the study medication intake. All AEs occurring following medication use were elicited by the investigator at visit 2. This study was designed to assess the efficacy and safety of a combined pharmaceutical modality including SPr in patients commonly affected by moderate to severe migraine attacks. The primary efficacy end point was the proportion of patients experiencing headache-free response 2 hours after dosing. A sample size of 93 patients in each group was required to provide

at least 80% power under the assumption that 70% of patients given SPr would be headache free vs 50% of patients given sumatriptan plus placebo (SP) (two-sided, α = 0.05). Moreover, it was supposed that approximately 30% of the patients would not complete the trial during the study period. Accordingly, it was estimated that approximately a total of 242 subjects would be required to be enrolled. An intention-to-treat analysis (ITT) was used as the primary analysis. The ITT population included all subjects who underwent randomization and provided CYTH4 at least 1 postdose efficacy assessment. The safety population included all patients treated with the study medications and whose follow-up safety data were available. Information missing for any planned assessment was replaced by the last-observation-carried-forward methodology. Comparisons of demographics, baseline characteristics, and AEs after each treatment between groups were performed by descriptive statistics. Categorical variables were compared using chi-square test or the Fisher’s exact test, as appropriate. Odds ratios (OR) and corresponding two-sided 95% confidence interval (CI) were given for treatment comparisons.

It has high sensitivity compared to single RT-PCR Moreover, fiel

It has high sensitivity compared to single RT-PCR. Moreover, field samples in China can be tested by this method for virus

detection. Our results show that one-step multiplex RT-PCR is a high-throughput, specific, sensitive method for tobacco virus detection. “
“Tree peony (Paeonia suffruticosais) plants with yellowing symptoms suggestive of a phytoplasma disease were observed in Shandong Peninsula, China. Typical phytoplasma bodies were detected in the phloem tissue using transmission electron microscopy. The association of a phytoplasma with the disease was confirmed by polymerase chain reaction (PCR) using phytoplasma universal primer pair R16mF2/R16mR1 followed by R16F2n/R16R2 as nested PCR primer pair. The sequence analysis indicated that the phytoplasma associated with tree peony yellows (TPY) was an isolate

of ‘Ca. Phytoplasma solani’ belonging to the stolbur (16SrXII) group. This is the first report of a phytoplasma associated with tree peony. “
“Pistachio is an important crop in Iran, which is a major producer and exporter of pistachio nuts. The occurrence of a new disease of pistachio trees, characterized by the development of severe witches’ broom, stunted growth and leaf Autophagy inhibitor mw rosetting, was observed in Ghazvin Province. A phytoplasma was detected in infected trees by polymerase chain reaction (PCR) amplification of rRNA operon sequences. Nested PCR with primer pairs P1/P7 and R16F2n/R16R2 was used for specific detection of the phytoplasma in infected trees.

To determine its taxonomy, the random fragment length polymorphism (RFLP) pattern and sequence analysis of the amplified rRNA gene were studied. Sequencing of the amplified products of the phytoplasma 16S rRNA gene indicated that pistachio witches’ broom (PWB) phytoplasma is in a separate 16S rRNA group of phytoplasmas (with sequence homology 97% in Blast search). The unique properties of the DNA of the PWB phytoplasma indicate that Epothilone B (EPO906, Patupilone) it is a representative of a new taxon. “
“Symptoms of unknown aetiology on Rhododendron hybridum cv. Cunningham’s White were observed in the Czech Republic in 2010. The infected plant had malformed leaves, with irregular shaped edges, mosaic, leaf tip necrosis and multiple axillary shoots with smaller leaves. Transmission electron microscopy showed phytoplasma-like bodies in phloem cells of the symptomatic plant. Phytoplasma presence was confirmed by polymerase chain reaction using phytoplasma-specific, universal and group-specific primer pairs. Restriction fragment length polymorphism analysis of 16S rDNA enabled classification of the detected phytoplasma into the aster yellows subgroup I-C. Sequence analysis of the 16S-23S ribosomal operon of the amplified phytoplasma genome from the infected rhododendron plant (1724 bp) confirmed the closest relationship with the Czech Echinacea purpurea phyllody phytoplasma.

Materials and Methods: We used two immortalized human hepatocyte

Materials and Methods: We used two immortalized human hepatocyte cell lines,

TTNT1 6 cells (in which hTERT was introduced) and T5B cells (in which the SV40 large T antigen (LT) was introduced). We used a retrovirus vector to introduce V12H-Ras, effector-loop mutants of oncogenic Ras, V12H-RasT35S (S35), V12H-RasE37G (G37), and V12H-RasY40C (C40), and c-myc into TTNT16 cells, HBx-expressing TTNT16 selleck inhibitor cells, LT+small T antigen (ST)-expressing TTNT16 cells, and ST+hTERT-expressing T5B cells. We then used these cells to analyze cell proliferation, senescence, transformation, and stem-like features. Results: First, we evaluated whether HBx and Ras induced the tumorigenic transformation of the immortalized human hepatocytes. TTNT1

6 cells expressing wild-type Ras or C40, but not S35 or G37, showed a profound senescence-like phenotype after transfection. In contrast, the introduction of these two genes into TTNT16-HBx or TTNT16-LT+ST cells did not induce this senescence-like phenotype. Moreover, wild-type Ras, but not S35, G37, or C40, enabled HBx- or LT+ST-express-ing human hepatocytes to form large colonies in soft agar and large tumors in nude mice. Next, we analyzed the relationship between c-myc and/or HBx introduction and the tumorigenic transformation of immortalized human hepatocytes. C-myc activated the STAT3 pathway and induced tumorigenicity in HBx-or LT+ST-expressing immortalized human hepatocytes. Furthermore, c-myc and HBx co-expression induced stem cell-like features in immortalized human hepatocytes. Conclusions: Modification of the ZD1839 chemical structure PI3K pathway can overcome active onco-gene-induced senescence. In addition, immortalized human hepatocytes require the activation of the Raf and Ral-GEF pathways for tumorigenic transformation. Furthermore, the co-expression of HBx and c-myc introduced stem cell-like features during the tumorigenic transformation of these cells. Disclosures: Shuichi

Kaneko – Grant/Research Support: MDS, Co., Inc, Chugai Pharma., Co., Inc, Toray Co., Inc, Daiichi Sankyo., Co., Inc, Dainippon Sumitomo, Co., Inc, Aji-nomoto Co., Inc, MDS, Co., to Inc, Chugai Pharma., Co., Inc, Toray Co., Inc, Daiichi Sankyo., Co., Inc, Dainippon Sumitomo, Co., Inc, Ajinomoto Co., Inc, Bayer Japan The following people have nothing to disclose: Naoki Oishi, Seishi Murakami, Wang Xuyang Background: Fibrinogen like protein-1 (Fgl1) a liver expressed protein has been shown to be downregulated in human hepatocellular carcinoma. Reduction of Fgl1 expression in vitro also increases anchorage independent cellular proliferation of human hepatoma cell lines. Aims: To determine if targeted disruption of Fgl1 leads to enhanced carcinogenesis following exposure to diethynitrosamine (DEN) and phenobarbitol (PB). Methods: We generated a knockout mouse by the targeted disruption of Fgl1.