Y.M. Moon

Fatal spontaneous bacterial peritonitis by Leclercia adecarboxylata in a patient with hepatocellular carcinoma

lation, or absence of fragmental pericardium with appendage herniation. TTE or TEE, MRI and cardiac catheterisation are effective diagnostic modalities for left atrial aneurysms. TTE is a noninvasive and convenient diagnostic tool (8,11). The anatomical relationship between an aneurysm and adjacent structures can be studied thoroughly. TEE is an alternative diagnostic approach when image study of TTE is inadequate (11). Moreover, TEE is an excellent tool for identifying the presence of an intracardiac thrombus (2), and is beneficial when studying pulmonary venous return. Magnetic resonance imaging is another effective noninvasive imaging study. With three-dimensional reconstruction, an MRI provides further information regarding the extent and size of an aneurysm, its relationship to adjacent structures, presence of compression to the left ventricle, and abnormal pulmonary venous drainage (11,12,15). Cardiac catheterisation was previously used for the definitive diagnosis of left atrial aneurysms. Trans-septal catheterisation or pulmonary angiogram in the levophase show that contrast enters the atrium with aneurysmal sac (6,8,10,16). Although cardiac catheterisation may be unnecessary when echocardiographic or MRI studies diagnose left atrial aneurysm and provide sufficient preoperative information (11,12,15), excluding other cardiac abnormalities such as systemic-to-pulmonary shunt is prudent. Surgical intervention is recommended even for asymptomatic patients as major complications, such as tachycardia, heart failure, or peripheral systemic embolisms, can develop. Following resection of an aneurysm, most reported cases obtained good results (2,6,7,12,14,16). In this case, no anti-dysrhythmic agents were needed after surgery. In conclusion, aneurysm of the left atrium could be associated with the paroxysmal atrial flutter and should be suspected when chest radiography shows abnormal atrium bulging. TTE should be used as the initial step for differential diagnosis. Other imaging studies, including TEE, computer tomography and MRI, also provide useful information.

[Concurrent chemo-radiation therapy for advanced hepatocellular carcinoma with portal vein thrombosis].

BACKGROUND/AIMS Advanced hepatocellular carcinoma with portal vein thrombosis has a poor prognosis. This study was undertaken to evaluate the therapeutic effects of concurrent chemo-radiation therapy in advanced hepatocellular carcinoma with portal vein thrombosis. METHODS A total of 54 patients with advanced hepatocellular carcinoma (TNM stage IVa) were enrolled. Nineteen patients were treated with external beam radiotherapy (4,500 cGy/ 5 weeks) and intrahepatic arterial 5-FU infusion (500 mg on 1-5 day and 30-35 day, respectively) via implanted chemoport. The others were treated with intrahepatic arterial cisplatin infusion (80 mg/m(2)). RESULTS In patients treated with concurrent chemo-radiation therapy, response rates at 2nd and 6th months were 42.1% and 26.3%, respectively. In patients treated with intrahepatic arterial cisplatin therapy, response rates at 2nd and 6th months were 2.9% and 0%, respectively. The median survival time was 11.6 months in concurrent chemo-radiation therapy and 4.8 months in intrahepatic arterial cisplatin infusion therapy. Concurrent chemo-radiation therapy produced better response rates and longer survival time than those of intrahepatic arterial cisplatin infusion therapy (p<0.05). CONCLUSIONS Concurrent chemo-radiation therapy achieved favorable results in advanced hepatocellular carcinoma with portal vein thrombosis and can be considered as a treatment option for the management of advanced hepatocellular carcinoma.

A novel primer‐extension assay for the detection of a G to A mutation in the distal precore region of hepatitis B virus DNA

The roles of genetic heterogeneity of the hepatitis B virus (HBV) precore gene in the pathogenesis of HBV infection are unclear. Various methods have been used to detect nucleotide (nt) 1896 precore mutants. We established a new primer‐extension assay to facilitate the detection of these mutants. This assay is based upon the fact that there is no adenine in the distal precore region of wild‐type HBV. Polymerase chain reaction (PCR)‐amplified template DNA was denatured and annealed to the [γ‐32P]‐labelled primer. During primer extension in the presence of DNA polymerase and dCTP, dGTP, dTTP and ddATP, the reaction terminates if there is a nucleotide A. When mixtures of different ratios of wild‐type and nt 1896 precore mutants were analysed in the primer‐extension assay, correlation between the percentage known amounts and the percentage measured amounts of nt 1896 precore mutants was excellent (r2=0.9669). When the primer‐extension assay and direct sequencing were compared in hepatitis B e antigen (HBeAg)‐positive and ‐negative chronic active hepatitis B patients, the primer‐extension assay detected a greater number of nt 1896 precore mutants than direct sequencing and thus most HBV infections were found to be mixed infections. In conclusion, the primer‐extension assay is a reliable and sensitive method for the detection of nt 1896 precore mutants.

Histologic Study of Chronic Active Hepatitis C; Comparison with Chronic Active Hepatitis B

Reports on the histologic findings of chronic active hepatitis C (CAH-C) have been rare, and the characteristic histologic findings of CAH-C have been not yet determined. To compare the differences in the histologic findings between CAH-C and chronic active hepatitis B (CAH-B) group, we analyzed the histologic findings of 19 patients with CAH-C, who had positive tests for HCV-antibody by EIA, and 19 patients with CAH-B who had negative tests for HCV-antibody but positive tests for HBsAg by RIA. Histologic features were analyzed between the CAH-C and CAH-B groups using a scoring system which is modified from Knodell’s histologic activity index-looking at portal inflammation, periportal necroinflammation, portal fibrosis, focal necrosis, regeneration, polyploid nuclear change, sinusoidal lymphocytic reaction and fatty change. Portal inflammatory cell infiltrations with prominent lymphocytes and follicular arrangement were more frequent in the CAH-C group (10 of 19 cases) than in the CAH-B group (5 of 19 cases). Severe sinusoidal lymphocytic reactions were also more prominent in the CAH-C group (11 of 19 cases) than in the CAH-B group (6 of 19 cases). However, periportal necroinflammation, portal fibrosis, focal hepatic necrosis, regeneration and polyploid nuclear changes were more prominent in the CAH-B group than in the CAH-C group. In conclusion, follicular portal inflammation and severe sinusoidal lymphocytic reactions were common histologic findings in serologically proven CAH-C when compared to CAH-B.

886 HTERT PROMOTER GENE POLYMORPHISM AND THE RISK OF HEPATOCELLULAR CARCINOMA (HCC) IN PATIENTS WITH CHRONIC HEPATITIS B

Background and Aims: Many investigations have revealed that a low recurrence rate of hepatocellular carcinoma (HCC) is associated with high serum albumin levels in patients; therefore, high levels of serum albumin comprise a major indicator of a favorable prognosis. The mechanism inhibiting the proliferation of HCC has not yet been elucidated, so we investigated the effect of serum albumin on HCC cell proliferation. Methods: Hep3B was cultured in MEM with no serum or containing 5 g/dl bovine albumin. As the control samples, Prionex (Polysciences, Inc.) was added to generate the same osmotic pressure as albumin. After 24-hour incubation, the expressions of afetoprotein (AFP), p53, p21, and p57 were evaluated with real-time PCR using total RNA extracted from the liver. Protein expressions and the phosphorylation of Rb (retinoblastoma) were determined by Western blot analysis using total protein extracted from the liver. For flow cytometric analysis of the cell cycle, FACS analysis was performed. The percentages of cell cycle distribution were evaluated by PI staining, and all samples were analyzed employing FACScalibur (BD) with appropriate software (ModFit LT, BD). Results: The mRNA levels of AFP relative to Alb(−): Alb(−), Alb(+), and Prionex, were 1.0.7±0.2 (p < 0.001 for Alb(−)), and 1±0.3, respectively. The mRNA levels of p21 were 1, 1.58±0.4 (p = 0.007 for Alb(−) and p=0.004 for Prionex), and 0.8±0.2, respectively. The mRNA levels of p57 were 1, 4.4±1.4 (p = 0.002 for Alb(−) and Prionex), and 1.0±0.1, respectively. The protein expression levels of Rb were similar within all culture media. The phosphorylation of P807/811 and P780 of Rb protein was reduced in Alb(+). More cells in the G0/G1 phase and less cells in S and G2/M phases were obtained in Alb(+) than in Alb(−) (G0/G1: 45.2, 49.5, 44.7%; G2/M: 44.3, 41.5, 43.0%; S: 10.5, 7.4, 17.2%, Alb(−), Alb(+), Prionex, respectively). The same results were obtained in HepG2. Conclusions: The presence of albumin in serum reduces the phosphorylation of Rb proteins and enhances the expression of p21 and p57, following an increase in the G0/G1 cell population, and suppresses cell proliferation. These results suggest that albumin itself suppresses the proliferation of hepatocellular carcinoma.

The expression of G1 cyclins and RB-E2F and the effect of vitamin E on hepatic stellate cells activated by acute CCL4 induced hepatic injury

K.S. Lee, K.J. Lee, K.H. Ham C.Y. Chon, Y.M. Moon Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea. Hepatic stellate cells(HSC) play a key role in hepatic fibrogenesis. Therefore HSC activation model is needed to find some agents to block HSC activation. We evaluated HSC activation and proliferation, G1-S cyclins expression, Rb-E2F expression and the effect of vitamin E during acute CCh induced hepatic injury. Methods: Sprague-Dawtey rats(100g) each received a single intraperitoneal injection(IP) of CCh in mineral oil at a dose of 2ml/kg body weight(Group I and II). In addition, animals received IP of 200mg/kg vitamin E(Group II) daily, 2 days before CCI4 administration until the sacrifice. 5 animals(each time, each group) were sacrificed at Oh, 8h, 16h, 32h, 48h and 60h after CCh injection and 30pCi of [~H]Thymidine was administered 4h before sacrifice. Blood samples were obtained to check serum ALT level. Liver were promptly removed and a piece was fixed and embedded in paraffin for aSMA immunohistochemical(IMH) staining and count aSMA positive cells in 400x HPF. HSC were isolated from fresh fiver by non0erfusion method and single step density Nycodenz gradient. Nuclei were prepared for gel shift assay(E2F). DNA(10#g) were prepared from HSC to check [~H]Thymidine uptake by using B counter. Protein(10pg: Rb, CDK2, CDK4 / 2Ozg: cyclin D1, cyclin E) were prepared from HSC for western blot. Results: The serum ALT level changed 74-+20.7, 170-+54.3, 258+ 83.5, 1178-+381.3, 274-+174.0, 92-+74.0 in group I and 64-+21.9, 152-+58.9, 156-+97.6, 576-+141.5, 70-+35.4, 62+55.4 in group II(peaked at 32h: P<0.01 / decrease in group I]: P<0.06). [°H]Thymidine uptake(c0m) changed 76.4 +-14.7, 76.6-+19.7, 78.8-+23.8, 529.2-+284.8, 299.0-+161.6, 179.6-+63.9 in g_+roup I and 71.6-+19.9, 90.4-+9.6, 85.0-+2A.0, Z23.0-+86.3, 171.2-+47.8, 127.8 19.3 in group II(oeaked at 32h: P<0.01 / decrease in group I/: P<0.05). IMI-I for aSMA showed 0, 0, 2.0-+0.8, 49.2-+6.8, 31.2-+13.6, 21.8-+10.1 in group I and 0, 0, 0, 14.8-+1.4, 18.1-+9.1, 12.2-+6.2 in group N(peaked at 32h: P<0.01 / decrease in group 1]: P<0.01). In western blot, cyclin D1 and Rb showed bands at 32h and cyclin E, CDK2 and CDK4 showed bands at 16h, 32h, 48h. All bands were diminished in group II. In gel shift assay, E2F showed shifted bands at 16h, 32h, 48h and NFkB showed shifted band at 32h. All shifted bands were diminished in group II. Conclusion: The serum ALT level, HSC proliferation and activation were peaked at 32h after CCI4 administration and significantly decreased in vitamin E treated group. G1-S cyclins, CDK2, CDK4, Rb showed bands at 1611, 32h, 48h or 32h after CC14 administration and diminished in vitamin E treated group. E2F showed shifted bands at 16h, 32h, 4811 and NFkB showed shifted band at 32h. All shifted bands were diminished in vitamin E treated group.