A functional polymorphism in pre-microRNA-196a-2 contributes to the susceptibility of hepatocellular carcinoma in a Turkish population: a case-control study.

MicroRNAs (miRNAs) are an abundant class of small nonprotein-coding RNAs with posttranscriptional regulatory functions as tumour suppressors and oncogenes. Aberrant expression and structural alteration of miRNAs are thought to participate in tumourigenesis and cancer development. It has been suggested that the presence of single-nucleotide polymorphisms in precursor miRNAs (pre-miRNAs) can alter miRNA processing, expression, and/or binding to target mRNA and represent another type of genetic variability that can contribute to the development of human cancers. Recent studies have indicated that the miR-196a-2 rs11614913 (C→T) polymorphism could alter mature miR-196a-2 expression and target mRNA binding. To determine the association of the miR-196a-2 rs11614913 polymorphism with the risk of hepatocellular carcinoma (HCC) development in a Turkish population, a hospital-based case-control study was designed consisting of 185 subjects with HCC and 185 cancer-free control subjects matched for age, gender, smoking and alcohol status. The genotype frequency of the miR-196a-2 rs11614913 polymorphism was determined by using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. Our data shows that the CC genotype of the miR-196a-2 rs11614913 polymorphism is associated with increased risk of HCC development in this Turkish population (OR = 2.41, 95% CI: 1.30-4.50, P = 0.005). Furthermore, according to stratified analysis, a significant association was observed between the homozygote CC genotype and HCC risk in the subgroups of male gender (OR = 3.12, 95% CI: 1.53-6.34, P = 0.002) and patients with hepatitis B virus (HBV)-related HCC (OR = 2.88, 95% CI: 1.33-6.22, P = 0.007). Because our results suggest for the first time that the miR-196a-2 rs11614913 polymorphism may be a genetic susceptibility factor for HCC (especially in the male gender and HBV-infected patients) in the Turkish population, further independent studies are required to validate our findings in a larger series, as well as in patients of different ethnic origins.

Plasma lipids, tumor parameters and survival in HCC patients with HBV and HCV.

INTRODUCTION AND AIMS: Hepatocellular carcinoma (HCC) is a consequence of chronic liver disease, particularly from hepatitis B or C and increasingly from obesity and metabolic syndrome. Since lipids are an important component of cell membranes and are involved in cell signaling and tumor cell growth, we wished to evaluate the relationship between HCC patient plasma lipids and maximum tumor diameter and other indices of HCC human biology. METHODS: We examined prospectively-collected data from a multi-institutional collaborative Turkish HCC working group, from predominantly HBV-based patients, for plasma lipid profiles, consisting of triglycerides, total cholesterol, LDL-cholesterol (LDL) and HDL-cholesterol (HDL) and compared these with the associated patient maximum tumor diameter (MTD), portal vein thrombosis, alpha-fetoprotein (AFP) and also with patient survival. RESULTS: We found that both low HDL (p=0.0002) and high LDL (p=0.003) levels were significantly associated with increased MTD, as well as in a final multiple linear regression model on MTD. The combination of low HDL combined with high HDL levels were significant in a regression model on MTD, PVT and an HCC Aggressiveness Index (Odds Ratio 12.91 compared to an Odds Ratio of 1 for the reference). Furthermore, in a Cox regression model on death, the HDL plus LDL combination had a significantly higher Hazard Ratio than the reference category. CONCLUSIONS: Low plasma HDL, high plasma LDL and especially the combination, were significantly related to more aggressive HCC phenotype and the combination was significantly related to a higher Hazard Ratio for death.

Serum levels of gamma-glutamyl transpeptidase in relation to HCC human biology and prognosis.

BACKGROUND AND AIM: Hepatocellular carcinoma (HCC) biomarkers are limited, as even the best studied, alpha-fetoprotein (AFP), is elevated in no more than 50% of HCC patients. The aim was to evaluate several serum liver function tests in relation to survival and tumor characteristics in a large cohort of Turkish HCC patients. METHODS: We retrospectively examined the serum levels of gamma glutamyl transpeptidase (GGT) in relation to patient survival. RESULTS: Kaplan-Meier analysis showed that only GGT and albumin amongst liver function tests, were significantly associated with survival. Survival worsened with increase in GGT levels semi-quantitatively. Increase in GGT levels was also found to significantly correlate with an increase in maximum tumor diameter from 4.5 to 7 cm, a 20-fold increase in serum alpha-fetoprotein level, an increase in tumor multifocality from 20 to 54% of patients, and a doubling in percent of patients with portal vein thrombosis (PVT) from 20 to 40%. Serum GGT levels also showed significant survival differences for patients with low AFP levels. A doublet combination of serum GGT with albumin levels was associated with higher hazard ratios in a Cox regression analysis, as compared with single parameter GGT. The combination parameter pair was also prognostically useful in the low-AFP patient subcohort and was associated with significant differences in patient tumor characteristics. CONCLUSIONS: Serum GGT levels and especially combination serum GGT plus albumin levels, were significantly associated both with HCC patient survival and tumor aggressiveness characteristics, regardless of AFP levels in a large Turkish cohort. This might be especially useful since the majority of HCC patients do not have elevated levels of AFP.

Relationship between functional polymorphism in the Aurora A gene and susceptibility of hepatocellular carcinoma.

Aurora A is considered a potential cancer susceptibility gene owing to overexpression or amplification of Aurora A gene that causes centrosome dysfunction, chromosome instability, tumourigenic transformation and checkpoint abnormalities. Functional coding region polymorphism F31I in the Aurora A gene has recently been shown to be associated with several human cancers, but its association with hepatocellular carcinoma (HCC) has yet to be investigated. Genetic polymorphism of Aurora A was investigated in 128 confirmed subjects with HCC and 128 cancer-free control subjects matched on age, gender, smoking and alcohol consumption by using a polymerase chain reaction-restriction fragment length polymorphism assay. Allele and genotype associations of Aurora A F31I polymorphism with HCC susceptibility were observed in comparisons between the patient and control samples (respectively; P = 0.005, P = 0.012). The proportion of the genotypes containing I31 allele in patients with HCC (39.8%) was significantly higher than that in patients without HCC (22.7%) (P = 0.003). The distribution F31I genotype was significantly associated with increased risk of HCC (P = 0.003, odds ratio = 2.26, 95% confidence interval = 1.31-3.90 for FI + II genotypes vs FF genotype). Our results suggest for the first time that the Aurora A F31I polymorphism may be a genetic susceptibility factor for HCC.

Infectious complications in the early postoperative period in liver transplant patients.

Liver transplantation (LTx) is a technically well established procedure in acute and in end-stage liver diseases. However, opportunistic infections remain one of the important complications in short and long-term outcome of LTx patients. Bloodstream and pulmonary infections are the major cause of death in the first year following liver transplantation. Due to extended use of chinolons and third generation cephalosporines there is a shift towards multidrug-resistant bacteria including methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecium, and extended b-lactamase-producing gram negative rods. Fungal infections are mainly due to Candida spp. Viral infections, such as with cytomegalovirus, human herpesvirus 6, herpes simplex virus, and Epstein-Barr virus infections are another major cause of morbidity in patients receiving solid organ transplants, including liver transplant patients. Studies of infection following LTx are necessary to improve management and to provide a better outcome after LTx. This review focuses on the most important bacterial, fungal and viral infections in LTx patients.

Expression of c-kit protooncogen in hepatitis B virus-induced chronic hepatitis, cirrhosis and hepatocellular carcinoma: has it a diagnostic role?

AIM: There are more than 350 million people worldwide chronically infected with hepatitis B virus (HBV), who are at high risk for the development of hepatitis, cirrhosis and hepatocellular carcinoma (HCC). Because of the conflicting results about c-kit expression in HCC and the key role played by c-kit in gastrointestinal stromal tumours (GIST) and other solid tumours, the aim of this study was to determine c-kit expression in the course of hepatitis B infection. MATERIALS AND METHODS: Paraffin-embedded tissues in Cukurova University Faculty of Medicine Department of Pathology between January 2002 and February 2006 were searched restrospectively to investigate this issue. We performed immunohistochemistry on biopsies of 125 patients with HBV infection, grouped as: mild, moderate and severe hepatitis, cirrhosis and HCC, 25 patients in each of them, using anti c-kit monoclonal antibody. The severity of parenchymal inflammation and of interface hepatitis was semiquantitatively graded on a haematoxylin and eosin stained paraffin sections. Additionally, 50 more HCC, formed on HBV basis, were studied to determine the prevalence of c-kit overexpression. RESULTS: In cirrhotic liver, lower intensity of staining and rarely c-kit positivity were present. The greatest number of the c-kit positivity and higher intensity of staining was found in the livers of patients with severe hepatitis and HCC. In chronic hepatitis B infection, the staining intensity was parallel with the grade and stage of the disease. In the areas where fibrosis was seen, c-kit positivity was rare or absent. In the HCC specimens, c-kit positivity appeared both inside and around the cancerous nodes. C-kit expression was observed in 62 of 75 HCC tissue specimens (82%) (p < 0.001). CONCLUSIONS: C-kit positivity was observed in the mitotic, proliferating and also dysplastic hepatic cells. These results suggest that c-kit expression may be used as an early diagnostic indicator for HBV induced HCC.

Mortality-lowering effect of systemic corticosteroid therapy in severe tetanus.

In a double-blind therapeutic trial, 32 patients suffering from severe tetanus were given prednisolone and 31 a placebo in addition to standard treatment. Mortality was lower in the prednisolone-treated group (31%) than in the placebo-treated group (55%). The women had more severe symptoms and a higher mortality than the men. The results suggest that the addition of a corticosteroid to standard treatment will lower mortality in tetanus, although meticulous nursing care is still the most important factor in treatment.

Hepatitis C virus genotypes in patients with hepatocellular carcinoma.

OBJECTIVES: Epidemiological studies have suggested a strong association between chronic hepatitis C virus (HCV) infection and the development of hepatocellular carcinoma (HCC). It has also been suggested that there is a relationship between HCV genotypes and the development of cirrhosis and HCC. To investigate the possible role of HCV genotypes in the development of HCC, we studied HCV genotypes in 22 HCV-seropositive patients with histologically proven cirrhosis of the liver and HCC. METHODS: Anti-HCV antibodies were detected by second-generation enzyme-linked immunosorbent assay (ELISA). Serum HCV-RNA was detected by nested reverse transcription-polymerase chain reaction (RT-PCR) using primers derived from the highly conserved 5'-untranslated region. The HCV genotype was determined by nested RT-PCR using type-specific primers derived from the core region. RESULTS: Anti-HCV and HCV-RNA were detected in all patients with HCC. HCV genotyping was achieved in all of them. All patients had genotype 1b HCV. CONCLUSION: These findings suggest that HCV remains in replication and genotype 1b HCV is the predominant type in our HCV-seropositive patients with HCC.

Expression of mesenchymal, hematopoietic, and biliary cell markers in adult rat hepatocytes after partial hepatectomy.

BACKGROUND AND PURPOSE: It has been suggested that liver regeneration can occur either by differentiated adult hepatocytes which retain the capability for several rounds of replication or by hepatic progenitor cells, depending on the number of hepatocytes lost. We sought to study the differentiation potential of hepatocytes following partial hepatectomy (PH) in rats. METHODS: Using immunohistochemistry and confocal microscopy we studied the distribution of cytokeratin 7 (CK7), CK19, vimentin, desmin, CD34, and c-kit among adult rat liver hepatocytes after PH at various times just after hepatectomy and after 8, 16, 24, 36, 48, and 60 hour and 6 and 16 days. RESULTS: Vimentin, c-kit, and desmin positivity were observed in regenerating hepatocytes in the early stages. Desmin and vimentin staining were also demonstrated in stellate cells. Staining enhancement in stellate cells progressed from day 3 to day 6. No liver sections were stained positive for CD34, CK19, or CK7. CONCLUSION: After PH, mature hepatocytes revealed their potential to regain the markers that they do not express when they are quiescent. This result supported the plasticity and differentiation potential of adult hepatocytes during regeneration.

Interferon-induced vitiligo in a patient with chronic viral hepatitis C infection.

Vitiligo is an autoimmune disease characterized by depigmentation of the skin due to destruction of melanocytes. Interferons have been used for the treatment of chronic hepatitis C and some malignancies. We report interferon alpha-2a-induced vitiligo in a male patient with chronic active hepatitis C. All skin lesions disappeared completely without requiring therapy after discontinuation of interferon. This case suggests that vitiligo may be developed during interferon therapy as a side effect.

Endoscopic management of biliary hydatid disease.

OBJECTIVE: To determine the effect of endoscopic sphincterotomy in the management of biliary hydatid disease. DESIGN: A case study between January 1992 and December 1994. SETTING: A university-affiliated hospital in Adana, Turkey. PATIENTS: Five patients with biliary hydatid disease, in which the cyst had ruptured into the biliary tree. The follow-up ranged from 3 to 12 months. INTERVENTIONS: Endoscopic sphincterotomy. MAIN OUTCOME MEASURES: Morbidity, morality and recurrence of the disease. RESULTS: All patients underwent successful endoscopic sphincterotomy, including removal of daughter cysts. During the follow-up period, ultrasonography and laboratory investigations showed complete cure in all patients. There were no complications due to endoscopic sphincterotomy. CONCLUSION: Endoscopic sphincterotomy is the treatment of choice for the management of hydatid cysts that have ruptured into the biliary tract causing obstructive jaundice.

Endoscopic retrograde cholangiopancreatography in the diagnosis and management of choledochal cysts.

Choledochal cysts are an uncommon anomaly of the biliary system manifested by cystic dilatation of the extra or intrahepatic biliary tree or both. It is most frequently found in Orientals and in females. Endoscopic retrograde cholangiopancreatography is a valuable imaging technique in the diagnosis of choledochal cysts in adults. Additionally, in selected cases, a choledochocele may be effectively managed by endoscopic sphincterotomy. We present clinical and endoscopic findings of six adult patients with choledochal cysts. Clinical symptoms were characterized by abdominal pain, jaundice and cholangitis. Associated hepatobiliary pathologic findings included cholelithiasis, recurrent acute pancreatitis, gallbladder carcinoma, Cystolithiasis, choledocholithiasis, biliary stricture and hepatic abscess.

Presentation and role of peritoneoscopy in the diagnosis of tuberculous peritonitis.

This study represents the clinical and laboratory features of 135 tuberculous peritonitis cases in whom peritoneoscopic investigation was used routinely. Disease was more common in women than men (1.5:1) and was most frequently encountered in the third and fourth decades in life. The most common presenting symptoms were abdominal distension (96%), abdominal pain (82%), weight loss (80%), weakness (76%), loss of appetite (73%) and fever (69%). The most common physical findings were ascites (96%), fever (75%) and abdominal tenderness (43%). One hundred and twenty-nine cases (95.5%) showed exudative type tuberculous peritonitis with variable amounts of ascites and filmy adhesions. In six patients (4.5%) the disease was of the plastic (dry) type. Peritoneoscopic investigations of 139 patients suggested tuberculous peritonitis but four cases showed histologically proven malignancy (3%). Laparoscopic diagnoses of the remaining cases were confirmed by histology (97%). The laparoscopic appearance of scattered yellowish-white nodules, approximately 1-5 mm in size, on the peritoneal surfaces, and filmy adhesions were suggestive of tuberculous peritonitis. A non-fatal colon perforation occurred as a major complication. After antituberculous therapy patients were followed for at least 1 year. Peritoneoscopy with simultaneous biopsy is the ideal and most accurate diagnostic modality in the diagnosis of tuberculous peritonitis.

Circulating IL-2 and IL-10 in chronic active hepatitis C with respect to the response to IFN treatment.

BACKGROUND: The importance of circulating immunoregulatory cytokines in response to IFN treatment and the change of in vivo production of these cytokines during interferon (IFN) treatment are not well known. We aimed to determine whether pretreatment serum levels of IL-2 and IL-10 are predictive of the response to IFN treatment and to investigate if treatment response or nonresponse has any effect on the circulating levels of these cytokines. PATIENTS AND METHODS: 37 patients (18 responders and 19 non-responders) with chronic hepatitis C virus (HCV) infection who received IFN-alpha2b for 6 months were studied. Responders were defined by complete alanine aminotransferase (ALT) normalization and loss of HCV RNA as detected by bDNA assay while patients who had elevated ALT levels and positive HCV RNA after 6 months were considered as nonresponders. RESULTS: Genotype distribution, ALT and HCV RNA levels were similar in responders and nonresponders. A significant number of patients with chronic hepatitis C (20/37 = 54%) had elevated IL-2 levels while IL-10 levels were not different from controls. No difference in baseline cytokine levels was observed between responders and non-responders. In the posttreatment serum samples some patients lost their detectable IL-2 or IL-10; some patients developed detectable cytokine levels after treatment irrespective of the treatment response. CONCLUSION: These results suggest that active liver injury in chronic hepatitis C is associated with increased circulating Th1 cytokine IL-2 but not with Th2 cytokine IL-10 and that circulating levels of these cytokines do not predict the response to IFN treatment. There is no constant and regular change in circulating levels of these cytokines under IFN treatment with respect to treatment response.

Risk factors for hepatocellular carcinoma in Turkey.

The contribution of hepatitis B, hepatitis C, and excess alcohol intake to the development of hepatocellular carcinoma in Turkey was assessed. The study was conducted through a questionnaire sent to seven major medical referral centers in different regions of Turkey and is based on 207 patients seen in the period 1994-1997. Of the seven centers, two were located in West Turkey (54 patients), two were in Central Turkey (85 patients), and two were in south and southeast Turkey (68 patients). In 196 of the 207 patients (94.7%), there was a history of chronic liver disease, and in 180 patients (87%) liver cirrhosis was documented. Of the 207 patients, 116 (56%) had hepatitis B, 48 (23.2%) had hepatitis C, and 33 (15.9%) had a history of excess alcohol intake. Anti-delta testing was available in 69 of 116 patients with hepatitis B, and anti-HDV was positive in 13 of these patients (13/69, 18.8%). Of the 33 patients with a history of heavy alcohol intake, 18 had concomitant chronic viral hepatitis infection, and alcohol alone was the etiology of hepatocellular carcinoma in only 15 cases (7.2%). The distribution of etiologic factors was not homogenous in different geographical regions in Turkey. In central, south, and southeastern Turkey, the predominant etiology of hepatocellular carcinoma was hepatitis B, whereas in western Turkey the impact of hepatitis B, hepatitis C, and alcohol was similar. This study indicates that hepatitis B virus infection is the leading cause of hepatocellular carcinoma in Turkey, followed by hepatitis C infection and alcoholic liver disease.