Clinical and Molecular Hepatology

"PCR"

Original Article

Steatotic liver disease

Enhanced A-FABP expression in visceral fat: potential contributor to the progression of NASH: Min Yong Yoon, Jun Mo Sung, Chang Seok Song, Won Young Lee, Eun Jung Rhee, Jun Ho Shin, Chang Hak Yoo, Seoung Wan Chae, Ja Yeon Kim, Wook Jin, Yong Kyun Cho; Korean J Hepatol 2012;18(3):279-286.
Published online September 25, 2012; DOI: https://doi.org/10.3350/cmh.2012.18.3.279

Abstract
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Background/Aims

Adipose tissue is an active endocrine organ that secretes various metabolically important substances including adipokines, which represent a link between insulin resistance and nonalcoholic steatohepatitis (NASH). The factors responsible for the progression from simple steatosis to steatohepatitis remain elusive, but adipokine imbalance may play a pivotal role. We evaluated the expressions of adipokines such as visfatin, adipocyte-fatty-acid-binding protein (A-FABP), and retinol-binding protein-4 (RBP-4) in serum and tissue. The aim was to discover whether these adipokines are potential predictors of NASH.

Methods

Polymerase chain reaction, quantification of mRNA, and Western blots encoding A-FABP, RBP-4, and visfatin were used to study tissue samples from the liver, and visceral and subcutaneous adipose tissue. The tissue samples were from biopsy specimens obtained from patients with proven NASH who were undergoing laparoscopic cholecystectomy due to gallbladder polyps.

Results

Patients were classified into two groups: NASH, n=10 and non-NASH, n=20 according to their nonalcoholic fatty liver disease Activity Score. Although serum A-FABP levels did not differ between the two groups, the expressions of A-FABP mRNA and protein in the visceral adipose tissue were significantly higher in NASH group than in non-NASH group (104.34 vs. 97.05, P<0.05, and 190.01 vs. 95.15, P<0.01, respectively). Furthermore, the A-FABP protein expression ratio between visceral adipose tissue and liver was higher in NASH group than in non-NASH group (4.38 vs. 1.64, P<0.05).

Conclusions

NASH patients had higher levels of A-FABP expression in their visceral fat compared to non-NASH patients. This differential A-FABP expression may predispose patients to the progressive form of NASH.

Citations

Citations to this article as recorded by

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Abstract: Oral

Localization of Hepatitis B Virus DNA in Hepatocellular carcinoma by PCR In Situ Hybridization: 조성원, 함기백, 이은섭, 윤미영, 김진홍, 이상인, 박광화, 심철; Korean J Hepatol 1995;1(1):110-110.

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Abstract: Poster

HCV 감염이 의심되는 대상에 있어서 혈청 HCV RT - PCR 의 의의: 서정일, 이헌주, 이채훈; Korean J Hepatol 1995;1(1):123-123.

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Original Articles

The Detection of Hepatitis G Virus RNA by RT - PCR in Various Liver Diseases: Kwang Hyub Han , Won Choi , Young Nyun Park , Young Woong Hwang , Wang Sik Ryu , Eun Sin Park , Kwan Sik Lee , Chae Yoon Chon , Young Myoung Moon , Chan Il Park; Korean J Hepatol 1997;3(2):123-132.

Abstract
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Background/Aims
Recently, nucleotide sequences from a novel virus, termed hepatitis G virus (HGV), were identified in serum from a patient with cryptogenic hepatitis and suggested as agent of non A-E hepatitis. HGV has been isolated from patients with various liver diseases but clinical implications of this new agent remain largely unresolved. In Korea, the etiology of substantial fraction of hepatitis has remained undefined and there has been no report concerning HGV. Methods: To determine the infection rate of HGV, RT-PCR of 5 UTR of HGV was performed, and to understand the clinical implication of HGV, medical records of 115 patients with various liver diseases were reviewed. Of 115 patients, 63 were male and 52 were female. Their mean age was 44 years (19-74) and their mean AST and ALT were 121.3+278.7 IU/L and 172.2+253.3 IU/L, respectively. Of 115 patients, 58 (50.4%) had no specific cause of liver diseases, 37 (32.2%) were infected with hepatitis B and/or C virus and 20 (17.4%) had non-viral identifiable liver diseases. Results: 1. HGV RNA was detected in 15 (13.0%) patients of 115 patients. 2, Among the 15 HGV RNA positive cases, 7 were male and 8 were female. Their mean age was 48 years (19-72) and their mean AST and ALT were 71.9+45.2 IU/L, 97.4+66.8 IU/I respectively. 3. HGV RNA was detected in 8(13.8%) of 58 patients without obvious causes of their liver diseases and in 7 (18.9%) of 37 patients infected with HBV and/or HCV. However, HGV RNA was not detected fram 20 patients with non-viral liver diseases such as alcoholic liver diseases, autoimmune hepatitis, PBC, or fatty liver. 4. HGV RNA was detected in 5 (19.2%) of 26 patients with acute hep- atitis, in 6 (9.4%) of 64 patients with chronic hepatitis, in 1 (14.3%) of 7 patients with liver cirrhasis, and iB 3 (27.3%) Of 11 pafients with hepatocellular caIcinoma. 5. These was no slatistically significant difference in sex, age, history of transfusion, serum ALT level, etiologies and status of liver diseases between HGV RNA positve and negative group. Conclusions: the prevalence of HGV infection is quite high among the patients who have no specific cause of acute or chronic liver diseases and HGV can be coinfected with HBV and/ar HCV infection in Korea.

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Identification of eplication-Competent Dimeric Human Hepatitis B Viral Using PCR Screening Technique: Seok Hyun Nam, Ph.D.1, Sung Won Cho, M.D.2; Korean J Hepatol 1999;5(3):184-189.

Abstract
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Background/Aims
Transfection of the hepatitis B virus (HBV) genome requires the cloning of tandem HBV sequences inserted into a plasmid vector, which is usually screened for by the restriction enzyme digestion of plasmid minipreparation from at least a dozen of bacterial colonies. The aim of this study was to develop a simple alternative screening method for bacterial colonies harbouring tandem HBV sequences by a PCR. Methods: A set of polymerase chain reaction (PCR) primer was designed to detect the bacterial colonies harbouring "head to tail" dimeric HBV DNA. PCR which amplifies the head to tail junction site of two tandem HBV molecules was performed. Results: PCR products with appropriate size (1.2kb) were obtained. The accurate detection by PCR screening technique was confirmed by enzyme digestion. Conclusions: These results suggest that PCR screening technique is a simple and rapid method for the identification of bacterial colonies containing tandem HBV sequences. (Korean J Hepatol 1999;5:184－189)

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Identification of the Gene associated with Hepatocellular Carcinoma using Differential Gene Expresion: Jeong Yeob Song,Jeong Hee Choi,Kwang Jae Lee,Byung Moo Yoo,Ki Baik Hahm,Jin Hong Kim,Sung Won Cho; Korean J Hepatol 2001;7(3):265-272.

Abstract
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Background/Aims
It has been acknowleged that diverse factors such as Hepatitis B or C virus, alcohol, food carcinogens, and environmental or genetic factors are involved in hepatocellular carcinogenesis. In the molecular biologic aspect, suppression of tumor suppressor gene or amplification of oncogene, abnormal regulation of cell cycle-related proteins, abnormal apoptosis mechanism, and diverse growth factors are reported to be factors that contribute to hepatocellular carcinogenesis. In this study, the genetic difference between hepatocellular carcinoma tissue and surrounding non-hepatocellular carcinoma tissue has been investigated to identify genes that are deleted, diminished, amplified, or newly developed in hepatocellular carcinoma using differential gene expression.Method:We studied each of 12 biopsy samples of hepatocellular carcinoma and surrounding non-hepatocellular carcinoma tissues obtained during surgical resections. Random arbitrarily primed-polymerase chain reaction(RAP-PCR) was applied for differential gene expression. The genes that are deleted, diminished, or amplified, newly developed in hepatocellular carcinoma are cloned, sequenced, and then identified by BLAST search, some genes are characterized by eletrophoresis motility shift assay(EMSA) and in situ hybridization. Results:We identified the various, diverse genes classified as tumor suppressor genes, oncogenes, growth factor genes, and some kinds of transcription factors. Some of these genes were identified to be repressed, deleted or diminished, others were amplified, or newly developed in hepatocellular carcinoma tissues.Conclusions:RAP-PCR is a good method in the identification of the gene associated with hepatocellular carcinoma. The result in this study shows that so many genes are different between hepatocellular carcinoma and surrounding non- hepatocellular carcinoma tissues, and that the genes related with hepatocellular carcinogenesis may be predicted. Further studies are necessary for analyzing the relationship between the identification of the gene associated with hepatocellular carcinoma and the diverse factors involved in hepatocellular carcinogenesis. (Korean J Hepatol 2001;7:265-272)

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Distribution of HBV Genotypes in Patients With Chronic HBV Infection in Korea: Soong Hwan Lee, M.D., Sung Hee Han, M.D., Seung Chul Cho, M.D., Byung Joo Roh, M.D., Joo Hyun Sohn, M.D., Duck An Kim, M.D. *, Dong Hoo Lee, M.D., and Choon Suhk Kee, M.D.; Korean J Hepatol 2001;7(4):373-380.

Abstract
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Background
/ Aims : Choronic HBV infection is associated with a wide spectrum of clinical manifestations, including asymptomatic carrier state, choronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. Genotypically, HBV genomes have been calssified into seven groups, designated A to G. Several studies have suggested recently that HBV genotypic difference influence the severity of liver disease and clinical outcomes. The distribution of HBV genotypes in Korea and its clinical relevance are poorly understood. We investigated the prevalence of HBV genotypes in Korea and the association between the distinct genotypes and the severity of liver disease. Methods : A total of 214 HBV-DNA positive serum samples, were used for the genotyping. All patients were HBV-bDNA positive chronic HBsAg carries. 199 patients were histologically verified with liver cirrhosis(6), chronic hepatitis(192) and fatty liver(1). The other patients were clinically diagnosed with liver cirrhosis(13) or hepatocelluar carcinoma(2). HBV genotype was determined by PCR using type-specific primers. Results : Genotyping was possible in all patients. Out of 214 patients, 213(99.5%) were HBV genotype C. Only one(0.5%) was genotype A, The patient with genotype A had minimal hepatitis as diagnosed by liver biopsy. Conclusions : These results indicate that almost all chronic HBV infections are genotype C in Korea. HBV grnoytpic difference therefore dose not influence the clinical outcome of HBV infection in Korea. Because genotype C may be associated with more severe liver disease, the predominance of genotype C in Korea may result in more severe outcomes than in other countries where other genotypes are predominant. (Korean J Hepatol 2001;7 :373 - 380)