中文English
ISSN 1001-5256 (Print)
ISSN 2097-3497 (Online)
CN 22-1108/R

2016 No. 3

Display Method:
Editorial
Metabolic syndrome and fatty liver
Fan JianGao, Yan ShiYan
2016, 32(3): 407-410. DOI: 10.3969/j.issn.1001-5256.2016.03.001
Abstract:
There is a close relationship of metabolic syndrome( MS) with chronic liver disease,especially fatty liver. There have been many studies on the clinical features,pathogenesis,diagnosis,and treatment of MS with fatty liver,but controversy still exists. This article reviews the latest progress and difficulties in the clinical studies of MS and related fatty liver,alcoholic liver disease,and chronic viral hepatitis,so as to help physicians improve the management of MS and fatty liver.
Therapeutic guidelines
An interpretation of recommendations for expert consensus on the diagnosis and treatment of primary sclerosing cholangitis(2015)
Han Ying, Zhu JiangYi
2016, 32(3): 420-422. DOI: 10.3969/j.issn.1001-5256.2016.03.003
Abstract:
Recommendations for APASL Clinical Practice Guidelines: management of hepatitis B (updated in 2015)
Zhu Peng, Tang Yi, Wang YuMing
2016, 32(3): 423-428. DOI: 10.3969/j.issn.1001-5256.2016.03.004
Abstract:
Recommendations for EASL clinical practice guidelines: liver transplantation(2015)
Shi YingHong, Tian MengXin, Fan Jia
2016, 32(3): 429-431. DOI: 10.3969/j.issn.1001-5256.2016.03.005
Abstract:
Discussions by experts
Nonalcoholic fatty liver disease and hepatocellular carcinoma
Li LiangPing
2016, 32(3): 432-436. DOI: 10.3969/j.issn.1001-5256.2016.03.006
Abstract:
As the etiology of hepatocellular carcinoma( HCC) has been changing,the incidence of HCC related to nonalcoholic fatty liver disease( NAFLD) is gradually increasing in developed countries in Europe and America and some countries in Asia. This article introduces the close association between NAFLD and HCC,risk factors,clinicopathological features,and prevention and screening,and points out that although the incidence of NAFLD is not as high as that of hepatitis B- or hepatitis C- related HCC,there are a large absolute number of NAFLD patients,especially the high- risk patients with diabetes and obesity,or liver fibrosis / cirrhosis,due to a huge base number of NAFLD patients. NAFLD- related HCC is commonly seen in the elderly with various comorbidities and a poor prognosis. This article also points out that the prevention should focus on the effective treatment of NAFLD. The strict screening of high- risk population is the strategy for the diagnosis of early- stage HCC. At present,the sensitivity of alpha- fetoprotein is relatively low,and imaging examinations including computed tomography are the main screening methods; however,there are no measures for early warning of NAFLD- related HCC.
Nonalcoholic fatty liver disease and serum uric acid
Xu BeiBei, Wang BingYuan
2016, 32(3): 437-441. DOI: 10.3969/j.issn.1001-5256.2016.03.007
Abstract:
Nonalcoholic fatty liver disease( NAFLD) is considered the manifestation of metabolic syndrome( MS) in the liver. Besides glucose and lipid metabolic disorders,the level of serum uric acid( SUA) is also associated with the progression of NAFLD. This article reviews the research achievements in the association between SUA and NAFLD and points out that SUA can independently predict the risks of MS,type 2 diabetes,and cardiovascular disease in both healthy people and patients. Its mechanism may be that SUA increases the expression of reactive oxygen species( ROS) / thioredoxin- interacting protein( TXNIP) through inducing ROS,and then it activates the NLR pyrin domain containing 3 inflammasome and induces the secretion of interleukin. Both basic and clinical research show that the drugs reducing SUA can inhibit the TXNIP pathway,reduce the blood glucose level,and alleviate liver ROS,inflammation,steatosis,and fibrosis. This article suggests that SUA may be a promising therapeutic method for NAFLD and needs further basic and clinical research.
Role of cell senescence in development and progression of nonalcoholic fatty liver disease
Zhang Jing, Wang BingYuan, Shi JunPing
2016, 32(3): 442-445. DOI: 10.3969/j.issn.1001-5256.2016.03.008
Abstract:
Nonalcoholic fatty liver disease( NAFLD) is a clinicopathological syndrome characterized by fatty degeneration of hepatocytes due to nonalcoholic reasons. Liver cirrhosis and hepatocellular carcinoma are two important outcomes of NAFLD. Cell senescence is a relatively stable state in which cells deviate from the normal cell cycle and experience an irreversible loss of proliferative capacity,which is an objective law in cell life. Research shows that cell senescence is closely associated with the progression of liver fibrosis and the development and progression of hepatocellular carcinoma in NAFLD,and provides a new perspective for the prevention and treatment of liver fibrosis and hepatocellular carcinoma.
Research advances in susceptibility genes and their role in the pathogenesis of nonalcoholic fatty liver disease
Xuan ShiYing, Yuan Chen, Lu LinLin, Xin YongNing
2016, 32(3): 446-452. DOI: 10.3969/j.issn.1001-5256.2016.03.009
Abstract(2129) PDF (1559KB)(1042)
Abstract:
Currently the incidence of nonalcoholic fatty liver disease( NAFLD) is increasing,and the age of onset is getting younger worldwide,resulting in a heavy economic burden for both individuals and the society. Since NAFLD is closely related to heredity,metabolism,and the environment,genetic factors play an important role in the development and progression of NAFLD. With the development and wide application of the techniques from the genome- wide association studies,new research advances have been achieved in the susceptibility genes of NAFLD. This review summarizes the related research findings at home and abroad,and investigates the pathogenic factors for NAFLD and related mechanisms with a focus on the polymorphisms of susceptibility genes.
Original articles_ Fatty liver and metabolic syndrome
Prevalence of nonalcoholic fatty liver disease and its prognostic factors
Ni ManMan, Zhang YingQiu, Chen YueXiang, Zhang PingPing, Ceng Xin
2016, 32(3): 453-458. DOI: 10.3969/j.issn.1001-5256.2016.03.010
Abstract:

Objective To investigate the prevalence,natural history,and causes of death of nonalcoholic fatty liver disease( NAFLD),as well as related influencing factors. Methods A total of 833 retired cadres and staff members who underwent physical examination in Shanghai Changzheng Hospital and Shanghai 85 Hospital of the PLA from January 1 to December 31,2011 and received follow- up visits in either hospital every year since 2011 were enrolled as study subjects,and were divided into NAFLD group( 459 patients who were diagnosed with NAFLD before December 31,2011) and control group( 374 patients without liver or biliary diseases). The patients' clinical data were collected,including body height,body weight,systolic pressure,diastolic pressure,blood biochemical parameters,presence or absence of diabetes,hyperlipidemia,cerebrovascular and cardiovascular diseases,and malignant tumor,and smoking and drinking,and the death time and causes of death were clarified for the patients who died. The prevalence and natural course of NAFLD and related risk factors and prognostic factors were analyzed in this population. The t- test was applied for comparison of continuous data between groups,the chi- square test was applied for comparison of categorical data between groups,the multivariate binary logistic regression was applied to analyze the risk factors for the pathogenesis of NAFLD,and the multinomial logistic regression was applied to analyze the influencing factors for aggravation or alleviation of NAFLD. Results The patients in NAFLD group accounted for 55. 1% of all subjects,and the proportion of male patients was higher than that of female patients( 58. 0% vs 46. 7%,χ2= 4. 962,P = 0. 026). Compared with the control group,the NAFLD group had significantly higher body mass index( BMI),systolic pressure,diastolic pressure,alanine aminotransferase( ALT),fasting blood glucose,serum uric acid,and triglyceride( TG),a significantly higher proportion of male patients,and significantly higher proportions of patients with hypertension,diabetes,metabolic syndrome,newly- onset diabetes,and newly- onset metabolic syndrome( all P < 0. 05),as well as a significantly lower high- density lipoprotein cholesterol level( t =- 9. 818,P < 0. 001). Of all 374 patients in the control group,64( 17. 1%) had newly- onset NAFLD,and the incidence rate of newly- onset NAFLD showed no significant difference between male and female patients( 18% vs 15%,χ2= 0. 349,P > 0. 05); BMI and TG were the independent risk factors for newly- onset NAFLD( OR =1. 304,95% CI: 1. 153- 1. 474,P < 0. 001; OR = 1. 509,95% CI: 1. 080- 2. 166,P = 0. 016). At the end of follow- up in NAFLD group,235 patients( 51. 2%) achieved a stable disease,40( 8. 7%) achieved improvement,144( 31. 4%) achieved remission,and 40( 8. 7%) experienced aggravation. The increase in BMI increased the risk of aggravation of NAFLD( OR = 1. 163,95% CI: 1. 028- 1. 316,P = 0. 016),while reductions in BMI and serum uric acid were the protective factors against NAFLD( OR = 0. 849,95% CI: 0. 781-0. 923,P < 0. 001; OR = 0. 997,95% CI: 0. 995- 1. 000,P = 0. 038). Ten patients in NAFLD group died( including 5 patients with malignant tumor and 2 patients with cerebrovascular and cardiovascular diseases),and 6 patients in the control group died( including 2 patients with malignant tumor,no patients with cerebrovascular and cardiovascular diseases),and the risk of death showed no significant difference between the two groups( P > 0. 05). Conclusion The prognosis of NAFLD is relatively good in people who underwent physical examination,and BMI,TG,and serum uric acid are the factors affecting the prognosis of such patients. The major causes of death in NAFLD patients are malignant tumor and cerebrovascular and cardiovascular diseases,rather than end- stage liver disease.

Clinical significance of Fibro Touch,ultrasound,and computed tomography in diagnosis of fatty liver disease: a comparative analysis
Li JingBo, Liu Shu, Wen Bo, Gao Nan, Wang BingYuan
2016, 32(3): 459-462. DOI: 10.3969/j.issn.1001-5256.2016.03.011
Abstract:

Objective To investigate the clinical significance of image- guided transient elastography Fibro Touch with fat attenuation parameter( FAP) in the diagnosis of fatty liver disease. Methods A total of 1080 persons who underwent physical examination in The First Affiliated Hospital of China Medical University from January to December,2014 were enrolled,and liver FAP,the results of color Doppler ultrasound for the liver,and liver / spleen CT ratio on computed tomography( CT) scan for the liver were collected and analyzed. The Kruskal- Wallis H test was applied for comparison of continuous data between groups,and the Kappa consistency coefficient and receiver operating characteristic curve were used to investigate the consistency between FAP and the results of color Doppler ultrasound for the liver or liver / spleen CT ratio on CT scan for the liver in the diagnosis of fatty liver disease and their diagnostic values. Results Liver ultrasound found 336 patients( 31. 11%) without fatty liver disease and 744 patients( 68. 89%) with fatty liver disease; with reference to the results of ultrasound,the sensitivity and specificity of Fibro Touch were 80. 5% and 75. 3%,respectively. Liver CT found 821 patients( 76. 02%)without fatty liver disease( grade 0) and 259 patients with fatty liver disease,among whom 216 had mild fatty liver disease( grade 1),34 had moderate fatty liver disease( grade 2),and 9 had severe fatty liver disease( grade 3); with reference to the results of liver CT,the sensitivity and specificity of Fibro Touch were 95. 4% and 47. 0%,respectively,and the sensitivity and specificity of ultrasound were 93. 1%and 36. 8%,respectively. As for the diagnosis of fatty liver disease,the results of Fibro Touch had a higher consistency with ultrasound results( Kappa coefficient = 0. 597,P < 0. 0001),and a lower consistency with CT results( Kappa coefficient = 0. 113,P < 0. 0001). Conclusion Fibro Touch has the advantages of high accuracy,easy operation,noninvasiveness,and low cost,and therefore it is a promising imaging method for the diagnosis of fatty liver disease.

Current status of nonalcoholic fatty liver disease and related risk factors: an analysis of an enterprise in Xi'an
Zhou Juan, Niu ChunYan, Wang Dan, Zhao HongXin, Yan YingLi, Yan Rong
2016, 32(3): 463-466. DOI: 10.3969/j.issn.1001-5256.2016.03.012
Abstract:

Objective To investigate the current status of nonalcoholic fatty liver disease( NAFLD) in an enterprise in Xi'an,China,and to provide a scientific basis for further studies on NAFLD in Shanxi. Methods A total of 415 employees who underwent health examination in The First Affiliated Hospital of Xi'an Medical University from June to July,2014 were enrolled. All the patients underwent physical examination,abdominal ultrasound,and serological examination,and completed related questionnaires. The t- test was applied for comparison of continuous data,and the Pearson correlation analysis was applied for correlation analysis. Results Of all the 415 patients,86( 20. 7%)had NAFLD,consisting of 70 patients( 16. 9%) with mild NAFLD and 16 patients( 3. 8%) with moderate NAFLD. The population with an age of 50- 58 years had the highest prevalence rate( 30. 8%,16 /52). Of all the 86 patients with NAFLD,57( 66. 3%) had obesity,and76( 88. 4%) had abdominal obesity. The employees with moderate NAFLD had a significantly higher alanine aminotransferase( ALT) level,a significantly higher aspartate aminotransferase( AST) level,a significantly larger abdominal circumference,and a significantly higher body mass index( BMI) than those with mild NAFLD( t =- 4. 140,- 3. 512,- 4. 405,and- 5. 051,respectively,all P < 0. 05). In the employees with NAFLD,BMI and abdominal circumference were correlated with AST level( r = 0. 283 and 0. 271,both P < 0. 05) and ALT level( r = 0. 325 and 0. 316,both P < 0. 05). Most employees in this enterprise had a bad eating habit,a sedentary lifestyle,and a lack of physical exercise. Conclusion This enterprise has a high prevalence rate of NAFLD. BMI and abdominal circumference of patients with NAFLD have certain clinical values in predicting the degree of fat deposition in the liver,and are correlated with AST and ALT levels.

Original articles_Viral hepatitis
Effects of serum levels of chemokines on sustained virological response in patients with HBeAg-positive chronic hep-atitis B after treatment with pegylated interferon α-2b
Li Yan, Wang Xia, Cheng LiWei, Pan XiuCheng, Dan Ben, Geng Jian, Liu RuiXia, Fu JuanJuan, Li Li
2016, 32(3): 467-471. DOI: 10.3969/j.issn.1001-5256.2016.03.013
Abstract:
Objective To investigate the changes in serum levels of interferon( IFN)- γ- inducible protein 10( IP- 10),monokine induced by IFN- γ( Mig),and chemokine regulated upon activation,normal T cell expressed and secreted( RANTES) in patients with HBe Ag- positive chronic hepatitis B( CHB) undergoing pegylated interferon( PEG- IFN) therapy,and to explore their predictive values for the efficacy of PEG- IFN therapy. Methods Fifty- three patients with HBe Ag- positive CHB who received PEG- IFN α- 2b therapy in our hospital from February 2012 to December 2013 were enrolled as subjects. Forty- six out of the fifty- three patients were followed up for 24 weeks after 48 weeks of treatment. According to the incidence of sustained virological response( SVR),these patients were divided into A group( SVR,n = 17) and B group( NO SVR,n = 29). Serum levels of IP- 10,Mig,RANTES,hepatitis B virus( HBV) DNA,HBs Ag,and alanine aminotransferase( ALT) in peripheral venous blood were measured at baseline,after 12,24,and 48 weeks of treatment,and at 24 weeks after treatment. Comparison of continuous data between two groups and within the groups was made by t test and paired t test,respectively. Correlation was analyzed using the Pearson correlation coefficient. Between- group comparison of categorical data was made by χ2test. Results Compared with the B group,the A group had significantly higher baseline serum levels of IP- 10 and Mig and a significantly lower level of HBs Ag( t = 2. 696,P < 0. 05; t = 2. 963,P < 0. 05; t = 2. 401,P < 0. 05). The baseline levels of IP- 10 and Mig were positively correlated with the baseline level of ALT( r = 0. 570,P < 0. 05; r = 0. 317,P < 0. 05). Compared with the baseline levels,patients had significantly reduced serum levels of HBV DNA,HBs Ag,ALT,and Mig after 12 weeks of treatment( t = 2. 126,P <0. 05; t = 2. 217,P < 0. 05; t = 2. 376,P < 0. 05; t = 2. 776,P < 0. 05) and 24 weeks of treatment( t = 2. 635,P < 0. 05; t = 2. 453,P <0. 05; t = 2. 627,P < 0. 05; t = 2. 803,P < 0. 05). There were significant differences in levels of IP- 10 and Mig between the SVR group and non- SVR group before and after 12 and 24 weeks of treatment( all P < 0. 05). In the A group,the levels of Mig after 12 and 24 weeks of treatment were significantly lower than the baseline level( t = 3. 061,P < 0. 01; t = 3. 105,P < 0. 01). In the B group,the serum level of Mig after 24 weeks of treatment was significantly lower than the baseline level( t = 2. 632,P < 0. 01). The baseline serum levels of IP-10,Mig,and HBs Ag had predictive values for SVR( all P < 0. 05). In the 46 patients,the incidence of SVR in patients with baseline levels of Mig higher than 80 pg / ml( 12 /20) was significantly higher than that in patients with baseline levels of Mig lower than 80 pg / ml( 5 /26)( 60. 0% vs 19. 2%,χ2= 8. 06,P < 0. 01); the incidence of SVR in patients with baseline levels of IP- 10 higher than 120 pg / ml( 12 /25) was significantly higher than that in patients with baseline levels of IP- 10 lower than 120 pg/ml( 5 /21)( 48. 0% vs 23. 8%,χ2= 3. 86,P < 0. 05). Conclusion The baseline serum levels of IP- 10 and Mig are both predictors of the incidence of SVR in patients with HBe Ag- positive CHB undergoing PEG- IFN therapy,and Mig provides superior prediction.
Correlation of HBV DNA load with liver pathology,liver function,and serological markers of liver fibrosis in children with chronic hepatitis B
Qin XiaoMei, Fan ChenYing, Li ShuangJie, Peng HuiYun
2016, 32(3): 472-475. DOI: 10.3969/j.issn.1001-5256.2016.03.014
Abstract:
Objective To investigate the correlation of HBV DNA load with liver pathology,liver function,and serological markers of liver fibrosis in children with chronic hepatitis B( CHB). Methods A total of 79 children with a confirmed diagnosis of CHB who were admitted to Liver Research Center of Children's Hospital of Hunan Province from July 2010 to June 2015 were selected,and according to the serum HBV DNA load,they were divided into low load group( 103< HBV DNA load≤105copies / ml,n = 8),medium load group( 105< HBV DNA load ≤107copies / ml,n = 54),and high load group( HBV DNA load > 107 copies / ml,n = 17). The degree of liver pathological changes and serum levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),procollagen type Ⅲ( PC Ⅲ),laminin( LN),hyaluronic acid( HA),and type Ⅳ collagen( Ⅳ- C) were compared between the groups. The Kruskal- Wallis rank sum test was applied for comparison of continuous data between groups,and Kendall's tau- b rank correlation analysis was applied for correlation analysis. Results The three groups mainly had a liver inflammation grade of G1,which accounted for 75%,74. 1%,and 64. 7%,respectively,and the high load group had an increased proportion of G2 patients( 35. 5%). The three groups mainly had a fibrosis stage of S1,which accounted for 75. 0%,72. 2%,and 70. 6%,respectively,and there were S4 patients in the low load group( 12. 5%). HBV DNA load was not correlated with liver inflammation grade or fibrosis stage( r = 0. 069 and- 0. 047,both P > 0. 05). The medium load group had higher serum levels of ALT and AST than the low load group,and the high load group had higher serum levels of ALT and AST than the medium load group; serum ALT level showed a significant difference between the three groups( χ2= 5. 37,P = 0. 048). The medium load group had lower serum levels of HA,LN,PCⅢ,and Ⅳ- C than the low load group,and the high load group had lower serum levels of HA,LN,PCⅢ,and Ⅳ- C than the medium group; serum PCⅢ level showed a significant difference between the three groups( χ2= 6. 26,P = 0. 044).Conclusion In children with CHB,HBV DNA load is not significantly correlated with liver pathology. A comprehensive analysis of HBV DNA load,liver function,and serological markers of liver fibrosis in clinical practice in combination with liver biopsy can achieve an objective and accurate evaluation of the conditions of children with CHB.
Classification tree model analysis of influencing factors for hepatocyte steatosis in patients with chronic hepatitis B
Wang Peng, Zhang ZhiQiao, Kang KaiFu, Wu GuoBiao
2016, 32(3): 476-479. DOI: 10.3969/j.issn.1001-5256.2016.03.015
Abstract:
Objective To investigate the influencing factors for hepatocyte steatosis in patients with chronic hepatitis B( CHB) and the high- risk population by classification tree model analysis,and to establish a simple method to assess the risk of hepatocyte steatosis in CHB patients. Methods The clinical data and pathological results of the CHB patients who underwent liver biopsy in Department of Infectious Diseases,The First People's Hospital of Shunde,from January 2006 and December 2014 were collected. The classification tree model was applied to analyze the influencing factors for hepatocyte steatosis,and index value curve,misclassification matrix,and error of estimation were applied for overall evaluation of classification results of the classification tree model. Results The influencing factors for hepatocyte steatosis in CHB patients were body mass index( BMI),total cholesterol,and low- density lipoprotein,and the most important factor was BMI. This classification tree model had a sensitivity of 84. 3%,a specificity of 81. 5%,an accuracy of 82. 9%,and an error of estimation of 0. 171,suggesting that this model was well fitted. Conclusion Classification tree model analysis shows that the pathogenesis of hepatocyte steatosis in CHB patients is closely related to the influencing factors BMI,total cholesterol,and low- density lipoprotein. A simple classification method is established based on these factors to evaluate the risk of hepatocyte steatosis in CHB patients. It is necessary to conduct further clinical studies to investigate the clinical value of this method.
Genotypes and resistance loci in patients with chronic hepatitis B achieving virologic breakthrough in treatment with lamivudine combined with adefovir after developing drug resistance in treatment with lamivudine alone
Yang DongQiang, Chen Fang, Kang Yi, Ding GangQiang, Xiao ErHui, Peng Zhen, Shang Jia
2016, 32(3): 480-483. DOI: 10.3969/j.issn.1001-5256.2016.03.016
Abstract:
Objective To investigate the genotypes and resistance loci in patients with chronic hepatitis B( CHB) achieving virologic breakthrough in the treatment with lamivudine( LAM) combined with adefovir( ADV) after developing drug resistance in the treatment with LAM alone. Methods A total of 89 CHB patients who were admitted to Henan Provincial People's Hospital and treated with LAM and ADV as the antiviral therapy after developing drug resistance in the treatment with LAM alone from June 2010 to June 2013 were enrolled,and their serum samples were collected. Quantitative real- time PCR was used for hepatitis B virus( HBV) detection,and sequencing was applied to determine the genotype and detect the resistance loci rt N236 T,rt A181 V,rt M204 V,and rt L180 M. The t- test was applied for comparison of continuous data between groups,and the chi- square test was applied for comparison of categorical data between groups. Results LAM rt M204 V mutation was detected in all the 89 patients after treatment with LAM alone. After the combination therapy with LAM and ADV,ADV rt N236 T mutation was detected in 9 patients,ADV rt A181 V mutation in 5 patients,and rt N236 T + rt A181 V mutation in 8 patients,with an overall ADV resistance rate of 24. 7%( 22 /89). As for HBV genotyping,82 patients had genotype C,and rt A181 V + rt N236 T mutation was detected in 8 patients; 7 patients had genotype B. Conclusion Compared with the patients with HBV genotype B,the patients with genotype C are more susceptible to rt A181 V + rt N236 T mutation during the combination therapy with LAM and ADV after developing drug resistance in the treatment with LAM alone.
Effects of HBV X protein on expression and promoter methylation of p16 tumor suppressor gene
Kang YanHong, Li Wei, Zhan WeiLi, Shang Jia, Yang Lin
2016, 32(3): 484-487. DOI: 10.3969/j.issn.1001-5256.2016.03.017
Abstract:
Objective To explore the effects of hepatitis B virus X protein( HBx) on the expression and promoter methylation of the p16 tumor suppressor gene,and to investigate the epigenetic role of HBx in the development and progression of hepatitis B virus( HBV)- associated hepatocellular carcinomas( HCC). Methods Experiments were performed in the human hepatoblastoma cell line Hep G2,Hep G2 cells expressing green fluorescent protein( Hep G2 / GFP),and Hep G2 cells stably expressing GFP- HBx fusion protein( Hep G2 / GFP- HBx).Western blot was used to determine the expression levels of the p16 protein in Hep G2 cells,Hep G2 / GFP cells,and Hep G2 / GFP- HBx cells. Hep G2 / GFP- HBx cells were treated with a universal inhibitor of DNA methyltransferase( DNMT),5- aza- 2'- deoxycytidine( 5-aza- 2'- d C). Methylation- specific polymerase chain reaction( MSP) was used to determine the promoter methylation of the p16 tumor suppressor gene in Hep G2 cells,Hep G2 / GFP cells,and Hep G2 / GFP- HBx cells treated with or without 5- aza- 2'- d C. Multiple- group comparison was made by analysis of variance. Results According to the results of Western blot,Hep G2 / GFP- HBx cells had a significantly