HBV-DNA Copies Research Articles

Enhanced immunity and antiviral effects of an HBV DNA vaccine delivered by a DC-targeting protein.

DNA vaccine targeting delivery to DC represents one effective strategy to improve the immunogenicity of the vaccine. In a previous study, we developed a novel DC-targeting recombinant protein that can deliver plasmid DNA to DCs by an electrostatic coupling effect and can thus improve the uptake efficiency of DCs, improving the expression of plasmid DNA in DCs. In this study, we coupled the protein with the HBV DNA vaccine pSVK-HBVA and investigated whether the immunogenicity and antiviral ability of the vaccine can be improved in HBV transgenic mice. The results show that a stronger specific immune response can be induced in mice after immunization with the coupling vaccine. The HBV DNA copy number and circulating antigen HBsAg in the serum of HBV transgenic mice were significantly decreased. Therefore, this study has demonstrated that the DC-targeting protein has the ability to improve the immunogenicity and the antiviral activity of the HBV DNA vaccine pSVK-HBVA. These findings indicate that this DC-targeting protein can be a potential method for the delivery of DNA vaccines directly to DCs.

Low-level expression of microRNA-375 predicts poor prognosis in hepatocellular carcinoma.

MicroRNAs are predicted to play fundamental roles in the tumorigenesis of hepatocellular carcinoma (HCC). MiR-375 is frequently downregulated in HCC and acts as a tumor suppressor by targeting multiple oncogenes. The objective of this study was to evaluate miR-375 expression and its relevance to the prognosis of HCC. MiR-375 expression was measured in cancerous tissues using qRT-PCR and dichotomized based on a median cutoff. The association between miR-375 expression and clinicopathological parameters and prognosis was subsequently determined. Expression levels of miR-375 were detected in a cohort of 38 HCC patients who underwent curative surgery. No significant correlations were observed between miR-375 expression and clinicopathological parameters, such as gender, age, performance status, preoperative serum AFP level, histological grade, HBV-DNA copy number, ascites, cirrhosis, tumor size, number of tumor nodules, and macrovascular invasion. However, miR-375 expression differs across CLIP scores significantly (p < 0.05). A trend toward poorer disease-free survival (DFS) was observed in patients with lower miR-375 expression compared to those with higher miR-375 expression (p = 0.307). Multivariate analysis demonstrated that low miR-375 expression was an independent prognostic predictor for progression (p = 0.032, risk ratio 3.273). Subgroup analysis revealed that low expression of miR-375 was significantly associated with adverse DFS in patients with poorly differentiated histology, higher serum AFP level (≥400ng/ml), and advanced tumor stage (CLIP score 1∼3) (p = 0.017, 0.009, and 0.024, respectively). Our study demonstrates that miR-375 expression is significantly correlated with DFS and may be a potential prognostic biomarker of disease progression in HCC.

Efficacy of telbivudine in the treatment of chronic hepatitis b and liver cirrhosis and its effect on immunological responses.

This study was aimed to evaluate the long-term effects of telbivudine (LdT) in the treatment of chronic hepatitis B (CHB) and HBV-related liver cirrhosis (LC) and to observe the changes of immunological responses during LdT treatment. Clinical data of 80 CHB and 28 HBV-related LC patients who were administered with LdT for 108 weeks and followed up were retrospectively analyzed. The liver function indicators including ALT, AST and γ-GT, HBV DNA copy number in serum and the rates of hepatitis B e antigen (HBeAg) seroconversion were analyzed before and 12, 24, 36, 48, 60, 72, 84, 96 and 108 weeks after LdT treatment in CHB and LC groups. Four serum fibrosis-related markers, including hyaluronic acid (HA), human laminin (LN), human type IV collagen (IV-C) and human N-terminal procollagen III peptide (PC-III), were detected before and after LdT treatment in LC group. The results showed favorable viral suppression and biochemical responses after treatment with LdT for 12 weeks, and a high rate of virological and biochemical control was maintained during the course of 108-week treatment in both CHB and LC groups. The four fibrosis-related markers, especially HA and LN, were down-regulated to some degrees in LC group. Moreover, LdT treatment led to the fluctuation of the circulating interferon-γ (IFN-γ) and interleukin-10 (IL-10) levels at different time points in CHB group. It was concluded that LdT could favorably lead to the virological suppression and biochemical remission. Besides, IFN-γ and IL-10 may represent a suitable and effective predictor of responsiveness during LdT therapy.

Impact of HBV DNA copies on clinical effects of chemotherapy combined with antiviral therapy in tumor patients with hepatitis B

Impact of HBV DNA copies on clinical effects of chemotherapy combined with antiviral therapy in tumor patients with hepatitis B

Analysis of HBV-DNA in Hepatitis B Patients with HBeAg Negative in JiLin Area

Objective: To illustrate the relationship between serum HBV-M and HBV-DNA in HBeAg-negative HBV patients. Methods: Serum isolated from HBV patients. FQ-PCR was used to detect the levels of HBV-DNA. ELISA was used to detect the serum HBV-M. Results: The positive rate of HBV-DNA was much higher in HBeAg-positive HBV patients (98.6%) than in HBeAg-negative HBV patients (48.7%). Among the different HBV-M patterns, the positive rate of HBV-DNA with positive HBsAg, HBeAb, and HBcAb was 59.4%; the positive rate of HBV-DNA with positive HBsAg and HBcAb was 32.3%; the positive rate of HBV-DNA with positive HBsAb, HBeAb, and HBcAb was 30.4%; the positive rate of HBV-DNA with positive HbeAb and HBcAb was 13.6%; the positive rate of HBV-DNA with positive HBcAb was 7.7%; the positive rate of HBV-DNA with positive HBsAb and HBcAb was 33.3%. Conclusion: HBV-DNA does still replicate in HBeAg negative HBV patients, and copies of HBV-DNA is a sensitive monitoring marker of monitoring HBV replication for those persons infected HBV.

Duck HBV Dna Copy Numbers in Isolated Hepatocyte Nuclei Vary Dramatically and Decline during Entecavir Therapy

We aimed to develop a quantitative assay to measure duck HBV (DHBV) DNA in single hepatocyte nuclei from DHBV-infected animals and to observe intranuclear DHBV DNA kinetics undergoing entecavir (ETV) therapy. DHBV DNA in isolated nuclei was amplified by quantitative real-time PCR. Liver tissues from chronically-infected ducks with or without ETV treatment were assessed. Cell cycle phases were defined with flow cytometry in single nuclei. We successfully established a quantitative assay to measure intranuclear DHBV DNA in single nuclei with high specificity, sensitivity and acceptable interassay variations. The intranuclear viral DNA copy numbers varied dramatically (2-204 copies/nuclei) in 11 ducks with active viral replication. Average intranuclear DHBV DNA copies from individual animals (7.57-57.67 copies/nuclei) significantly correlated with total intranuclear (rs=0.955, P<0.001) and serum (rs=0.745, P=0.008) viral DNA levels. The median intranuclear DHBV DNA copies in virus-positive nuclei were greater in gap 0/1 than those in gap 2/mitosis and synthesis phases (P<0.001). Median intranuclear viral DNA copies in virus-positive nuclei decreased from 21 to 6 (P<0.001) under 14-19 weeks of ETV therapy. However, subsequently, further reductions were not achieved in four animals after extended 16 week treatment (6 versus 11, P=0.034). Intranuclear DHBV DNA levels varied significantly, which could be partially attributed to effects of cell cycle phases, and could be decreased by ETV therapy.

Abnormal platelet kinetics are detected before the occurrence of thrombocytopaenia in HBV‐related liver disease

Thrombocytopaenia is a frequent feature in patients with HBV-related liver disease. Its underlying mechanism is not fully understood. Multiple factors might contribute to the development of thrombocytopaenia. In this study, we investigated the reticulated platelets (RP), glycocalicin (GC), serum thrombopoietin (TPO) and platelet glycoprotein (GP) in different stages of the disease. One hundred and fourteen patients with HBV-related liver disease (30 with chronic hepatitis B (CHB), 20 patients in Child A without thrombocytopaenia, 19 patients in Child A with thrombocytopaenia, 45 in Child B/C with thrombocytopaenia) and 25 normal controls (NC) were enrolled. Liver cirrhosis (LC) was classified according to modified Child-Turcotte-Pugh (CTP) score. Serum TPO levels and GC were measured by ELISA. RP and platelet glycoprotein (GP) expression were detected by flow cytometry. The TPO levels of patients with LC were significantly lower than that of controls, even in patients of Child A without thrombocytopaenia group. Serum TPO level was positively correlated (r = 0.65, p < 0.01) with serum albumin in Child B/C group. Both the RP percentages and the glycocalicin index (GCI) levels were significantly higher in patients groups including CHB and Child A without thrombocytopaenia than that of normal controls. A negative correlation existed in HBV DNA copies and the GPs% in patients with CHB and Child A without thrombocytopaenia. Abnormal platelet production, destruction and platelet-specific glycoproteins levels were detected before the occurrence of thrombocytopaenia in HBV-related liver disease, indicating that multiple mechanisms might play roles in thrombocytopaenia in HBV-infected patients.

Analysis of relationship of HBV PreS1 antigen, anti-HBc IgM, DNA load and genotypes in hepatitis B patients

To explore the relationship of HBV PreS1 antigen, anti-HBc IgM, DNA load and genotypes, and the significance for clinical diagnosis and prognostic. Enzyme linked immune-sorbent assay was used to test the HBV serum markers of HB patients; HBV-DNA copies was detected by time fluorescence quantitative PCR; using nested PCR to amplify the S fragment of HBV genome, then sequence and make blast with HBV standard sequences to ascertain genotypes. Make comprehensive analysis of these indexes. 355 serum specimens of acute or chronic HB patients were collected. The positive rates of HBV PreS1-Ag and HBV-DNA in model I (positive for HBeAg) were 80.2% and 73.7% respectively, which both higher than other models. The abnormal rate of ALT and AST were higher in PreS1-Ag positive group than negative, as well as in anti-HBc IgM positive group. There are 4 samples is genotype B (2.9%), 76 genotype C (55.9%) and 56 genotype D (41.2%). Positive rate of HBeAg and HBV-DNA of genotype C samples were both higher than which of genotype B and D. PreS1-Ag and Anti-HBe-IgM indexes are of great value to viral hepatitis B early diagnosis, HBV replication surveillance and prognostic evaluation; the major HBV genotypes in Henan province are C and D, and the positive rate of HBeAg and HBV-DNA were both higher in genotype C HBV infection population than genotype B and D.

The naturally occurring YMDD mutation among patients chronically infected HBV and untreated with lamivudine: a systematic review and meta-analysis.

BackgroundSeveral recent reports have demonstrated that tyrosine (Y)-methionine (M)-aspartic acid (D)-aspartic acid (D) (YMDD) motif mutations can naturally occur in chronic HBV patients without antiviral treatment such as lamivudine therapy. This paper aims to assess the overall spontaneous incidence and related risk factors of YMDD-motif mutations among lamivudine-naïve chronic HBV carriers, so as to provide some clue for clinical treatment of hepatitis B.Methodology/Principal FindingsChinese and English literatures were searched for studies reporting natural YMDD mutations among untreated chronic HBV patients from 2001 to 2010. The incidence estimates were summarized and analyzed by meta-analyses. Forty-seven eligible articles from eight countries were selected in this review (13 in English and 34 in Chinese). The pooled incidence of YMDD-motif mutation among untreated chronic HBV patients from eight countries was 12.21% (95% CI: 9.69%–14.95%). China had an incidence of 13.38% (95% CI: 10.90%–16.07%) and seven other countries had an incidence of 9.90% (95% CI: 3.28%–19.55%), respectively. Lamivudine therapy would increase the risk of mutations 5.23 times higher than the untreated patients. A higher HBV DNA copy number was associated with increased incidence of natural YMDD mutation. No significant difference was found in YMDD mutation incidence between groups of different gender, age, HBeAg status, patients' ALT (alanine aminotransferase) level, and between the groups of HBV genotype B and C.ConclusionsThe YMDD-motif mutations can occur spontaneously with a relatively high incidence in CHB patients untreated with lamivudine. These mutations might be the consequence of accumulated base mismatch due to the nature of viral polymerase. More fundamental and clinical studies are needed to clarify the influence of YMDD mutations in hepatitis B progression and antiviral treatment.

The clinical value of hepatitis B virus Pre-S1 antigen detection

Objectives To explore the clinical significance of detection of HBV Pre-S1 antigen in the diagnosis of Hepatitis B. Methods 254 blood samples with positive HBV markers detected by ELISA were divided into three groups based on different modes. Pre-S 1 antigen, HBV DNA, and ALT were detected in each group and then were comparatively analyzed. Results The detection rates for PreS1 antigen and HBV-DNA were 77.5％ and 93.1％ in the group with HbsAg ( + ), HBeAg ( + ), and HBcAb ( + ), 45.7％ and 60.1％ in the group with HBsAg ( + ), HBeAb ( + ), and HBcAb ( + ); and 0％ and 14.3％ in the group with HBsAg (-), HBeAb ( + ), and HBcAb ( + ). The positive rate of PreSl was 64.4％ ( 116/180 ) and A LT abnormality rate was 61.1％ ( 110/180 ) in 180 samples with a HBVDNA copy number of greater than 500IU/ml, and they were 35.1％ and 33.8％ in 74 samples with a copy number of less than 500IU/ml, respectively. The positive of Pre-S1 antigen was increased with an elevation in HBV DNA viral load, but the ALT abnormality rate was not linearly associated with the increase in HBV DNA viral load. Conclusions HBV Pre-Sl antigen has a higher consistency with HBV-DNA, which has an important clinical value in the diagnosis of type B hepatitis and can provide more reliable referrence for judging viral replication in vivo and efficacy of treatment. Key words: Hepatitis B virus; Pre-Sl antigen; HBV DNA viral load; ALT

Heat impact upon the infectivity of hepatitis B virus in serum

This article was to explore the impact of temperature on hepatitis B virus infectivity. HBV positive serum with a HBV DNA titer of 1.33 × 10(8) copies/ml was aliquots into 23 Ep tubes with 1.5 ml, 100 µl in one tube.15 tubes were incubated at 37, 56 and 65°C for 0, 30, 60, 120 and 600 minutes, respectively. The other 8 tubes were incubated at 98°C for 0, 5, 10 and 30 minutes, respectively. Post-treated serum at all time points were selected to infect HepG-2 cell. When 18 hours after infection, these cells were extensively washed with phosphate buffered saline. Cells were harvested after the addition of fresh culture medium to culture cells for 48 hours. HBV DNA was detected by FQ-PCR. HBV DNA was detected in cells that were infected by serum at 37°C and 56°C for 30, 60, 120 and 600 minutes, respectively. The titers for the cells incubated at 37°C were (4.85 ± 1.71) × 10(5), (3.85 ± 1.76) × 10(5), (1.67 ± 0.67) × 10(5), (7.86 ± 1.03) × 10(4) copies/ml, and those for the cells incubated at 56°C were (4.01 ± 0.16) × 10(5), (9.77 ± 0.97) × 10(4), (6.36 ± 0.65) × 10(4), (5.05 ± 0.24) × 10(3) copies/ml at different incubation time points. For the cells incubated at 65°C for 60 and 120 minutes, HBV DNAs were (5.15 ± 7.28) × 10(3) and (7.56 ± 10.60) × 10(2) copies/ml, respectively, which were much lower than those in the controls cells ((6.79 ± 1.48) × 10(5) copies/ml). The results of HBV DNA were different (F = 104.4, P < 0.001) in groups treated with different temperature, and results of HBV DNA were also different (F = 144.0, P < 0.001) in groups processed for different period of time. Temperature and processing time had interaction (F = 23.6, P < 0.001). After heating at 98°C for 10 minutes and boiling for 5 minutes, the HBV DNA copy number ((3.02 ± 4.26) × 10(2), (4.31 ± 6.09) × 10(2) copies/ml) in infected cells decreased by about 10 folds than that in the control group ((6.79 ± 1.48) × 10(5) copies/ml). HBV DNAs were not detected in cells that were infected by serum which was heated at 98°C for 30 minutes and boiled for 10 minutes. The infectivity of HBV serum in vitro was relatively stable at low temperature, and it would lose its infectivity in short period of time at high temperature.

The study on significance of detecting the hepatitis B virus large proteins in public health physical examination

It was to explore the application of the hepatitis B virus large proteins (HBV-LP) in public health physical examination through clinical testing of HBV-LP in Catering practitioners, students and soldiers of New requisition etc. 709 samples of HBV infection people were collected in public health physical examination and all collected samples were detected. ELISA was used to detect HBV serum markers (HBV-M) and HBV-LP. Real-time PCR was applied to detect quantity of HBV DNA. (1) The detectable results of HBV-LP and HBV DNA were no statistical significance in 709 samples. (2) The detectable results of HBV-LP and HBV DNA were also no statistical significance in different models of HBV infection patients. The values of chi2 and P were respectively 2.67, 0.60, 0.00 and 0.10, 0.44, 1.00. S/A of HBV-LP and the copies of HBV-DNA had a positive relationship (r = 0.959, P < 0.05). HBV-LP could reflect the level of hepatitis B virus replication and be used to judge the level of hepatitis B virus replication in people of public health physical examination. HBV-LP may be worth using widely in physical examination.

The expression and role of Toll receptor 4 in renal tubular epithelial cells in hepatitis B virus infection

To explore the expression and role of Toll receptor 4 (TLR4) in human proximal tubular epithelial cell line HK-2, infected by HBV. The serum of HBV DNA copies between 10(7) - 10(8)/ml was collected. Before and after infected by HBV DNA positive serum, the HK-2 cells' morphology and the expression of α-smooth muscle actin (α-SMA) were observed by microscopy and immunofluorescence, and the effects of different concentrations of lipopolysaccharides (LPS, TLR4-stimulating factor) and CLI-095 (TLR4 Inhibitor) on the proliferation rate of HK-2 cells were observed by MTT assays. After HBV serum and 10 µg/ml LPS and 5 µl/ml CLI-095 acted on HK-2 cells, TLR4 protein expression was measured by immunofluorescence and Western-blotting assay, and HBsAg and HBeAg in cell culture medium were detected by ELISA, and HBV DNA copies by fluorescence quantitative PCR. The longer HBV infected HK-2 cells, the more irregular of the cells' shape, the fewer number of the cells were left. But compared with HBV infected after 24 hours, α-SMA was more expressed after HBV infected 12 hours.After infected by HBV serum in 24 hours, HK-2 cells' proliferation rate was positively correlation in a dose range of LPS, but was negatively correlated with the CLI-095 (P < 0.05). The levels of HBsAg and HBeAg in cell culture medium were largest when the LPS concentration was at 10 µg/ml and CLI-095 at 5 µg/ml. The expression of TLR4 significantly increased in HK-2 cells treated with LPS compared with those with CLI-095, but HBV DNA levels and HBsAg and HBeAg expression levels were lower. HBV infection may promote cell transdifferentiation and cell injury. The stimulation of HK-2 infected with HBV by LPS may upregulate the expression of TLR4 and reduce the copies of HBV DNA.

Correlation between HBV serological markers and HBV-DNA

Objective To analyze the correlation between the lovel of HBV semiogloat markers and HBV-DNA.Mothods Serum samples of 800 patients from Xinyi Pepole's Hospital clinic department suffering HBV were detected by ELISA and HBV-DNA were detected by FQ-PCR.then analyze the correlation between HBV serological markers and HBV-DNA.Results The HBV-DNA was detoeted in some HBV serological markerg groups and each group was different in the result.HBsAg positive.such as HBsAg,HBeAg and Anti-HBc positive samples has higher amount of copies of HBV-DNA;the low level of HBV-DNA copies were detected in the group of HBsAg.HBeAg or Anti-HBe,Anti-HBo lmsitive; and the HBV-DNA copies rate is 2.98%in the single HBsAg positive.Among 120 HBe Ag positive samples.there were 118 HBV-DNA positive,the positive rate of HBV-DNA was 98.33%and the level of HBeAg positive wail correlated to HBV-DNA(r=0.993).Conclusion Both the detection of HBeAg and HBV-DNA cab reflect the active level of duplicate of HBV-M.The HBV-DNA level showed all obvious correlation to the HBeAg contend in Bera. Key words: HBV-M; HBV-DNA; Correlation

Activated IL-23/IL-17 pathway closely correlates with increased Foxp3 expression in livers of chronic hepatitis B patients

BackgroundFoxp3 protein plays a critical role in mediating the inflammatory response and can inhibit the proinflammatory IL-23/IL-17 pathway. However, the molecular interplay of Foxp3 and the IL-23/IL-17 pathway in patients with chronic hepatitis B (CHB) remains unclear. To this end, we analyzed the expression patterns of Foxp3- and IL-23/IL-17 pathway-related proinflammatory cytokines in 39 patients with acute-on-chronic liver failure, 71 patients with CHB and 32 healthy controls.ResultsFoxp3 expression was found to be elevated in and mainly expressed by the CD4+ T cell sub-population of peripheral blood mononuclear cells and liver tissues of patients with hepatitis B. The intrahepatic expression of Foxp3 strongly correlated with the copies of HBV DNA and the concentration of surface antigen, HBsAg. IL-23/IL-17 pathway-related proinflammatory cytokines were also found to be significantly increased in patients' liver tissues, as compared to healthy controls. Moreover, Foxp3 expression was strikingly correlated with the production of these cytokines in liver tissues of CHB patients.ConclusionsThe closely-correlated increase of Foxp3 and IL-23/IL-17 pathway activity in HBV-infected livers suggests that the proinflammatory IL-23/IL-17 pathway had not been effectively suppressed by the host immune machinery, such as Treg (Foxp3) cells. Constitutive activation of the IL-23/17 pathway, thus, may support the chronic hepatitis B state.

Clinical study of telbivudine on HBeAg-positive chronic hepatitis B

Objective To investigate the curative effect of telbivudine versus lamivudine in chronic hepatic B with a therapy of 48 weeks. Methods 94 patients who were diagnosed chronic hepatitis B were investigated. The therapeutic group and the control group (n = 47) were respectively assigned to receive 600mg of telbivudine and 100mg of lamivudine in 48 weeks. Copies of HBV-DNA、serum markers、alanine transarninase(ALT) level were tested before therapy and 4 weeks、12 weeks、24 weeks、48 weeks in the the course of treatment. The HBV-DNA conversion、HBeAg conversion and the normal rate of the ALT were compared. Results The copies of HBV-DNA decreased as the treatment progressing,especially at the 4 thweek. Conversion rate of the HBV-DNA of the telbivudine group was 72.3% ,which was higher than the lamivudine group 42.6%. There is a significant difference between two groups (x2 = 8.530,P ＜ 0.05). 95.7% of ALT in the telbivudine group revived to normal level versus 91.5% of the lamivudine group became normal. There is no significant difference between two groups (P ＞ 0.05). At the end of treatment,the rate of the HBeAg converting to negative in the telbivudine group is 38.3% versus the lamivudine group 31.9%, but there is no significant statistical difference between two groups(x2 = 0.420 ,P ＞ 0.05). The frequence of seroconversion of HBeAg in the telbivudine group is 23.4% versus the lamivudine group 17.0%, but no statisticall difference between two groups (x2 = 0. 594, P ＞ 0.05). Conclusion It was more effective to treat the HBeAg positive chronic hepatitis with telbivudine than lamivudine,and lower frequence of seroconversion of the HBV-DNA、HBeAg. Key words: Hepatitis B,chronic; Telbivudine

Clinical significance of measuring hepatitis B virus large surface protein in serum

To explore the significance of serum hepatitis B virus large protein( HBV-LP), HBV-DNA and markers of hepatitis B virus (HBV-M)in the diagnosis of viral replication. Serum HBV-DNA level was quantitatively detected using PCR Real-time polymerase chain reaction, HBV-LP was detected by enzyme-linked immunosorbent assay (ELISA) and HBV markers expression were measured by chemiluminescence immunoassay method in 1886 cases of seurm. The results of hepatitis virus large protein (HBV-LP) detection and the detection results of HBV-DNA was no significant difference (chi2 = 1.142, P > 0.05). HBV-DNA logarithm of copies and A vaule of HBV-LP was a positive correlation (r = 0.487, P < 0.01). HBV-DNA copies of different groups was significantly different from HBV-LP A values (F = 7.772, P < 0.01). The results of HBV-LP and HBV-DNA detected in different patterns of HBV-M were not significantly different. In 36 healthy people,the detecting results of HBV-DNA and HBV-LP are negative. There is a good correlation between the copies of HBV-DNA and the levels of HBV-LP. HBV-LP expression can reflect the replication of HBV.

Inhibition of HBV replication by small interference RNA in HepG2.2.15 cells

Objective Virus-specific RNA interference (RNAi) is a powerful inhibitor of gene expression and replication of HBV. It is known to have high efficiency, specificity, and few side effects. We wanted to evaluate the effects of siRNA silencing HBV replication on the growth of hepatocellular carcinomatic(HCC) cells to find out an ideal method for treatment of HCC. Methods We transfected siRNA into HepG2.2. 15 cells (HCC cell inserting HBV gene) and detected the HBsAg and HBV DNA copies for evaluating the inhibitory effects of siRNA. Then we evaluated cell growth and self-renewal ability after transfection of siRNA by MTT. Results The HBsAg level and HBV DNA copies were reduced after the transfection of siRNA, the highest inhibition rate was 83.9%,while the inhibition rate of HBV DNA copies reached 73. 4%. The siRNA group's growth ability and self-renewal rate were lower than the control group in 5 days. Conclusion siRNA can effectively inhibit HBV replication and expression in HepG2.2.15 cells and silencing HBV replication can inhibit HepG2.2.15 cell's growth and self-renewal. Key words: Carcinoma hepatocellular; Hepatitis B virus; siRNA; HepG2.2. 15

Relationship between hepatitis B virus DNA and hepatitis B virus large envelope protein

To investigate the relationship between HBV DNA and hepatitis B virus large envelope protein (HBV-LP) in patients with chronic hepatitis B. Serum HBV DNA was detected by RT-PCR and the HBV-LP was detected by enzyme linked immunosorbent assay in 320 serum samples collected from patients with chronic hepatitis B. There were no significant difference between positive rate of HBV-LP and that of HBV DNA in different HBeAg patterns (P>0.05). Serum HBV-LP levels were closely correlated with HBV DNA copies (r=0.949). Serum HBV-LP is a reliable serological marker that can reflect the replication of HBV.

Do Liver IL-12 Levels Predict Sustained Response to IFN-α Therapy in Children with Chronic Hepatitis B?

The aim of this study is to investigate the immunoregulatory role of interleukin-12 and interferon-gamma in children with chronic hepatitis B who are treated with interferon-alpha therapy. The patients were divided into 2 groups: Group I included 16 children with naive chronic replicative hepatitis B infection, and Group II included 6 children who are inactive hepatitis B virus (HBV) carriers. Group I received interferon-alpha subcutaneously (10 mU/m(2)/dose), 3 times a week during 4 months. Initial serum alanine aminotransferase (ALT) levels, hepatitis B serologic markers, serum interleukin-12 and interferon-gamma levels were measured. In Group I, laboratory tests were re-evaluated in the second and fourth months. Liver biopsy was performed in all patients and samples were used for tissue interleukin-12 level evaluation and histopathological examination. Hepatic activity index (HAI) and serum interferon-gamma were significantly higher in Group I (P < 0.05). Initial tissue interleukin-12 levels in Group I were low but a significant increase was observed at the fourth month (P < 0.05). While responder patients in Group I had marked elevation of tissue interleukin-12 levels, nonresponders did not reveal considerable changes at the fourth month evaluation. A negative correlation was found between serum HBV-DNA copies and interferon-gamma levels prior to therapy (P < 0.01, r: -0.66). The analysis of cytokine levels with serum transaminases demonstrated a positive correlation between the tissue interleukin-12 levels at the fourth month and serum ALT levels at the beginning and second month of the therapy (r: 0.77, P < 0.05 and r: 0.92, P < 0.05, respectively). This is the first study emphasizing the relationship between tissue cytokine levels and therapy success. Understanding the course of chronic hepatits B in the pediatric population will help us to clarify some debates on the treatment.