OBJECTIVETo observe the Chinese medicine constitution types and human leukocyte antigen (HLA)-DQA1 gene polymorphism in patients with hepatitis B (HB) virus infection in Chinese Han population of Zhejiang Province, for exploring the roles of constitution factor in pathogenesis of HB.

METHODSA total of 240 subjects, including 120 biopsy-proven chronic HB (CHB), 60 HB asymptomatic carrier (ASC) and 60 resolved from HBV infection spontaneously (RHBS) were studied. Their Chinese medicine constitution type was judged by Wangqi's classification, and their genotype of HLA-DQA1 was detected by polymerase chain reaction sequence specific primer for comparing the difference between groups in distribution frequency (DF) of constitution types and genes.

RESULTS(1) As compared with the RHBS group, DF of yin-deficiency constitution and phlegm-dampness constitution in the CHB group was significant higher (20.0% vs. 6.7% and 12.5% vs. 1.7%), and that of placid constitution was significant lower (11.7% vs. 31.7%), showing statistical significance between groups (OR = 3.5, 95% CI: 1.16-10.60; OR = 8.4, 95% CI: 1.09-65.42; OR = 0.161, 95% CI: 0.076-0.34; all P < 0.05). (2) As compared with the ASC group, DF of damp-heat constitution was significant higher (24.2% vs. 6.7%, P < 0.05, OR = 4.462, 95% CI: 1.49-13.36), and that of placid constitution was significant lower (11.7% vs. 45.0%, P < 0.01, OR = 0.285, 95% CI: 0.13-0.62) in the CHB group. (3) As compared with RHBS group, DF of HLA-DQA1 * 0201 allele in CHB group was significant higher (38.3% vs. 5.8%, P < 0.01, OR = 10.04, 95% CI: 4.48-22.48); and that of HLA-DQA1 * 0102 allele was significant lower (9.6% vs. 36.7%, P < 0.01, OR = 0.183, 95% CI: 0.10-0.32). (4) As compared with ASC group, DF of HLA-DQA1 * 0201 allele in CHB group was significant higher (38.3% vs. 7.5%, P < 0.01, OR = 7.667, 95% CI: 3.7-15.87), and that of HLA-DQA1 * 0102 allele was significant lower (20.0% vs. 9.6%, P < 0.01, OR = 0.424, 95% CI: 0.23-0.79).

CONCLUSIONBoth Chinese medicine constitution and HLA-DQA1 gene polymorphism show connection with the outcomes of HB virus infection in Chinese Han population, but the real association between them is required for further study.

Adolescent ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Body Constitution ; Carrier State ; virology ; Female ; Gene Frequency ; Genotype ; HLA-DQ alpha-Chains ; genetics ; Hepatitis B ; diagnosis ; genetics ; virology ; Hepatitis B virus ; Heterozygote ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Polymorphism, Genetic ; Young Adult

OBJECTIVETo observe the correlation between constitution of yin deficiency syndrome (YDS) and polymorphism of HLA-DQA1/treatment response of Peg-lFNalpha therapy in HBeAg positive chronic hepatitis B (CHB) patients, and to explore constitution of Chinese medicine (CM) in response of interferon therapy.

METHODSTotally 120 HBeAg positive CHB patients who were treated with Peg-IFNalpha were enrolled, and assigned to YDS group (59 cases) and non-YDS group (61 cases) according to classification of CM constitutions. All patients were subcutaneously injected with Peg-IFNalpha-2b (1.0 microg/kg body weight) or Peg-IFNalpha-2a (180 microg), once per week. Effective efficacy was primarily judged when complete response (CR) or partial response (PR) was obtained at month 6. Those with CR or PR completed 1 year therapeutic course. HLA-DQA1 gene types were detected by polymerase chain reaction sequence specific primers (PCR-SSP). The distribution difference of CM constitutions in patients with CR or PR and their inter-group HLA-DQA1 allele frequency were compared.

RESULTSDifferent treatment responses of Peg-IFNalpha were observed in CHB patients of two different CM constitutions. The ratio of CR + PR was 61.0% (36/59) in YDS group, obviously lower than that in NYDS group [78.7% (48/61), P < 0. 05]. Patients with CR had a lower allele frequency of HLA-DQA1 * 0501 than those with no-response [14.8% (8/54) vs. 30.6% (22/72)] with statistical difference (P < 0.05). Patients with CR had a higher allele frequency of HLA-DQA1 * 0601 than those with no-response [18.5% (10/54) vs. 5.6% (4/72)] with statistical difference (P < 0.05). The allele frequency of HLA-DQA1 * 0301 was lower in YDS group than in non-YDS group [2. 5% (3/118) vs. 9.8% (12/122)] with statistical difference (P < 0.05). The allele frequency of HLA-DQA1 * 0501 was higher in YDS group than in non-YDS group [33.9% (40/118) vs. 18.9% (23/122)] with statistical difference (P < 0.05). Yet statistical significance was lost after adjustment (Pc > 0.05 for both).

CONCLUSIONSBoth constitutions of CM and HLA-DQA1 gene polymorphism af- fect HBeAg positive CHB patients' response to Peg-INFalpha. Constitutions of YDS and HLA-DQA1 * 0501 was not favorable to response, their association needed to be further studied.

Antiviral Agents ; therapeutic use ; Gene Frequency ; HLA-DQ alpha-Chains ; genetics ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; drug therapy ; genetics ; Humans ; Interferon-alpha ; therapeutic use ; Medicine, Chinese Traditional ; Polyethylene Glycols ; therapeutic use ; Polymorphism, Genetic ; Recombinant Proteins ; therapeutic use ; Remission Induction ; Yin Deficiency ; genetics

OBJECTIVETo study on the correlation between chronic asymptomatic HBV carriers (ASC) of yin asthenia constitution and genotypes of HLA-DRB1 and HLA DQA1 alleles.

METHODSTotally 105 ASC were assigned to two groups according to their constitutions, i.e., the yin asthenia group (47 cases) and the non-yin asthenia group (58 cases). The genotypes of HLA-DRB1 and HLA DQA1 alleles were determined using PCR-SSP.

RESULTSThe gene frequency of HLA-DRB1 * 09 allele and HLA-DQA1 * 0301 allele (being 12.1% and 19.1%) were obviously lower in the yin asthenia group than in the non-yin asthenia group (being 27.8% and 39.7%, P < 0.05). The gene frequency of HLA-DRB1 * 11 allele and HLA-DQA1 * 0501 allele were obviously higher in the yin asthenia group (being 12.1% and 28.7%) than in the non-yin asthenia group (4.3% and 9.5%), showing statistical difference (P < 0.05, P < 0.01).

CONCLUSIONSHLA-DRB1 * 09 allele and HLA-DQA1 * 0301 allele might be the molecular bases for non-yin asthenia patients with ASC. HLA-DRB1 * 11 allele and HLA-DQA1 * 0501 allele might be the molecular bases for yin asthenia patients with ASC.

Adolescent ; Adult ; Carrier State ; Constitution and Bylaws ; Female ; Gene Frequency ; Genotype ; HLA-DQ alpha-Chains ; genetics ; HLA-DRB1 Chains ; genetics ; Hepatitis B, Chronic ; genetics ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Young Adult

OBJECTIVEEstablish the model of mouse with chronic hepatitis B virus (HBV) and nonalcoholic fatty liver disease (NAFLD).

METHODSTake 100 HBV transgenic, BALB/c mice of 4 weeks old, with each gender half. Then pick out 70 mice in one group to feed high-fat feed and the rest to feed normal feed. At the end of week 16, random kill 10 mice of high-fat, then liver tissue and serological detection target identification model is established in this paper. After that, divide the mice into model group and comparison group with 30 mice in each group. Feed model group with high-fat feed, comparison group with normal feed and normal group with normal feed till week 72 (including previous 16 weeks). Kill 10 mice of each group at the end of week 24, 48 and 72 respectively, fully automatic biochemical instrument detection of serum ALT, AST, TC, TG, FBG, fluorescence quantitative PCR method to detect HBV-DNA, chemiluminescence detection of HBsAg, liver biopsy after HE staining to evaluate histology change, observe mice model of dynamic evolution.

RESULTS(1) Feed high fat feed after 16 weeks, mice's weight, serum ALT, AST, TC, TG, FBG and blood biochemical indicators increased, HBV-DNA positive, liver HE staining obviously big blister fatty degeneration of liver cells and within the lobule lymphocytes infiltration, NAFLD activity score (NAS) getting close to NASH, the model of chronic HBV carries with NAFLD mouse built successfully. (2) The TC and TG values of model group in each period were higher than that of comparison group and normal group. (3) In week 24 and 72, HBV-DNA values of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05). (4) In week 48 and 72, NAS of each group are obvious different from the other two groups and the difference can be applied to statistical significance (P < 0.05).

CONCLUSIONS(1) Chronic HBV carries with NAFLD mice model can be established by HBV transgenic mice fed by high fat feed. (2) NAFLD accelerates the liver disease of the mice carrying HBV to some extent.

Animals ; Disease Models, Animal ; Fatty Liver ; complications ; pathology ; virology ; Female ; Hepatitis B virus ; genetics ; isolation & purification ; physiology ; Hepatitis B, Chronic ; complications ; pathology ; virology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Non-alcoholic Fatty Liver Disease

OBJECTIVETo explore the correlation of serum hepatitis B surface antigen (HBsAg) level and hepatic tissue pathological staging in the chronic hepatitis B infected persons.

METHODSCollect the clinical data of 272 cases who are HBsAg-positive more than 6 months and accepted hepatic biopsy in our hospital. Detect serum HBsAg quantification, ALT, HBV DNA, complete blood count, hepatic tissue pathological staging, grouping the cases according to the stage of inflammation and the fibrosis degree respectively. Observe serum HBsAg quantification, HBV DNA and the stage of inflammation and the fibrosis degree. Analyse the correlation between HBsAg quantification and HBV DNA.

RESULTSThe correlation of serum HBsAg level and HBV DNA is notable. Serum HBsAg level is a variable affecting hepatic tissue pathological stage significantly.

CONCLUSIONSSerum HBsAg level is a marker having higher specificity and sensitivity to diagnose the hepatic fibrosis.

Adolescent ; Adult ; DNA, Viral ; blood ; Female ; Hepatitis B Surface Antigens ; analysis ; blood ; immunology ; Hepatitis B virus ; immunology ; Hepatitis B, Chronic ; blood ; immunology ; pathology ; virology ; Humans ; Liver ; chemistry ; immunology ; pathology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the effect of extract of ginkgo biloba leaf (EGb) during the formation of HBV-related hepatocellular carcinoma (HCC).

METHODS99 HBV transgenic mice were randomly divided into control group, high-dose group, low-dose group. High-dose group and low-dose group were intraperitoneal injected 35mg/(kg x d) and 17.5 mg/(kg x d) of the shuxuening injection. Control group without special treatment. The serological markers and immunohistochemical markers in liver tissue will be done at the first 12 months and 18 months.

RESULTS(1) HBV transgenic mice can be found HCC at the 18 months. The incidence of HCC was lower in high-dose group and low-dose group, there was statistically different among the three groups. (2) The semi-quantitative scoring of liver HBx expression was highest in the control group at the 12 months. The semi-quantitative scoring of liver HBx, p53 and Bcl-2 expression was highest in the control group at the 18 months. They all appeared statistically different among the three groups. (3) Spearman correlation analysis showed that HCC incidence and liver tissue HBx, p53, Bcl-2 expression was a certain degree of positive correlation, r was 0.536, 0.487 and 0.403, P < 0.05.

CONCLUSIONEGb can reduced the incidence of the HCC with HBV transgenic mice. The reason may be that the EGb can reduce liver HBx, p53, Bcl-2 protein expression in the HBV transgenic mice.

Animals ; Carcinoma, Hepatocellular ; drug therapy ; etiology ; genetics ; prevention & control ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Ginkgo biloba ; chemistry ; Hepatitis B ; complications ; drug therapy ; Humans ; Liver Neoplasms ; drug therapy ; etiology ; genetics ; prevention & control ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic

OBJECTIVETo investigate the association between HLA-DQA1 gene polymorphism and the outcomes of hepatitis B virus infection in Chinese Han population.

METHODSA total of 180 consecutive patients with biopsy-proven hepatitis B virus infection (120 patients with chronic hepatitis B and 60 patients with asymptomatic HBV carrier) and 60 subjects who resolved from HBV infection spontaneously were studied. Genotype of human leukocyte antigen(HLA)-DQA1 was detected by polymerase chain reaction sequence specific primer(PCR-SSP).

RESULTS(1) The frequency of HLA-DQA1 * 0201 allele in chronic hepatitis B group was significant higher than the frequency in resolved from HBV infection spontaneously group (38.3% vs. 5.8%, P < 0.001, A = 10.04, 95% CI: 4.48-22.48). The frequency of HLA-DQA1 * 0102 allele in chronic hepatitis B group was significant lower than the frequency in resolved from HBV infection spontaneously group (9.6% vs. 36.7%, P < 0.001, A = 0.183, 95% CI: 0.10-0.32). (2) The frequency of HLA-DQA1 * 0201 allele in chronic hepatitis B group was significant higher than the frequency in asymptomatic HBV carrier group (38.3% vs. 7.5%, P < 0.01, A = 7.667, 95% CI:3.7-15.87). The frequency of HLA-DQA1 * 0102 allele in chronic hepatitis B group was significant lower than the frequency in asymptomatic HBV carrier group (20% vs. 9.6%, P < 0.01, A = 0.424, 95% CI: 0.23-0.79).

CONCLUSIONHLA-DQA1 gene polymorphism may play an important role in the outcomes of hepatitis B virus infection in-Chinese Han population. The HLA-DQA1 * 0102 allele could keep individuals away from HBV infection, and HLA-DQA1 * 0201 allele could aggravate persistant infection of HBV and hepatic inflammatory.

Adult ; Asian Continental Ancestry Group ; ethnology ; genetics ; Female ; Genome-Wide Association Study ; HLA-DQ Antigens ; genetics ; HLA-DQ alpha-Chains ; Hepatitis B ; ethnology ; genetics ; virology ; Hepatitis B virus ; physiology ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Young Adult

OBJECTIVETo investigate the beneficial effects of Rhein (RH) on hepatic progression in hepatitis B virus (HBV)-transgenic mice with nonalcoholic steatohepatitis induced by a high-fat (HF) diet.

METHODSA mice model of HBV chronic infection concomitant with liver steatosis was induced by a HF diet in 4-week old HBV-transgenic mice for 16 weeks (n = 130). Thereafter, the mice were divided randomly into control group (back to normal chow), model group (continuing HF diet), RH group [continuing HF diet and administering with 120 mg/(kg x d) RH by gavage] and Essentiale group [continuing HF diet and administering with 69.2 mg/(kg x d) Essentiale by gavage] with 30 mice in each, and were sacrificed at the end of 24-week and 48-week respectively. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglyceride (TG) and fasting plasma glucose (FPG) were measured by an automatic biochemical analyzer, and serum HBV-DNA was determined with qPCR. Hepatic histology was evaluated by HE staining with a light microscope.

RESULTS(1) An histological change composed of steatosis, lymphocytes intralobular infiltration and ballooning was observed after 48 weeks feeding of HF diet, in part mimicking that of NASH patients as evidenced by a NAFLD activity score (NAS) of 3.58 +/- 1.44 points. (2) Histologically, the NAS of model group was higher than that of control group at both time points. RH failed to lessen NAS whereas Essentiale improved the NAS at 48-week. (3) Serum levels of TC, TG and FPG were significantly different between 4 groups at 24-week, with a comparable low value in both RH and Essentiale group. A similar change was evident at 48-week. (4) In terms of HBV viral load, a significantly lower level in Essentiale group than the others was observed at both time points.

CONCLUSIONHF diet feeding is able to induce a mouse model of HBV chronic infection concomitant with NASH. RH is effective in alleviating the glucose and lipid metabolism but ineffective in improving the hepatic histology in this model, in contrast, backing to normal chow achieved a better effect in this aspect.

Animals ; Anthraquinones ; administration & dosage ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Disease Progression ; Fatty Liver ; complications ; etiology ; metabolism ; prevention & control ; Female ; Glucose ; metabolism ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; complications ; metabolism ; virology ; Humans ; Lipid Metabolism ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Non-alcoholic Fatty Liver Disease

OBJECTIVETo investigate the expression of F4/80, NF-kappaB, p-AKT, AKT in the liver of nonalcoholic fatty liver disease (NAFLD) mice. To determine the role of Kupffer cells (KCs) in the development of NASH (non-alcoholic steatohepatitis), and understand the pathogenic mechanism of NASH.

METHODSFive C3H/HeN mice fed with normal diet were served as controls, while fifteen fed with high fat, high fructose, high fat combined fructose diet respectively for 16 weeks were as NAFLD mice models. The liver inflammation and hepatic damage were examined, and the expression of F4/80, NF-Kb, p-AKT, AKT and the content of lipid in the liver were also detected.

RESULTSChronic intake of high fat and 30% fructose solution caused a significant increase in hepatic steatosis in animals in comparison to water controls. Liver F4/80 and NF-kappaB were significantly higher in high fat and high fat combined fructose diet fed mice than that in controls (P < 0.01, P < 0.01), F4/80 protein were higher in high fat diet treated mice than those in fructose and high fat combined fructose groups (P < 0.01, P < 0.01). Markers of insulin resistance (e. g, hepatic phospho-AKT, AKT) were only altered in fructose-fed or high fat combined fructose animals (P < 0.01, P < 0.01).

CONCLUSIONHigh fat and fructose diet may induce NAFLD in C3H/HeN mice. Kupffer cells and signal pathway proteins were activated, and they may play key roles in the initiation and progression of NASH.

Animals ; Diet, High-Fat ; adverse effects ; Fatty Liver ; etiology ; immunology ; metabolism ; Female ; Fructose ; adverse effects ; Humans ; Kupffer Cells ; immunology ; Lipid Metabolism ; Liver ; immunology ; metabolism ; Male ; Mice ; Mice, Inbred C3H ; NF-kappa B ; immunology ; Non-alcoholic Fatty Liver Disease ; Oncogene Protein v-akt ; immunology ; Signal Transduction

OBJECTIVETo investigate the effect of RNA interfering TLR4 signal pathway on phagocytosis of Kupffer cells.

METHODSRAW2647 mice mononuclear macrophage leukemia cells were observed. The tested group was interfered by Tlr4-mus-1567 RNA which had the best result confirmed by QPCR, cells interfered by Negative Control RNA as NC group, and normal cell as control. We perform the phagocytosis test on each group.

RESULTSThe tested group has lower phagocytes percentage than control (17.67% +/- 3.51% vs 32.00% +/- 3.00%, P < 0.01), and lower phagocytic index (46.33% +/- 7.51% vs 82.00% +/- 6.08%, P < 0.01).

CONCLUSIONSDecreased phagocytic activity was observed on Kupffer cells by RNA interference.

Animals ; Kupffer Cells ; immunology ; Mice ; Phagocytosis ; RNA Interference ; Signal Transduction ; Toll-Like Receptor 4 ; genetics ; immunology