Improvement situation on indexes of the zebrafish disease model of non-alcoholic fatty liver disease with FGF21 analogues / 中华肝脏病杂志

Objective: To detect the therapeutic efficacy of FGF21 analogues on the zebrafish model of non-alcoholic fatty liver disease. Methods: A zebrafish model of non-alcoholic fatty liver disease was established by providing the normal diet fed to wild-type zebrafish three times daily. PF-05231023 was administered exogenously at a final concentration of 0.5 μmol/L. Body length, body weight, triglycerides, and other indexes were measured after 20 days. Pathological changes were evaluated in liver tissue sections by HE staining. Quantitative PCR was used to identify expressional changes in genes related to lipid metabolism, endoplasmic reticulum stress, and inflammation. Results: QPCR and immunofluorescence staining results showed that FGF21 was highly expressed in the zebrafish model group. The addition of the FGF21 analogue PF-05231023 significantly reduced the body length and body weight (P < 0.01), and the triglyceride content (P < 0.05) in the zebrafish model group. The liver HE staining results showed that PF-05231023 had alleviated the large and tiny bullae fat, lesions, and others in the zebrafish model group. The quantitative PCR results demonstrated that PF-05231023 reduced the expression of lipogenic factors (P < 0.01), inflammatory-related factors (P < 0.001), and genes related to endoplasmic reticulum stress (P < 0.05), but raised lipid-oxidation-related factors (P < 0.05) in the zebrafish model group. The addition of PF-05231023 reduced oleic acid-induced lipid and triglyceride levels in HepG2 cells. Conclusion: FGF21 analogue addition can improve indexes in the zebrafish disease model of non-alcoholic fatty liver disease.

Effect of Compound Zhajin Granule on Toll-like Receptor 4 Signaling Pathway in Nonalcoholic Steatohepatitis Mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Compound Zhajin Granule (CZG) on Toll-like re-ceptor 4 (TLR4) signaling pathway in high-fructose corn syrup induced NASH mice.</p><p><b>METHODS</b>Thirty 6-week-old male C3H mice were divided into the high fat and high fructose (HFHFr) group (n = 20) and the control group (n = 10) according to body weight. Mice in the HFHFr group ate high fat diet and drank 20% fructose water, while those in the control group ate common diet and drank common water. After 8 weeks mice in the HFHFr group were divided into two group according to body weight, the HFHFr group and the CZG group, 10 in each group. Mice in the CZG group were fed with high fat forage and 20% fructose water, and administered with 50 mL/kg 12. 8% CZG (prepared by hawthorn, Radix Curcumae, Alisma Orientale, Fritillaria Thunbergii, Silybum Marianum, peach seed in the ratio of 3:1.5:1.5:2:1.5:2:1) by gastrogavage. Mice in the HFHFr group were fed in the same way and daily administered with equal volume of distilled water by gastrogavage. Sixteen weeks later all mice were sacrificed. Body weight, liver wet weight, liver function, and lipid metabolism were detected. Pathological changes of liver tissues were assessed by HE staining, oil red O staining, and Masson staining. Expressions of TLR4, myeloid differentiation factor 88 (MyD88), tumor necrosis factor-alpha (TNF-α) were detected using immunohistochemical staining and real-time fluorescent quantitative PCR.</p><p><b>RESULTS</b>Body weight, alanine aminotransferase (ALT), aspartate aminotransferase (AST) were obviously lower in the CZG group than in the HFHFr group (P < 0.05); oil red O stained area and density were decreased more in the CZG group than in the control group. HE staining showed ballooning inflammation was reduced more in the CZG group than in the HFHFr group. Masson staining was negative. Positive rates of TLR4 and MyD88 and mRNA expressions were significantly lower in the CZG group than in the HFHFr group (all P < 0.05).</p><p><b>CONCLUSION</b>CZG could significantly inhibit TLR4 signaling pathway of liver in NASH mice.</p>

The effect of RNA interfering TLR4 signal pathway on phagocytosis of Kupffer cells / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the effect of RNA interfering TLR4 signal pathway on phagocytosis of Kupffer cells.</p><p><b>METHODS</b>RAW2647 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.</p><p><b>RESULTS</b>The 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).</p><p><b>CONCLUSIONS</b>Decreased phagocytic activity was observed on Kupffer cells by RNA interference.</p>

Activation of Kupffer cell and related signal pathway proteins in the liver of high fat and high fructose diet induced NAFLD mice / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Five 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.</p><p><b>RESULTS</b>Chronic 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).</p><p><b>CONCLUSION</b>High 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.</p>

Protective effects of rhein on hepatic progression in HBV-transgenic mice with nonalcoholic steatohepatitis induced by a high-fat diet / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>A 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.</p><p><b>RESULTS</b>(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.</p><p><b>CONCLUSION</b>HF 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.</p>

Model building-up and observation on the mouse carried chronic hepatitis B and nonalcoholic fatty liver disease / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Establish the model of mouse with chronic hepatitis B virus (HBV) and nonalcoholic fatty liver disease (NAFLD).</p><p><b>METHODS</b>Take 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.</p><p><b>RESULTS</b>(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).</p><p><b>CONCLUSIONS</b>(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.</p>

Aspartate-ornithine granules in the treatment of nonalcoholic steatohepatitis: a multiple-dose parallel controlled clinical trial / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic efficacy and safety of aspartate-ornithine granules in patients with nonalcoholic steatohepatitis (NASH).</p><p><b>METHODS</b>Seventy-two patients with NASH were included in this multiple-dose parallel controlled clinical trial and received a 12-week course of aspartate-ornithine granule treatment at either high-dose (6 g bid po; n = 38) or low-dose (3 g bid po; n = 34). Clinical efficacy was assessed by monitoring data from urinalysis, serologic tests (alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), and triglyceride (TG)), and abdominal computed tomography (CT) scan. Safety was assessed by occurrence of adverse events (fatigue, anorexia, abdominal distension, nausea, and vomiting). Statistical analyses were conducted to determine the significance of differences between parameters before (baseline) and after treatment.</p><p><b>RESULTS</b>After 12 weeks of treatment, the liver and spleen CT ratios in both the high-dose group (0.89 +/- 0.19) and the low-dose group (0.80 +/- 0.15) were significantly higher than at baseline (S = 329, P less than 0.0001 and S = 246, P less than 0.0001); the overall improvement was more robust in the high-dose group (52.63%) than in the low-dose group (38.23%) (Z = -2.1042, P less than 0.05). After 6 and 12 weeks of treatment, the serum ALT levels in both the high-dose group and the low-dose group were significantly lower than at baseline (6 weeks: S = 324.5, P less than 0.0001 and S = 223, P less than 0.0001; 12 weeks: S = 370.5, P less than 0.0001 and S = 297.5, P less than 0.0001); the overall improvement was more robust in the high-dose group (79.0%) than in the low-dose group (53.0%) (Z = -2.0533, P less than 0.05). Similar trends were seen for the serum levels of AST and GGT after 6 and 12 weeks of treatment (all P less than 0.01) and serum levels of TG after 12 weeks of treatment. The rate of adverse reactions was low and similar between the two groups (high-dose: 4.8% and low-dose: 4.4%; all gastrointestinal).</p><p><b>CONCLUSION</b>Aspartate-ornithine granule therapy was an effective and safe treatment of nonalcoholic steatohepatitis, with the higher dose of 6 g bid po providing more robust clinical benefit without affecting the safety profile.</p>

Three-year efficacy and side effect of adefovir dipivoxil for the treatment of the old patients with chronic hepatitis B virus infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate 3-year antiviral efficacy and side effect of adefovir dipivoxil (ADV) on the old patients with hepatitis B chronic infection.</p><p><b>METHODS</b>31 HBeAg-negative chronic hepatitis B virus infected old patients (include 8 patients with chronic hepatitis B and 23 patients with liver cirrhosis) with serum HBV DNA levels > 1000 copies/ml, and ALT > 2 times the upper limit of normal, without company with other liver diseases, cancer, renal dysfunction, and autoimmune disease. All the patients were treated with ADV orally (10 mg once daily) for 36 months. HBV DNA and biochemical and blood routine indexes were checked after treated.</p><p><b>RESULT</b>Serum total bilirubin, direct bilirubin, alamine aminotransferase, aspartate aminotransferase and load of HBV DNA decrease significantly after therapy (P < 0. 001). Other biochemical indexs and blood routine are no significant changes (P > 0.05).</p><p><b>CONCLUSION</b>The way to treat with ADV is safe and effective for old patients with chronic hepatitis B virus infection.</p>

Genotypes and evolution characteristics of three patients with poor response to initial treatment of Lamivudine and Adefovir dipivoxil for hepatitis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To understand the genotype characteristics and its evolution of patients with poor response to initial combined treatment of Lamivudine and Adefovir dipivoxil for chronic hepatitis B.</p><p><b>METHODS</b>We detected the HBV genotypes of three patients-S1, S2, S3, who with poor response to initial treatment of Lamivudine and Adefovir dipivoxil for chronic hepatitis B over 12 months by the application of cloning and sequencing method at the time point of baseline,4 weeks after treatment, 12 weeks, 24 weeks, 48 weeks, 60 weeks. 25 clones were randomly selected to identify and sequence at each time point.</p><p><b>RESULTS</b>The total number of clones from 3 patients with poor response to initial combined treatment of Lamivudine and Adefovir dipivoxil for chronic hepatitis B at each time point was 398. About patient S1 at baseline, genotype C accounting for 8.3%, genotype B, for 91.7%, so genotype B was in dominant (22/24). But genotype C has gradually developed to 100% after treatment for 60 weeks. About patient S2 and S3, genotype B was the only type at baseline. However type B has gradually "drift" to type C during treatment. When treatment for 60 weeks, type C has taken the absolute advantage 75% for S2, and 100% for S3.</p><p><b>CONCLUSIONS</b>The cloning and sequencing can represent the overall genotype level better. HBV genotype has performed the evolution trend that genotype has drifted from B to C during long-term drug pressure, which is the main reason for poor response to initial combined treatment of Lamivudine and Adefovir dipivoxil for chronic hepatitis B.</p>

Impact of liver steatosis on antiviral effects of pegylated interferon-alpha in patients with chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the impact of hepatic steatosis on virologic response in chronic hepatitis B (CHB) patients treated with pegylated interferon-alpha (PEG-IFNa).</p><p><b>METHODS</b>Ninety-six naive patients positive for hepatitis B e antigen (HBeAg) and with biopsy-proven CHB were administered PEG-IFNa-2a or PEG-IFNa-2b for 48 weeks. Virologic response (HBeAg clearance and hepatitis B virus (HBV) DNA less than 5 log10 copies/ml) and biochemical response (alanine transaminase (ALT) normalization) were compared between patients with (n=34) and without (n=62) steatosis.</p><p><b>RESULTS</b>The HBV DNA titer in the steatosis group was significantly lower than that of the non-steatosis group (6.961.27 vs. 7.541.28 log10 copies/ml; t=2.161, P=0.033). After 48 weeks of PEG-IFNa treatments, there was no significant difference in HBeAg seroconversion or the percentage of undetectable HBV DNA (less than 3 log10 copies/ml) between steatosis and non-steatosis patients. However, the steatosis patients presented with a significantly lower complete response rate (virologic response plus biochemical response) compared to non-steatosis patients (26.5% vs. 48.4%; x² =4.373, P=0.037). Of the 45 CHB patients with undetectable HBV DNA after 48 weeks of treatment, seven did not achieve ALT normalization. The rate of patients with non-biochemical response was significantly higher in the steatosis group than in the non-steatosis group (33.3% vs. 6.67%; P=0.032).</p><p><b>CONCLUSION</b>Hepatic steatosis does not affect the virologic response, but does affect the biochemical response in CHB patients treated with PEG-IFNa for 48 weeks.</p>

Value of delta model for end-stage liver disease in evaluating the prognosis of liver failure patients with hepatitis B virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the prognostic value of the model for end-stage liver disease (MELD) and deltaMELD in liver failure patients infected with hepatitis B virus.</p><p><b>METHODS</b>Based on prospective study design, 98 hospitalized cases were studied and followed up for 24 weeks. The clinical data were recorded. We calculated the score of MELD and deltaMELD, and also compare the score between the survival group and death group. Using ROC curve plotting obtained the better decisive threshold. The case fatality rate were compared at different time points which the patients were classified by the best critical value of MELD and deltaMELD. We draw the Kaplan-Meier survival curve of different group and analyse the change of survival rate by log-rank analysis.</p><p><b>RESULTS</b>52 of 97 patients died and 46 survive during 24 weeks of followup. There was significant difference between the two groups for MELD and deltaMELD (P < 0.01). The case fatality rate in group which MELD > or = 23 was obviously higher than in that MELD < 23. The rate in group which deltaMELD > 4.5 was obviously higher than in that deltaMELD < 4.5 (P < 0.001). The area under curve (AUC) for the twelfth and 24th week's prognosis judgment of deltaMELD (0.823, 0.815) was larger than that of MELD (0.680, 0.684) (P < 0.05). Survival analyses (Kaplan-Meier) indicated that there were significant differences in cumulative survival rates among the groups which were grouped by optimization critical value ( P = 0. 000).</p><p><b>CONCLUSIONS</b>The scoring system of MELD also applied to the forecasting of prognosis for severe hepatitis B patients in China. The accuracy of deltaMELD to predict the prognosis was higher than that of MELD. The combination of MELD and deltaMELD showed good clinical practical value.</p>

Establishment and identification of non-alcoholic fatty liver disease in chronic hepatitis B virus infected mice / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To establish and identify an animal model of non-alcoholic fatty liver disease in chronic HBV infected mice.</p><p><b>METHODS</b>Transgenic mice with sustaining HBV production were established by microinjection of ocyte. Then they were randomly assigned into 4 groups (male control, male NAFLD model, female control and female NAFLD model) and treated with high fat diet (2% cholesterol, 10% lard, 88% forage) and common forage, respectively. NAFLD-related physical indexes, liver and kidney function, glucose and lipid metabolism were investigated at the time points of 8 weeks, 16 weeks and 24 weeks. Meanwhile, HBV type, serum levels of HBV DNA and HBeAg, immunohistochemical staining of hepatic HBsAg were detected. The establishment of NAFLD was evaluated by serum levels of total cholesterol (TC), triglycerides, glucose, aspartate aminotransferase (AST), alkaline phosphatase (ALP), alanine aminotransferase (ALT), gamma-glutamyltransferase (GGT), etc. Histological changes were also analyzed by HE, oil red O and Masson's trichrome staining. The status of CHB was assessed on the basis of immunohistochemistry and real-time PCR.</p><p><b>RESULTS</b>The body and liver weights, liver index in HBV transgenic mice were significantly increased in regardless of the gender of HFD feeding, and the levels of serum ALT, AST, ALP, GGT, TBIL, TBA, TC, TG, HDL-C, LDL and FBG were higher in HFD groups as compared with the control mice. Lipid droplets, cytologic ballooning and liver steatosis could be observed in most lobules of HFD groups after 8-week administration, fatty degeneration of hepatocytes, patch necrosis, mild to moderate chronic inflammatory infiltration were also observed in some of HFD feeding, reflecting the emerge of steatohepatitis. At the time point of 24-week perisinusoidal fibrosis and local fibrosis occurred in HFD groups. Immunohistochemical staining and real-time PCR analysis of the liver tissues showed positive signal of HBsAg in all groups of mice, although no significant difference was documented.</p><p><b>CONCLUSION</b>Our study suggests that animal model of NAFLD can be established in HBV transgenic mice and provide a nice animal model for further studies on NAFLD with chronic hepatitis B infection.</p>

Efficacy and safety of lamivudine plus adefovir combination therapy and entecavir monotherapy for chronic hepatitis B patients / 中华肝脏病杂志

To compare the efficacy and safety of Lamivudine (LAM) plus Adefovir dipivoxil (ADV) combination therapy and Entecavir (ETV) monotherapy for chronic hepatitis B patients. 120 patients with chronic hepatitis B managed in a single-centre clinical practice (median 96 weeks) were split into 2 cohorts, one was treated with de-novo combination Lamivudine (100 mg/day) plus Adefovir (10 mg/day) (LAM+ADV), the other with Entecavir (0.5 mg/day) monotherapy. Serum levels of ALT, creatinine, HBsAg, HBeAg and HBV viral load, together with genotypic resistence were analyzed at 0, 12, 24, 48, 96 weeks, respectively. HBV DNA was determined by real-time PCR. HBsAg and HBeAg were assessed by chemiluminescence. Serum levels of ALT and creatinine were detected by automatic biochemical analyzer. HBV genotypic resistence was tested by direct sequencing. (1) At the time point of 96 weeks, a total of 99 patients (51 cases in combination therapy cohort and 48 case in monotherapy cohort) were compared. The baseline characteristics as for HBV viral load, median age, serum levels of ALT and creatinine were compatible between combination therapy cohort and monotherapy cohort. (2) The rates of HBV DNA values is less than 300 copies/ml and HBV DNA values is less than 1000 copies/ml had no significant difference between LAM + ADV and ETV cohorts by the 12 and 24 weeks (P more than 0.05). (3) At the time point of 48 weeks, the rates of HBV DNA is less than 1000 copies/ml, HBeAg seroconversion, and ALT normalization were similar in both cohorts, though the rate of HBV DNA values is less than 300 copies/ml was obviously higher in combination therapy cohort than that of monotherapy cohort (90.7% vs 76%, P values is less than 0.05). (4) At the time point of 96 weeks, the rates of HBV DNA values is less than 300 copies/ml (96.1% vs 79.2%), HBV DNA values is less than 1000 copies/ml (98% vs 87.5%) and the HBeAg seroconversion (41.7% vs 16.7%) were markedly higher in combination therapy cohort than those of monotherapy cohort statistically (P values is less than 0.05 for all). The mean values of decreases for HBV viral loads and HBsAg levels were smilar in both cohorts at 48 and 96 weeks. (5) Elevated serum creatinine not be found in both cohorts at the end of treatment. (6) No virological breakthrough occurred in combination therapy cohort at the end of treatment. Four patients in monotherapy cohort were found with virological breakthrough at 96 weeks and three cases among were confirmed to be of variants associated with ETV resistance (rtL180M + T184L + M204V). Present study suggests that Lamivudine plus Adefovir dipivoxil de-novo combination therapy was more efficacious than Entecavir monotherapy for CHB patients and the tolerance is compatible.

P53 expression in liver tissue of patients with chronic hepatitis B and its influencing factors / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To observe p53 expression in liver tissue of patients with chronic hepatitis B and its influencing factors.</p><p><b>METHODS</b>17 cases HBeAg-negative chronic hepatitis B patients and 31 cases HBeAg-positive chronic hepatitis B patients were divided into 2 groups.</p><p><b>RESULTS</b>(1) HBeAg-negative chronic hepatitis B patients were older, mostly male and HBV DNA lower. These three indicators between two groups patients appeared statistical difference. Serum markers were no statistical difference between two groups patients except Glo. (2) Pathological inflammation and fibrosis Staging were no statistical difference between two groups patients. p53 expression positive rate and p53 expression semi-quantitative scoring in liver tissue were no statistical difference between the two groups. (3) Logistic regression analysis showed that only liver fibrosis staging (S) is a risk factor for p53 expression. Compared with the S0-1, p53 expression increased by 3.9 times the rate of positive in S > or = 2.</p><p><b>CONCLUSION</b>Liver fibrosis staging in patients with chronic hepatitis B is a risk factor for p53 positive expression in liver.</p>

The effect of extract of ginkgo biloba leaf during the formation of HBV-related hepatocellular carcinoma / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the effect of extract of ginkgo biloba leaf (EGb) during the formation of HBV-related hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>99 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.</p><p><b>RESULTS</b>(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.</p><p><b>CONCLUSION</b>EGb 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.</p>

To investigate the relationship of alcohol intake and hepatocellular carcinoma among patients with hepatitis B virus infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the relationship of alcohol intake and hepatocellular carcinoma among patients with hepatitis B virus infection.</p><p><b>METHODS</b>A total of 553 patients with HCC and 160 control subjects affected with hepatitis B virus were recruited. Serum virology, serum biochemistry, as well as demographic information were studied. Finally, risk factors were selected by stepwise Logistic regression analyse. Odds ratios (ORs) were estimated for each risk factor. According to alcohol intake, HCC patients were divided into three groups,then to observe the differences between them.</p><p><b>RESULTS</b>Elevated AST, GGT, ALP and AFP levels were seen more frequently in the HCC case groups compared to control group (P < 0.05). Multivariate analysis revealed that heavy alcohol use, smoking, positive family history of liver cancer is associated with HCC development among patients with hepatitis B virus infection. Significantly increased risk was found among patients for heavy alcohol use [A = 2.66 (2.01-3.50)] and for smoking [A = 2.51 (1.66-3.80)] and for positive family history of liver cancer [A = 1.64 (1.04-2.59)]. Compared to patients who did not have alcohol use, elevated GGT and ALP were seen more frequently in patients who had alcohol use either mild or heavy (P < 0.05).</p><p><b>CONCLUSIONS</b>Heavy alcohol use, smoking, positive family history of liver cancer is positive correlation with HCC development among patients with hepatitis B virus infection in China. In patients with hepatitis B virus infection who also has history of heavy alcohol, the most risk factor of HCC is hepatitis B virus infection, not alcohol.</p>

Dynamic expression of TGF-beta1/Smad protein in CCl4-induced liver fibrosis and its significance in rats / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the dynamic expression of TGF-beta1/Smad protein in rats with liver fibrosis induced by carbon tetrachloride (CCl4), to study mechanism of TGF-beta1/Smad signaling and the relationship between its transduction and liver fibrosis.</p><p><b>METHODS</b>Fifty healthy male SD rats were randomly divided into two groups: normal control group and model group. Experimental liver fibrosis was induced by subcutaneous injection of CCl4. After six weeks and nine weeks, histopathological changes and degrees of fibrosis were observed by optical microscopy. Meanwhile, the expression of TGF-beta1, TP3R-I, Smad2/3 and Smad7 proteins was detected by immunohistochemistry.</p><p><b>RESULTS</b>(1) Pathological observation of hepatic specimens: hepatic tissue of model group rat had inflammation and fibrosis in different degrees. By comparing with the degrees of inflammation and fibrosis model groups were more severe than normal control group (P < 0.05). (2) Changes of protein levels about TGF-beta1/Smad: the expressions of TGF-beta1, TbetaR-I, Smad2/3 and Smad7 in rat hepatic tissue were detected with immunohistochemistry techniques. The expressions of the four items in model group were higher than those of normal control group (P < 0.01). In fibrosis model group, there exist considerable positive correlations among expressions of TGF-beta1, TbetaR-I, Smad2/3, Smad7 and degrees of fibrosis in livers (P < 0.05 or 0.01).</p><p><b>CONCLUSION</b>There is close relation between the level of TGF-beta1, TbetaR-I, Smad2/3, Smad7 and the different liver fibrosis grades due to CCl4. The up regulation of TGF-beta1, TbetaR- I, Smad2/3 and Smad7 in liver tissue is involved in the progression of hepatic fibrosis.</p>

One-year combination therapy de novo of adefovir dipivoxil and lamivudine for decompensated cirrhosis related to HBV / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and potential renal impairment of one-year combination therapy de novo of adefovir dipivoxil (ADV) and lamivudine (LMV) for decompensated cirrhosis related to HBV.</p><p><b>METHODS</b>A total of 36 patients with decompensated cirrhosis related to HBV, nobody had nucleos(t)ide analogs (NAs) treatment history, were recruited and were divided into two group (control group and observation group) randomly. A monotherapy of LMV (100 mg per day) was selected to individuals in control group (n = 18), in contrast, a combination therapy de novo of ADV (10 mg per day) and LMV (100 mg per day) was applied to those in observation group (n = 18). Basic approaches including liver protection, symptom-driven intervention, and supporting therapy, were given to all of the individuals. A course of one year was applied to all. Liver function, Child-Pugh score, serum creatinine (sCr) level, virological response (VR) rate, and virological breakthrough rate were observed pro- and post- treatment, differences between the two populations were analysed statistically.</p><p><b>RESULTS</b>(1) The averages of gender, age, HBeAg status, HBV viral load, sCr level, and Child-Pugh score were all compatible in the two groups at baseline (P > 0.05 for all). (2) At the endpoint of treatment, none of deaths was reported. Comparing with the status before treatment in each group itself, liver function, Child-Pugh score, and viral load were improved statistically (P < 0.01 for all), especially in observed group (P < 0.01 for all variables, vs control group), as for VR rate, result is significant superior to that of control group too (88.89% vs 66.67% , P < 0.05). (3) Virological breakthrough occurred to none in observed group and three cases (16.67%) in control group, all of them were confirmed to be rtM204V variant in the following detection of direct sequencing. (4) Elevated level of sCr didn't arised at the end of treatment in two groups.</p><p><b>CONCLUSION</b>Present study reveals that in populations with decompensated cirrhosis related to HBV, one-year combination therapy de novo of ADV and LMV is superior to monotherapy of LMV, and the renal safety is favorable within one year.</p>

Analysis of 96 cases with novel H1N1 influenza A / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To review the epidemiologic and clinical characteristics of 96 cases with novel H1N1 influenza A, and improve the diagnosis and treatment level of novel H1N1 influenza A.</p><p><b>METHODS</b>96 cases of novel H1N1 influenza A admitted to the isolation wards from Oct 20 to Sep 23, 2009 were studied. Their epidemiologic, clinical, laboratory, and radiologic characteristics were analyzed.</p><p><b>RESULTS</b>The median age of the 96 patients was 26.52 +/- 10.62 years (range, 5 to 60 years). Sixty-four of the 96 patients had a close contact with novel H1N1 influenza A patients. The main symptoms included fever 100%, cough 86.4% , sore throat 66.6% and myalgia 32.3%.</p><p><b>CONCLUSION</b>The clinical presentation of novel H1N1 infection is largely indistinguishable from that of seasonal influenza. Combines both a symptom complex with the epidemiological investigation and laboratory characteristics can improve the accuracy of diagnosis of novel H1N1 influenza A.</p>

Association of chinese medicine constitution and human leukocyte antigen-DQA1 gene polymorphism with outcomes of hepatitis B virus infection / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>A 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.</p><p><b>RESULTS</b>(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).</p><p><b>CONCLUSION</b>Both 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.</p>