Objective To investigate the relationship between HBV DNA load with the serological markers(HB-M)HBsAg, HBeAg,HBeAb,HBcAb in the persons infected by chronic hepatitis B virus (HBV)in Lanzhou.Methods The real-time fluores-cent quantitative PCR was used to detect the HBV DNA load and the double antibody sandwich chemiluminesent immunoassay was used to measure the serum HBsAg,HbeAg,HbeAb and HbcAb levels in 724 cases of HBVinfection.Results The HBsAg level was positively correlated with the HBV DNA load in chronic HBV infection in Lanzhou area(r=0.342,P <0.05),there was an ob-vious positive correlation between HBeAg and HBV DNA load(r=0.463,P <0.05),the HBeAb level and HBV DNA load had the negative correlation (r=-0.227,P =0.001),the HBcAb level and HBV DNA load had no significantly correlation (r=-0.062, P =0.366).Conclusion There is obvious positive correlation between HBV DNA load with HBsAg,HBeAg in chronic HBV infec-tion in Lanzhou area,which indicating that the observation by combining HBsAg and HBeAg with HBV DNA can judge the infec-tious degree of the patients more accurately.

Objective To optimize the formative technology for Zhuqi Fugan Granules. Methods By using orthogonal experiment design, and setting parameters of granules as determination standards, concrete density, excipient types and dosage, and consentration of wetting agent were optimized to determine the optimal conditions. Results The optimal forming technology was as follows:The optimal ratio of starch to micrite cellulose was 3∶2;the optimal density of extration was 1.20;the ratio of concrete to excipient was 1∶1;the optimal concentration of wetting agent was 80% of ethyl alcohol. Zhuqi Fugan Granule enjoyed good mobility and with a critical relative humidity of 67%. Conclusion Parameters of Zhuqi Fugan Granules prepared by this method are satisfying, with stable properties and low moisture absorbtion. This preparation technology is simple and feasible.

Objective To investigate the correlation between the liver fibrosis markers laminin(LN),hyaluronan acid(HA),col-lagen Ⅳ(CⅣ)and procollagen type Ⅲ N-terminal peptide(PⅢNP)with the replication of hepatitis B virus(HBV)DNA in the pa-tients with chronic HBV in Lanzhou.Methods Real-time PCR and chemiluminescent immunoassay(CLIA)were used to quantita-tively measure serum HBV-DNA and liver fibrosis markers in 724 cases of HBV infection in Lanzhou with the different clinical classifications and the natural history stage as the background.Results The values of the liver fibrosis indicators and HBV-DNA level in all clinical types were higher than normal levels,and reached to maximum in the liver cirrhosis goup;CⅣ and PⅢNp were highest in the HBeAg (+)CHB group;according to the hepatitis B natural history stage,the PⅢNP and CⅣ values reached the peak value in the immune clearance phase,moreover the CⅣmean value in the HBeAg and HBV-DNA (+)group was higher than that in the HBV-DNA and HBeAg (-)group.4 indicators in the HBeAg (+)CHB group were higher than those in the HBeAg (-)CHB group,but only HA had statistical difference(P <0.01 ).Conclusion There is certain correlation between CⅣ with HBV-DNA and HBV-M,indicating that which can indirectly reflect the degree of liver damage and viral replication level.

Objective To explore changes of levels of interferon‐γ(IFN‐γ) ,interleukin‐32(IL‐32) and interleukin‐6(IL‐6) in pe‐ripheral blood and the correlation between peripheral IFN‐γ,IL‐32 ,IL‐6 and liver function level and hepatitis B virus(HBV) DNA load in HBV carriers .Methods Sixty HBV carriers ,including 39 cases of chronic HBV carriers and 21 cases of inactive hepatitis B surface antigen(HBsAg) carriers ,and 50 healthy individuals were collected .Serum levels of IFN‐γ,IL‐32 and IL‐6 ,the amount of HBV DNA and liver function were detected ,and clinical correlations were analysed .Results Compared with the control group ,ser‐um levels of IFN‐γ,IL‐32 and IL‐6 of chronic HBV carriers and inactive HBsAg carriers were significantly increased (P<0 .05) . Compared with chronic HBV carriers with low amount of HBV DNA loads and high amount of HBV DNA loads ,chronic HBV car‐riers with medial HBV DNA loads had higher serum levels of IFN‐γ,IL‐32 and IL‐6 ,but no statistically significant differences were observed(P>0 .05) .There was positive correlation between IL‐32 level and ALT level (r=0 .32 ,P<0 .05) ,and negative correla‐tion between IL‐32 level and ALB level(r= -0 .27 ,P<0 .05) .Conclusion IFN‐γ,IL‐32 and IL‐6 may play important roles in chro‐nic HBV infection ,the levels of IFN‐γ,IL‐32 and IL‐6 could be used as important indicators to assess the severity of inflammation in HBV carriers .

Objective To establish the quality standard for Jinyin Lidan Oral Liquid. Methods Artemisiae Scopariae Herba, Taraxaci Herba, Aurantii Fructus and Schisandrae Chinensis Fructus in Jinyin Lidan Oral Liquid were identified by TLC. The contents of naringin and neohesperidin were determined by HPLC. The HPLC separation was performed on Phenomenex C18 column (4.6 mm × 250 mm, 5 μm) with the mobile phase of methanol-0.1% phsophonic acid solution and gradient elution. The UV detection wavelength was 254 nm;flow rate was 1.0 mL/min;column temperature was 25 ℃. Results The results of TLC showed that relevant spots were clear, with strong specificity, without interference of negative sample. The calibration curves for naringin and neohesperidin were in good linearity in the range of 0.56-5.60μg (r=0.999 9) and 0.38-3.80μg (r=0.999 8), respectively. The average recoveries were 99.86%and 98.95%with RSD of 2.04%and 2.45%, respectively. Conclusion The qualitative and quantitative determination method is simple, accurate and reliable, with good stability and reproducibility, and can be used for the quality control of Jinyin Lidan Oral Liquid.

Objective To establish the quality standard of Jiangzhi Fugan Capules. Methods TLC was used to the qualitative identification of Paeoniae Radix Rubra and Salviae Miltiorrhizae Radix et Rhizoma. The contents of Paeoniflorin and Tanshinone ⅡA were determined by HPLC. The HPLC separation was performed on Phenomenex C18 column (4.6 mm×250 mm, 5 μm), with methanol-0.1% phsophonic acid (31∶69, 75∶25) as mobile phase, and the flow rate was 1.0 mL/min. Results The results of TLC showed that relevant spots were clear without interference against the negative sample. The calibration curves for Paeoniflorin and Tanshinone ⅡA were found to be liner within the range of 0.448-4.48μg, 0.057 6-0.576μg, respectively. The correlation coefficients were 0.999 4 and 0.999 5, respectively. The average recoveries were 99.93%and 99.75%, with RSD of 1.98%(n=9) and 1.70%(n=9), respectively. Conclusion The method is accurate, reliable, stable, rapid, and reproducible, and can be used for the quality control and evaluation of Jiangzhi Fugan Capules.

Objective:To investigate the optimal water extraction process of Jinyin Lidan oral liquids. Methods:An orthogonal de-sign was used in the extraction optimization using the yield and content of chlorogenic acid,narigin and neohesperidin as the evaluation indices and the volume of water,extraction times,extraction time and soaking time as the influencing factors. The contents of chloro-genic acid,narigin and neohesperidin were detected by HPLC to screen the best extraction technology. Results:The best water extrac-tion conditions were as follows:the amount of the added water was 8 times of the medicinal materials,the soaking time was 1 h,and the decoction was carried out twice with the duration of 1. 5 h for each. Conclusion:The process is stable and reasonable,which pro-vides research foundation for the production and quality control.

Objective To investigate the changes and clinical correlation of IL-4 in patients with HBV infection.Methods Sixty cases with chronic asymptomatic HBV carriers,60 cases with chronic hepatitis B,60 cases with liver cirrhosis,60 cases with hepato-cellular carcinoma and 50 healthy controls were collected for serum.ELISA was used for detection of cytokine IL-4 levels;The a-mount of HBV DNA was measured by fluorescence quantitative PCR;Liver function was tested by automatic biochemical analyzer. Results Compared with the healthy controls[(1.64±0.17)ng/mL],IL-4 levels of patients with CHB,LC and HCC were signifi-cantly increased[(4.18±0.48),(4.71±0.42),(3.62±0.31)ng/mL,P <0.05].LC group have the highest IL-4 levels,while ASC was the lowest.Compared with the ASC group,IL-4 levels of patients with CHB,LC and HCC were significantly increased(P <0.05).Compared with the HCC group,IL-4 levels of patients with LC were significantly increased(P <0.05).IL-4 level was posi-tively correlated with TBIL levels in LC patients(r=0.529,P <0.01),while the IL-4 level was positively correlated with ALT and TBIL level in HCC patients(r=0.263,0.323,P <0.05).Conclusion IL-4 may play an important role in chronic HBV infection, the levels of IL-4 can be used as an important indicator to assess the severity of chronic hepatitis B.

Objective To investigate the value and clinical significance of CD4+CD25+ regulatory T cells (Treg) ,inteleukin‐18 , interferon‐γand transforming growth factor‐βin patients with hepatitis B virus (HBV) infection .Methods A total of 175 patients with HBV infection were divided into chronic type B hepatitis (CHB) group and chronic asympotomatic HBV carrier (ASC) group , which were further divided into hepatitis B e antigen (HBeAg) positive and negative groups .CD4+ CD25+ Treg ,cytokines levels and liver function were measured .Healthy subjects were enrolled into control group .Results Proportion of CD4+CD25+ Treg were without significant difference among healthy controls ,HBeAg(+ ) and HBeAg(-) ASC groups ,and HBeAg(+ ) and HBeAg(-) CHB groups (P>0 .05) .Compared with control group ,cytokines levels were significant higer in CHB and ASC group (P<0 .05) . CD4+CD25+ Treg level was significant positive correlation to alanine aminotransferase and ratio of aspartate aminotransferase to plateles in HBeAg(+ )ASC group (P<0 .05) .Conclusion CD4+CD25 + Treg and related cytokines could play important roles in the course of CHB ,while CD4+CD25+ Treg expressing might be correlated with inflammatory degree of hepatitis .

Cholestatic liver disease is a common disease that causes bile flow dysfunction due to various reasons. The etiology of cholestatic liver disease is complexed, and therapeutic drugs are extremely limited. To date, ursodeoxycholic acid is the only FDA-approved drug for treating primary biliary cirrhosis, whereas its efficacy is limited to early stage of the disease, therefore novel drugs are urgently needed. Nuclear receptors become therapeutic hotspot target in cholestasis since these receptors play a key role in regulating bile acid homeostasis. Peroxisome proliferator-activated receptor (PPAR) is an important nuclear receptor involved in regulating multiple mechanisms of cholestasis in vivo. It can improve intrahepatic cholestasis by inhibiting bile acid synthesis, reducing bile acid toxicity, affecting the expression of bile acid metabolic enzymes and transporters, and can play an anti-inflammatory, anti-oxidation and anti-fibrosis role. A number of studies have shown that PPAR agonists represented by fibrates alone or in combination can improve liver function indexes, inflammatory factors and fibrosis markers in patients with cholestasis. This review analyzes and summarizes the lastest advances in the molecular mechanism of PPAR as a therapeutic target for cholestasis and drug treatment in development or have been used in clinical.