OBJECTIVE:To optimize the water extraction process for Shenshu yangwei capsule.METHODS:The decoction conditions were optimized by orthogonal design with table L9(34)taking the yield of Hesperidin as index.RESULTS:The optimal watering extraction conditions of Shenshu yangwei capsule were as follows:adding 8 times amount of water,then extracting for 3 times with 1 h for each time.CONCLUSION:The water extraction technology of Hesperidin is simple,with complete extraction and high yield of Hesperidin,thus applicable for industrialized large-scale production.

Objective To compare the difference of the Cif gene expression and the inflammation markers (such as WBC and CRP et al)between mucoid and non-mucoid Pseudonmonasaeruginosa,combining with the clinical data.Methods Real-time RT-PCR was used to determine the expression level of Cif in Pseudonmonas aeruginosa and clinical data were collected to discuss the relationship among the number of WBC,the level of CRP and the Cif gene expression.Results The expression level of Cif gene in mucoid Pseudonmonas aeruginosa had a significantly higher than the non-mucoid Pseudonmonas aerugi-nosa,which was statistically significant (t=2.09,P<0.05).Specimen data collected from patients with the infection of mu-coid or non-mucoid Pseudonmonas aeruginosa showed that the number of WBC and the level of CRP were not significantly different (t=0.65,0.60,P>0.05).The data also showed that the cystic fibrosis transmembrane conductance regulator in-hibitory factor (Cif)gene expression level had little correlation with the number of WBC (a=0.095,R2=0.029).Conclusion The expression of Cif gene may works for the biofilm formation of mucoid Pseudonmonas aeruginosa,which maybe a new virulence factor to Pseudonmonas aeruginosa.

Objective To investigate the correlation of hepatitis B patients in Fuzhou between resistance patterns in HBV P region and genotype,HBeAg,the hepatitis B process.Methods This was a retrospective study.The serum and clinic data of 1 115 hepatitis B patients were collected from the inpatient and outpatient Center for Liver Diseases in First Affiliated Hospital of Fujian Medical University between October 2011 and January 2015.HBV DNA was extracted and sequenced using the Sanger method to detect HBV genotype and resistance mutations in P region,HBeAg and HBeAb concentration were detected by chemiluminescent assay.The relationship between P region resistance mutations pattern,HBV genotype,serum HBeAg and the hepatitis B process was analyzed.The x2-test was used to compare the resistance rate and positive rate.Results There were significant differences between 14 kinds of resistance loci and the genotype distribution(x2 =30.788,P =0.004),the C/B genotype ratio of three common resistance loci (rtM204V/I,rtL180M,rtA181T/V) were 85/82,49/25 and 27/9,respectively,which in genotype C was higher than genotype B.The resistance ratio of hepatocellular carcinoma,liver cirrhosis,chronic hepatitis B,hepatitis B carriers was 31.4％ (11/35),37.6％ (65/173),27.3％ (146/535) and 21.8％ (43/197),respectively,which showed significant difference between the four clinical diagnosis (x2 =11.858,P =0.008).The highest percentage of resistance was liver cirrhosis,followed by hepatocellular carcinoma and chronic hepatitis B.There was significant difference in the distribution of HBV genotype between HBeAg (+) group and HBeAg (-) group (x2 =11.093,P =0.001),the HBeAg positive rate in genotype C [37.53％ (295/786)] was higher than in genotype B[35.62％ (280/786)].However,the total resistance rate between HBeAg (+) group and HBeAg(-) group was not significantly different[23.7％ (136/573) and 24.6％ (52/211),respectively,x2 =0.07,P =0.791].Conclusions HBV genotype was related to the resistance rates,HBeAg levels and the progress of hepatitis B.The resistance rate and HBeAg positive rate of genotype C were higher than those of genotype B,and clinical outcomes were worse in genotype C.HBV resistance rates and HBeAg levels were related to the progress of hepatitis B,the higher the resistance rates,the worse clinical outcomes.

Objective:To investigate the effect of Cyr61 on the proliferation of Fibroblast-like Synoviocytes (FLS) in rheumatoid arthritis (RA).Methods:Cyr61 expression in synovial tissues (ST) and FLS was examined using Real-time PCR,Western blot and immunohistochemistry simultaneously.FLS were isolated from synovial tissue of RA patients and cultured in vitro.The proliferation of FLS stimulated with synovial fluid (SF) was determined by 3 H-TdR incorporation.Cyr61 protein level in RA SF was detected by ELISA.Results:Cyr61 was over expressed in ST,FLS of RA patients while hardly examined in normal individuals and osterarthritis (OA) patients.Meanwhile,Cyr61 protein level was elevated in SF,which promoted the proliferation of RA FLS.This proliferation was abrogated by knockdown the Cyr61 gene of FLS or neutralizing monoclonal antibody against human Cyr61.Moreover,inflammatory factor IFN-γ and TNF-α up-regulated the expression of Cyr61.Conclusion:These results indicate that the elevated level of IFN-γ and TNF-α in RA SF can promote the proliferation of the FLS derived from RA patients by increasing expression of Cyr61,suggesting that Cyr61 may play an important role in the development of RA.

Objective To investigate the epidemiological and molecular virological characteristics in HBV-infected patients with copositive HBsAg and anti-HBs.Methods HBV serological markers were analyzed in 52 070 specimens.The epidemiological characteristics of HBsAg and anti-HBs simultaneously positive patients (the experimental group) and HBsAg positive and auti-HBs negative patients (the control group) were compared.The S protein of HBV coding region was amplified by semi-nested PCR and sequenced.The statistical differences between the two groups were compared in different gene regions,genotypes and different clinical diagnosis.Results HBsAg was positive in 20.40％ (10 621/52 070) of all specimens.In the patients with positive HBsAg,2.48％ (263/10 621) was positive anti-HBs.The prevalence of co-positive HBsAg and auti-HBs was higher in aged 0 to 9 years and greater than or equal to 80 years than that in other age,and the prevalence of positive HBsAg and negative anti-HBs was completely opposite.The mutation rate of S protein in the experimental group was significantly higher than that in the control group (1.52％ vs 0.81％,P ＜0.01) with the mutation in the major hydrophilic region (MHR) (1.68％ vs 0.57％,P ＜0.01).The mutation rates of S protein of HBV carriers,chronic hepatitis B (CHB) patients and patients with liver cirrhosis (LC) in the experimental group were significantly higher than those in the control group (1.47％ vs 0.65％,1.28％ vs 0.84％,2.21％ vs 0.44％,P ＜0.05,respectively),except for the patients with hepatocellular carcinoma (HCC) (1.97％ vs 2.21％,P ＞ 0.05).Conclusion Co-positive HBsAg and anti-HBs in HBV-infected patients was more common in HBsAg positive patients aged 0 to 9 years and greater than or equal to 80 years than the others.Coexistence of HBsAg and anti-HBs in HBV-infected patients may relate to immune escape caused by mutation of S protein (mainly MHR).The mutation rates of S protein in the two groups of patients,co-positive HBsAg and anti-HBs and the positive HBsAg combined with negative anti-HBs,were associated with the stage of liver disease.

Objective The study aims to investigate the associationbetweencholesterol 7α-hydroxylase (CYP7A1) gene polymorphism and different clinical outcomes after Hepatitis B virus (HBV)infection in Fujian Han population and lay a foundation for understanding the mechanisms of genesis anddevelopment of HBV-related diseases.Methods Case-control study was conducted.586 patients of HBVpersistent infection without antiviral therapy and 225 HBV rehabilitation patients (35-55 years old) werecollected from May 2015 to June 2016 in the Liverish Center of First Clinical College of Fujian MedicalUniversity.The group of HBV persistent infection without antiviral therapy included 246 patients with chronichepatitis B, 177 patients with hepatitis B-related cirrhosis, and 163 patients with hepatitis B-related liver cancer.The rs3824260, rs4738687and rs8192871 loci of CYP7A1 gene were detected by improved multipleligase detection reaction (iMLDR).Logistic regression analysis and chi-square test were used to analyze thegenotyping results.Results Three SNPs ( single nucleotide polymorphisms ) of CYP7A1 gene wereselected and compared between HBV persistent infection group and HBV rehabilitation group and betweenchronic hepatitis B subgroup, liver cirrhosis subgroup and liver cancer subgroup.After adjustment for factorsincluding age andgender, there was no significant difference in the distribution of rs3824260 genotype amongthe groups(χ2 =1.565,P =0.459), however,the frequency of allele C in HBV rehabilitation group wassignificantly higher than in HBV persistent in fectiongroup for men (χ2 =4.365,P =0.037), whereas thefrequency of rs3824260 CC and CT was more likely to be observed in liver cancer group than in non -livercancer group (chronic hepatitis B subgroup and liver cirrhosis subgroup ) for women (χ2 =5.768,P =0.012;χ2 =10.130,P =0.001).The frequency of rs4738687 GG genotype was more likely to be observed innon-liver cancer group than in liver cancer group (χ2 =4.403,P =0.041;χ2 =6.940,P =0.009).Theresults of gender stratification showed that there were significant differences in the distribution of rs 4738687among the HBV persistent infection groups for men (χ2 =10.697,P =0.030), however, there was nosignificant difference in the distribution of rs4738687 among the HBV persistent infection groups for women(χ2 =4.627,P =0.329), and there was no significant difference in the distribution of genotype frequencyand allele frequency among all groups(χ2 =0.489,P =0.792).There was no significant difference after sexstratification either (χ2 =1.282, P =0.526;χ2 =1.565,P =0.465) .Conclusions These findingssuggested that CYP7A1 gene polymorphism was related todifferent clinical outcomes in Fujian Hanpopulation.The rs3824260 mutation had a certain gender preference and the mutation allele was detected ina higher proportion in male patients.Male HBV patients with rs3824260 C allele had more chance ofswitching to rehabilitation.The rs4738687 was likely to be related to the occurrence of liver cancer in FujianHan population, and GG genotype may delay the occurrence and development of liver cancer especially in themale group.The rs8192871 was not found to be related to the different clinical outcomes of HBV infection.