OBJECTIVE: To establish an HPLC-MS/MS method for the determination of blood-bivalirudin concentration. METHODS: Following a SPE method, the analytes were separated on a C18 column interfaced with a triple-quadrupole tandem mass spectrometer using positive electrospray ionization. RESULTS: The calibration linear rang of blood-bivalirudin concentration was 25-20000 μg · L-1 (r = 0.9969). The between-day and within-day RSD% were less than 6.63%. The recoveries were more than 80%. CONCLUSION: The rapid, sensitive and reliable HPLC-MS/MS method can be applied to monitor blood- bivalirudin profile.

Objective To understand the clinical features of hepatitis B virus(HBV)/hepatitis C virus(HCV)coinfected patients with different virological profiles.Methods The clinical data of 186 patients with HBV/HCV coinfection from May 1999 to May 2010 in the Third Affiliated Hospital of Sun Yat-Sen University were analyzed retrospectively.The demographic data,epidemiological data,laboratory results and pathological index were analyzed.The statistical analysis was done using t test and chi square test.Results A total of 186 patients were divided into 4 groups:66(35.5%)in HBV DNA(-)/HCV RNA(-)group,8(4.3%)in HBV DNA(+)/HCV RNA(+)group,68(36.6%)in HBV DNA(+)/HCV RNA(-)group and 44(23.7%)in HBV DNA(-)/HCV RNA(+) group.The gender composition,complication incidence,transmission among drug users,alanine aminotransferase(ALT)level,total bilirubin(TBil)level,prothrombin activity(PTA)and hapatitis B e antigen(HBeAg)negative rate were all significantly different among four groups(F or x2=11.578,8.451,11.738,2.669,5.102,4.254 and 18.413,respectively;all P<0.05).In groups of HCV RNA(-)and HCV RNA(+),the proportions of patients infected through drug abuse were 49.3%and 23.1%,respectively(x2=9.987,P:0.002)and blood transfusion transmission were 29.9%and 46.2%,respectively(x2=4.412,P=0.036).When HBV DNA was negative,the median ALT levels in HCV RNA(-)and HCV RNA(+)patients were 177 U/L and 62 U/L,respectively(t=2.200,P<0.05),median TBil levels were 133 μmol/L and 20μmol/L,respectively (t=3.608,P<0.05)and PTA were 70.6%±27.7%and 83.3%±27.8%,respectively(t=-1.982,P<0.05).The HBeAg negative rate was not affected by HCV RNA levels(t=0.707,P>0.05).The HBeAg negative rate in HBV DNA(-)patients was 85.5%,which was higher than that in HBV DNA(+)patients(59.2%)(x2=16.393,P<0.05).Conclusions HBV DNA(+/-)/HCV RNA(-)profile were major components in HBV/HCV confection.HBV DNA level is related to disease progression and prognosis,but not relate to disease severity.Liver function damage and disease severity are aggravated with HCV RNA level decreases.HBV DNA level is related to HBeAg negative rate,while HCV RNA level is not related to HBeAg seroconversion rate.

It is now well established that inflammation plays an important role in the development of numerous chronic metabolic diseases including insulin resistance (IR) and type 2 diabetes (T2DM). Skeletal muscle is responsible for 75% of total insulin-dependent glucose uptake; consequently, skeletal muscle IR is considered to be the primary defect of systemic IR development. Our pre- vious study has shown that rutaecarpine (Rut) can benefit blood lipid profile, mitigate inflammation, and improve kidney, liver, pan- creas pathology status of T2DM rats. However, the effects of Rut on inflammatory cytokines in the development of IR-skeletal muscle cells have not been studied. Thus, our objective was to investigate effects of Rut on inflammatory cytokines interleukiri (IL)-1, IL-6 and tumor necrosis factor (TNF)-α in insulin resistant primary skeletal muscle cells (IR-PSMC). Primary cultures of skeletal muscle cells were prepared from 5 neonate SD rats, and the primary rat skeletal muscle cells were identified by cell morphology, effect of ru- taecarpine on cell proliferation by MTT assay. IR-PSMC cells were induced by palmitic acid (PA), the glucose concentration was measured by glucose oxidase and peroxidase (GOD-POD) method. The effects of Rut on inflammatory cytokines IL-1, IL-6 and TNF-α in IR-PSMC cells were tested by enzyme-linked immunosorbent assay (ELISA) kit. The results show that the primary skeletal muscle cells from neonatal rat cultured for 2-4 days, parallel alignment regularly, and cultured for 7 days, cells fused and myotube formed. It was shown that Rut in concentration 0-180. 0 μmol x L(-1) possessed no cytotoxic effect towards cultured primary skeletal muscle cells. However, after 24 h exposure to 0.6 mmol x L(-1) PA, primary skeletal muscle cells were able to induce a state of insulin resistance. The results obtained indicated significant decrease (P < 0.05 to P < 0.001) IL-1, IL-6 and TNF-α production by cultured IR-PSMC cells when incubating 24 hours with Rut, beginning from 20 to 180.0 μmol x L(-1). IL-1, IL-6 and TNF-α in the Rut treated groups were dose-dependently decreased compared with that in the IR-PSMC control group. Our results demonstrated that the Rut promoted glucose consumption and improved insulin resistance possibly through suppression of inflammatory cytokines in the IR-PSMC cells.
Animals ; Cell Proliferation ; drug effects ; Cytokines ; metabolism ; Female ; Glucose ; metabolism ; Indole Alkaloids ; pharmacology ; Inflammation ; metabolism ; Insulin Resistance ; Male ; Muscle, Skeletal ; cytology ; drug effects ; metabolism ; Quinazolines ; pharmacology ; Rats

Objective To retrospectively investigate the relapse rates of different duration of extended consolidation after withdrawal of nucleos(t) ide analogues (NAs) treatment in patients with chronic hepatitis B (CHB) who met NAs cessation criteria.Methods 102 CHB patients discontinued treatment according to NAs cessation criteria or extended duration of consolidation therapy after meeting the cessation criteria.30 patients meeting the cessation criteria were Group A.72 patients extending consolidation therapy after meeting the cessation criteria were Group B.Based on different duration of extended consolidation therapy,72 patients were divided into 3 groups.Patients with a duration of extended 12 months after meeting NAs cessation criteria were Group B1.Patients with a duration of extended 24 months were Group B2.Patients with duration of extended 36 months were Group B3.After cessation of NAs treatment,the cumulative relapse of different group was calculated by the Kaplan-Meier method.The cumulative relapses between the selected groups were analyzed with Log-rank test.Results The cumulative relapse rates after 6,12,18,24,36 and 48 months after cessation of NAs treatment were 52.3％,70.0％,74.3％,76.7％,82.4％ and 88.4％ in Group A;24.0％,38.8％,40.6％,43.3％,43.3％ and 43.3％ in Group B;respectively.The relapse rate of Group B was much lower than that of Group A.The cumulative relapse rates after 6,12,18,24,36 and 48 months after cessation of NAs treatment were 35.0％,48.2％,51.9％,57.9％,57.9％ and 57.9％ in Group B1;18.1％,30.7％,30.7％,30.7％,30.7％ in Group B2;7.1％,21.4％,21.4％,21.4％ in group B3;respectively.The relapse rate of Group B1 was the highest,the following was of Group B2,and of Group B3 was the lowest one.The total amount of relapse in Group A,B1,B2 and B3 was 26,21,5 and 3 respectively.Conclusions A longer duration of extended consolidation therapy after meeting NAs cessation criteria may contribute to the lower relapse rates.

Objective To determine the value and the optimal cutoff point of waist circumference (WC) in screening diabetes mellitus (DM) and to provide evidence for DM prevention and identifying population at risk in mid-western rural areas of Shandong province.Methods A sample consisting 16 341 rural residents was selected and studied.All participants were physically examined on height,weight,WC and fasting plasma glucose (FPG).Oral glucose tolerance test (OGTT) was performed for subjects with FPG valued from 6.1 to 7.0 mmol/L.DM was defined according to the criteria set by WHO in 1999.Area under the curve (AUC),sensitivity,specificity and Youden index were computed based on the receiver operating characteristic (ROC) curve analysis.Optimal cutoff point was determined by the maximum of Youden index.Results The prevalence rates of DM for males and females increased along with the rise of WC (trend test X2=72.01,122.65,P＜0.01 ).It appeared significantly higher in those with WC 85 cm in females and≥80 cm in males,with those WC ＜85 cm for females and ＜80 cm for males,in particular.AUCs were 0.639 and 0.655 for males and females respectively and both had significant differences (t=7.22,11.07,P ＜0.01 ).However,the AUCs did not show significant difference (t=0.70,P ＞ 0.05) between males and females.The Youden index reached maximum when WC approached 85 cm for females (24.90%) and 80 cm for males (24.39%).The sensitivity and specificity were 58.04%and 66.86%for males,and 67.08%and 57.31%for females.Conclusion WC seemed to be an effective indicator for screening the DM.The optimal cutoff point of WC would be 85 cm for females and 80 cm for males in screening DM and defining the population at risk in this area.

Objective To investigate the efficacy of combined treatment of PEG IFNα-2a and recombinant hepatitis B vaccine in CHB patients with HBeAg positive.Methods 75 CHB patients with HBeAg positive were enrolled into this study.45 patients received the monotherapy of pegylated IFNα-2a (group A),and 30 patients were treated with PEG IFNα-2a combined with recombinant hepatitis B vaccine (group B).The two groups were compared clinical features,such as ALT,HBsAg levels and HBeAg seroconversion rates,HBV DNA suppression,at different time point(At 0,24,48,72 week).Results At week 0,levels of aminotransferases,HBsAg and HBV DNA were not statistically significant between the two groups(P ＞0.05).But the level of HBeAg in group B was much more than that in group A.This diversity show statistical significance(P ＜ 0.05).During week 24 to week 48,rates of aminotransferases normalization HBsAg seroconversion HBeAg seroconversion,and HBV DNA suppression were also not statistically significant between group A and B (P ＞ 0.05).At the 72 week of follow up,levels of aminotransferases,HBeAg seroconversion rate and HBsAg levels were not statistically significant among the two groups (P ＞0.05),but the negative conversion rate of HBV DNA drop in group B was much more than that in group A,the difference was statistically significant (P =0.032).Conclusion The combined treatment of PEG IFNα-2a and recombinant hepatitis B vaccine in CHB patients with HBeAg positive can improve the negative conversion rate of HBV DNA 72 weeks after the end of the 48 week of treatment,but wasn't associated with HBeAg seroconversion and HBsAg levels.

Objective To investigate short-term efficacy and security of transplantation of human umbilical cord mesenchymal stem cells (HUCMSCs)in the course of treatment of decompensated cirrhosis.Methods Ninety-six Hepatitis B patients with decompensated cirrhosis were enrolled,among which 50 were sbject to transplantation of HUCMSCs in addition to routine therapy (the treatment group) and the rest patients were underwent routine therapy(the control group).We observed the efficacy and security at 2,4,6 and 12 week of the treatment course.Results The symptoms of atigue,anorexia,bloating and edema were greatly relieved in both groups after 4 weeks,and sustained remission after 4-12 weeks.The overall survival rate was 96％ after 12 weeks of the treatment group,2 patients were died after 8 and 12 weeks of the HUCMSCs transplantation due to hepatic encephalopathy repecticely.Compared to the baseline,ALT,AST,TBil and ALB(albumin) were improved after 2,4,6 and 12 weeks in both groups(P ＜ 0.05),and the averages of ALB(albumin) of treatment group were higher than the control group.PT(prothrombin time) and PTA(prothrombin time activity) were improved after 4,6,12 weeks in the treatment group (P ＜ 0.05),however,there were no differences in the control group during the 4,6 and 12 weeks(P ＞0.05).During the course of HUCMSCs tranplantation,the security was satisfied and no special side effects were observed.Conclusion The transplantation of HUCMSCs for therapy of Hepatitis B patients with decompensated cirrhosis is a safety and effective therapy,and can improved ALB,PT,PTA,liver function and clinical symptoms within a short period,which is an alternative method of treatment recommended.

Objective To investigate whether the combination therapy of pegylated IFNα-2a plus adefovir dipivoxil (ADV) improve the efficacy of the treatment in CHB patients with HBeAg positive or not. Methods 57 CHB patients with HBeAg positive received 48-week pegylated IFNα-2a therapy were enrolled into this study. If serum HBV DNA levels exceeded 1000 copies/ml at week 24, the patients were assigned to group A (pegylated IFN-α2a plus ADV, 21 cases) or group B (pegylated IFNα-2a only, 14 cases);otherwise, they received the unceasing monotherapy of pegylated IFNα-2a (group C, 22 cases). Results At week 48, HBeAg seroconversion rates were 23.8% , 28.6% and 63.6% (A vs C,P=0.014), but rates of aminotransferases normalization and HBV DNA suppression (< 1000 copies/ml) were not statistically significant among three groups. But during week 24 to week 48, rates of HBeAg seroconversion, aminotransferases normalization and HBV DNA suppression were also not statistically significant between group A and B. But amplitude of DNA drop in group A was much more than that in group B (2.60±1.37 vs 0.86±2.09, P=0.005). Conclusion An ADV add-on therapy in pegylated IFNα-2a treatment seems able to improve the inhibition of HBV DNA in chronic hepatitis B patients with HBeAg positive. It requires a large, double-blind, randomized clinical trial to further provent.

Objective To investigate the efficacy of nucleot (s)ide analogues therapy in patients with HBeAg-negative cirrhosis in China. Methods 111 patiens with HBeAg-negative cirrhosis were divided into antiviral group (58 cases, 25 entecavir, 19 adefovir dipivoxil, 13 lamivudine, 1 telbivudine) and control group (53 cases, supportive and symptomatic treatment). These two groups were matched for demography, liver function and Child-Push score. Results At the 96th week, the rate of ALT normalization and HBV DNA drop (lg copies/ml) in antiviral group were higher than those in control group(P<0.05). The rates of HBV DNA negative (< 500copies/ml) were 88.7% (47/53) and 32.5% (13/40) , respectively (P < 0.05). There were no differences in the rates of developing HCC and undergoing variceal bleeding between antiviral group and control group (P>0.05 ). 15.4% patients with lamivudine treatment emerged YMDD mutations. 10.5% patients with adefovir dipivoxil treatment emerged virologic breakthrough and hepatitis flare during the second year. 2 patients (3.5%) in treatment group and 6 patients (11.5%) in control group died of liver failure or variceal bleeding or HCC (P>0.05). Conclusions Neucleot (s) ide analogues are effective in suppressing HBV replication in patients with HBeAg-negative cirrhosis, but the impact of which on the mortality and complications of cirrhosis should be prolongly observed. For continuing treatment, the neucleot(s) ide analogues with strong effective and low resistance are the first choices to prevent viral mutation and drug resistance.

Objective To indentify the relation between hepatic ceils apoptosis and the lesion of liver tissue in acute toxic lethal hepatitis. Methods 60 Wistar mice were randomly divided into normal control, model group and treatment group. Normal control and model group were pretreated by portal vein injection of normal saline, the treatment group was pretreated by portal vein injection of BCL-Xl adenoviruses. The mice of model group and treatment group were received an injection of D-gain and LPS to establish fulminant hepatic failure models 7 days after pretrement. To observe BCL-Xl expression, serum ALT,AST, hepatocyte apoptosis rate, and mortality rate of the three groups. Results The BCL-Xl expression was higher in treatment group than in model group ;6 hours after fulminant hepatic failure models were established, the serum ALT, AST level of treatment group was lower than model group; The hepatoeyte apoptosis rate of treatment group was lower than model group. The death rate of treatment group was lower than model group. Conclusion In fulminant mice hepatic failure models,the hepatocyte apoptosis rate has a positive correlation with death rate, the overexpression of BCL-Xl can decrease the hepatocyte apoptosis rate and the death rate.