Six compounds were isolated from Glycyrrhiza yunnattettsis. on the basis of physico-chemical properties and spectroscopic analysis, their structures were identified as homopterocarpiu (Ⅰ ), medicarpin (Ⅱ ), formononetin (Ⅲ ), 4' -methoxy-4-hydroxy challone (Ⅳ ), glyyunnansapogeuin B (Ⅴ ) and ?-sitosterol (Ⅵ ). Ⅰ~Ⅴ are fouud for the first time in this species,

Objective To investigate the characteristics of secondary infections in hepatitis B patients with acute-on-chronic liver failure(ACLF)and its impact on the prognosis. Methods Infection sites, clinical and etiological characteristics were retrospectively reviewed in 186 hospitalized patients with ACLF from the Third Affiliated Hospital of Sun Yat-sen University during January to December 2007. Logistic regression was used to analyze risk factors of secondary infections. Results In 186 patients with ACLF, 160 patients(86.0%) were complicated with infections, and the common sites of infections were abdominal cavity, biliary tract, lung and intestinal tract. The rates of secondary infections were higher in patients with serum albumin(Alb)≤30 g/L, total bilirubin(TBil)>342 μmol/L, prothrombin time(PT)>28 seconds, and those complicated with one or more complications(χ~2=5.4, 7.3, 21.3 and 14.7, P<0.05). The fatality rates of patients with and without infections were 74. 5%(119/160) and 42.3%(11/26), respectively, and the difference was of statistical significance(χ~2=10.9,P=0.000). Patients with multi-organ infections had a higher fatality rate(79.8%, 79/99)than those with one organ infections(65.6%, 40/61), and the difference was also significant(χ~2=4.0, P=0.045). Conclusion Patients with ACLF are liable to infection, and the severity is closely related with the prognosis.

Background and purpose:At present,the treatment of metastatic breast cancer has remained diff icult for patients who have failed after the use of anthracycline and taxane,so the treatment options for those with advanced metastatic breast cancer are very limited.Therefore,pemetrexed combined with cisplatin in the treatment of anthracycline and taxane failed advanced metastatic breast cancer patients,its effi cacy and adverse reactions were studied.Methods:10 patients with advanced metastatic breast cancer patients received pemetrexed and cisplatin chemotherapy,pemetrexed 500mg/m2 intravenous infusion,cisplatin 75mg/m2 intravenous drip,repeated every 3 weeks.Evaluation of response and adverse reactions were practiced every 2 cycles.Results:10 patients were evaluable,CR 1 patient,PR 4 patients,SD 2 patients,PD 3 patients.With a median follow-up of 6 months(4-18 months),8 patients survived and 2 patients died.The median time to progression was 4 months(3-14 months),the median survival time was 8 months(4-18 months).The most common adverse reaction was fatigue,white blood cells decrease and the digestive tract reactions,followed by oral ulcers.Conclusion:Pemetrexed combined with cisplatin in the treatment of anthracycline and taxane treatment failed patients with advanced metastatic breast cancer could improve survival,adverse reactions could be tolerated.

Objective To study programmed death-1 (PD-1) and T-cell immunoglobulin mucin3 (Tim-3) co-expression on peripheral blood mononuclear cells (PBMC) and hepatitis B virus (HBV)-specific CD8+ T cells in patients with chronic HBV infection and its correlation with liver inflammatory activities.Methods One hundred and sixty subjects with chronic HBV infection who visited the outpatient and inpatient Department of Infectious Diseases in the Third Affiliated Hospital of Sun Yatsen University were enrolled,including 63 cases of mild or moderate CHB (MCHB),31 of severe CHB (SCHB),55 of acute-on-chronic liver failure (ACLF) and 11 inactive HBsAg carriers.Twenty healthy subjects were enrolled as controls.Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect HBV DNA,enzyme-linked immunosorbent assay (ELISA) to measure HBV serological markers,and flow cytometry to detect human leukocyte antigen (HLA)-A2 and determine the expression of PD-1 and Tim-3 on PBMC and HBV-specific CD8+ T cells.Cell counts of Tim-3+,PD-1+,and Tim-3+/PD1+ PBMC and HBV-specific CD8+ T cells were compared among different groups,and their correlation with commonly used inflammatory activity indicators were studied.Analysis of variance and Kruskal-Wallis test were used for measurement data with gaussian distribution and skewed distribution,respectively.Spearman correlation analysis was used to assess the association between Tim-3/PD-1 expression and inflammatory activity indicators.Results The frequency of Tim-3+/PD-1+ PBMC was 0.25 ％ in ACLF group (P=0.0049),0.24 ％ in SCHB group (P-0.0025) and 0.13％ in MCHB group (P=0.0006),which were significantly higher than that in control group (0.03％),and the frequency of Tim-3 /PD-1-PBMC in the three groups were significantly lower than that in control group (P =0.0000),but the differences between HBsAg carriers (0.10％) and controls (0.03％) were not statistically significant (P=0.28).Among 160 subjects,78 were HLA-A2 positive.The frequency of Tim-3+/PD-1+ HBV-specific CD8+ T cells was 68.72％±17.21％ in ACLF group,and 59.66％± 19.25％ in SCHB group,which were significantly higher than that in HBsAg carrier group (33.93％ ± 10.80％,P=0.0000,P=0.0054).The frequency of Tim-3 /PD-1-HBV-specific CD8+ T cells in ACLF group was 2.80％,which was significantly lower than that in HBsAg carrier group (27.24％,P=0.0004).The frequency of Tim3+/PD-1+ HBV-specific CD8+ T cells was positively correlated with alanine aminotransferase (r=0.26,P=0.022),aspartate aminotransferase (r=0.28,P=0.012) and total bilirubin levels (r=0.36,P=0.001); and inversely correlated with albumin level (r=-0.35,P=0.002) in serum.Furthermore,it was positively correlated with international normalized ratio (INR,r=0.34,P =0.045) and model for end-stage liver disease score (r=0.43,P=0.027) in ACLF group.Conclusions Co-expressions of Tim-3 and PD-1 on PBMC and HBV-specific CD8+ T cells are significantly upregulated in patients with CHB,and correlated with liver inflammatory activities.These findings indicate that Tim-3 and PD-1 co-expression may involve in disease activity and liver failure in CHB.

Objective To analyze the relationship between HBsAg level and HBV DNA level as well as illness severity in patients with HBV-related acute-on-chronic liver failure (ACLF).Methods One hundred and nineteen patients with chronic hepatitis B (CHB group) and 98 patients with ACLF(ACLF group) were enrolled.HBsAg and HBeAg were assayed with Roche electrochemical luminescence method.HBV DNA was quantified using a real-time polymerase chain reaction (PCR) assay.HBsAg and HBV DNA levels were compared between two groups and between HBeAg-positive and HBeAg-negative patients in ACLF group respectively,also the correlationbetween HBsAg and HBV DNA was studied.Results The proportions of HBeAg-negative patients were 68.4％(67/98) and 42.9％(51/119) in ACLF group and CHB group respectively,and there was significant difference between two groups (P ＜0.01).There was no significant difference in HBV DNA between two groups (P ＞ 0.05).HBV DNA in HBeAg-positive patients was higher than that in HBeAg-negative patients in two groups(P ＜ 0.05).There was no significant difference in HBsAg between HBeAg-positive patients and HBeAg-negative patients in ACLF group (P ＞ 0.05 ),but they were higher than that in HBeAg-positive patients in CHB group (P＜ 0.05).HBsAg was correlated to ALT,AST in HBeAg-positive patients (P ＜ 0.05).No significant correlation was found among HBsAg and HBV DNA as well as biochemical changes (P＞ 0.05).There was no significant difference in the ratio of different HBsAg levels among the patients of different HBV DNA in ACLF group (P＞ 0.05).Conclusion The level of HBsAg does not directly correlate with serum HBV DNA level,and has no directly correlation with the severity of the disease in patients with HBV-related ACLF.

Objective To investigate whether the spleen diameter,serum albumin and periphefial blood cells might be as non-invasive predictive indicators for the presence of esophageal varices(EV)in patients with liver cirrhosis.The predictive values of these parameters to the large esophageal varices were evaluated.Methods OBe hundred and sixty-seven patients with liver cirrhosis underwent endoscopic examination.Among them,127 patients(mild in 41,medium in 38,severe in 48)were found with EV and 40 patients without(NEV).The diameters of portal vein and spleen vein,the sizes of spleen and the ratio of platelet count/spleen size were examined by Doppler ultrasound.The platelet count and the level of albumin were calculated.Results The average of age,diameter of portal vein and spleen vein,and sizes of spleen were higher in EV group than those in NEV group,while the platelet count,the level of albumin and the ratio of platelet count/spleen size in EV groups were lower than those in NEV group.Multifactor analysis revealed that the index related to serious EV were the blood platelet count(＜70×109/L),the ratio of platelet count/spleen size(＜1.0)and albumin level (＜35 g/L).Conclusions The degree of EV in patients with liver cirrhosis were paralleled with the degree of portal hypertension.The patients who present with platelet count＜70×109/L,or platelet count/spleen size＜1.0 or albumin＜35 g/L should be considered as EV,and endoscopic examination is needed.

OBJECTIVETo study the morphologic and growing alterations of oral cancer cell line Tca8113 before and after cocultured with tumor stromal fibroblasts (TSF) and normal stromal fibroblasts (NSF) respectively, and evaluate the influence of mesenchymal cells on tumor cells.

METHODSTSF and NSF were isolated and cultured. To observe the morphologic change of Tca8113 cells after cocultured with TSF and NSF respectively.

RESULTSWhen cocultured with NSF, the Tca8113 cells proliferated as rapidly as monocultured to form colonies, while the NSF proliferated slowly to form pieces and then joined each other to form network. The NSF network segmented and surrounded the colonies of cancer cells so that the cancer cells shrank, turn round, broke away from the bottom and floated into the medium. The cancer cells proliferated actively but they were elbow out entirely in the end. TSF proliferated slowly when cocultured with cancer cells, projected several branched protrusions. The cancer cells proliferated along the two sides of protrusions of TSF, or projected short protrusions to connect the body or protrusions of TSF, and overlaid the protrusions gradually, finally, cover the body. In the end, TSF melt away, and the cancer cells took on the figure of TSF.

CONCLUSIONThe results do suggest that, oral cancer cell line Tca8113 are restrained when coculture with NSF, but are promoted when with TSF.

Objective To evaluate the effects of nucleoside/nucleotide analogue treatment on immunoglobulin and complement in patients with chronic hepatitis B (CHB).MethodsA total of 157 CHB patients were recruited and divided into CHB group,liver cirrhosis (LC) group and severe hepatitis B (SHB) group.There were 50 patients who received oral antiviral treatment (lamivudine 100 mg/d,or entecavir 0.5 mg/d,or telbivudine 600 mg/d).Serum levels of complement 3 and 4 (C3,C4),C-reaction protein (CRP),hemolytic complement (CH50),immunoglobulin G,M,A (IgG,IgM,IgA),hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) were detected by enzyme-linked immunosorbent assay (ELISA) or immunoturbidimetry.Hepatitis B virus (HBV) DNA was quantified by real-time polymerase chain reaction (RT-PCR) before and 1,2,3 and 4 weeks after nucleoside antiviral therapy.Comparison of means was done by t test and Mann-Whitney test.The correlation was analyzed by Pearson correlation coefficient test.ResultsSerum IgA and IgM levels of SHB and LC patients were significantly higher than those of CHB patients (P＜0.01).Levels of C3,C4,CH50 and CRP were significantly different among three groups.Levels of C3,IgM,IgG and HBV DNA in HBeAg positive patients were significantly different from those in HBeAg negative patients.There was a statistically significant difference of IgA,IgM,C3 and CH50 levels between patients with high HBV DNA level and low HBV DNA level in HBeAg-positive patients.While in the HBeAg-negative patients,only the IgA level was significantly different with HBV DNA levels.After anti-viral treatment,immunoglobulin and HBV DNA levels were all decreased in three groups,while the serum complement level was increased compared to baseline,and the differences became significant at week 4 of treatment. HBV DNA level was negatively correlated with C3 (r=-0.78,P=0.021) and HBeAg titer was positively correlated with C3 (r=0.87,P=0.015).ConclusionsThe immunoglobulin,CRP,C3,C4,and C H50 could reflect the inflammatory activity in liver.The changes of C3 level can predict the efficacy of antiviral therapy.

Objective To study the relationship between programmed death-1 (PD-1)/programmed death-1 ligand (PD-L1) expressions and serum hepatitis B virus (HBV) DNA levels in chronic hepatitis B (CHB) patients. Methods A total of 137 CHB patients and 10 healthy controls were enrolled in the study. The peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples. HBV-specific cytotoxic T lymphocyte (CTL) was expanded in vitro in 64 human leucocyte antigen (HLA)-A2 positive patients. Flow cytometry was used to detect HLA-A2 type,expressions of PD-1/PD-L1 on PBMCs and PD-1 on HBV specific CTL. Interferon gamma (IFN-γ)was measured by commercial enzyme-linked immunosorbent assay (ELISA) kits. PD-1/PD-L1expressions on PBMCs, HBV-specific CTL and IFN-γ level in PBMC culture medium were compared among patients with different baseline HBV DNA levels. Ten hepatitis B e antigen (HBeAg) positive patients were treated with telbivudine for 24 weeks. The above mentioned parameters were determined and compared before and after the antiviral treatment. Independent-samples t test were used to compare means between two groups and one-way A NOVA were used to compare means among multigroups. We used the pearson corretation test to assess corretation significance. Results The PD-1 and PD-L1 expressions on PBMCs in patients with baseline HBV DNA＜3 lg copy/mL, 3-6 lg copy/mL and ＞6 lg copy/mL were all significant higher than those in healthy control group, but no statistical differences were found. PD-1 expressions on HBV-specific CTL in the three CHB patient groups were (69.3±11.2)%, (76.5±9. 1)% and (78.0±11.7)%, respectively. However, PD-1 expression on HBV-specific CTL was higher, while the frequency of HBV-specific CTL cells was lower in HBV DNA ＞6 lg copy/mL group compared to HBV DNA＜3 lg copy/mL group. The above parameters, including expressions of PD-1 and PD-L1, the frequency of HBV-specific CTL and its PD-1 expression were not significantly different between HBeAg-positive group and HBeAg-negative group. Compared with baseline, PD-1 and PD-L1 expression decreased obviously accompanying with increase of HBV-specific CTL cells frequency and IFN-γ level after 12 weeks and 24 weeks of telbivudine treatment. Conclusions PD-1 expression on HBV-specific CTL correlates with serum HBV DNA level, but not HBeAg status in CHB patients. Suppression of HBV replication can reduce PD-1/PD-L1 expressions and partially restore HBV specific CTL function.

Objective To evaluate the therapeutic efficacy and its related factors of entecavir treatment for patients with acute on chronic hepatitis B liver failure (ACHBLF).Methods One hundred and eight patients with ACHBLF were enrolled and divided into entecavir group (n=53) and control group (n=55).HBV DNA level, liver function and 48-week survival rate were observed, and C ox regression model was established to identify the factors which may affect the efficacy of entecavir treatment.Results Totally 70 patients died in the study and 66 died within 12 weeks.The statistical difference on cumulative survival rate between two groups was observed from the third week on (χ2=5.357, P ＜ 0.05).The 48-weekcumulative survival rate in entecavir group was 47.2% (25/53), while that in the control group was 23.6%(13/55) (χ2=7.432, P ＜ 0.01).In entecavir group, for patients aged ＜ 40 with serum bilirubin level ＜513 μnol/L and international normalized ratio (INR) ＜ 2.5, the fatality rates decreased 74.9%, 75.3%and 76.0%, respectively.Conclusions Entecavir may improve the survival rate of patients with ACHBLF.Age, serum bilirubin level and INR are major factors related to the therapeutic efficacy.