Mechanisms of Apoptosis Induced by FTY720 in Multiple Myeloma Cell Line U266 / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of FTY720 on apoptosis in multiple myeloma cell line U266 and to clarify the molecular mechanism of apoptosis induced by FTY720.</p><p><b>METHODS</b>U266 cells were treated with 2.5, 5, 10 and 20 µmol/L of FTY720 for 24 hours, the apoptotic rates were tested by flow cytometry with Annexin-V-FITC/PI staining. Then U266 cells were treated with 20 µmol/L FTY720 for 0, 6, 16 and 24 hours, the apoptotic rates were tested. U266 cells were treated with DMSO and FTY720 separately and then were stained with DAPI for 5 min. Drop the cells to the slides and cover the slide with the glass. The cells were observed by fluorescence microscopy. U266 cells were treated with 5 µmol/L FTY720 or together with different doses of Z-VAD-fmk (12.5, 25, 50 µmol/L), a pancaspase inhibitor, for 24 hours, then the cell viability was tested by CCK-8. U266 cells were treated with 2.5, 5, 10 and 20 µmol/L of FTY720 for 24 hours, the expression of cleaved caspase-3 was tested by Western blot. U266 cells were treated with 0, 5, 10 and 20 µmol/L of FTY720 for 24 hours, the expressions of MCL-1, survivin, BCL-2, BID, BAX, BAK, P-ERK were tested by Western blot.</p><p><b>RESULTS</b>The apoptotic rate increased in U266 cells treated with FTY720 and showed the characteristic of time-dependent and dose-dependent manner. Karyopyknosis and nuclearfragmentation could be observed in U266 cells treated with FTY720 after being stained with DAPI under fluorescent microscope. The same effect was not observed in the cells treated with DMSO. Z-VAD-fmk could rescue the apoptosis in U266 cells treated with FTY720 in dose-dependent manner. The expression of MCL-1, survivin and BCL-2 decreased in U266 cells treated with FTY720. The cleavage of BID could be observed in U266 cells treated with FTY720. FTY720 had no effect on the expression of BAX, BAK and P-ERK.</p><p><b>CONCLUSION</b>FTY720 can induce the apoptosis in U266 cells, the apoptosis was Caspase-3-depended. The apoptosis induced by FTY720 is due to the decrease of MCL-1, survivin and BCL-2, which are the inhibitors of apoptosis. Meanwhile, the apoptosis was also due to the activation of BID, which is pro-apoptotic protein.</p>

Inhibition of silent information regulator-1 in hepatocytes induces lipid metabolism disorders and enhances hepatitis C virus replication / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role of the host-encoded silent information regulator 1 (SIRT1) on hepatocytes' lipid metabolism under conditions of hepatitis C virus (HCV) infection and assess its potential effects on virus replication in vitro.</p><p><b>METHODS</b>The Huh-7.5 human hepatocyte cell line was used as the control group and Huh-7.5 cells stably expressing the HCV replicon (Huh7.5-HCV) were used as the experimental group. Effects of interferon (IFN) treatment and activation of SIRT1 by resveratrol were also observed. The mRNA and protein expression levels of SIRT1 were detected by real time (q)PCR and western blotting. Effects on SIRT1 protein activity were tested by measuring the levels of reactive oxygen species (ROS) and the nicotinamide adenine dinucleotide (NAD+)/beta-nicotinamide adenine dinucleotide, reduced (NADH) by flow cytometry and chromatometry, and the levels of triacylglycerol (TG), total cholesterol (TC), and fatty acid beta oxidation rate by enzymatic analysis and liquid scintillation counting. Effects on mRNA expression of SIRT1 downstream lipid-metabolism genes were measured by qPCR.</p><p><b>RESULTS</b>The Huh7.5-HCV cells had a significantly higher level of ROS (3.8+/-0.5 vs. Huh-7.5: 1.0+/-0.2; t = 12.736, P less than 0.01) but significantly lower levels of NAD+/NADH (0.03+/-0.01 vs. 0.12+/-0.03; t = 6.971, P less than 0.01), SIRT1 activity (0.3+/-0.1 vs. 1.0+/-0.2, 0.9+/-0.2, F = 6.766, P less than 0.01), SIRT1 mRNA (0.4+/-0.1 vs. 1.0+/-0.3, 0.9+/-0.2, F = 5.864, P less than 0.01), and SIRT1 protein (0.3+/-0.1 vs. 0.8+/-0.2, 0.9+/-0.2, F = 5.419, P less than 0.01). The lower levels of SIRT1 in Huh7.5-HCV cells accompanied decreased phosphorylation of the forkhead box O1 (FoxO1), which not only up-regulated the downstream genes of SREBP-1c, FAS, ACC, SREBP-2, HMGR and HMGS (which increased fatty acid synthesis) but also down-regulated the downstream genes of PPAR and CPT1A genes (which decreased fatty acid beta oxidation). IFN treatment restored all of the aforementioned changes. Resveratrol-induced SIRT activation improved the perturbations in lipid metabolism pathways, as evidenced by an increase in fatty acid beta oxidation and a decrease in TG and TC synthesis, as well as inhibited HCV replication.</p><p><b>CONCLUSION</b>HCV may decrease the NAD+/NADH ratio in hepatocytes, leading to a down-regulation of SIRT1 activity and expression and perturbing the downstream expression profile of lipid metabolism-related factors, ultimately causing lipid metabolism disorders and establishing a permissive intracellular environment for HCV replication.</p>

Analysis on Hantaan virus in hemorrhagic fever patients with renal syndrome in Heilongjiang / 中国地方病学杂志

Objective To separate and amplify Hantaan virus(HV)in serum of hemorrhagic fever patients with renal syndrome(HFRS)in Heilongjiang,and look for its difference from intemational standard type strain(76-118strains).Methods HVs of different phase in the 8erum of 50 HFRS patients were separated and amplified by RTnested-PCR,its products were analyzed the amplified by sequencing.Results Detectable rate of HV in the patients serum was 36.36%(8/22)in 7 days after onset,it Was 13.04%(3/23)in patients having an onset 8 days to 14 days earlier,5 cases were not detectable 15 days after onset.Comparing the sequence of HV S gene fragment,sample 1,9,18,31,37,38,44 strain had a homology of 90.24%,86.72%,89.97%,89.16%,86.45%,87.26%and 89.43%to 76-118 strains,respectively.Conclusions The positive rate is the highset in 7 days after onset.Nucleotide sequence difference exists between pathogenic strain of Heilongjiang's HV and international standard strain,indicating that not only hosts but also locations can affect HV.

Retrospective analysis of epidemiologicai and clinical features of patients with acute brucellosis in Harbinand nearby counties over the past twenty years / 中国地方病学杂志

Objective To analyze the changes of epidemiological and clinical features of patients with acute brucellosis over the past twenty years.Methods The epidemiological and clinieM data of patients with acute brucellosis in Harbin and counties around from 1982-1984 and 2002-2004 in our hospital were retrospeclively analyzed,and the routes of infection,epidemiologieal area,clinical manifestations,laboratory examination,time distribution and the treatment for relapse of different groups were compared.Results One hundred and sixty-five cases were collected.including 79 cases in 1982-1984 and 86 cases in 2002-2004.There were significant differences in routes of infection(X2＝11.758,P＜0.01)and epidemiological area(X2＝8.903,P＜0.05)between two groups of patients based on epidemiologieal data.Clinical manifestations were significantly different in two groups.such as sweating 74.7%(59/79)and 50.0%(43/86)(X2=10.629,P＜0.01);hepatomegaly 21.5%(17/79)and 9.3%(8/86)(X2＝4.780,P＜0.05);orchiditis,25.3%(20/79)and 8.1%(7/86)(X2＝8.877,P＜0.01);headache,20.3%(16/79)and 7.0%(6/86)(X2=6.281,P＜0.05);the distribution of fever severity(X2＝11.671,P＜0.01)and type(X2＝29.946,P＜0.01);the distribution of total white blood cells(X2=11.550,P＜0.01);rates of thrombocytopenia,2.5%(2/79)and 19.8%(17/86)(X2＝12.005,P＜0.01);and rates of liver dysfunction,21.5%(17/79)and 54.7%(47/86)(X2＝19.037,P7＜0.01).Relapse rate at 3-week 25.0%(6/24)was higher than that 6.5%(4/62)at 6-week(X2＝5.793,P＜0.05).Conclusion The epidemiologieal and clinical feature of acute burgeri infection has changed over the past twenty years due to the different strain of Bacterium.

The impact of fatty liver and its related factors on the efficacy of antiviral therapy in patients with chronic hepatitis C / 中华传染病杂志

Objective To investigate the impact of fatty liver and its related factors on the effi- cacy of antiviral therapy in patients with chronic hepatitis C.Methods ninety-eight patients with chronic hepatitis C were treated with 180?g peginterferon?-2a plus ribavirin.Hepatitis C virus (HCV) genotypes were measured by enzyme-linked immunosorbent assay (ELISA) and fatty liver were detected by ultra sonography.The body mass index (BMI),waist to hip ratio (WHR) and ho- meostasis model assessment of insulin resistance (HOMA-IR) were calculated.The serum HCV RNA level was determined by polymerase chain reaction (PCR) and serum insulin level was detected by chemiluminescence analysis.The impact of fatty liver and its related factors on the efficacy of antiviral therapy were analyzed by multivariate Logistic regression analysis.Results Of 98 patients with chro- nic hepatitis C,35 (35.7%) were genotype 1,41 (41.8%) were genotype 2,13 (13.3%) were gen- otype 3,9 (9.1%) were undetermined genotype.The incidence of fatty liver in HCV infection of genotype 1,2,3 and undetermined genotype was 11.4%,9.8%,38.5% and 11.1%,respectively (X~2=7.83,P

Comparative study on dendritic cells stimulated with HBsAg or HBcAg in patients with chronic hepatitis B / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study activation of dendritic cells (DC) isolated from peripheral blood monocytes of patients with chronic hepatitis B after stimulation with HBsAg or HBcAg.</p><p><b>METHODS</b>DCs were isolated from peripheral blood monocytes of patients with chronic hepatitis B. DCs were impulsed with HBsAg and HBcAg separately before their maturation. The expression levels of DC surface molecule were analyzed by using flow cytometry and the ability of DC to induce T lymphocyte proliferation was evaluated by a liquid scintillation counter and the amount of interleukin-12 (IL-12) in mixed lymphocytic (IL-12) in mixed lymphocytic reaction (MLR) was measured by using ELISA.</p><p><b>RESULTS</b>The expression rate of CD86 significantly increased to (29.20 +/- 5.18)% on DC after loading with HBcAg as compared with those after loading with HBsAg (76.19 +/- 3.90)% and controls (62.37 +/- 4.24)%, P>0.01. The ability of DC after loading with HBcAg to induce T lymphocyte proliferation (34,326 +/- 3088 cpm) was significantly higher than that of DC after loading with HBsAg (20,306 +/- 2897 cpm) and controls (3454 +/- 409 cpm), P greater than 0.01. The amount of IL-12 in MLR of DC after loading with HBcAg was (348 +/- 42.8) ng/L, which was significantly higher than those of DC after loading with HBsAg (226 +/- 30.6) ng/L and controls (116 +/- 15.6) ng/L, P greater than 0.01.</p><p><b>CONCLUSION</b>Human dendritic cell stimulated with HBcAg could more efficiently present antigen and induce specific T cell immune response.</p>

Changes of immunocytes in livers of chronic hepatitis C patients treated with IFN alpha-2b and ribavirin / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the immunocytodynamic changes in the livers of chronic hepatitis C patients treated with IFN alpha-2b and ribavirin and to find new bases for an effective immune regulation therapy.</p><p><b>METHODS</b>Forty-two chronic hepatitis C patients were treated with combined IFN alpha-2b and ribavirin and their peripheral blood and liver tissues were collected before the treatment for analyses. After the treatment, peripheral blood and liver tissue specimens were obtained from only 11 patients. All the specimens were exposed to three monoclonal antibody fluorescence dyes, and the CD45+ cells with triple colors were analyzed using flow cytometry.</p><p><b>RESULTS</b>Compared to the control groups, the positive rates of CD56+, CD57+, CD161+ cells in the livers of those with chronic hepatitis C sharply decreased (Probability value less than 0.01), and CD56+T cells had decreased mildly; CD28 from the CD56+T cells decreased mildly, but the expression of CD152 increased (P<0.05); the positive rates of CD83+CD1a+ cells had decreased mildly, and the positive rates of CD80+CD11c+ and the CD86+CD11c+ cells significantly decreased (P<0.01). After the treatment, the CD56+, CD161+, CD56+T, CD161+T, CD80+CD11c+, CD86+CD11c+ cells in the responding group increased.</p><p><b>CONCLUSION</b>Combined interferon alpha-2b and ribavirin treatment can improve the suppressed cell immunity function.</p>

Observation of clinical efficacy of preventing liver injury of lamivudine on hepatitis B virus carriers with anti-tuberculosis drugs / 中华传染病杂志

Objective To evaluate the clinical efficacy of lamivudine in preventing liver injury induced by anti-tuberculosis drugs in hepatitis B virus(HBV)carriers.Methods One hundred and ten HBV carriers treated with anti-tuberculosis drugs were randomly divided into lamivudine group and control group.Patients in both groups were treated with conventional anti-tuberculosis drugs (isoniazid,rifampicin,pyrazinamide,streptomycin or ethambutol)for 6-8 months.However, patients in lamivudine group were treated with lamivudine 100 mg orally dairy concomitantly.Before and after treatment,the clinical manifestation,liver function and serum HBV DNA level of patients were evaluated.Statistical analysis was performed using t test and x~2 test.Results During 6-8 months of treatment,the incidence rate of liver injury was 9.1% in lamivudine group,while it was 38.2% in control group(P0.05).Conclusion Lamivudine is effective and safe in reducing liver injury induced by anti-tuberculosis drugs in HBV carriers.

Retrospective analysis of epidemiological and clinical features of 86 patients with acute brucellosis / 中华传染病杂志

Objective To investigate epidemiological and clinical features of patients with acute brucellosis.Methods The epidemiological,clinical,laboratory and treatment data of patients diagnosed as acute brucellosis during 2002 to 2004 in our hospital were retrospectively analyzed. Results Fifty-one patients had a history of close contact with sheep or cows with brucellsis.Twenty- seven patients had drunk milk or eaten instant boiled mutton.The transmission routes were unknown in 8 patients.All the patients had fever and most had low-grade fever.Fifty-five patients had irregu- lar fever and 20 patients had intermittent fever.The most common manifestations were fever(86/86), fatigue(63/86),sweating(43/86),arthralgia(68/86),orchiditis(7/86),hepatomegaly(8/86),sple- nomegaly(7/86)and headache(18/86).Forty-seven patients had liver dysfunction and 17 patients had thrombocytopenia.Eighty patients recovered and 6 patients relapsed after combination therapy with rifampicin,sulfamethoxazole and quinolone.Conclusion The changes in epidemiological and clinical features of patients with acute brucellosis should be noticed.