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>

Preliminary clinical experience on radiofrequency catheter ablation of right-sided accessory pathway guided by Ensite-NavX navigation / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the feasibility and methodology of radiofrequency catheter ablation (RFCA) guided by 3D navigation system (Ensite-NavX) for right atrioventricular accessory pathway.</p><p><b>METHOD</b>Thirty-three cases of right accessory pathway atrioventricular reentrant tachycardia including 16 cases in right free wall, 3 in right middle septum, 14 in right posterior septum; 23 cases of dominant accessory pathway and 10 cases of concealed were treated by RFCA guided by NavX navigation. NavX navigation modeling method or spatial localization method was exploited to locate target positioning.</p><p><b>RESULT</b>All patients were successfully ablated without serious complications. Among them, 25 cases were operated without exposure to X-ray, 7 patients were exposed for several seconds to verify catheter position, 1 case in right free wall was ablated under X-ray combined with Swartz sheath ablation.</p><p><b>CONCLUSION</b>Nonfluoroscopy or less fluoroscopy RFCA for right atrioventricular accessory pathway with Ensite-NavX is safe and feasible, modeling or spatial orientation method are helpful to locate the ablation target positioning.</p>

Nested real-time quantitative polymerase chain reaction assay for detection of hepatitis B virus covalently closed circular DNA / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Successful treatment of hepatitis B can be achieved only if the template for hepatitis B virus (HBV) DNA replication, the covalently closed circular HBV DNA (cccDNA) can be completely cleared. To date, detecting cccDNA remains clinically challenging. The purpose of this study was to develop a nested real-time quantitative polymerase chain reaction (PCR) assay for detecting HBV cccDNA in peripheral blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (MMNCs).</p><p><b>METHODS</b>Based on the structural differences between HBV cccDNA and HBV relaxed circular DNA (rcDNA), two pairs of primers were synthesized as well as a downstream TaqMan probe. Blood and bone marrow samples were collected from hepatitis B patients and healthy controls. To remove rcDNA, samples were incubated with mung bean nuclease and the resultant purified HBV cccDNA was then amplified by nested real-time fluorescence quantitative PCR. The cccDNA levels were calculated using a positive standard.</p><p><b>RESULTS</b>The nested real-time fluorescence quantitative PCR method for HBV cccDNA was successful, with a linear range of 3.0 × 10(2) copies/ml to 3.9 × 10(8) copies/ml. Of the 25 PBMC samples and 7 MMNC samples obtained from chronic hepatitis B or liver cirrhosis patients, 3 MMNC samples and 9 PBMC samples were positive for HBV cccDNA, while all of the 21 PBMC samples from healthy controls were negative.</p><p><b>CONCLUSION</b>The nested real-time fluorescence quantitative PCR may be used as an important tool for detecting cccDNA in hepatitis B patients.</p>

Comparison of cryoablation and radiofrequency ablation for treating atrioventricular nodal reentrant tachycardia / 中华心血管病杂志

<p><b>OBJECTIVE</b>To compare the efficacy and safety between cryoablation (Cryo) and radiofrequency (RF) ablation for treating patients with atrioventricular nodal reentrant tachycardia (AVNRT).</p><p><b>METHODS</b>Patients with AVNRT (n = 304) were divided into Cryo group (n = 67) and RF group (n = 237). The procedure success rate, complete slow pathway block rate, atrioventricular block rate and relapse rate were compared between two groups.</p><p><b>RESULTS</b>There was no statistically difference between 2 groups in the success rate (Cryo group 98.5% vs RF group 97.0%, P = 0.820), complete slow pathway block rate (Cryo group 98.5% vs RF group 91.6%, P = 0.088), atrioventricular block rate (Cryo group 0 vs RF group 2.5%, P = 0.413), relapse rate (Cryo group 0 vs RF group 1.7%, P = 0.643). But Cryo group had more advantage than RF group.</p><p><b>CONCLUSION</b>Efficacy and safety were comparable between cryoablation and radiofrequency ablation for treating patients with AVNRT.</p>

Establishment and application of nested real-time quantitative polymerase chain reaction assay for detection of hepatitis B virus covalently closed circular DNA / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish a nested real-time quantitative polymerase chain reaction (PCR) assay for detection of hepatitis B virus covalently closed circular DNA in PBMC( peripheral blood monocyte) and MMNC (marrow monocyte).</p><p><b>METHODS</b>Based on the structural differences between HBVcccDNA and HBV rcDNA, two pairs of specific primers spanned the gap of the positive and negative chains and a specific TaqMan probe situated downstream were designed. To remove rcDNA, cccDNA was processed by Mung Bean Nuclease,and then amplified by nested real-time quantitative PCR using a pair of outer primers and a pair of inner primers. According to the standard preparation, cccDNA levels of specimen were calculated.</p><p><b>RESULTS</b>We have established a nested real-time fluorescent quantitative PCR method for HBV cccDNA successfully, and the linear range is from 5.0 x 10(2) to 3. 9 x 10(7) copies per milliliter. Of the 25 PBMC samples and 7 MMNC samples of the chronic hepatitis B or liver cirrhosis patients, 3 MMNC samples and 9 PBMC samples were HBV cccDNA positive, while all of the 21 healthy donator blood PBMC samples were negative.</p><p><b>CONCLUSIONS</b>The nested real-time fluorescent quantitative PCR method may be applied to detect HBVcccDNA level in PBMC and MMNC. HBVcccDNA can be detected in PBMC and MMNC.</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

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.

Plasma exchange treatment using MELD scoring system improve the prognosis of fulminant Hepatitis in Chinese patients / 中华传染病杂志

Objective To study prognosis of patients with fulminant hepatitis after plasma ex- change treatment using model for end-stage liver disease(MELD)scoring system.Methods 160 pa- tients were randomly divided into plasma exchange group and control group,and MELD score was calculated according to the original formula for each patient.The efficacy of plasma exchange was as- sessed by mortality and improvement in biochemical parameters and MELD score.Results The levels of total bilirubin(TBIL),INR and MELD score of patients whose MELD scores were between 30 and 40[TBIL,(379.4?40.4)?mol/L; INR,2.5?0.2; MELD,30.8?3.8]were lower than before PE treatment[TBIL,(509.7?64.6)?mol/L;INR,3.5?0.3;MELD,37.3?3.5].The levels of TBIL and INR and MELD score of patients whose MELD scores were higher than 40 [TBIL,(595.6?61.5)?mol/L;INR,3.8?0.4;MELD,39.8?3.5]were lower than before PE treatmem [TBIL, (650.4?66.3)?mol/L;INR,4.4?0.6;MELD,45.2?4.2].The mortality of patients in PE group with MELD score from 30 and 40 was 50.0%,while it was 86.7% in control group,showing significant differ- ence between PE group and control group(P＜0.01).The mortality of patients with MELD scores higher than 40 was 91.2% in PE group and 100% in control group,showing no significant difference between these two groups(P＞0.05).Conclusions Plasma exchange treatment can decrease the serum TBIL level, INR and MELD score of patients with fulminant hepatitis and improve liver function.Compared with the control group,plasma exchange can significantly decrease the mortality of patients in PE group with MELD score from 30 to 40,but no effect on patients with MELD score higher than 40.