Objective To study the mRNA expression of apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (APOBEC3G) in the peripheral blood mononuclear cells (PBMC) in patients with chronic hepatitis C (CHC) and its regulation by exogenous interferon-α (IFN-α).Methods Twenty-eight CHC patients were recruited as case group and 14 healthy subjects were recruited as control group.APOBEC3G mRNA level (the ratio of APOBEC3G mRNA to housekeep geue 18s rRNA) in PBMC was determined by TaqMan real-time polymerase chain reaction (RTPCR).APOBEC3G mRNA levels were also dynamically measured in CHC patients treated with pegylated interferon (IFN)-α 2a at week 0,2,4,12,24,36 and 48 of treatment,and the plasma levels of IFN-α were simultaneously detected by enzyme-linked immunosorbent assay (ELISA).The data were analyzed by t test and analysis of variance using SPSS 11.0 software.Results The level of APOBEC3G mRNA in PBMC of CHC patients before treatment was 1.60× 10-4 ± 1.35 × 10-4,which was significantly lower than healthy controls 6.20 × 10-4 ±1.30 × 10-4 (t=3.147,P=0.003).The expressions of APOBEC3G mRNA were upregulated at week 12,24,36 and 48 of IFN treatment,which were 5.69×10-3±1.61×10-2,1.01×10-2±2.15×10-2,2.01×10-2±3.75×10-2 and 2.45× 10-2 ±4.08× 10-2,respectively,and all higher than that of pretreatment (F=3.46,5.67,10.27 and 25.65,respectively; P=0.042,0.030,0.010 and 0,respectively).IFN-α level in plasma were increased with treatment and reached the plateau at week 2 of the treatment until the end of observation.Conclusion Hepatitis C virus infection may be one of the reasons of APOBEC3G downregulation.

Objective To explore whether the mitochondrial toxicity markers of peripheral blood mononuclear cells (PBMC)are of significance in monitoring mitochondrial toxicity during highly active antiretroviral therapy (HAART)in patients with acquired immune deficiency syndrome (AIDS).Methods Mitochondrial DNA (mtDNA),mitochondrial thymidine kinase (TK2 )and p53-inducible ribonucleotide reductase small subunit 2 (p53R2 )were selected as mitochondrial toxicity markers.The expression changes of theses markers of PBMC in 22 AIDS patients were detected by real time quantitative polymerase chain reaction (q-PCR)at baseline,48 weeks and 96 weeks after initiation of the treatment. All the patients received stavudine/zidovudine and lamivudine as the mainstay of the HAART regimen. Independent-samples t test was used.Results The relative expression level of mtDNA in patients before HAART was 3.27 ± 0.94,and decreased to 2.16±0.85 at week 48 and 1 .66±0.66 at week 96, respectively.The differences were both significant compared with the level prior to the treatment (t =-3.90,P <0.01 and t =-6.29,P <0.01 ,respectively).The relative expression level of TK2 before HAART was 0.37 ±0.13,and increased to 1 .01 ±0.25 at week 48 and 2.13 ±0.61 at week 96 of the treatment.After pairwise comparisons of the three pairs of data (pre-HAART vs week 48 of the treatment,pre-HAART vs week 96 of the treatment and week 48 vs week 96 of the treatment),the differences were all significant (t = 10.77,8.00 and 3.56,respectively;all P < 0.01 ).The relative expression level of p53R2 was 0.86±0.39 before HAART,but gradually increased to 2.36 ±1 .14 and 7.73±0.65 ,respectively,at week 48 and week 96 of the treatment.The differences in p53R2 levels among three groups after pairwise comparison were all significant (t=3.27,12.26 and 13.25,respectively;all P < 0.01 ).Conclusions The expression levels of mtDNA,TK2 and p53R2 in PBMC could change significantly during HAART in AIDS patients,which might be used as indexes for monitoring mitochondrial toxicity.

Objective To evaluate the efficacy and safety of HLA identical and haploidentical related allogeneic hematopoietic stem cell trans?plantation in the treatment of acute myeloid leukemia(AML)and myelodysplastic syndrome(MDS). Methods A total of 21 patients with AML and 8 patients with MDS who underwent allogeneic hematopoietic stem cell transplantation in our hospital from April 2011 to April 2016 were ana?lyzed retrospectively,including 16 cases of HLA?identical allogeneic HSCT,10 cases of haploidentical allogeneic HSCT,and 3 cases of syngeneic HSCT. BUCY2 or TBI plus CY ± chemotherapeutic agents was the regular conditioning regimen. No graft versus host disease(GVHD)prophylax?is was required for syngeneic HSCT,but cyclosporine in combination with methotraxate was essential for allogeneic HSCT,cyclosporine,methotrax?ate,antithymocyte globulin,mycophenolate mofetil and glucosteroids for haploidentical HSCT. Results All patients achieved fully donor?originat?ed hematopoiesis. Two patients died of severe acute GVHD within 100 days post HSCT. Acute GVHD with gradeⅡ?Ⅳoccurred in 23.1%(6/26) patients,chronic GVHD in 50%patients,therapy and relapse?relevant mortality was 4/29(13.8%)and 6/29(20.7%)cases within a median follow?up of 23(1?60)months,respectively. Two?year overall survival and leukemia free survival rates are 68.09%( 95%CI:45.77%?82.78%)and 60.22%(95%CI:38.19%?76.55%),respectively. High risk AML is still the main challenge to long?term leukemia free survival. Conclusion HLA identical and haploidentical allogeneic HSCT for AML and MDS is safe ,effective and feasible. Minimal residual disease monitoring and pre?ventative as well as preemptive intervention is necessary for improving prognosis of high risk AML.

Objective To detect the change of hepatitis C virus (HCV)RNA in the peripheral blood mononuclear cells (PBMC)and serum of patients with chronic hepatitis C (CHC)during treatment with peg-interferon α-2a (Peg IFNα-2a)plus ribavirin (RBV),and to analyze the clinical significance of HCV RNA detection in PBMC.Methods The peripheral blood samples of 20 CHC patients who visited Department of Infectious Diseases in Guangzhou No.8 People′s Hospital from June 2013 to December 2014,were collected during treatment with Peg IFNα-2a+RBV at different time points (week 0,2,4, 12,24,36 and 48).Serum and PBMC were separated.Accurate fluorescence quantification assay (Cobas TaqMan real time polymerase chain reaction[PCR])was used to detect HCV RNA level in serum,while real-time PCR and nest-PCR were applied to detect HCV RNA in PBMC.Categorical data were analyzed byχ2 test.Results Accurate fluorescence quantification of serum HCV RNA showed that HCV RNA level decline rapidly after treatment (F = 148.06,P < 0.01 ),and 18 patients achieved HCV RNA undetectable at week 12 of treatment.The positive rate of nest-PCR was higher than real-time PCR (all P <0.01).Comparison of HCV RNA levels in serum and PBMC from 20 cases found that,the clearance rate of HCV RNA in PBMC was postponed.Two patients whose HCV RNA in PBMC kept detectable relapsed at week 24 after end of treatment.Conclusions HCV RNA can be detected in PBMC of CHC patients and the positive rate of nest-PCR is higher than real-time PCR.Antiviral therapy is effective on HCV both inside and outside PBMC,but the clearance rate of HCV RNA in PBMC is postponed compared with that in serum.Slow clearance of HCV in PBMC may be a risk factor for relapse after end of treatment.

Objective To study the impact of HIV and hepatitis C virus ( HCV ) infection on peripheral expression of antiviral protein A3G and plasma IFN-αlevels.Methods Untreated patients with chronic hepatitis C(HCV infection group, n=43), AIDS(HIV infection group, CD4 +T<200 cells/μL, n=45) and HIV/HCV co-infection (CD4 +T<200 cells/μL, n=45) were recruited in the study, and 23 healthy subjects were also enrolled as controls.A3G mRNA in peripheral blood mononuclear cells (PBMC) was measured by quantificational real-time PCR, and plasma IFN-αlevel was determined by enzyme linked immunosorbent assay (ELISA).Rank-sum test and Spearman rank correlation analysis were performed. Results A3G mRNA levels in HIV infected group, HIV/HCV co-infected group, HCV infected group and healthy control group were 4.89 (0.59), 4.85 (0.71), 3.89 (1.08) and 3.69 (0.81) lg copies/mL, respectively.A3G mRNA levels in HIV infected group and HIV/HCV co-infected group were much higher than those in healthy control group (Z=-6.306 and -6.280, P<0.01) and HCV infected group (Z=-7.358 and -7.275, P<0.01).Plasma IFN-αlevels in HIV infected group, HIV/HCV co-infected group, HCV infected group and healthy control group were 2.79 (1.25), 2.05 (1.29), 2.32 (1.84) and 2.16 (2.19) pg/mL, respectively.Plasma level of IFN-αin HIV infected group was higher than that in the HIV/HCV co-infected group (Z=-2.332, P<0.05), but no significant difference was observed among other groups (all P>0.05).There was no significant correlation between plasma IFN-αlevel and A3G mRNA expression (rs =0.04, P>0.05), and the levels of A3G mRNA and IFN-αshowed no correlation with HIV RNA and HCV RNA (all P>0.05).Conclusions A3G is highly expressed in PBMCs from HIV infected patients, and it may not be affected by the infection of HCV.A3G mRNA is not closely correlated with IFN-α, and it has not significant influence on HIV RNA and HCV RNA replication.

Objective To investigate the epidemiologic and clinical features of human immunodeficiency virus (HIV)/hepatitis B virus (HBV)co-infected patients.Methods Patients who confirmed with HIV infection and received highly active anti-retroviral therapy (HAART)at Guangzhou Eighth People′s Hospital were enrolled.HIV/HBV co-infected patients and HIV mono-infected patients were screened and their epidemiological and clinical features were analyzed before HAART.Comparison of the levels of alanine transaminase (ALT),aspartate transaminase (AST),CD4 + T lymphocyte and HIV RNA between the two groups were conducted.The data were statistically analyzed by chi-square test and nonparametric test.Results One hundred and sixty-five out of 1 218 (13.5 %)patients were hepatitis B surface antigen positive.The median ALT and AST levels of HIV mono-infected patients were 29 U/L and 34 U/L respectively,which were both higher than HIV/HBV co-infected patients (22 U/L and 25 U/L, respectively)(Z = - 4.270 and Z = - 5 .780,respectively,both P = 0.000 ).The median CD4 + T lymphocyte count of HIV/HBV co-infected patients was significantly lower than that of HIV mono-infected patients (Z = -2.980,P =0.003 ).The CD4 + T lymphocyte count was lower in hepatitis B e antigen (HBeAg)positive patients than HBeAg negative patients (Z =-2.660,P =0.008).The median CD4 + T lymphocyte count in patients with HBV DNA≥5 lg copy/mL was significantly lower than those with HBV DNA<5 lg copy/mL (Z = -2.311 ,P =0.021 ).The proportions of positive HBV DNA, HBV DNA≥5 lg copy/mL,abnormal ALT and AST in 54 patiens with CD4 + T lymphocyte counts <50/μL were 81 .5 %,66.7%,44.4% and 53.7%,respectively.All were significantly higher than patients with CD4 + T lymphocyte count≥50/μL(χ2 =6.159,P =0.046 ;χ2 =6.618,P =0.037 ;χ2 =7.144,P =0.028 andχ2 =9.586,P =0.008,respectively).Conclusions The prevalence of HBV/HIV co-infection is high in this study.The CD4 + T lymphocyte counts in HIV/HBV co-infected patients are lower,especially in patients with HBeAg positive and high HBV DNA level.The CD4 + T lymphocyte counts are associated with HBV DNA replication levels.

Objective To study the relationship between APOBEC3G mRNA level in peripheral blood mononuclear cells (PBMC) and serum hepatitis C viral RNA level in patients with chronic hepatitis C infection. Methods TaqMan real-time fluorescence relative quantitative polymerase chain reaction (RT-PCR) was used to quantify APOBEC3G mRNA levels in PBMC from 49 patients with chronic hepatitis C (CHC) and 31 healthy subjects. The relationship between APOBEC3G mRNA level and hepatitis C virus (HCV) viral load was analyzed. SPSS11. 0 statistics software was used for t test and regression analysis. Results APOBEC3G mRNA level in CHC patients [(1.5×10-5±1.9×10-5 ) copy/mL] was significantly lower than that [( 5. 2 × 10-5 ± 5. 5 × 10-5 ) copy/mL] in the healthy control subjects (t=-3.005, P＜0.01). While APOBEC3G mRNA level was not related with HCV viral loads (r=-0.082, P＞0.05). Conclusion HCV has an inhibitive effect on APOBEC3G expression, whereas APOBEC3G doesn't affect HCV replication directly in vivo.

Objective To compare the plasma hepatitis C virus(HCV)RNA levels detected by the fully automated viral load detection system(COBAS TaqMan)and the national real-time quantitative polymerase chain reaction(PCR)kit,and to investigate the clinical application value of these two methods in clinical practice.Methods A total of 168 serial plasma samples collected from 26 patients with chronic hepatitis C(CHC)before and at week 2,4,12,24,36 and 48 of antiviral treatment were detected by both COBAS Taqman 48 analyzing system and the national real-time quantitative PCR kit.The results of two methods were compared by chi square test and t test.Resnlts Both COBAS and national kit showed great positive detecting results when HCV RNA≥1×104IU/mL(at week O),and the virus load value detected by national kit was significantly higher than that detected by COBAS(t=2.05,P<0.05).However,when HCV RNA<1×104(at week 2-48),the positive rate of HCV detected by COBAS was significantly higher than that detected by national kit (t=3.66,P<0.01).At week 4 of treatment,the rapid virological response(RVR)rate was 46.2 % (12/26)detected by COBAS,while that was 88.5%(23/26)detected by national kit,and the difference was significant(x2=10.575,P<0.01).At week 12 of treatment,the complete early virological response(cEVR)was 95.7%(22/23)detected by COBAS,while that was 100%(17/17)detected by national kit,and the difference was not significant(x2=0.726,P>0.05).Conclusions The national TaqMan real-time quantitative PCR kits could be used to screen the suspected cases of HCV infecrion and to diagnose CHC cases with high HCV virus load.COBAS detection is more sensitive in cases with low HCV virus load and in on-treatment monitor during anti-HCV therapy.

Objective To explore the diagnostic value of Talaromyces marneffei (T.marneffei)-specific mannose glycoprotein Mp1p antigen for T.marneffei infection in acquired immune deficiency syndrome (AIDS) patients.Methods All cases were recruited in this study from January 2012 to June 2015 in Guangzhou No.8 People′s Hospital, including 184 AIDS patients with T.marneffei infection confirmatively diagnosed by culture, and 205 controls including 176 AIDS patients without T.marneffei infection and 29 health controls.Double antibody sandwich enzyme linked immunosorbent assay and fluoroimmunoassay combined with double-antibody sandwich were both utilized to detect serum Mp1p antigen levels, and their sensitivity and specificity for diagnosing T.marneffei infection in patients with AIDS were analyzed.x2 test and t test were used for statistical analysis.Results The ratio of males to females and age of the study group were both comparable to those of the control group (x2=0.019, P=0.889;t=1.810,P=0.07, respecitvley).The sensitivities of double antibody sandwich enzyme linked immunosorbent assay and fluoroimmunoassay combined with double-antibody sandwich were 82.07%(151/184) and 83.15%(153/184), respectively (x2=0.076, P=0.783).The specificities were 93.17%(191/205) and 92.68%(190/205), respectively (x2=0.037, P=0.847).The accuracy values were 87.92%(342/389) and 88.17%(343/389), respectively (x2=0.012, P=0.912).The false positive rates were 6.83%(14/205) and 7.32%(15/205), respectively.The false negative rates were 17.93%(33/184) and 16.85%(31/184), respectively (x2=0.049, P=0.829).The positive predictive values were 91.52%(151/165) and 91.07%(153/168), respectively (x2=0.021, P=0.886).The negative predictive values were 85.27%(191/224) and 85.97%(190/221), respectively (x2=0.045, P=0.832).The Kappa values were 0.83 and 0.80, respectively.Conclusion Detection of serum Mp1p antigen of T.marneffei possesses high specificity and sensitivity, which may be utilized for rapid and early diagnosis of T.marneffei infection in patients with AIDS.

Objective To explore the pathogen spectrum, drug resistance rate and clinical characteristics of pneumonia caused by non-tuberculous mycobacteria (NTM) in acquined immuno-deficiency syndrome (AIDS) patients.Methods The clinical data of 31 hospitalized AIDS patients with bronchoalceolar lavage flind (BALF) culture confirmed NTM pulmonary disease in Guangzhou No.8 People′s Hospital from January,2008 to February,2015 were retrospectively analyzed, including pathogen spectrum, drug resistance rate and clinical characteristics.The clinical characteristics and drug resistance were compared between Mycobacterium avmm-intracellulare complex (MAC) pneumonia and the non-MAC pneumonia, and t test and chi-square test were used.Results Of the 31 AIDS patients,28 were male and 3 were female, with the mean age of 40.9 years old.The 31 NTM strains were consisted of 14 MAC strains and 17 non-MAC strains (including 4 M.kansasii strains,3 M.lentiflavumstrains, 2 M.szulgai strains, 2 M.yongonense strains etc).There was no significant difference between two groups in sex ratio, mean age, clinical manifestations, laboratory tests and treatment outcome (all P>0.05).The major clinical manifestations included fever, productive cough, weight loss, anemia and low CD4+ count (<50/μL).Most patients showed thoracic lymphadenectasis and patchy shadows in lungs, and few patients had millet shadows and pericardial effusion.Compared with non-MAC strains, MAC strains had higher drug resistant rate of moxifloxacin (10/14 vs 4/17), levofloxacin (14/14 vs 8/17), and clarithromycin (11/14 vs 7/17).More extensively drug resistance strains were seen in non-MAC strains compared with MAC strains (11/14 vs 7/17).Conclusions MAC is the most common pathogen of NTM pulmonary disease in AIDS patients.The clinical features of pneumonia caused by MAC and non-MAC are similar, but drug resistance of MAC strains are more severe.