RNA methyltransferase is responsible for transferring methyl and resulting in methylation on the bases or ribose ring of RNA, which existed widely but mostly remains an open question. A recombinant protein PH1948 predicting RNA methyltransferase from Pyrococcus horikoshii OT3 has been crystallized. The crystals of selenomethionyl PH1948 belong to space group C2, with unit-cell parameters a=207.0 A, b=43.1 A, c=118.2 A, b=92.1 degrees , and diffract X-rays to 2.2 A resolution. The V(M) value was determined to be 2.8 A3/Da, indicating the presence of four protein molecules in the asymmetric unit.
Crystallization ; methods ; Molecular Conformation ; Pyrococcus horikoshii ; enzymology ; X-Ray Diffraction ; tRNA Methyltransferases ; analysis ; chemistry

Objective To estimate the clinical features of hepatitis B virus (HBV) and hepatitis C virus (HCV) co-infection among acquired immune deficiency syndrome (AIDS) patients and the interaction of lamivudine (3TC) contained antiretroviral therapy (ART) with hepatitis virus replication.Methods From 2004 to 2010,199 human immunodeficiency virus (HIV)/HBV coinfected patients admitted to Zhongnan Hospital of Wuhan University were enrolled,including 76 cases of HIV/HBV/HCV triple infection and 123 cases of HIV/HBV dual infection.Hepatitis B surface antigen (HBsAg) and hepatitis B core antibody (anti-HBc) were detected routinely.HBV DNA,HCV RNA before and after ART with 3TC and incidence of end-stage-liver-diseases in two groups were compared.Categorical data were analyzed by chi-square test,and measurement data were compared by t test.Results Positive rates of HBV DNA in HIV/HBV and HIV/HBV/HCV coinfection group before treatment were 45.5 ％ (56/123) and 25.0 ％ (19/76),respectively (x2 =8.429,P=0.004).The levels of HBV DNA in the two groups before treatment were (5.61±1.88) lg copy/mL and (4.70±1.84) lg copy/mL,respectively (t=2.589,P=0.003).After ART with 3TC,detectable rate of HBV DNA in HIV/HBV/HCV group decreased to 9.2％ (7/76),which was significantly lower than pretreatment (x2 =6.681,P=0.010),but serum HCV RNA increased significantly from 56.6％ (43/76) pretreatment to 72.4％ (55/76) post-treatment (x2 =4.136,P=0.042).The incidence of end-stage-liver-diseases in HIV/HBV/HCV co-infected group was significantly lower than that of HIV/HBV dual infection group (18.8 per 1000 person years vs 42.1 per 1000 person years; x2 =4.459,P =0.035) during an average of 5.6 years of follow up.Conclusion It is possible that there are interactions between HBV and HCV when the two viruses are co infected.The timing of patient enrollment might be an impact factor on study results.

The clinical data of 459 patients,who were first diagnosed as HIV/HBV co-infection from January 2007 to December 2013,were retrospectively analyzed.Among all patients,there were 89 cases with CD4 ＜ 50/μl,134 cases with CD4 50-200/μl and 236 cases with CD4 ＞ 200/μl,when HIV infection was diagnosed.In these three groups with different CD4 levels,the HBV DNA positive rates were 49.3％ (37/75),50.5％ (54/107) and 33.7％ (66/196);the HBV viral load were (6.37 ± 1.71) log10 copies/ml,(5.82 ± 1.86) log10 copies/ml and (4.36 ± 1.64) log10 copies/ml;the rates of abnormal liver function were 29.2％ (26/89),29.1％ (39/134) and 10.6％ (25/236);the occurrence rates of end-stage-liver-diseases were 16.9％ (15/89),14.9％ (20/134) and 5.1％ (12/236);the mortality rates were 10.1％ (9/89),9.7％ (13/134) and 3.8％ (9/236),respectively.The HBV DNA positive rates,HBV viral load,the rates of abnormal liver function,the occurrence rates of end-stage-liver-diseases and the mortality rates in CD4 ＞ 200/μl group were lower than that in CD4 ＜ 50/μl group and 50-200/μl group.The results suggest that for HIV and HBV co-infection patients,HBV replication level and prognosis of liver diseases are associated with CD4 + T lymphocyte count.

RNA methyltransferase is responsible for transferring methyl and resulting in methylation on the bases or ribose ring of RNA, which existed widely but mostly remains an open question. A recombinant protein PH1948 predicting RNA methyltransferase from Pyrococcus horikoshii OT3 has been crystallized. The crystals of selenomethionyl PH1948 belong to space group C2, with unit-cell parameters a=207.0 (A),b=43.1 (A), c= 118.2 (A), β=92.1°, and diffract X-rays to 2.2(A) resolution. The VM value was determined to be 2.8(A)3/Da, indicating the presence of four protein molecules in the asymmetric unit.

Objective To understand major risk factors associated with end-stage liver disease (ESLD) among patients with human immunedeficiency virus (HIV)/hepatitis B virus (HBV) co-infection.Methods Patients with HIV/HBV co-infection were followed-up and factors related to ESLD were analyzed using logistic regression model to estimate odds ratios (ORs) for them. Results A total of 255 patients with HIV/HBV co-infection were investigated, with an incidence of ESLD of 19. 2% ( 49/255 ). Major risk factors associated with ESLD among patients with HIV/HBV co-infection included count of CD4 below 200 cells/μl at baseline, HIV RNA load decreasing to the lower limit of its detection level within six months after antiretroviral treatment (ART), abnormal of serum activities of transaminase (ALT or AST), longer persistently positive of HIV RNA and HBV DNA, and use of lamivudine-based ART, with OR of 6. 503,14. 456, 0. 049, 1. 814, 1. 536 and 0. 012, respectively. Conclusions Lower CD4 count, abnormal serum transaminases, persistent replication of HIV and HBV all are closely related to ESLD in patients with HIV/HBV co-infection. Therefore, lamivudine-based ART should be of choice for patients with HIV/HBV coinfection to decrease incidence of ESLD.

ObjectiveTo explore the epidemiologic features and distribution pattems of hepatitis C virus (HCV) genotype infection among HIV positive former blood donors (FBDs) and transfusion recipients in Hubei province.Methods597 serum samples from HIV-positive patients in Hubei were collected and examined for anti-HCV by enzyme-linked immunosorbent assay ( ELISA ).Reverse transcription nested polymerase chain reaction (RT-nested PCR) amplification and DNA sequencing were used to evaluate the HCV core regions.ResultsThe prevalence rates of HCV in HIV positive FBDs and transfusion recipients were 76.5％ (205/268) and 57.4％ (189/329) respectively.HCV genotypes 1b (92.8％,90/97) and 2a (7.2％,7/97 ) were detected.ConclusionsBlood donation and blood transfusion are the major modes of HIV-HCV co-infection in Hubei province.The prevalence of HCV in HIV positive transfusion recipients is lower than that in HIV positive FBDs.HCV genotypes 1b and 2a are the predominant strains among HIV-positive FBDs and transfusion recipients.

Objective To analyze the incidence,mortality and risk factors of death in human immunodeficiency virus (HIV) and hepatitis C virus (HCV) co-infected patients with combined antiretroviral therapy (cART).Methods A total of 427 HIV/HCV co-infected patients admitted to Zhongnan Hospital of Wuhan University or local disease prevention and control canters from January 2003 to December 2010 were enrolled in the study.The demographic and clinical data of patients were retrospectively studied.Cox progressive regression model was used for data analysis,and Kaplan-Meier method was used to evaluate the effect of end-stage liver diseases on the death.Results of 427 HIV/HCV co-infected patients,53 ( 12.4％ ) died during the follow-up,in which 28 (52.8％) died of liver-related diseases.Male gender ( RR =2.63,P =0.05 ),infection via blood transfusion ( RR =2.15,P =0.04),baseline CD4 + T cells ＜50 cells/μL ( RR =2.83,P =0.02),HIV RNA≥ 104copies/mL at the end of follow-up (RR =2.79,P =0.00 ) and complicated with end-stage liver disease ( RR =7.79,P =0.00) were significantly related to the death.Duration of cART ＞ 5 years is a protective factor for the death ( RR =0.03,P =0.00).Themortality of patients complicated with end-stage liver diseases was 52.7％ ( 29/55 ).Conclusion Liver disease-related death has become the leading cause of death in HIV/HCV co-infected patients,and patients with end-stage liver diseases are of high risk of death.

Objective To investigate the incidence and risk factors of end-stage liver disease (ESLD) in human immunodeficiency virus (HIV) and hepatitis C virus (HCV) co-infected patients after antiretroviral therapy (ART).Methods The demographic and clinical data of HIV-HCV coinfected patients in Zhongnan Hospital,Wuhan University and local Centers for Disease Control and Prevention (CDC) from Jan 2003 to Dec 2010 were analyzed retrospectively. Single factor and multiple factor Logistic regression were used to analyze the correlation between the variables and incidence of ESLD,such as baseline age ≥40,male,previous blood transfusion history,duration of HCV persistent infection,hepatitis B virus (HBV) co-infection (HBsAg positive),HIV RNA≥-1 ×104 copy/mL at last visit,HCV RNA≥ 1.× 105 copy/mL at last visit,CD4 count ＞ 200 / μL at last visit,alanine aminotranferase (ALT) ≥ 2 × upper limit normal (ULN) at last visit,ART containing nevirapine (NVP),follow-up duration,ART duration＞5 years and HCV genotype 1b.The effect of ESLD on the survival of HIV-HCV co-infected patients was analyzed by Kaplan-Meier method.Results Totally 427 HIV-HCV co-infected patients were followed up with average of 3.7 years. Fifty-five patients (12.9％) developed ESLD,and 52 patients (12.2％) died.Factors independently associated with ESLD included baseline age≥40 (OR=2.385,P=0.039),ALT ≥2× ULN (OR=16.374,P=0.000),HBV-coinfection (OR=2.507,P=0.042),duration of ART ＞ 5 years (OR=3.232,P=0.010),and CD4 count ≥200/μL (OR=0.364,P=0.011).The cumulative mortality of HIV-HCV co-infected patients with ESLD was 50.9％,whereas that of HIV-HCV co-infected patients without ESLD was 6.5％ (P=0.000).Conclusion In the ART era,ESLD is common among HIV-HCV co-infected patients in China,which is responsible for reducing the survival time of the patients.

Objective To investigate the influence of hepatitis B virus (HBV) infection on efficacy of combined antiretroviral therapy (cART) in patients with acquired immunodeficiency syndrome (AIDS).Methods Seventy-eight subjects with human immunodeficiency virus (HIV)/HBV co-infection and 156 subjects with HIV mono-infection were included.CD4+ T cell count,HIV viral load,HBV-markers and liver functions were routinely tested.The differences in survival rate,as well as immunological and virological responses between the two groups (HIV/HBV co-infection group and HIV mono-infection group) during cART were compared.Categorical data were compared by Chisquare test,measurement data were compared by t test,and measurement data with abnormal distribution were compared by Mann-Whitney test.Results At month 42 of cART,HIV RNA levels and CD4+ T cell counts of the two groups were comparable.However,at month 48,54 and 60 of cART,the immunological and virological responses of HIV/HBV co-infection group were less favorable than those of HIV mono-infection group.At each time point of month 12,24,36,48 and 60 of cART,3 out of 13 subjects with HIV/HBV co-infection maintained hepatitis B e antigen (HBeAg)loss; the HBeAg seroconversion rates were 32.1％ (9/28),50.0％ (14/28),53.6％ (15/28),64.3％ (18/28) and 71.4％ (20/28),respectively (x2 =10.189,P=0.037) ; HBV DNA negative rates were 95.1％ (39/41),82.9％ (34/41),68.3％ (28/41),43.9％ (18/41) and 43.9％ (18/41),respectively (x2 =29.982,P=0.000); liver dysfunction rate was 32.1 ％ (25/78),51.4％ (38/74),33.8％ (22/65),47.9％ (23/48) and 6.7％ (3/45),respectively (x2 =28.053,P=0.000).Mortalities in HIV/HBV co-infected and HIV mono-infected individuals were 24.4％ (19/78) and 5.1 ％ (8/156),respectively (x2 =18.841,P＜0.01).Sixteen out of the 19 deaths (84.2 ％) in HIV/ HBV co-infected subjects died of end stage liver diseases.Conclusions HBV co-infection diminishes the long term efficacy of cART.End stage liver diseases are the primary cause of death in HIV/HBV co-infected subjects during cART.

Objective To investigate the influence of safe sex education and antiretroviral therapy (ART) on human immunodeficiency virus (HIV) antibody sero-conversion status among HIV-discordant spouses.Methods Totally 1 258 HIV/acquired immunodeficiency syndrome (AIDS) cases and their spouses were enrolled and the related information was collected during 2005 to 2007.The HIV negative spouses were tested for HIV antibody once every 6 months.The effects of safe sex education and ART on sero-conversion status of HIV-discordant spouses were analyzed.The date were analyzed by x2 test.Results Without any intervention,505 out of 1 258 HIV/AIDS spouses were positive for HIV test,with the HIV natural spousal transmission rate of 40.1％.Among 442 blood source and 816 sexual source index HIV cases,HIV was positive in 103 and 402 of their spouses,respectively,with the HIV transmission rates between couples of 23.3％ and 49.3 ％,respectively.HIV transmission rate in sexual source group was higher than that in blood source group (x2 =80.421,P＜0.01).Among 608 male and 650 female index HIV cases,HIV was positive in 333 and 172 of their spouses,respectively,with the HIV transmission rates between couples of 54.8％ and 26.5％,respectively.Rate of HIV transmission from men to women was higher than that from women to men (x2 =104.770,P＜0.01).Among 753 HIV discordant couples,only 5 spouses had HIV sero-conversion (0.7％) after comprehensive intervention measures during 5 years of follow-up period.Among 31 HIV/AIDS patients who were only accepted safe sex education,3 of their spouses were HIV positive (9.7％).The transmission rate was lower than that in patients without any intervention (40.1％) (x2 =11.760,P＜0.01).Among 722 HIV/AIDS patients received ART,2 of their spouses were HIV positive (0.3 ％),which was lower than the transmission rate (9.7％) in ART-na(i)ve group (x2=39.821,P＜0.01).Conclusion Safe sex education should be implemented throughout the health management of HIV/AIDS patients and their spouses.If possible,early ART should be carried out to reduce the risk of HIV transmission between married couples.