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

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.

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.

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 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 explore the clinical characteristics, pathological features and prognostic factors of patients with acquired immune deficiency syndrome (AIDS) related lymphoma (ARL).Methods The clinical characteristics, treatment regimen and survival status were retrospectively analyzed.At a ratio of 1∶2, 106 general non-Hodgkin lymphoma (NHL) cases were included after matching for demography and clinical characteristics with 53 ARL patients.Chi-square test was used for statistical analysis.Overall survival was analyzed using Kaplan-Meier curves.Cox regression was used for multivariant analysis.Results The predominant pathologic type of ARL was diffuse large B cell lymphoma (60.4%, 32/53).B cell lymphoma accounted for 88.7% (47/53) and T cell lymphoma accounted for 11.3% (6/53).Patients in ARL group (62.3%, 33/53) had lower proportion of receiving radiotherapy or chemotherapy compared with patients in general NHL group (94.3%, 100/106) (χ2=26.58, P<0.05).ARL group had lower hepatitis B surface antigen (HBsAg) positivity rate compared with general NHL group (1.9% vs 26.4%, χ2=14.26, P<0.05).No other characteristic was found significantly different between these two groups.The survival time of ARL and general NHL patients was (6.0±1.3) months and (48.0±10.0) months, respectively (t=8.13, P<0.01).The 1-year, 2-year, 3-year and 5-year survival rates of ARL patients were 39.6%, 32.7%, 27.7% and 20.1%, respectively, while those of general NHL patients were 79.2%, 56.8%, 42.4% and 25.0%, respectively.Of the 33 ARL patients and 100 general NHL patients who received anti-NHL treatment, the 1-year survival rates were 60.6% and 83.0%, respectively (χ2=4.040, P=0.043), the 2-year survival rates were 53.5% and 60.5%, respectively (χ2=0.003, P=0.096), the 3-year survival rates were 48.1% and 45.9%, respectively (χ2=0.288, P=0.59), the 5-year survival rates were 39.1% and 27.5%, respectively (χ2=0.798, P=0.372).Multivariate analysis revealed that anti-NHL therapy and international prognostic index score were independent predictors for prognosis (both P<0.05).Conclusions Diffuse large B cell lymphoma is the predominant pathologic type of ARL.ARL patients has significantly lower survival rate compared with general NHL patients.Combination of anti-HIV therapy and anti-lymphoma therapy in individuals with ARL can prolong their survival time.

AIM:To observe related biological parameters of 3 minutes dark-room provocative test in patients with laser peripheral iridectomy(LPI) in the fellow eyes of acute primary angle-closure (APAC) by ultrasound biomicroscopy (UBM).To explore the risk factors in primary angle closure suspect(PACS) patients with progressive angle closure after LPI.METHODS: Seventy-eight eyes of APAC patients without peripheral anterior synechia were selected.Each eye underwent 3 minutes dark-room provocative test after LPI.Anterior segment parameters, including anterior chamber depth (ACD), anterior chamber angle open distance500 (AOD500), peripheral iris thickness (PIT), iris convex (IC), the position of iris insertion and trabecular-ciliary process distance (TCPD), and the number of positional angle closure(NPAC) were observed and analyzed by statistic methods.RESULTS:Patients with APAC were examined by UBM after LPI and 26 eyes(33%) occurs at least one positional angle closure,19 eyes(24%)were positive in 3 minutes dark-room provocative test among them.It occurs a positive relationship between the elevation intraocular pressure and the number of positional angle closure in dark-room provocative test(r=0.84, P<0.01).AOD500, IT and IC were significantly changed from normal light to darkroom between positional angle closure positive group and positional angle closure negative group(all P<0.01).In single factor analysis, AOD500(P=0.003), IT(P=0.012), IC(P=0.043), TPCD(P=0.015), the position of iris insertion(P=0.024) were correlative factors of positive results.In multiple-factor analysis, only IT(P=0.011), TPCD(P=0.009), iris root attachment points(P=0.02) were independent risk factors of positive results.CONCLUSION:A certain proportion of patients with PACS after LPI appeared positional angle closure in a dark room.Peripheral iris hypertrophy, anterior displacement of the ciliary body and iris root attachment points are vital risk factors.Long-term follow-up study and intervention treatment are required in these patients after LPI.

· AIM: This study was designed to evaluate the effectiveness of anterior lens capsule inclusion in combined trabeculectomy and cataract surgery in preventing scar formation of the filtering bleb in rabbit model.· METHODS: Twerty-Four eyes (12 rabbits) with glaucoma model were studied, anterior lens capsule inclusion in trabeculectomy with the small-incision cataract surgery were performed on all right eyes (experimental group) and no implants were applicator in trabeculectomy with the small-incision cataract surgery on all left eyes (control group). These operated eyes were followed up from day 1 to 6 months postoperatively. Intraocular pressure (IOP) was measured and filtering blebs were observed after surgery. Other main outcome measures: cornea、conjunctiva、formation of the anterior chamber, anterior chamber depth、inflammatory reaction、achievement ratio of operation and complications were analyzed. On week 1, 2, 4, 12 and 24 after surgery the animal were killed in batch. Tissue was harvested from the bleb area and was made pathological section. HE staining、light microscope and micro photo analysis technique were applied to observe the cytological and histopathologic characteristics of the filtering tunnels.· RESULTS: There were significant differences between the two groups on IOP (1, 2, 4 weeks)、filtration bleb, achievement ratio of operation and complications. In experimental group, at the first month postoperatively, anterior lens capsule absorption started with inflammatory characteristics. The peak of inflammatory reaction occurred 1 week after operation and all the cells in the filtrating tunnel disappeared 6month after surgery. The fibroblast proliferation in control group occurred at I week and the filtrating tunnel closed with angiogenesis at 1 month after surgery. Fibroblast proliferation started 1week after surgery with no statistical difference during the time course (P ＞0.05). Significant statistical differences were observed by comparing the fibro blasts numbers per unit area in the filtrating tunnel in experimental group and those in control groups (P＜0.05).· CONCLUSION: Anterior lens capsule was totally absorbed at 6 months postoperatively. Anterior lens capsule inclusion in trabeculectomy with cataract surgery can possibly control intraocular pressure effectively, long-term sustainability of functional filtration bleb, inhibition of the proliferation of fibroblasts and opening of the filtrating pathway in the experimental animal models were satisfied. Compared to the control group, anterior lens capsule application has less complication.

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.

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.