Distribution of subtypes of pol gene in HIV-1 epidemic strains in Guangxi Zhuang Autonomous Region, 2010- 2012 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the distribution and proportion of subtypes of pol gene in HIV-1 epidemic strains in Guangxi Autonomous Region.</p><p><b>METHODS</b>152 HIV-1 patients were enrolled from 11 cities in Guangxi Autonomous Region from 2010 to 2012 by convenient sampling. Inclusion criterias were listed as the fdlowing: HIV-1 infection was confirmed by Western blot, HIV-1 viral load >1 000 copies/ml, > 18 year-old, and without any serious illnesses. 5 ml of peripheral blood samples were obtained from each patient. The viral RNA was isolated from plasma and used for amplification of full-length pol gene by nested RT-PCR. The amplified products were sequenced. After editing and modification, all sequences were characterized for preliminary subtyping by genotyping and confirmed with phylogenetic tree constructed by MEGA 5.03 software. The recombinant identification of 2 unknown recombinant strains was determined by RIP and jpHMM at GOBICS.</p><p><b>RESULTS</b>Among 152 patients, 137 full-length pol genes were successfully amplified and 127 HIV-1 subtypes were identified. The distribution and proportion of subtypes was summarized as the following 71 cases of CRF01_AE, accounting for 55.9% (71/127), 38 CRF08_BC, 29.9% (38/127), 13 CRF07_BC, 10.2% (13/127), and 3 B (B'), 2.4% (3/127), 2 unknown recombinant strains, 1.6% (2/127). In 11 cites of Guangxi Autonomous Region, subtype CRF01_AE was the dominant strain. Among heterosexual transmitted patients and drug abusers, the proportions of subtype CRF01_AE were 67.4% (58/86) and 34.1% (14/41), respectively. There was a significance different in the distribution of CRF01_AE in different routes of transmission (χ(2)=15.07, P<0.001). In age 21- 35, age 36- 60 and age>60 groups, the proportions of CRF01_AE was 43.6% (17/39), 57.6% (38/66), 77.3% (17/22), and CRF08_BC was 43.6% (17/39), 28.8% (19/66), 9.1% (2/22), respectively, the difference in proportions was significant(χ(2)=8.48, P= 0.014). The patterns of two unknown recombinant strains were found to be CRF01_AE/B (B') and CRF01_AE/C/B(B'), respectively.</p><p><b>CONCLUSION</b>CRF01_AE was the dominant HIV-1 subtype in Guangxi Autonomous Region from 2010 to 2012, with heterosexual transmission as its main spreading route. The two unknown recombinant strains in Guangxi Autonomous Region were reconstructed by subtype CRF01_AE and CRF_BC.</p>

Investigation of the HIV-1 drug resistance in Guangxi during 2009 to 2012 / 中华微生物学和免疫学杂志

Objective To investigate the HIV-1 drug resistance in Guangxi during 2009 to 2012 and to analyze the correlations between drug resistance and HIV-1 subtypes.Methods Patients with human immunodeficiency virus infection or acquired immune deficiency syndrome ( HIV/AIDS) were randomly re-cruited from different areas in Guangxi.HIV-1 RNA was extracted from blood samples of the subjects and converted into complementary DNA ( cDNA) by using reverse transcription.The pol gene was amplified and sequenced.Subtyping analysis was performed by using the online analysis tool of Genotyping in combination with the MEGA 5.03 software.The HIV resistance mutations were determined and scored with the use of Stanford HIV Drug Resistance Database.Results A total of 196 pol gene sequences were obtained from 103 antiretroviral therapy (ART)-treated subjects (52.55%) and 93 ART-na?ve subjects (47.45%).The 196 pol gene sequences were classified into four subtypes including CRF01_AE, CRF08_BC, CRF07_BC and B, accounting for 48.47%, 44.90%, 6.12%and 0.51%, respectively.The HIV drug resistance rates in sub-jects with and without ART were 10.68% and 7.53%, respectively.Among the 196 subjects, 14 cases showed low level of drug resistance, 3 cases showed moderate level of drug resistance and 4 cases showed high level of resistance.Only one case was resistant to both nucleoside reverse transcriptase inhibitors ( NR-TIs) and non-nucleoside reverse transcriptase inhibitors ( NNRTIs) .The resistance rates of the 196 cases to protease inhibitor (PIs), NRTIs, NNRTIs, and integrase inhibitors (INs) were 6.63%, 3.06%, 11.22%and 8.67%, respectively.The frequencies of PIs-related mutations in subtypes CRF01_AE, CRF07_BC and CRF08_BC were 6.32%, 41.67% and 2.27%, respectively.Most of the PI-related A71V/T mutations were identified in strains belonging to subtype CRF07_BC, accounting for 75% of all A71V/T mutations found in the 196 strains.The NNRTI-related E138A mutations only appeared in strains belonging to subtype CRF08_BC.Conclusion The drug resistance rate among patients with HIV-1/AIDS in Guangxi was higher than the average level in China.The drug resistance rates varied with the subtypes of HIV-1 strains.

The phylogenetic evolution and genetic variations of gag gene among the prevalent human immunodeficiency virus-1 strains in Guangxi region / 中华传染病杂志

Objective To study the phylogenetic evolution and genetic variations of gag gene among the prevalent human immunodeficiency virus (HIV )‐1 strains in Guangxi Zhuang Autonomous Region . Methods Plasma samples of 158 HIV‐1 infected patients in Guangxi area were collected during October 2011 to March 2012 .The gag gene fragments of HIV‐1 were amplified by reverse transcription/nested‐polymerase chain reaction and then sequenced .MEGA 5 .03 was utilized to construct phylogenetic tree and to calculate the genetic distances and selection pressures (globle ω) of gag gene and its coding regions . The comparisons between two groups were tested by Student′s t test ,and the comparisons of multiple groups were tested by one‐way ANOVA .Results A total of 140 amplification products of gag gene were obtained from 158 samples .Four subtypes of HIV‐1 were found ,including CRF01_AE (80 ,57 .1% ) , CRF08_BC (46 ,32 .9% ) ,CRF07_BC (10 ,7 .1% ) ,and subtype B (B′) (4 ,2 .9% ) .The genetic distances of gag gene of the above subtypes were 0 .036 ± 0 .001 ,0 .031 ± 0 .002 ,0 .043 ± 0 .003 and 0 .102 ± 0 .006 ,respectively ,with statistical significance (F=220 .62 ,P<0 .01) .The p17 and p24 coding regions suffered negative selection pressure (globleω<1) .Neither the globle ω in p17 region nor that in p24 region had significant differences among different subtypes (F=0 .761 ,P=0 .469 and F=0 .037 ,P=0 .964 , respectively ) . Conclusion CRF01_AE is the major subtypes of HIV‐1 in Guangxi Zhuang Autonomous Region .The coding regions of gag gene are relatively conserved during evolution .Changes of HIV‐1 prevalence ,however ,may affect the genetic variation of gag gene ,which should be continuously monitored .

Effects of alcohol on HCV replication and its mechanism / 中华微生物学和免疫学杂志

Objective To investigate the effects of alcohol on hepatitis C virus( HCV) replication and type I interferon signaling pathway in human hepatocytes .Methods Primary hepatocytes were treated with different concentrations of alcohol , and then infected with HCV .The infected cells were collected to measure the level of HCV RNA .The alcohol-treated hepatocytes were also collected to detect the expression of HCV Core, IFN-α, IFN-β, IRF-7, suppressor of cytokine signaling SOCS-2 and SOCS-3 at mRNA and protein levels by real-time quantitative PCR and ELISA or Western blot , respectively .Results Alcohol treatment enhanced HCV infection and replication in primary hepatocytes at concentrations higher than 10 mmol/L in a dose-dependent manner (P<0.05).Treatment with 40 mmol/L of alcohol significantly reduced the expression of IFN-α, IFN-βand IRF-7 at mRNA and protein levels , and increased the expression of SOCS-2 and SOCS-3 at mRNA and protein levels .Conclusion Alcohol treatment could damage the host in-nate immunity in human hepatocytes and promote HCV replication by reducing the expression of type Ⅰinter-feron ( IFN-αand IFN-β) and IRF-7 and increasing the expression of negative regulators including SOCS-2 and SOCS-3.These results demonstrated that the impairment of innate immunity in liver of alcohol abusers might contribute to the enhancement of HCV infection and result in poor therapeutic effect of IFN -α.