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Objective To sequence and analyze the near full-length genome of an HIV-1 subtype G strain identified in Guangxi,China.Methods The demographic information of an individual infected with HIV-1 subtype G strain was investigated,whose peripherial blood was collected.Viral RNA in plasma was extracted.The near full-length genome of HIV was amplified in two halves using RT-nested-PCR.The PCR products were purified and sequenced.Phylogenetic analysis was made using MEGA6 software.Results A near full-length genome of 8847 bp was obtained.In the neighbor-joining tree,the strain clustered with subtype G references,as supported by the high Bootstrap value (100%).The closest phylogenetic relationship was found between our strain and another subtype G strain (JN106043)previously identified in Guangxi,which was supported by the genetic distance (5%)and high Bootstrap value (100%).Conclusion The strain identified in the study might have originated from subtype G strains in Guangxi,suggesting that subtype G has spread locally in Guangxi.Further surveillance of subtype G epidemic in Guangxi is necessary.The near full-length subtype G strain genome will provide information for the surveillance of HIV in Guangxi.
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Objective To observe the effect of Fufei Gushen Decoction on the BODE index, an index for body mass index(BMI), airflow obstruction, dyspnea, and exercise capacity, in severe and extremely severe chronic obstructive pulmonary disease (COPD) patients with lung-kidney deficiency interweaved with phlegm and blood stasis at stable stage. Methods Eighty qualified COPD patients were randomly divided into treatment group and control group, 40 cases in each group. Both groups were given inhalation of Seretide (Salmeterol Xinafoate and Fluticasone Propionate Powder for inhalation) , and the treatment group was given oral use of Fufei Gushen Decoction additionally. The treatment for the two groups lasted for 3 months. Before and after treatment, BMI, the percentage of forced expiratory volume in one second of the predicted value (FEV1%) , dyspnea index of modified British Medical Research Council (MMRC), and exercise performance index of 6-min walking test (6MWT) in the two groups were observed. Results (1) After treatment, FEV1%, MMRC dyspnea index, 6MWT scores and BODE index overall scores of severe and extremely severe patients in the treatment group were much improved(P 0.05). MMRC dyspnea index, 6MWT scores and BODE index overall scores of severe and extremely severe patients inthe control group were much improved (P 0.05).(2) Except for BMI, the parameters of FEV1%, MMRC dyspnea index, 6MWT scores and BODE index overall scores of the treatment group were much improved as compared with those of the control group after treatment(P < 0.05). Conclusion Fufei Gushen Decoction combined with inhalation of Seretide exerts certain effects on decreasing the BODE index scores, relieving symptoms, and improving pulmonary function, exercise performance and the quality of life of COPD patients with lung-kidney deficiency interweaved with phlegm and blood stasis at stable stage.
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Objective To compare the efficacy of highly active anti-retroviral treatment (HAART) with and with AZT. Methods Data of 235 HIV patients accepted from Oct. 2010 to Nov. 2013 who took AZT (133) or TDF (102) containing regimen as first-line HAART are analyzed retrospectively. CD4+ T cell counts acted as the base line after 12 months of HAART. Increase in CD4+ T cell count number after the HAART, virological failures and drug resistance were compared between the two groups. Results The two groups had comparable baseline CD4+T cell count, gender ratio, and HIV transmision mode; after 12 months of HARRT, no statistically differences were found between the two groups with regard to CD4 + T cell count and increase in CD4+ T cell count after the 12-month HAART (P > 0.05); AZT-containing group had more virological failure (3/0). Meanwhile AZT-containing group had one thymidine analog mutation (TAMs) which confers resistance to AZT (P > 0.05 for both). Conculsion The two HAARTs have same immunological effects; AZT-containing group exhibits 2.2% viralogial failure, but its direct relationship with AZT has not been confirmed.
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Objective To determine whether Morphine has the ability to enhance HIV-1 replication in MT2 and Macrophage in vitro and assess the influence of Naloxone on Morphine2s effect.Methods MT2 cells were randomly assigned into 4 groups: (1) Morphine treatment for MT2 group, (2) Morphine+Naloxone co-treatment for MT2 group, (3) Naloxone treatment for MT2 group and (4) MT2 Control;Macrophages were also randomly assigned into 4 groups: (5) Morphine treatment for Macrophage group, (6) Morphine+Naloxone co-treatment for Macrophage group, (7) Naloxone treatment for Macrophage group and (8) Macrophage Control. Group (2), (3), (6) and (7) were pre-treated with 10-8 mol/L Naloxone for 0.5 h, and then group (1) and (2) were treated with 10-12, 10-10 and 10-8 mol/L Morphine for 24 h;group (5) and (6) were disposed of 10-10 mol/L Morphine for 24 h.All 8 groups were added in HIV-1 viral strain with 50% tissue culture infective dose(TCID50).P24 antigen in MT2 cells culture supernatant at day 3, 4, 5 and 6, and in Macrophages culture supernatant at day 4, 6, 8, 10 and 12 after infection were determined with ELISA.Student2s t-test and ANOVA were used to compare the differential expression in different groups, and repeated measures ANOVA was used to compare the increasing or decreasing expression of p24 antigen in morphine treatment groups than that in the control group at different time points.Results On the 3rd day of infection with HIV-1 in MT2 cells, the expression of p24 antigen in 10-12, 10-10 and 10-8mol/L dose of group (1) were (4.44?.30), (5.59?.25) and (4.60?.24) ng/ml respectively, compared to control[(1.93?.05) ng/ml, t= 14.15, 24.74 and 19.14, all P<0.01].On the 4th day, 10-12, 10-10 and 10-8mol/L dose of group (1) resulted in a significant increase of p24 antigen expression [(24.30?.66), (31.73?.17) and (26.02?.37) ng/ml]in culture supernatants compared to control[(8.03?.09) ng/ml, t=10.59, 34.92 and 81.2, all P<0.01].On the 5th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (1) were (56.30?.26), (81.77?.49) and (63.66?.57) ng/ml respectively, compared to control [(15.30?.91) ng/ml, t= 45.83, 43.51 and 30.07, all P<0.01].On the 6th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (1) were (150.70?.97), (243.09?.93) and (173.72?.73) ng/ml respectively, compared to control [(41.01?.84) ng/ml, t= 21.09, 39.02 and 29.55, all P<0.01].The enhanced multiple of p24 antigen expression in three doses of morphine treatment group compared to control increased with HIV-1 infected MT2 cells time, trend analysis of repeated measurements showed statistically significant time effect (F=842.18, P<0.01). On the 4th day of infection with HIV-1 in Macrophage cells, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (5) were (0.68?.15), (0.87?.41) and (0.75?.09) ng/ml respectively, compared to control [(0.60?.01) ng/ml, t= 7.27, 11.06 and 3.02, all P<0.05]. On the 6th day, 10-12, 10-10 and 10-8 mol/L dose of group (5) resulted in a significant increase of p24 antigen expression[(1.64?.57) , (2.07?.12 ) and (1.75?.17) ng/ml]in culture supernatants compared to control [(1.16?.07) ng/ml, t=8.93, 11.3 and 5.45, all P<0.01].On the 8th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (5) were (6.31?.17), (8.81?.34) and (7.19?.11) ng/ml respectively, compared to control [(3.84?.45) ng/ml, t=8.83, 15.11 and 12.42, all P<0.01]. On the 10th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of Morphine treated group were (32.30?7.55), (50.74?7.55) and (39.74?.56) ng/ml respectively, compared to control [(17.55?.86) ng/ml, t= 13.65, 17.84 and 36.69, all P<0.01].The enhanced multiple of p24 antigen expression in three doses of group (5) compared to control increased with HIV-1 infected Macrophage cells time, trend analysis of repeated measurements showed statistically significant time effect (F=135.58, P<0.01).Conclusions Morphine has the ability to enhance HIV-1 replication in MT2 cell and Macrophage. This Morphine-mediated increase of p24 antigen expression can be blocked by Naloxone.
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BACKGROUND: To investigate the distribution of CCR5-Delta32, CCR2-64I, and SDF1-3'A alleles in Guangxi province Zhuang population. METHOD: A total of 180 healthy individuals and 150 HIV-positive patients were recruited. All individuals were genotyped for CCR5 polymorphism using polymerase chain reaction (PCR), and CCR2 and SDF1 polymorphism using PCR restriction fragment length polymorphisms (PCR-RFLPs). RESULTS: No CCR5-Delta32 mutation was found. CCR2-64I frequency was 24.7% in the healthy group and 20.3% in the HIV-positive group, and SDF1-3'A was 30.6% and 29.3% in the 2 groups, respectively. There were no significances in the frequencies of CCR2-64I (chi(2) = 1.795, P = .180) and SDF1-3'A (chi(2) = 0.116, P = .733) between the 2 groups. CONCLUSIONS: The protective CCR5-Delta32 mutation is absent in the Zhuang population, but CCR2-64I and SDF1-3'A are relatively common and seem not to confer protection against HIV-1 infection. The results suggest that the Zhuang people may have a similar genetic susceptibility to HIV-1 infection with most other Chinese ethnic groups.
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Quimiocina CXCL12/genética , Infecciones por VIH/genética , Receptores CCR2/genética , Receptores CCR5/genética , Adolescente , Adulto , Estudios de Casos y Controles , China/epidemiología , Femenino , Frecuencia de los Genes , Genotipo , Infecciones por VIH/epidemiología , Heterocigoto , Homocigoto , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Adulto JovenRESUMEN
Objective To compare the genetic barriers to development of primary mutations related to drug resistance to protease inhibitors (PI), nucleioside reverse transcriptase inhibitors ( NRTI ), and non-nucleioside reverse transcriptase inhibitors ( NNRTI ) among human immunodeficiency virus (HIV)-1 CRF01_AE, CRF07_BC, and CRF08_BC strains, and to understand the difference of varying patterns of drug resistance related mutations within these subtypes. Methods One hundred and ninety naive HIV-positive subjects from Nanning City and Liuzhou City, Guangxi Zhuang Autonomous Region, were recruited. Peripheral blood samples were collected from all participants. HIV-1 RNAs were extracted from plasma, and the pol regions were amplified and sequenced. Sequences were subjected to phylogenetic analysis to determine the subtypes of HIV-1 isolates. Nucleotide transitions and transversions were counted for each primary mutation in these sequences. According to the phenomena that transitions occur on average 2. 5 times frequently than transversions, each transition was scored as 1, and each transversion scored as 2. 5. The sum of the scores for a particular substitution was calculated, and this value was taken as the genetic barrier to development of this mutation. Then, the differences of genetic barriers among the subtypes were assessed by Kruskal-Wallis test and Nemenyi test. Results A total of 123 sequences of CRF01_AE,CRF07_BC and CRF08_BC strains were selected. CRF08_BC had a lower genetic barrier for T/S69Dsubstitution than CRF01_AE and CRF07_BC (χ2 =107. 501, P<0.01), while CRF01_AE and CRF07_BC had lower genetic barriers for V118I and L210W substitution than CRF08_BC. In addition,CRF07_BC had a decreased genetic barrier for V106M compared with CRF01_AE and CRF08_BC.Conclusions In the presence of the same selective pressure, subtypes CRF01_AE and CRF07_BC may be more likely to develop V118I and L210W substitution than CRF08_BC. However, CRF08_BC may be more likely to develop T/S69D substitution than CRF01_AE and CRF07_BC. Meanwhile, CRF07_BC may be easier to develop V106M substitution than CRF01_AE and CRF08_BC.