Incidence and factors associated with hepatitis B surface antigen seroclearance in patients co-infected with HBV/HIV during antiretroviral therapy in Guangdong, China / 中华医学杂志(英文版)

BACKGROUND@#Hepatitis B surface antigen (HBsAg) clearance is vital for a functional cure of hepatitis B virus (HBV) infection. However, the incidence and predictors of HBsAg seroclearance in patients co-infected with HBV and human immunodeficiency virus (HIV) remain largely unknown in Guangdong, China.@*METHODS@#Between 2009 and 2019, patients co-infected with HBV/HIV undergoing antiretroviral therapy (ART) in Guangzhou Eighth People's Hospital affiliated to Guangzhou Medical University were retrospectively reviewed with the endpoint on December 31, 2020. The incidence and risk factors for HBsAg seroclearance were evaluated using Kaplan-Meier and multivariate Cox regression analyses.@*RESULTS@#A total of 1550 HBV/HIV co-infected patients were included in the study, with the median age of 42 years and 86.0% (1333/1550) males. Further, 98.3% (1524/1550) received ART containing tenofovir disoproxil fumarate (TDF) plus lamivudine (3TC). HBV DNA was examined in 1283 cases at the last follow-up. Over the median 4.7 years of follow-up, 8.1% (126/1550) patients achieved HBsAg seroclearance, among whom 50.8% (64/126) obtained hepatitis B surface antibody, 28.1% (137/488) acquired hepatitis B e antigen seroconversion, and 95.9% (1231/1283) undetectable HBV DNA. Compared with patients who maintained HBsAg positive, cases achieving HBsAg seroclearance showed no differences in age, gender, CD4 + T cell count, alanine aminotransferase (ALT) level, or fibrosis status; however, they presented lower HBV DNA levels, lower HBsAg levels, and higher rates of HBV genotype B at the baseline. Multivariate analysis showed that baseline HBsAg <1500 cutoff index (COI) (adjusted hazard ratio [aHR], 2.74, 95% confidence interval [95% CI]: 1.48-5.09), ALT elevation >2 × upper limit of normal during the first six months after receiving ART (aHR, 2.96, 95% CI: 1.53-5.77), and HBV genotype B (aHR, 3.73, 95% CI: 1.46-9.59) were independent predictors for HBsAg seroclearance (all P <0.01).@*CONCLUSIONS@#Long-term TDF-containing ART has high anti-HBV efficacy including relatively high overall HBsAg seroclearance in HBV/HIV co-infected patients. Lower baseline HBsAg levels, HBV genotype B, and elevated ALT levels during the first six months of ART are potential predictors of HBsAg seroclearance.

Impact of low level viremia on the antiviral effect of human immunodeficiency virus/acquired immunodeficiency syndrome patients received anti-retroviral therapy / 中华传染病杂志

Objective:To investigate the impact of low level viremia (LLV) on the prognosis of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients received anti-retroviral therapy (ART).Methods:From January to December 2015, the HIV/AIDS patients with LLV received ART over one year were recruited in Guangzhou Eighth People′s Hospital, Guangzhou Medical University (LLV group). Patients with viral load (VL) less than 50 copies/mL were matched at ratio of 1∶1 according to gender, age and the transmission route were included in the control group (suppression group). The LLV group was divided into three subgroups according to VL (LLV-1 subgroup was 50-200 copies/mL, LLV-2 subgroup was 201-400 copies/mL, and LLV-3 subgroup was 401-1 000 copies/mL). The influence of LLV on the antiviral response during the following three years was investigated.The Wilcoxon signed rank test, Kruskal-Wallis test and chi-square test were used for statistical analysis.Results:One hundred and thirty-seven patients were enrolled in the LLV group, of whom 111 were males and 26 were females, with age of (39.5±13.5) years old. At the same time, 137 patients were included in the suppression group. There were 93 cases in LLV-1 subgroup, 25 cases in LLV-2 subgroup and 19 cases in LLV-3 subgroup. There were no significant differences in the CD4 + T lymphocyte counts and CD4 + /CD8 + T lymphocyte counts ratios between LLV group and suppression group before ART (both P>0.05). During the three-year follow-up, the cumulative number of viral failures in LLV group (7.3%(10/137)) was significantly higher than that in the suppression group (1.5%(2/137)) ( χ2=5.578, P=0.018). Virological failure occurred in eight patients (8.6%) in the LLV-1 subgroup, two patients (8.0%) in the LLV-2 subgroup, and no patients in the LLV-3 subgroup. There was no statistical significance in the incidence of virological failure among all the subgroups ( P>0.05). At one, two, three years follow-up, the CD4 + T lymphocyte counts increased in both LLV group and suppression group without statistical differences (all P>0.05), and the CD4 + /CD8 + T lymphocyte counts ratios in each LLV group were lower than that in the suppression group ( Z=-3.183, -2.094 and -2.312, respectively, all P<0.05). At one, two, three years follow-up, There were no significant differences in CD4 + /CD8 + T lymphocyte counts ratios among the LLV-1, LLV-2 and LLV-3 subgroups (all P>0.05). Conclusion:HIV/AIDS patients with LLV having received ART over one year are more likely to develop virological failure and delay the recovery of immune function, which requires early relevant interventions.

Clinical characteristics and pathogenic spectrum of pulmonary filamentous fungal infection in acquired immunodeficiency syndrome patients in Guangdong Province / 中华传染病杂志

Objective:To investigate the clinical characteristics and pathogen spectrum of acquired immunodeficiency syndrome (AIDS) patients complicated with pulmonary filamentous fungal infection in Guangdong Province, so as to provide evidences for improving the diagnosis and treatment.Methods:A total of 143 AIDS patients with pulmonary filamentous fungal infection hospitalized in Guangzhou Eighth People′s Hospital, Guangzhou Medical University from January 2016 to December 2018 were included. The filamentous fungi cultured in bronchoalveolar lavage fluid of these patients were identified with morphological and molecular biological methods. And their clinical characteristics were analyzed. Nonparametric Kruskal-Wallis H test and chi-square test were used for statistical analysis. Results:Among the 143 patients, 116(81.1%) had fever, 104(72.7%) had cough, 83(58.0%) had expectoration, and 59(41.3%) had anhelation. The CD4 + T lymphocyte count was 22.0(9.3, 60.8) cells/μL and 118(82.5%) cases were below 100.0 cells/μL. The white blood cell counts decreased in 52(36.4%) cases and increased in 18(12.6%) cases, anemia was found in 109(76.2%) cases, platelet count decreased in 29(20.3%) cases. Sixty-four (44.8%) cases were positive for galactomannan test. Chest computed tomography showed diffuse infection of both lungs in 114(79.7%) cases, miliary changes in 12(8.4%) cases, pleural effusion in 44(30.8%) cases, and enlargement of pleural and (or) mediastinal lymph nodes in 45(31.5%) cases. After receiving antifungal therapy, 124 (86.7%) cases were cured or improved, and 19 (13.3%) cases were discharged automatically or died of disease deterioration. Among the 143 strains of filamentous fungi, there were 56 strains of Aspergillus species pluralis (39.2%, including 24 strains of Aspergillus fumigatus), 37 strains of Talaromyces marneffei ( T. marneffei) (25.9%), 22 strains of Penicilium species pluralis (15.4%), and 28 strains of other genera of filamentous fungi (19.6%). The median CD4 + T lymphocyte counts in patients infected with Aspergillus species pluralis, T. marneffei, Penicilium species pluralis and other genera were 24.5, 15.0, 53.5 and 22.0 cells/μL, respectively, and the difference was statistically significant ( H=11.282, P=0.010). The proportions of AIDS patients with different pulmonary filamentous fungal infection of CD4 + T lymphocyte count ≤50.0 cells/μL in descending order were T. marneffei group (89.2%(33/37)), Aspergillus species pluralis group and other genera group (67.9%(38/56), 67.9%(19/28)), and Penicillium species pluralis group (54.5%(12/22)), and the difference was statistically significant ( χ2=9.296, P=0.026). Conclusions:The clinical manifestations of pulmonary filamentous fungal infection in AIDS patients in Guangdong Province are not specific. The pathogenic spectrum contains various genera, and T. marneffei and Aspergillus fumigatus are dominant, which could be correlated with CD4 + T lymphocyte count.

Failed detection of the full-length genome of SARS-CoV-2 by ultra-deep sequencing from the recovered and discharged patients retested viral PCR positive

Over 10 percent of recovered and discharged patients retested positive for SARS-CoV-2, raising a public health concern whether they could be potential origins of infection. In this study, we found that detectable viral genome in discharged patients might only mean the presence of viral fragments, and could hardly form an infection origin for its extremely low concentration.

Prognostic factors for COVID-19 pneumonia progression to severe symptom based on the earlier clinical features: a retrospective analysis

Approximately 15-20% of COVID-19 patients will develop severe pneumonia, about 10 % of which will die if not properly managed. Earlier discrimination of the potential severe patients basing on routine clinical and laboratory changes and commencement of prophylactical management will not only save their lives but also mitigate the otherwise overwhelmed health care burden. In this retrospective investigation, the clinical and laboratory features were collected from 125 COVID-19 patients, who were classified into mild (93 cases) or severe (32 cases) groups according to their clinical outcomes after 3 to 7-days post-admission. The subsequent analysis with single-factor and multivariate logistic regression methods indicated that 17 factors on admission differed significantly between mild and severe groups, but that only comorbid with underlying diseases, increased respiratory rate (>24/min), elevated C-reactive protein (CRP >10mg/liter), and lactate dehydrogenase (LDH >250U/liter), were independently associated with the later disease development. Finally, we evaluated their prognostic values with the receiver operating characteristic curve (ROC) analysis and found that the above four factors could not confidently predict the occurrence of severe pneumonia individually, but that a combination of fast respiratory rate and elevated LDH significantly increased the predictive confidence (AUC= 0.944, sensitivity= 0.941, and specificity= 0.902). A combination consisting of 3- or 4-factors could further increase the prognostic value. Additionally, measurable serum viral RNA post-admission independently predicted the severe illness occurrence. In conclusion, a combination of general clinical characteristics and laboratory tests could provide high confident prognostic value for identifying potential severe COVID-19 pneumonia patients. SummaryWith our successful experience of treating COVID-19 patients, we retrospectively found that routine clinical features could reliably predict severe pneumonia development, thus provide quick and affordable references for physicians to save the otherwise fatal patients with the limited medical resource.

An exploratory randomized, controlled study on the efficacy and safety of lopinavir/ritonavir or arbidol treating adult patients hospitalized with mild/moderate COVID-19 (ELACOI)

BackgroundAntiviral therapies against the novel coronavirus SARS-CoV-2, which has caused a global pandemic of respiratory illness called COVID-19, are still lacking. MethodsOur study (NCT04252885, named ELACOI), was an exploratory randomized (2:2:1) controlled trial assessing the efficacy and safety of lopinavir/ritonavir (LPV/r) or arbidol monotherapy for treating patients with mild/moderate COVID-19. FindingsThis study successfully enrolled 86 patients with mild/moderate COVID-19 with 34 randomly assigned to receive LPV/r, 35 to arbidol and 17 with no antiviral medication as control. Baseline characteristics of the three groups were comparable. The primary endpoints, the average time of positive-to-negative conversion of SARS-CoV-2 nucleic acid and conversion rates at days 7 and 14, were similar between groups (all P>0.05). There were no differences between groups in the secondary endpoints, the rates of antipyresis, cough alleviation, or improvement of chest CT at days 7 or 14 (all P>0.05). At day 7, eight (23.5%) patients in the LPV/r group, 3 (8.6%) in the arbidol group and 2(11.8%) in the control group showed a deterioration in clinical status from moderate to severe/critical(P =0.206). Overall, 12 (35.3%) patients in the LPV/r group and 5 (14.3%) in the arbidol group experienced adverse events during the follow-up period. No apparent adverse event occurred in the control group. ConclusionsLPV/r or arbidol monotherapy present little benefit for improving the clinical outcome of patients hospitalized with mild/moderate COVID-19 over supportive care. FundingThis study was supported by project 2018ZX10302103-002, 2017ZX10202102-003-004 and Infectious Disease Specialty of Guangzhou High-level Clinical Key Specialty (2019-2021).

Temporal dynamics in viral shedding and transmissibility of COVID-19

We report temporal patterns of viral shedding in 94 laboratory-confirmed COVID-19 patients and modelled COVID-19 infectiousness profile from a separate sample of 77 infector-infectee transmission pairs. We observed the highest viral load in throat swabs at the time of symptom onset, and inferred that infectiousness peaked on or before symptom onset. We estimated that 44% of transmission could occur before first symptoms of the index. Disease control measures should be adjusted to account for probable substantial pre-symptomatic transmission.

Changes of total HIV-1 DNA in peripheral blood mononuclear cells of HIV-1/HCV co-infected patients after HAART / 中华临床感染病杂志

Objective:To analyze the dynamic changes of total HIV-1 DNA in peripheral blood mononuclear cells (PBMC) following highly active antiretroviral therapy (HAART) in HIV-1/HCV co-infected patients.Methods:Thirty patients with HIV-1/HCV co-infection without anti-HCV treatment (co-infected group) and 42 HIV-1 infected patients with initial treatment (mono-infected group) admitted to Guangzhou Eighth People’s Hospital from May 2015 to December 2017 were retrospectively analyzed. The virological and immunological responses of the two groups at 12, 24, 48, 72 and 96 weeks after HAART, the changes of total HIV-1 DNA in PBMC and its relationship with peripheral blood HIV-1 RNA and T lymphocyte subsets were observed. SPSS 22.0 statistical software was used to analyze the data.Results:The plasma HIV-1 virus inhibition rate, CD4 + T cells and CD4 + /CD8 + ratio increased and the total HIV-1 DNA in PBMC decreased in both groups after HAART. The inhibition rate of HIV RNA at week 72 in co-infected group was significantly lower than that in the mono-infected group ( χ2=7.93, P<0.01). Compared with the mono-infected group, the CD4 + T cells at week 12, 24, 72 and 96 after HAART were lower in the co-infected group ( U=313.50, 329.00, 286.00 and 204.50, P<0.05 or <0.01). The CD4 + /CD8 + ratio at week 48 in the co-infected group was lower than that in the mono-infected group ( U=294.50, P<0.05). The total HIV-1 DNA of the co-infected group at baseline and week 12 was lower than that of the mono-infected group ( U=362.00 and 359.00, P<0.01 or <0.05). There was no significant correlation between total HIV-1 DNA in PBMC and HIV-1 RNA or CD4 + /CD8 + ratio in both groups ( P>0.05). There was no correlation between total HIV-1 DNA and CD4 + T cells in HIV-1/HCV co-infected group ( b=-0.001, P>0.05), but it had negative correlation in the mono-infected group ( b=-0.001, P<0.05). Conclusion:Total HIV-1 DNA in PBMC was significantly decreased after HAART in HIV-1/HCV co-infected patients. Co-infected with HCV may delay the decrease of total HIV-1 DNA after HAART in patients with HIV-1 infection.

Analysis on variation characteristics of HIV-1 Gp120 sequences in men who have sex with men in Guangzhou / 中华实验和临床病毒学杂志

Objective@#To analyze the variation characteristics of HIV-1 Gp120 sequences in men who have sex with men (MSM) in Guangzhou.@*Methods@#Plasma samples were collected from HIV-1 infected MSM before antiretroviral treatment. Viral RNA was extracted from plasma. Gp120 gene sequences were amplified by reverse transcription and nested-PCR using specific primers. Phylogenetic tree, length polymorphism, amino acid characteristics of V3 loop, co-receptors and signature amino acids were analyzed.@*Results@#The phylogenetic tree were divided into 4 clusters, and the most prevalent subtypes were CRF07_BC (34/61, 55.74%) and CRF01_AE (24/61, 39.34%). Majority of HIV-1 Gp120 sequences had 496-515 amino acids. Among five hypervariable regions, the V1 region had the highest levels of length polymorphism and V3 region had the lowest. The top four peptide of V3 loop were GPGQ (56/58, 96.55%). Most of the co-receptors HIV-1 strains used was CCR5(50/58, 86.21%)according to four methods of comprehensive prediction. There are four signature amino acids in CRF01_AE subtype strains, and the frequency of occurrence was 0.75-0.83; there are eight signature amino acids in CRF07_BC subtype strains, and the frequency was 0.74-0.94.@*Conclusions@#The length of Gp120 sequences in MSM in Guangzhou has a high polymorphism. The top four peptide of V3 loop, co-receptor and signature amino acid of V3 ring have formed unique patterns.

Analysis of antigen phenotypic epitopes variation in HBV Pre-S/S region in HIV/HBV co-infected patients / 中华实验和临床病毒学杂志

Objective@#To analyze the characteristic mutations of epitopes in HBV Pre-S/S region in HIV/HBV co-infected patients’ peripheral blood to provide basic data for studying the pathogenesis of HIV/HBV co-infection.@*Methods@#The chronic hepatitis B infected patients admitted to the Infectious Disease Center of the Eighth People′s Hospital of Guangzhou from January 2009 to December 2011 were enrolled into HIV/HBV co-infected group and HBV mono-infected group according to the result of HIV antibody detection respectively before treatment. HBV DNA in serum was extracted and Pre-S/S region of HBV DNA was amplified by nested-PCR. After sequencing of the obtained PCR products (direct sequencing), ContigExpress software was used for sequence splicing and BioEdit software was used for sequence alignment. With reference to the standard sequence of the matched genotype HBV, mutants of HBV Pre-S/S region in HIV/HBV co-infected group and HBV mono-infected group were analyzed respectively. Statistical analysis was performed by chi-square test with SPSS19.0 statistical analysis software.@*Results@#HBV Pre-S/S fragments were successfully amplified from 150 patients, including 90 cases of HIV/HBV co-infected group and 60 cases of HBV mono-infected group, with matched gender, age, genotype, HBeAg status, alanine aminotransferase (ALT), aspartate aminotransferase (AST). The result of analyzing mutants of HBV Pre-S/S region indicated that the incidence of mutation in all epitopes for cytotoxic T cells (CTL cells) was higher in the HIV/HBV co-infected group, and Pre-S2 aa1-15 epitope was significantly higher (χ2=6.964, P=0.008). The incidence of deletions in PreS2 aa1-15 epitope in HIV/HBV co-infected group (11.1%) was higher than HBV mono-infected group (3.3%) (χ2=2.959, P=0.085). In the B cell epitopes, the incidence of mutations in Pre-S2 aa1-26 in the HIV/HBV co-infected group was significantly higher than HBV mono-infected group (χ2=6.924, P=0.010), and there was no statistical significance between two groups in other B cell epitopes. No differences in helper T cell (Th cell) epitopes were found between the two groups.@*Conclusions@#Co-infection with HIV increased the CTL cell epitopes’ mutations in the HBV Pre-S/S region, especially the 5′ end epitope mutations in Pre-S2 region, which indicated that HBV mutation is related to the host immune status, and showed guiding information for further study on the pathogenesis of HIV/HBV co-infection

Discussion on optimal duration of pegylated interferon α combined with ribavirin for chronic hepatitis C in HIV-infected patients / 中华肝脏病杂志

Objective@#To investigate the optimal duration of pegylated-alpha interferon (Peg-INFα) combined with ribavirin (RBV) in treating chronic hepatitis C infection in human immunodeficiency virus (HIV)-infected patients.@*Methods@#A multicenter prospective study was conducted. The study subjects were divided into two groups; HIV/HCV co-infections (Group A, n = 158) and control with HCV-monoinfections (Group B, n = 60). All recruited patients received standard Peg-INFα plus RBV therapy. Group A was divided into 3 subgroups according to CD4+ cell counts: A1 subgroup, 79 cases, CD4+ counts > 350 cells /μl, who received anti-HCV before combination antiretroviral therapy(cART); A2 subgroup, 45 cases, CD4+ counts between 200 and 350 cells/μl, who did not start anti-HCV until they could tolerate cART well; A3 subgroup, 34 cases, CD4+ counts < 200 cells /μl, cART was administered first, and anti-HCV therapy was started when CD4+ counts > 200 cells/μl. The anti-HCV efficacy of two groups and 3 subgroups were compared. Statistical analysis for normal distribution and homogeneity of variance data was calculated by t-test and the counting data was analyzed by χ 2 test. The Mann-Whitney U test was used for non-normal data. A one-way analysis of variance (ANOVA) was used for the comparison of multiple groups, followed by SNK method. Multiple independent samples were used for non-parametric tests.@*Results@#There was no significant difference in age and baseline HCV RNA levels between groups and subgroups (P > 0.05). By an intent-to-treat (ITT) analysis, in Group A, the ratio of complete early virological response (cEVR) rate was 75.3% (119/158), the ratio of end of treatment virological response (eTVR) rate was 68.4% (108/158), and the ratio of sustained virological response (SVR) rate was 48.7% (77/158); in Group B, the ratio of cEVR rate was 93.3% (56/60), the ratio of eTVR rate was 90.0% (54/60), and the ratio of SVR rate was 71.7% (43/60); The therapeutic index of Group A were lower than those of Group B (P≤0.05). By per-protocol (PP) analysis, the ratio of cEVR rate in Group A [75.2% (88/112)] was still lower than that in Group B [93.3% (56/60)], but no significant differences were found in the ratio of eTVR rate and SVR rate between 2 groups (P > 0.05). Comparing the efficacy of subgroups (A1, A2 and A3) by ITT analysis, the ratios of cEVR rate were respectively 78.5% (62/79), 75.6% (34/45) and 67.6% (23/34); the ratios of eTVR rate were respectively 68.4%(54/79), 80.0%(36/45)and 52.9%(18/34); and the ratios of SVR rate were respectively 41.8%(33/79), 64.4%(29/45)and 44.1%(15/34). The ratio of eTVR in subgroup A2 was obviously higher than that in subgroup A3 and the ratio of SVR in subgroup A2 was statistically higher than that of subgroup A1(P≤0.05). However, by PP analysis, no significant differences of the therapeutic indexes were found among the respective subgroups (P > 0.05).@*Conclusion@#HIV-HCV co-infected patients would have better anti-HCV efficacy with Peg-INFα-2a plus RBV than HCV- monoinfected patients. The best time for initiating anti-HCV therapy in HIV-HCV co-infected patients is when CD4+ counts 200 cells/ μl.

Research on the causes of death associated with combined effects of HBV and HCV infection in patients with acquired immunodeficiency syndrome / 中华肝脏病杂志

Objective@#To investigate the combined effects of hepatitis B virus and hepatitis C virus (HBV/HCV) infection on the cause of death in patients with acquired immunodeficiency syndrome (AIDS).@*Methods@#The causes of death of 111 cases of AIDS with HBV/HCV (combined infection group) and 210 AIDS patients (single infection group) admitted to our hospital from 2012 to 2016 data were compared using chi-square test.@*Results@#There was no statistically significant difference in gender composition and age in the combined infection groups (P > 0.05). The main causes of death in the combined infection group were severe pneumonia (44.1%), end-stage liver disease (18.9%), and central nervous system infection (14.4%). The main causes of death in the single infection group were severe pneumonia (47.6%) and central nervous system infection (14.3%) and tumor (13.3%). There was no case of end-stage liver disease. The ratio of end-stage liver disease in the former group was significantly higher than that in the latter group (χ2 = 42.511, P < 0.001). The main cause of death in 12 HIV/HBV/HCV triple-infected patients was end-stage liver disease, accounting for 41.7%, which was significantly higher than 18.9% of end-stage liver disease in HIV/HBV or HIV/HCV dual infection (99 cases). And the difference was statistically significant (χ2 = 4.539, P = 0.033); however, the ratio of end-stage liver disease in 50 HIV/HBV co-infected patients and 49 HIV/HCV co-infected patients was 16.0% vs. 16.3%, respectively, and the difference was not statistically significant (χ2 = 0.002, P = 0.965). In the co-infected group, 36 patients had CD4+ cell counts >100/μl, the primary cause of death was end-stage liver disease, accounting for 38.2%. 75 patients with CD4+ ≤ 100/μl died due to end-stage liver disease, accounting for 9.3% and the difference was statistically significant (χ2 = 13.852, P < 0.05).@*Conclusion@#End-stage liver disease is the main cause of death in patients with AIDS combined with HBV or HCV, especially triplet infection and CD4+ cell count > 100/μl. An early diagnosis and treatment of HBV or HCV infection should commence as soon as possible.

Characterization and primary resistance mutations in gag-pol gene of human immunodeficiency virus-1 CRF07_BC strain in Guangdong Province / 中华传染病杂志

Objective To investigate the genetic variations of gag-pol gene in human immunodeficiency virus-1(HIV-1) CRF07_BC strain in Guangdong Province.Methods From February to September in 2015,plasma samples of 78 cases with HIV 1 CRF07_BC infection in Guangdong were collected before antiretroviral treatment.Viral RNA was extracted from plasma.Gene (gag and pol) sequences were amplified by reverse transcriptase and nested-PCR using specific primers.Phylogenetic tree,genic dispersion rate,nucleotide polymorphism,selection pressure and variation characteristics were analyzed.Results The main transmission route of the enrolled patients was homosexual transmission (80.77％,63/78).The gag pol gene phylogenetic tree was divided into two sub-clusters.The strains from different transmission routes were not in cluster.The average genetic dispersion rate and average entropy of gag gene were both higher than those of pol gene.The average genetic dispersion and average entropy of p17 and p6 regions of gag gene were both higher than those of p24.The average genetic dispersion and average entropy of pol gene were higher than those of rt region.The average ds/dn values of gag and pol genes were greater than one.Compared with the common HIV-related antigenic epitopes (A2,A11,B39,B60,Cw1,Cw3,Cw8),the cytotoxic lymphocyte (CTL) epitope mutations in the P17 region were more in the consensus of GAG region than those in the P24 region.The epitope conserved rates were 26.92％,0,1.28％,0,96.15％,82.05％,84.62％ and 98.72％,respectively.The drug resistance rate of pol gene was 2.56％ (2/78).Conclusions The gag and pol genes of CRF07_BC strain in Guangdong are all mutated.Diversity of gag gene is greater than that of pol gene,and gag gene variation is mainly in p17 and p6 regions.gag and pol genes are both affected by negative selection pressure.P17 protein CTL epitope variability is greater than P24 protein epitope.The prevalence of drug resistance mutation is lower than the threshold.It's important to monitor the spread of drug-resistant strains.

Non-nucleoside reverse transcriptase inhibitors (NNRTIs)resistance-related gene mutations among AIDS patients with antiviral treatment failure in Guangdong province 2015 / 中华临床感染病杂志

Objective To investigate the prevalence and characteristics of non-nucleoside reverse transcriptase inhibitors (NNRTIs)resistance-related gene mutations among the AIDS patients with virological suppression failure in Guangdong Province 2015.Methods Plasma samples from AIDS patients receiving highly active antiretroviral therapy for more than one year with viral loads > 1000 copies/mL from Guangdong province (except Shenzhen)were collected from January to December 2015.Total 612 HIV-1 gene fragments were amplified from plasma samples using self-developed lab method.Sub-genotypes were determined by phylogenetic tree according to the sequences,NNRTIs resistance-related mutations were determined in Stanford University HIV-1 Drug Resistance Database. The NNRTIs-resistance, the relationships of NNRTIs resistance-related mutations with baseline CD4 +T lymphocyte counts,transmission routes,antiviral regimens and HIV-1 genotypes were analyzed.SPSS 17.0 software was used to analyze the data.Results In 612 patients with virological suppression failure,the main NNRTIs resistance-related mutations were K103 (26.80%),Y181 (14.71 %),V179 (13.73%),G190 (11 .44%) and V106 (10.62%).The susceptibility rate of 310 patients (50.65%)to NNRTIs had changed,the highly resistant rate to nevirapine was 49.51 %,which was higher than that of efavirenz (43.14%),etravirine (5.56%) and rilpivirine (12.25%),respectively,and the differences were statistically significant (χ2 =5.00,296.3 and 198.0,all P 200 cells/μL was lower than that in those with baseline CD4 +T lymphocyte counts <200 cells/μL (χ2 =17.93,P <0.01 );the incidence rate of drug resistance was lower in intravenous drug abusers than that of sexually transmitted patients (χ2 =44.21 ,P <0.01 );while the incidence of drug resistance in patients receiving NVP-containing regimens was higher than that in those receiving EFV-containing regimens (χ2 =8.93,P <0.01 ),and the incidence rate was higher in patients with CRF01 _AE than that in those with CRF07_BC and CRF08 _BC (χ2 =8.46 and 8.47,P <0.01 ).Conclusions The results suggest that compliance education and follow-up should be strengthened in patients with high baseline CD4 +T lymphocyte counts and intravenous drug users,and patients with liver diseases should avoid using drugs containing NVP regimens.

Pathogenic spectrum, clinical features and drug resistance of pneumonia caused by nontuberculous mycobacteria in acquined immunodeficiency syndrome patients / 中华传染病杂志

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.

Diagnostic value of serum Mp1p antigen detection for Talaromyces marneffei infection in acquired immune deficiency syndrome patients / 中华传染病杂志

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.

Detection of hepatitis C virus RNA in the peripheral blood mononuclear cells in patients with chronic hepatitis C and its relationship with effect of anti-viral treatment / 中华传染病杂志

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.

Epidemiological and clinical features of human immunodeficiency virus/hepatitis B virus co-infected patients / 中华传染病杂志

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.

The mycobacterial species distribution in acquired immunodeficiency syndrome patients in Guangzhou / 中华传染病杂志

Objective To investigate the characteristics of mycobacteria species distribution in human immunodeficiency virus (HIV)-positive patients co-infected with mycobacteria in Guangzhou. Methods A total of 133 mycobacteria strains isolated from HIV-positive patients and 150 strains isolated from HIV-negative patients were included in this study. After DNA extraction of mycobacteria, mycobacteria species identification was performed by sequencing of multiple genes.Differences in the identified species were compared between patients with and without HIV infection and the correlation between CD4 + T cells level and the mycobacterial species distribution was analyzed.Chi-square test was used for statistical analysis.Results Of the 133 mycobacteria strains isolated from HIV-positive patients, 82 were identified as Mycobacterium tuberculosis complex (MTC ). Fifty-one were identified as nontuberculous mycobacteria (NTM),of which the main species was Mycobacterium avium complex (MAC,31/51).Of the 150 mycobacteria strains isolated from HIV-negative patients,126 were identified as MTC and 24 as NTM,of which the main species was Mycobacterium abscessus (9/24).In patients with CD4 + T cell counts ≤100/μL,the positive rate of mycobacteria was 75 .94%(101/133),93.55 %(29/31) of MAC and 85 .00%(17/20)of other NTM.When the CD4 + T cell counts >100/μL,the positive rate for mycobacteria were all obviously decreased.Conclusions The proportion of NTM infection is higher in HIV-positive patients than HIV-negative patients in Guangzhou. Among HIV-positive patients > the most prevalent NTM species is MAC, while Mycobacterium abscessus is the most common species in HIVnegative patients. Mycobacterial infection in acquired immunodeficiency syndrome patients is closely associated with low CD4+ cells level.

Impact of HIV/HCV co-infection on peripheral expression of A3G mRNA and interferon-α / 中华临床感染病杂志

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 +T0.05).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.