[Anti-prostate cancer effect of roemerine: An experimental study].

OBJECTIVE: To investigate the anti-prostate cancer (PCa) effect of roemerine in vitro and in vivo in the mouse model of PCa. METHODS: We detected the effects of roemerine on the proliferation, apoptosis and migration of PCa cells DU145, LNCaP, PC-3 and 22RV1, screened out the sensitive cell line and constructed a tumor-bearing model in mice for verification of the antitumor efficacy of roemerine in vivo. RESULTS: Roemerine inhibited the proliferation and migration of the DU145, LNCaP, PC-3 and 22RV1 cells and induced their apoptosis in different degrees, particularly those of the LNCaP cells. The average tumor weight was less in the roemerine intervention group (ï¼»1.99±0.95ï¼½ g) than in the control (ï¼»2.95±1.04ï¼½ g), the least in the high-dose roemerine (30 mg/kg) plus paclitaxel intervention group (ï¼»0.90±0.16ï¼½ g). The mean heart, liver, and kidney indexes were markedly lower in the roemerine (0.58±0.06, 6.20±0.42 and 1.49±0.33) than in the paclitaxel group (0.66±0.04, 6.99±0.72 and 1.95±0.34), while the mean spleen and thymus indexes were remarkably higher in the former (0.54±0.11 and 0.06±0.01) than in the latter (0.41±0.09 and 0.05±0.01). Pathological staining showed a lower degree of malignancy and metastasis in both the roemerine and the roemerine + paclitaxel intervention group than in the control, as well as a lower degree of visceral injury in the roemerine and roemerine + paclitaxel groups than in the paclitaxel group. CONCLUSIONS: Roemerine has some anti-PCa effect and alleviates adverse reactions in paclitaxel combination administration.

[Epidemiological survey on the hepatitis C virus and its genotyping analysis in Henan province in 2012].

OBJECTIVE: To investigate the prevalence and distribution of hepatitis C virus (HCV) genotypes in Henan province in 2012. METHODS: A total of 32 203 permanent residents (1 to 74 years old) in Henan were recruited using multi-stage random samping method from March to June 2012. All participants were asked to complete a questionnaire to collect demographic information, past medical history and the exposure history of risk factors. A blood sample of 5 ml was collected at the same time. The condition of anti-HCV and HCV RNA was determined through the ELISA test and nested RT-PCR. HCV RNA positive samples were further subject to the nonstructural protein 5 region (NS5B) gene amplification and sequencing. The sequence was amplified for the phylogenetic tree and genetic analysis. The differences of the positive rate of anti-HCV and HCV RNA and the HCV genetic subtype distribution in different respondents'characteristics were analyzed. RESULTS: Among 32 203 subjects, the overall positive rate of anti-HCV and HCV RNA were 0.48% (153/32 203) and 0.24% (78/32 203), in which men were 0.42% (65/15 634), and 0.23% (36/15 634), and women were 0.53% (88/16 569) and 0.25% (42/16 596). The differences between men and women were not statistically significant (χ(2) values were 2.26, 0.18, respectively, both P values > 0.05). The results of NS5B genotyping and molecular evolution analysis showed that there were six subtypes in the 71 HCV RNA positive samples.In those six subtypes, the proportion of genotypes 1b, 6a, 3a, 2a, 3b and 1a were 56.3% (40/71), 19.7% (14/71), 11.3% (8/71), 8.5% (6/71), 2.8% (2/71) and 1.4% (1/71), respectively. The HCV genetic subtypes of infestor were mainly present with two branches of 1b and 6a, and the two subtypes Bootstrap values were 0.95. CONCLUSION: The prevalence of HCV infection was high in Henan. The major HCV genotypes in patients with HCV infection were 1b and 6a.

[Association between amino acid mutations in Gag protein and human leukocyte antigen class I alleles in human immunodeficiency virus-1 B' infected Han Chinese people].

OBJECTIVE: To explore the human leukocyte antigen (HLA)-associated mutations in Gag protein of B' clade (human immunodeficiency virus-1) HIV-1 infected Han Chinese people and evaluate the impact of HLA associated Gag mutations on the disease progression of HIV infection. METHODS: A total of 95 B' clade HIV-1 infected Han Chinese cases were recruited. The gag sequences were amplified from viral RNA and sequenced directly. HLA-I genotypes were detected with the assay of polymerase chain reaction-sequence specific primer (PCR-SSP). HLA-associated mutations were identified and the relationships between HLA-associated mutations and CD4+ T cell counts or plasma viral loads analyzed. RESULTS: Forty-seven kinds of mutations at 28 sites (15, 18, 26, 30, 34, 46, 62, 67, 81, 84, 90, 102, 118, 121, 122, 125, 146, 147, 173, 176, 252, 357, 374, 376, 437, 470, 471, 478) of Gag protein were significantly associated with particular HLA class I allelotypes (P < 0.05). Among which, 9 sites (26, 30, 81, 84, 125, 146, 147, 357, 437) were located within 13 known cytotoxic T-lymphocyte (CTL) epitopes or flanking regions. The number of HLA-associated mutations was significantly associated with both CD4 T cell counts (r = -0.318, P = 0.002) and viral loads (r = 0.360, P = 0.003). CONCLUSION: HLA-associated mutations may have a significant impact on HIV disease progression in B' clade HIV-1 infected Han Chinese population.

[The primary HIV drug resistance in partial region of Henan province].

OBJECTIVE: To study the prevalence of primary HIV drug resistance in antiretroviral therapy (ART) areas of Henan province. METHODS: A total of 121 drug-naive long-term infected individuals and 154 patients with newly diagnosed from January 2011 to March 2012 were recruited, the questionnaires were surveyed and whole blood were collected to analyze the CD4(+)T cell counts and viral load. In-house method for genotypic resistance test was determined in those with viral load > 1000 copies/ml samples, the differences of demographic characteristics, immunological parameters and primary drug resistance were compared between the two groups. RESULTS: A total of 121 cases of long-term individuals who had infected (12.50 ± 3.21) years were mainly previous paid blood donors, and the age was (46.61 ± 9.32) years old. The infection route of the newly diagnosed were diversity, including blood, sexual transmission and others, the cases were 73, 73, 8, respectively, the confirmatory year was (0.91 ± 0.28) years, and average age was (22.21 ± 3.11) years old. The difference were statistically significant in the route of transmission, age and infection time from demographic analysis of the two groups (P < 0.05). The absolute M(P(25)-P(75)) counts of CD4(+)T lymphocytes of long-term group was 322 (217 - 422) cell/µl, which was lower than the newly diagnosed was 434(308 - 578) cell/µl (P < 0.05), and viral load was 4.0 (2.96 - 4.64) copies/ml, 3.77 (2.94 - 4.53) copies/ml, the difference was not significant (P > 0.05). The prevalence of primary drug resistance in long-term group and newly diagnosed was 5.79% (7/121), 9.09% (14/154), respectively, and the difference was statistically different (P < 0.05), and one PI-resistant strain was found in the newly diagnosed group. CONCLUSION: The primary drug resistant strains in untreated patients were found in Henan province of ART areas, and there was difference in degree of resistance between long-term infected individuals and newly diagnosed.

[Subtype analysis of gp41 gene of HIV-1 among men who have sex with men in Zhengzhou].

OBJECTIVE: This study aimed to investigate the subtype distribution of gp41 gene of human immunodeficiency virus-1 (HIV-1) among men who have sex with men (MSM) in Zhengzhou. METHODS: Thirty blood samples were collected from men who have sex with men infected by HIV. The complete gp41 gene was amplified by RT-PCR and nested-PCR and sequenced. All sequences were edited by Bioedit and subtyped with HIV sequence library US Los Alamos National Laboratory and online genotyping software provided by American National Center of Biotechnology Information. Phylogenetic analysis of gp41 gene was performed using the MEGA 3.1 software, and the genic dispersion rates among subtype of gp41 gene were analyzed. RESULTS: A total of eighteen gene sequences of HIV-1 gp41 gene were obtained from thirty men who have sex with men infected by HIV, which belonged to subtype CRF15-01B (50% (9/18)), CRF01-AE (22% (4/18)), CRF07-B (22% (4/18)) and B (6% (1/18)), respectively. The intersubtype HIV-1 strains aggregate with according reference strains. The genetic distance inter-subtype of subtype CRF15-01B, CRF01-AE and CRF07-B were 0.050 ± 0.007, 0.052 ± 0.009 and 0.082 ± 0.012, respectively. CONCLUSION: The prevalent subtypes of HIV-1 among among MSM in Zhengzhou was complicated and recombinant HIV-1 strains were the most prevalent strains.

[Study on the transmission of drug resistant human immunodeficiency virus-1 in Henan province].

OBJECTIVE: To survey the prevalence of HIV-1 drug resistance after five years antiretroviral treatment in Henan province. METHODS: Through the cross-sectional study, serum specimens of 69 HIV infected individuals that were 2 to 25 years old who were newly diagnosed according the WHO standard from November 2007 to August 2008 and did not receive antiretroviral treatment (ART) were collected. HIV-1 pol genetic mutations associated with drug resistance were identified with RT-PCR and interpreted. RESULTS: Out of 69 samples, 50 samples were successfully amplified and sequenced. Seven drug resistant mutation in reverse transcriptase region were detected and three mutations in protein region. In one specimen, a mutation (K103N) in reverse transcriptase was identified which caused high level resistance to NNRTIs, but no proteinase inhibitor mutation was found in protein region. According to the sampling and threshold surveillance criteria, the prevalence of drug resistant HIV-1 in Henan was less than 5%. CONCLUSION: The prevalence of drug resistant HIV-1 was still at low level in Henan. However, the proportion of resistant strains would be higher with the antiretroviral treatment. We should pay more attention to the transmission of resistant strains and continue the drug resistance surveillance.

The impacts of current antiretroviral therapy regimens on Chinese AIDS patients and their implications for HIV-1 drug resistance mutation.

This study sought to investigate the impacts of the antiretroviral (ARV) therapy regimens currently used in Chinese HIV-1-infected individuals. Seven hundred eighteen ARV-treated and treatment-naive HIV-1-infected individuals living in seven provinces were enrolled in 2005 by a multistage sampling approach according to a national cross-sectional survey program on HIV-1 drug resistance. All patients were investigated clinically, and CD4+ T cell counts and HIV-1 viral loads were measured while genotyping for drug resistance was determined by a home brew nested PCR. Viral inhibition in ARV-treated individuals was higher than that in ARV treatment-naive individuals. The overall prevalence of drug-resistant mutations was 37.8%. Higher frequencies of mutations in ARV-treated and drug withdrawal groups were found than in the ARV treatment-naive group (P<0.01). Of the four regimens currently used, the D4T/3TC/NVP regimen showed a higher-level viral inhibition. No statistical significance was found among the four regimens in drug-resistant mutations. The rate of resistance-associated mutations to non-nucleotide reverse transcriptase inhibitors (NNRTIs) was higher than that to nucleotide reverse transcriptase inhibitors (NRTIs) (P<0.01). The most common mutations conferring resistance to NNRTIs were K103N, Y181C and G190A, representing 56.5, 30.4 and 14.5%, respectively. Furthermore, higher viral inhibition and a lower rate of drug-resistant mutations were achieved in the good compliance group. This study revealed an efficient viral inhibition achieved with the current first-line regimens in China. Most of these regimens could rapidly result in emergence of drug-resistant mutations, suggesting that a second-line ARV therapy is urgently needed and that the compliance with treatment must be emphasized during long-term treatment.

[Preliminary study on the relationship between sow maternal behaviour during early lactation and polymorphism of PRLR gene].

PCR-RFLP technique was applied to analyze single nucleotide polymorphisms of PRLR gene in Minpig and Landrace to investigate the possible effect of PRLR gene on sow maternal behaviour. A Nae I-RFLP site was detected in the PRLP gene. The single nucleotide polymorphism, a T-->C transition at nucleotide 1,620 of the cDNA sequence, was a silent mutation. Least square analysis between the genotypes and the maternal behavioural traits showed that sows with genotype AB had a significantly higher frequency of lateral-lying-to- other-posture trait and percentage of sow-terminated nursing trait than sows with the AA and BB genotypes, although no significant differences were found in other behavioural traits. It is possible that allele A is the unfavorable allele for sow maternal behaviour.

[Prevalence and related factors of prostatitis-like symptoms in young men].

OBJECTIVE: To investigate the prevalence and related factors of prostatitis-like symptoms among young men. METHODS: The study was a cross-sectional survey of 2500 young men aged 18-30 years in the city of Weifang, and all of them completed a questionnaire on prostatitis. The univariate and multivariate logistic regression procedures were used to investigate the risk factors among the young men with chronic prostatitis-like symptoms. RESULTS: The valid response rate was 85% (n = 2125). Of the 128 subjects (6.02%) identified as having chronic prostatitis-like symptoms, the mean age was 21.8 years, the average pain score was 6.98 +/- 0.29, and the average voiding score was 3.77 +/- 0.25. Of the sampled population, 39 men had prostatitis-like symptoms with an index pain score of 8 or more. Significant risk factors include frequent masturbation, prolonged sitting, long-time fixed posture, cold environment, stress at home and work. CONCLUSION: The study suggested that chronic prostatitis-like symptoms are common among young men, and the urethritis history, frequent masturbation, prolonged sitting, long-time urine holding, cold environment, and stress at home and work might be significant risk factors.

[Efficacy of anti-HIV treatment and drug-resistance mutations in some parts of China].

OBJECTIVE: To evaluate the Virologic and Immunologic efficacy of HAART on Chinese HIV/AIDS patients and to assess the impact of of HAART on drug resistance mutations. METHODS: Three cohorts of Liaoning, Jilin and Henan province received three different regimens for 6 months respectively. Regimen of Liaoning cohort comprised Efavirenz + Indinavir (EFV + IDV), regimen of Jilin cohort comprised Stavudine + Didanosine + Efavirenz (d4T + ddI + EFV) and regimen of Henan cohort comprised Stavudine + Didanosine + Nevirapine (d4T + ddI + NVP). Viral load, CD4(+) T cell count and drug resistance genotype were detected on the three cohorts before and after treatment. Partial HIV-1 pol genes encoding protease and 1 - 220 amino acid of reverse transcriptase were amplified by RT-PCR and then automatically sequenced. All sequences were compared with the data of Stanford HIV Drug Resistance Database to assess resistance mutations against reverse transcriptase inhibitors (RTIs) and protease inhibitors (PIs). RESULTS: During observation of 6 months, viral suppression to undetectable level and Elevated CD4(+)T cell count efficacy were achieved on partial Chinese HIV/AIDS patients in each of the three different regimens, even in some patients with rather low CD4(+)T cell count baseline. Before HAART, no primary mutations against PIs and RTIs were detected on the three cohorts, except one patient in Liaoning cohort. But after HAART, drug resistance mutations against RTIs occurred on each of the three cohorts. K103N is the most common mutation against NNRTIs, which can cause high-level resistance to each of the available NNRTIs. Y181C is another common mutation occurred in Henan cohort, which causes crossing drug resistance and multi-drug resistance to NNRTIs. In addition, intermediate level and low level resistance against NRTIs caused by K65R and L74V can also be found, but less commonly. CONCLUSION: Treatment naive Chinese HIV/AIDS patients were sensitive to HAART. Expected virologic and immunologic efficacy of HAART were achieved on Chinese HIV/AIDS patients, but after the introduce of HAART, the high prevalence of drug resistance mutations against NNRTIs and NRTIs, crossing drug resistance and multi-drug resistance reminded us to pay more attention to the drug resistance mutations detection, treatment standardization, and to avoid drugs wasting and prevent the prevalence of drug resistance strains.

[Single nucleotide polymorphism analysis in sow mu opioid receptor gene exon III].

Stereotypies are the most common abnormal behaviours in sows. Stereotypies are repeated, relatively invariable sequences of movements which have no obvious purpose. Mu opioid receptor is a member of G protein-coupled receptor family, distributed in the pain transduction region in the brain and related emotion and behavior regions where influence the animal neural reaction and behavior. The possibility of the MOR gene as the candidate gene to affect the stereotyped behavior traits in sows was discussed in this study. The primer of sow MOR gene exon III partial sequence was designed to analyze single nucleotide polymorphisms by PCR -SSCP in Landrace, Yorkshire and Duroc breeds. Two polymorphisms were found, which was caused by a single nucleotide mutation of C to T and C to A at the positions of 1169 and 1226, respectively compared with the sequence in GenBank, but it was the silence mutation. The results of chi 2 test showed that the frequencies of genotypes resulted in different breeds were significantly different (P < 0.01). The least square analysis between the nucleotide acid mutant induced three genotypes and the stereotyped behavioral traits in Yorkshire showed that individuals with BB genotype have significant higher (P < 0.01) sham-chewing behavior than those with AA genotype, but not for bar-biting and standing. According to the above results, we can putatively draw the conclusion that MOR gene is probably the major gene affecting the sham-chewing behavioral traits or linked to the major gene.

[Single nucleotide polymorphism in exon partial sequence of sow kappa opioid receptor gene].

The possibility of the Kappa opioid receptor (KOR) gene as the candidate gene affecting the stereotyped behavior traits in sows was discussed in the current study. Based on the exon partial sequence of sow KOR gene, three pairs of primers were designed to analyze single nucleotide polymorphisms to develop PCR-SSCP method in Landrace, Yorkshire and Duroc breeds. One polymorphism, found in PCR product amplified with F1/R1, was caused by a single nucleotide mutation of C to T at the position of mRNA 105 compared with the sequence published in GenBank, which was a silence mutation. The results of chi2 test showed that the frequencies of genotypes in different breeds were different significantly (P<0.01). The least square analysis between three genotypes and the stereotyped behavioral traits showed that individuals with BB genotype had significantly higher (P<0.01) still standing behaviors than those with AA and AB genotype, but no difference were found in other behavioral traits. According to the above results, we may conclude that SNP of KOR gene has effect on the still standing behavioral trait of sows.

[Clinical significance of expression of PSA, hK2, PSMA in the peripheral blood of patients with prostate cancer].

OBJECTIVE: To find sensitive and specific micro-metastic markers for prostate cancer. METHODS: Using nested reverse transcription-PCR, we examined the expression of PSA, hK2 and PSMA mRNA in peripheral blood mononuclear cells of 51 patients with prostate cancer, 33 patients with benign prostate hyperplasia (BPH) and 32 normal young people. RESULTS: The expression rates of PSA, hK2 and PSMA mRNA were 52.9%, 43.1% and 64.7%, respectively in prostate cancer group, and 6.2%, 7.7% and 4.6%, respectively in control group (BPH patients and normal young people) with statistical significance (P < 0.01). Although the expression rate of PSA and hK2 mRNA increased with cancer progression, there was no statistical significance among patients in different stages. The expression rate of PSMA mRNA was higher than that of PSA and hK2 mRNA in each clinical stage. CONCLUSION: PSMA mRNA expression detected by nested RT-PCR is of greater value for the diagnosis, therapy choice and prognostic evaluation of prostate cancer patients.

[Single nucleotide polymorphism analysis in sow mu opioid receptor gene].

Mu opioid receptor (MOR) is a member of G protein-coupled receptor family, distributed in the pain transduction region in the brain and related to emotion and behaviour. This study was designed to investigate the Single Nucleotide Polymorphism (SNP) of Mu opioid receptor gene in various breeds,including duroc, landrace and Yorkshire. 5' UTR (untranslate region), coding region and 3' UTR of Mu opioid receptor gene were amplified by eight pairs of primers, and the Single Nucleotide Polymorphism (SNP) were detected by SSCP. Five polymorphisms were found (Genebank Accession number: AF521309). The results of chi(2) test showed that the frequencies of genotypes in different breeds were significantly different (P<0.01). The frequencies of mutation genotypes in Yorkshire were significantly higher than Duroc and Landrace. According to the above results, we can speculate the difference of the frequencies of genotypes may be the results of long term choice pressure.

[Epidemiological characteristics of hepatitis C in Henan province].

OBJECTIVE: To understand the epidemiological characteristics of hepatitis C in Henan province. METHODS: Multistage sampling was used to investigate a group of 1-75 year-old general population, living in 30 districts of 18 cities, Henan province. Blood samples were collected and tested for anti-HCV and hepatitis C virus(HCV), from April to July 2012. RESULTS: 32 203 persons were investigated. Among the general population aged 1 to 75 years old, the overall prevalence rates of anti-HCV and HCV RNA were 0.64% and 0.35% respectively. The prevalence rates of anti-HCV among males and females were 0.60%, and 0.68% respectively, with the rates of HCV RNA as 0.37%, and 0.33% respectively. The prevalence rates of anti-HCV and HCV RNA were increasing with age. The prevalence rates of anti-HCV and HCV RNA in urban area were 0.58% and 0.32%, and in rural area as 0.41%, and 0.19% respectively. CONCLUSION: After the implementation of comprehensive measures for HCV control and prevention, Henan province became low prevalent for HCV infection. Our data revealed that the overall prevalence remained low, but the epidemic was severe in some geographical regions in Henan province.

[A cross-sectional study on the prevalence of HIV drug resistance in patients receiving antiretroviral treatment in Shenqiu county, Henan province].

OBJECTIVE: To understand the prevalence of drug resistance in AIDS patients who had been receiving HAART in a long run, in Shenqiu county, Henan province. METHODS: This cross-sectional study included 120 HIV infected patients who began receiving ART (antiretroviral therapy) in 2003. Viral loads and CD(4)(+) T cells counts were measured, and In-house drug resistance test was performed in VL > 1000 copies/ml patients. RESULTS: 114 cases out of 120 patients had complete viral load data. Among them, 33 cases having viral loads less than 50 copies/ml, and the remaining viral loads showed an average of lg (4.09 ± 1.10) copies/ml. The average of CD(4)(+) T cell counts was (377 ± 218) cells/ml, with 64 (53.3%) cases showing their CD(4)(+) T cell counts higher than 350 cells/ml. In 67 patients, 58 of them showed genotypic resistance, and 40 cases showed reverse transcriptase inhibitors (RTIs) resistance. The ratios of nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) resistance were 53.4% (31/58) and 67.2% (39/58), respectively. There were no differences of drug resistance ratio in the three treatment programs. The highest drug resistance rates in NRTIs and NNRTIs were zidovudine, lamivudine, nevirapine. However, protease inhibitors (PIs) resistance variants were not found. CONCLUSION: The prevalence of drug-resistant strains seemed to be high in Shenqiu country, Henan province. Long-term follow-up monitoring strategy should be developed to optimize the timely treatment programs.

[Follow-up study on 84 AIDS patients having received the replaced therapy program for six months in one county of Henan, China].

OBJECTIVE: To analyze the situation of AIDS patients who had received replaced therapy program in Henan province. METHODS: 84 AIDS patients had been enrolled into the national free first-line antiretroviral treatment for more than 5 years and would soon be replaced with another antiretroviral treatment program, were selected to a follow-up program to be carried out six months later. Data on CD4(+) t-lymphocyte count, viral load and genotypic resistance were included in the study. RESULTS: The DDI+AZT+NVP treatment program was used by all the 84 patients at baseline. A replacement by 3TC + AZT + NVP (post first-line) in 31 patients and 3TC + TDF + LPV/r (second-line) in another 53 patients were taken place within a week. All the patients were followed for six months. RESULTS: showed that: all of the 84 patients appeared an amelioration of CD4(+) t-lymphocyte count median from the baseline of 374.00 cell/µl to 406.50 cell/µl (P = 0.005). Those patients who had changed to second-line treatment program also showed an improvement of CD4(+) t-lymphocyte count median from the baseline of 267.00 cell/µl to 365.00 cell/µl (P = 0.015), while patients who were on the post first-line program with their CD4(+) t-lymphocyte count mean did not show significant change as compared to the baseline (P = 0.158) data. All the 84 patients showed a decrease of virus load median from the baseline of 3.61 log(10)copies/ml to 0.00 log(10) copies/ml (P = 0.000). Both of the two types of patients who had been changed to different programs, had a lower virus load median in the end of the follow-up period (for post first-line: P = 0.007; for second-line: P = 0.000). 13 patients kept their viral load more than 1000 copies/ml, including 5 cases bore more than three thymidine analogue mutations (TAMs) a the end of the follow-up program. Another 4 patients had no resistance mutations detected and no significant variation of viral load (less than 3 times) in the pre- or post-surveys. CONCLUSION: AIDS patients who had received long-term first-line antiretroviral treatment program, showed an amelioration six months after changing of the treatment program. Timely and effective testing on drug resistance as well as the strengthening of the follow-up program still seemed to be the link to those patients who were receiving first-line treatment that should not be ignored.

[Study on HIV-1 drug resistance profile of 257 AIDS patients with failure on the first-line antiretroviral treatment in Henan].

OBJECTIVE: To understand the current situation related to genotypic resistance in patients receiving the first-line antiretroviral treatment, but with with virologic failure, in Henan province and to compare the patterns of drug resistance in patients from different areas. METHODS: 276 patients with virologic failure on first-line antiretroviral treatment were selected in three different areas of Henan, in 2010. CD4(+)T cells, virus load and genotypic resistance were measured and tested. Prevalence and mutations related to drug-resistant were analyzed. RESULTS: The overall prevalence of drug-resistance was 68.48% in 257 patients, with non-nucleoside reverse transcriptase inhibitor (NNRTIs) as 67.70%. Rate of nucleoside reverse transcriptase inhibitor (NRTIs) was 54.09%, and protease inhibitors (PIs) was 1.18%. The prevalence rates of drug-resistance in A, B and C groups were 82.35%, 97.47% and 52.80%, respectively, and withs significant differences (χ2=50.624, P=0.000). The Prevalence rates related to resistance of NNRTIs and NRTIs were also significantly different (χ2=48.771, P=0.000 and χ2=33.912, P=0.000). 26.46% of the samples had M184V/I mutation which was the highest NRTIs mutation among the 257 patients. The prevalence rates on resistance of A and B were 47.06% and 49.37%, higher than that of C (13.04%, χ2=39.905, P=0.000) followed by TAMs, TAMs-1 and TAMs-2 which were 8.56% and 4.28%. C had the lower prevalence of TAMs-1 than A and B (χ2=13.499, P=0.001). 40.47% of the samples harbored≥1 TAM, with T215Y/F having the most, as 33.85%. 31.13% of 257 patients appeared most NNRTIs mutation K103N in this study, with the prevalence rates also significant different (χ2=14.213, P=0.001) in the three areas. Two PIs mutations were detected in 257 patients: M46I/L, (1.17%) and V82F (0.39%). However, none was detected in area A. CONCLUSION: Different patterns of drug resistance were found in different areas of Henan province and should be treated differently. The work related to AIDS second-line antiretroviral therapy in Henan should be more opportune, rigorous and standardized.

[Men who have sex with men and its relationship with HIV-1 strains prevailing in the paid blood donors from Zhengzhou city, Henan].

OBJECTIVE: To clarify the genetic characteristics of human immunodeficiency virus (HIV) circulated in the population of men who have sex with men (MSM) in Zhengzhou, Henan and to analyze its relationship with HIV-1 prevailing in the paid blood donors (PBDs). METHODS: Thirty-one MSM who were confirmed as HIV positive individuals in 2010 together with 41 HIV-positive former PBDs were enrolled in the study. Information on related epidemiological characteristics and their plasma were collected. RT-PCR was used to amplify HIV-1 full length gag (1584 bp), pol (3147 bp) genes and partial env gene (C2V3 segment, 558 bp) followed by sequencing on those subjects. Online software available at LosAlamos HIV Database was used to identify the HIV subtypes based on the findings of the sequences. Phylogenetic tree was used to identify the possible relationship of transmission. RESULTS: Fifty-three full length gag, 38 full length pol and 48 partial env (C2V3) genes were collected from 72 participants. Among the 31 HIV(+) MSM individuals, 14 CRF01_AE strains, 5 CRF07_BC strains and 12 subtype B (1 subtype B and 11 B') strains were identified respectively. All of the 41 strains identified from former PBDs were infected by B' strains. The CRF01_AE strains identified in MSM showed a close relationship to those identified from both Hebei and Liaoning provinces. The CRF07_BC strains showed a close relationship with those from Shijiazhuang and Beijing cities. Among the 12 subtype B strains, 8 sequences grouped into 1 cluster with 1 sequence from the former PBDs. Two sequences grouped with 02HNseq4 suggested that B' had been prevailed in the MSM population might come from the former PBDs and were closely related to the strains identified in the MSM population. CONCLUSION: Complicated genetic background and multiple introductions of HIV in the MS population in Zhengzhou, were found. This was also the first report which noticed that the subtype B epidemic among Zhengzhou MSM was mainly originated from the B' among the former PBDs.

[Subtype and sequence analysis of gag gene of HIV-1 among men who have sex with men in Zhengzhou, Henan Province].

To investigate the subtype distribution of human immunodeficiency virus-1(HIV-1) infection among men who have sex with men (MSM) in Zhengzhou, Henan Province, forty blood samples were collected from HIV-1 carriers, who acknowledged to have sex with men. The complete gag gene was amplified by RT-PCR and nested-PCR and sequenced. All sequences were edited by BioEdit and subtyped by genotyping software. Phylogenetic analysis of gag gene were then performed using the MEGA 3.1 software, the gene distances were calculated by Distance program. There were three different HIV-1 subtypes including B, CRF01-AE and CRF07-BC present among twenty four MSMs in Zhengzhou. Genotyping results showed that 33.33% (8/24) were B, 41.67% (10/24) were CRF01-AE and 25% (6/24) were CRF07-BC, and subtype CRF01-AE had become the most prevalent HIV-1 subtype in Zhengzhou, Henan province. In conclusion, recombinant HIV-1 strains are circulating in Henan province and the epidemiology is complicated.