Analysis on the genetic characteristics of HIV-1 infected MSM individuals in Zhejiang province / 预防医学

Objective To analyze the subtypes and characteristics of HIV-1 among men who have sex with men (MSM) in Zhejiang province from the year 2004 to 2011.Methods Blood DNA/RNA was extracted from the MSM HIV -1 infected individuals and then HIV-1 gag and pol fragments were amplified by nested polymerase chain reaction (nested-PCR)or RT-PCR.The positive PCR products were sequenced and the obtained sequences were analyzed by phylogenetic inference.Results A total of 117 HIV-1 infected MSMindividuals were analyzed,covering 21 provinces according to their household registry.Totally,three major strains were identified including CRF01_AE (84.62%,n =99 ),B (5.98%,n=7)and CRF07_BC (5.13%,n=6). Moreover,3 novel 01B recombinant stains,1 CRF59_01B and 1 CRF08_BC were found.According to the neighbor-joining tree by pol fragments,there were more than 30 sub clusters with bootstrap value higher than 70% among the 84 CRF01_AE sequences.Conclusion CRF01_AE is the major HIV-1 infection strain among MSMin Zhejiang province.CRF59_01B and other novel 01B recombinant stains are first reported among MSM individuals in Zhejiang province.

Molecular epidemiological study on HIV/AIDS under the follow-up program in Zhejiang province in 2009 / 中华流行病学杂志

Objective To analyze the molecular epidemiological characteristics on HIV infectors/AIDS patients (HIV/AIDS) under a follow-up program in Zhejiang province in 2009.Methods 303 cases were randomly sampled.Information on the cases was collected and followed by genomic DNA extraction.Gag gene fragments were amplified by nested PCR,followed by sequencing and bio-informatic analysis.Results The rate of success for sequence acquisition was 74.3％ (225/303).Distributions of HIV subtypes were as follows:CRF01_AE (58.7％),CRF07_BC (13.8％),CRF08_BC (9.8％),B' (15.1％),C (1.8％),G (0.4％) and unassigned BC (unique recombinant form 0.4％).Results from the HIV BLAST analysis showed that the sources of strains with the highest homology involved in 10 provinces/municipalities (Liaoning,Guangxi,Yunnan,Henan,etc.) and five other countries (Thailand,Vietnam,India,South Africa and Libya).The CRF01 _AE phylogenetic tree was divided into four clusters.The sequences of HIV/AIDS with homosexual transmission showed a gathering in cluster 1,and mixing with those infected through heterosexual contact.Conclusion Circulating recombinant forms of HIV seemed to play a dominant role in Zhejiang province.Unique recombinant form and new subtype of HIV were found.People living with HIV under homosexual transmission and heterosexual transmission had a trend of interwoven with each other.Increase of both the diversity and complexity of HIV strains were also noticed in Zhejiang province.

Survey of HIV drug resistance threshold in Zhejiang province from 2009 to 2011 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To survey the prevalence of drug resistant HIV in Zhejiang province in 2009-2011.</p><p><b>METHODS</b>WHO truncated sequential sampling technique was adopted annually by using 63, 62 and 57 samples of newly diagnosed as HIV positive and aged 16-25 years in Hangzhou, Ningbo and Wenzhou from 2009 to 2011, respectively. RNA was prepared and HIV pol region was amplified by RT-PCR and nested PCR. Pol genetic mutation associated with drug resistance was analyzed.</p><p><b>RESULTS</b>The success rates for sequence acquisition of the survey were 82.5% (52/63), 95.2% (59/62) and 94.7% (54/57) from year 2009 to 2011, respectively, and the main subtype was CRF01_AE (68.5% (37/54)-71.2% (37/52)). A total of 4 surveillance drug-resistance mutation (SDRMs), 2 SDRMs and 2 SDRMs were found by analyzing the 47 sequences each year, sampled from year 2009 to 2010, respectively, indicating that the prevalence of drug resistant HIV stains was moderate in 2009, and low for the next two years (2010-2011). A total of 8 individuals with drug resistant HIV stains found in this study were all infected by sexual transmission, especially in homosexual transmission (6 cases), and the main subtype was CRF01_AE (7 cases). SDRMs for protease inhibitor (PI), nucleotide HIV-reverse transcriptase inhibitor (NRTI) and non-NRTI (NNRTI) (L90M, T215S and Y188L) were all found in one case.</p><p><b>CONCLUSION</b>The prevalence of drug resistant HIV stains in major areas with AIDS epidemic in Zhejiang province was low in 2009-2011.</p>

Molecular characteristics of HIV-1 CRF01_AE strains in Zhejiang province in 2009 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze the molecular epidemiological characteristics of HIV-1 subtype CRF01_AE strains being prevailed among HIV/AIDS in Zhejiang province in 2009.</p><p><b>METHODS</b>A total of 303 subjects were identified by stratified random sampling among HIV infected individuals in Zhejiang province in 2009. Gag fragments of the HIV-1 strains were amplified by nested polymerase chain reaction from the DNA extracted from whole blood of HIV-1 infected individuals. PCR products were sequenced and analyzed by phylogenetic method.</p><p><b>RESULTS</b>A total of 132 HIV-1 subtype CRF01_AE sequences were identified from the 225 samples that sequenced successfully, accounting for 58.67% (132/225). A total of 90.91% (120/132) CRF01_AE strains infected HIV/AIDS were transmitted mainly by sexual contacts. A total of 65.91% (87/132) of the cases infected by heterosexual route and 25.00% (33/132) by homosexual route. There were three main clusters in the phylogenetic tree. Pairwise DNA distance within three groups was 0.037 ± 0.011, 0.034 ± 0.008 and 0.047 ± 0.010, which has statistical significance (P < 0.05). Distribution of the sequence of homosexual behavior infected individuals was relatively concentrated in clusters one (96.97%, 32/33), and crossed with heterosexual behavior infectors, and presented the close relations with strains from Jiangsu province, Zhengzhou of Henan province, Liaoning province, Shijiazhuang of Hebei province.</p><p><b>CONCLUSION</b>The CRF01_AE strains were the dominant subtypes among HIV infected individuals. The majority of the CRF01_AE infected cases had high risk sexual behavior. The heterosexual infected cases were more than homosexual cases. The circulating status of CRF01_AE strains in homosexual population was relatively independent, but also had evidence of transmission from man who have sex with man to heterosexual population.</p>

Psychological Status of Stroke Patients during Rehabilitation / 中国康复理论与实践

@#Objective To understand the psychological status of stroke patients during rehabilitation. Methods 58 stroke patients were detected with Symptom Checklist (SCL-90) after 2 weeks. Results All the patients presented various psychological problems with various degrees after 2 weeks. There was no difference in types and degrees of psychological problem between hemorrhagic and ischemic stroke patients (P>0.05). The scores of depression, anxiety, terror, forced and interpersonal sensitivity were higher in serious limb paralysis patients than in mild ones (P<0.01). Conclusion The incidence of psychological problem is really high in patients after stroke. The more serious limb paralysis is, the more serious psychological problem is.

Relationship between strain subtypes and transmission of HIV infection within marriage-based immigrant women in rural area of Zhejiang province / 中华流行病学杂志

Objective To analyze the relationship between strain subtypes and transmission of HIV infection on marriage-based immigrant women,their spouses and children in rural area of Zhejiang province.Methods Marriage-based immigrant women with HIV infection,their HIV infected spouses and children in rural area in Zhejiang province,were selected as study objects.Analysis on genetic sequence and epidemiologic information was carried out.Subgenomic gag was amplified by nest-PCR analysis on the whole blood samples.Genetic subtype characterization and the source of HIV strains were analyzed.Relationships on sequences were also examined by phylogenetic tree analysis.Results Genetic sequences of 72 samples from HIV infected marriage-based immigrant women were obtained.The genetic subtypes comprised 21 CRF01_AE (29.2％),12CRF07 BC (16.7％),31 CRF08 BC (43.1％),6 B (8.3％),2 C (2.8％).HIV strains from 45 cases (62.5％) were similar to the prevalent HIV strains in the province where former census of marriagebased immigrant women were registered.In total,there were 26 (70.3％) cases from Yunnan province.84.7％ of the infected women had heterosexual behaviors before settling down in Zhejiang province.Genetic sequences of 17 pairs showed the same subtype between the couples and data from phylogenetic tree analysis supported the assumption of transmission linkage in the family.Conclusion The HIV subtype strains detected in those HIV infected marriage-based immigrant women in the rural area of Zhejiang province characterized with diversity,showing CRF08_BC and CRF01_AE were the main HIV strain subtypes.HIV infection originated mainly fiom Yunnan province and nearby regions.Heterosexual behaviors of the marriage-based immigrant women in the original region where they had their residence registration,seemed to be the primary high risk factors for these women.Surveillance and intervention programs on these marriage-based immigrant women and their family members should be improved.

Subtype and transmission of HIV strain in both HIV infected spouses in Zhejiang province / 中华流行病学杂志

Objective To analyse the subtype and transmission of HIV strain in both HIV infected spouses. Methods Reported both HIV infected spouses were selected as objects. Analysis on genetic sequence and high risk behaviors was carried out. Subgenomic gag was amplified by nest-PCR analysis of whole blood samples from objects. Genetic subtype characterization of HIV was identified and pairwise genetic distances were calculated. Sequence relationships were also examined by phylogenetic tree analysis. Results Genetic sequences of 46 pairs (92 cases) were obtained. The genetic subtype comprised 39 CRF01_AE(42.4%), 10 CRF07_BC(10.9%), 18 CRF08_BC (19.6%),18 B (19.6%) ,5 C (5.4%) and 2 CRF02_AG(2.2%). 44 pairs had the same subtype between the two partners, accounted for 95.7% of the total. 33 of the 41 pairs with phylogenetic tree analysis were found having epidemiological linkage in pair wise. Sexual behaviors of out-marriage were the main risk factors of CRF01_AE and CRF08_BC and CRF02_AG strains infection. Blood transmission was associated with B and CRF07_BC. Conclusion The HIV strains subtype detected in HIV infected spouses characterized with diversity. CRF01_AE was the main strain subtype. The main route of transmission to the spouses was through unprotected sexual contacts. Surveillance programs on HIV infected partner together with intervention between the spouses should be improved.

Genotypic drug-resistance of HIV-1 CRF01_AE in Zhejiang province, 2004-2007 / 中华流行病学杂志

Objective To study the genotypic drug-resistant mutation among treat-naive or treated patients infected with HIV-1 CRF01_AE in Zhejiang province during 2004-2007. Methods HIV-i pol amplicons (PR+RT) from 13 treated and 43 treat-naive patients were obtained by reverse transcription-polymerase chain reaction (RT-PCR). The sequences were analyzed for genotypic antiretroviral resistance through online tools (http://hivdb.stanford.edu). Results The median count of CD44+ T lymphocytes in 43treat-naive patients was 229 cells/mm3 and the median log10 viral load was 3.41. Some drug-resistant mutations were seen in these samples including amino acid 10, 46, 71, in the genes of protease (PR) and 103, 118, in the genes of reverse transcriptase (RT) whereas twenty-nine resistance mutations in the genes of PR and RT were obtained in the 13 treated patients (8/13, 61.5% ). The high prevalence of drug-resistant mutations was observed in patients who had been receiving HAART (hight active antiretroviral therapy). Among them, cross drug resistance was dominant. Correspondingly, the median counts of CD44+ T lymphocytes and the log10 viral load were 186 cells/mm3 and 3.91. Conclusion There was a low prevalence of genotypic drug-resistant mutations in treat-naive patients, but higher drug-resistant mutation in treated patients. More attention should be paid to the transmission of drug-resistant HIV strains and the antiretroviral therapy recipe should be adjusted correspondingly for the development of ART drugs, intervention as well as clinical therapy programs.

Rapid simultaneous detection of Salmonella and Shigella using modified molecular beacons and real-time PCR / 中华流行病学杂志

<p><b>OBJECTIVE</b>Dual detection of Salmonella and Shigella using modified molecular beacons and real-time PCR was developed. The established method was applied to rapid diagnosis of Salmonella and Shigella' food poisoning, and for routine monitoring programs.</p><p><b>METHODS</b>Two sets of primers were designed based on the core sequence of invA gene and ssaR gene published on GenBank to detect Salmonella, and ipaH gene were selected to detect Shigella. Three corresponding modified molecular beacons labeled with different fluorophors were designed. The molecular beacons and primer sets were tested against numerous strains from 55 different bacterial species. Then the two assays were combined to establish the dual real-time PCR assay, and were applied to the food poisoning diagnosis and surveillance.</p><p><b>RESULTS</b>For the modified molecular beacons-based dual real-time PCR assay, the sensitivity achieved was 69-93 fg/microl, 32-64 CFU/ml or 1-2 CFU/PCR reaction. There was no cross-reaction with other bacteria served as control. The dual real-time PCR assay was used to detect 134 Salmonella strains and 67 Shigella strains but no false signals were observed. 1100 food poisoning samples were tested with 569 Salmonella and 42 were Shigella identified by real time PCR. Among the positive samples, 551 were detected Salmonella and 41 were Shigella by traditional culture method. The overall test could be finished within 2 hours to one day starting from sample preparation.</p><p><b>CONCLUSION</b>The modified molecular beacons-based dual real-time PCR assay was rapid, sensitive, and specific. It could be applied to the rapid diagnosis of Salmonella and Shigella' food poisoning.</p>