Adult ; Antibodies, Anticardiolipin ; blood ; Antibodies, Antiphospholipid ; blood ; Autoantibodies ; blood ; Female ; Humans ; Male ; Middle Aged ; Phosphatidic Acids ; immunology ; Phosphatidylcholines ; immunology ; Phosphatidylethanolamines ; immunology ; Phosphatidylinositols ; immunology ; Phosphatidylserines ; immunology ; Reference Values

Objective To assess pertinent indicators of elderly-friendly city, so as to provide the basis for carrying out the related work. Methods There were 1 500 elderly people aged at and above 60 years old that were surveyed via questionnaires and structured interviews. Results The core indicators:79.8%reported that their neighborhood was convenient for walking;90.3% reported that bus, under-ground and taxi in Shanghai were suitable for all types of people ( including the physically handicapped, the blind, and the deaf);94.8%reported that there was no difficulty in the payment of housing;96.0%reported that the community had good atmosphere of respecting for the elderly; 56 .8% participated in community volunteer activities;6.7%participated in paid work;53.5%participated in social and cultural activities;99 .7% reported that they could obtain information on health and social services from the community;71.2%reported that they could obtain services for free or below market price when necessary at home.Some core indicators had obvious differences among different genders, age groups, culture, marital status, household types, income sources and income levels.Supplementary and optional indicators:98.6%thought there should be at least one item of house facilities for home-based care for the aged, while 75.6%thought there should be at least four items.26.3% reported to have participated in community team activities.55.2%reported to have participated at least one education or training project during the past one year.90.5% self-reported of good health conditions. Conclusion There is high coverage of barrier-free facility construction, good atmosphere of respecting for the elderly in the community, wide coverage of home-based care for the aged.The elderly people, to some extent, are involved in community activities and various types of training.However, there are still improvements of participation of the elderly in decision-making of local policies, and of senile transformation of the houses the elderly.At the same time, economic, social, demographical, cultural and other factors should be paid close attention to.

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.

OBJECTIVEUsing molecular epidemiology method to characterize human immunodeficiency virus type 1 (HIV-1) subtype CRF01 _ AE strains being prevailed in Zhejiang province.

METHODSGag fragments of the HIV-1 strains were amplified by nested-polymerase chain reaction (nPCR) from the DNA extracted from whole blood of HIV-1 infected individuals in Zhejiang province. PCR products were sequenced and analyzed by phylogenetic method.

RESULTS81 HIV-1 subtype CRF01 _ AE sequences were identified from the 192 samples that sequenced successfully. As one of the dominant subtypes in Zhejiang, CRF01 _ AE was transmitted mainly by heterosexual or homosexual contact in local residents. In migrants living in Zhejiang, CRF01 _ AE were transmitted mainly by heterosexual contact or injecting drug use. There were three main clusters in the phylogenetic tree which bootstrap value was larger than 60. We named the clusters with group MIX (47 sequences), group SEX (7 sequences) and group MSM (12 sequences) based on the transmission. Pairwise DNA distances in the gag region within the three groups and between CM240 were different (P = 0.000). Data through the analyses of deduced amino acid sequences from the three groups showed that several signature amino acid sites were distinct from the same positions of the subtype reference strains.

CONCLUSIONThe CRF01 _ AE strain prevailing in Zhejiang province was from several sources, transmitted by more than three different transmission routes, and becoming the main subtypes circulating in homosexual population in this study. More attention needs to be paid to the epidemic characteristic of CRF01 _ AE.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; methods ; Phylogeny ; Young Adult

OBJECTIVETo evaluate the potential usefulness of a multivariable model in predicting the response to immunosuppressive therapy (IST) in patients with aplastic anemia (AA), and its application to the clinical practice.

METHODSPB T cells subpopulation and BM T cells intracellular IFN-gamma and IL-4 were serially analyzed by flow cytometry (FCM) before and during treatment. HLA-DRB1 * 1501 phenotype was analyzed by PCR-SSP. The predictive potentials of different parameter combinations for clinical responsiveness were statistically assessed.

RESULTSIn all evaluated parameters, CD8+ cell intracellular IFN-gamma had the relatively best diagnostic value with sensitivity and specificity of 94.3% and 62.5%, and positive and negative predictive value of 84.6% and 83.3% respectively. Positive CD8+ cell intracellular IFN-gamma plus Tc1/Tc2 < 50 could increase the positive predictive value to 92.3%. A multivariable model consisting of absolute neutrophil count (ANC), BM T cell intracellular IFN-gamma, Tc1/Tc2 ratio and HLA-DRB * 1501 phenotype of the patients was finally established.

CONCLUSIONThe multivariable model is superior to each of the single parameters in terms of predictive power of IST therapeutic outcome, and its higher accuracy and the clinical application make it potentially useful in practice.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; drug therapy ; immunology ; Child ; Feasibility Studies ; Female ; HLA-DR Antigens ; immunology ; Humans ; Immunosuppression ; Immunosuppressive Agents ; therapeutic use ; Male ; Middle Aged ; Models, Statistical ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes ; immunology ; Treatment Outcome

OBJECTIVETo compare the immunophenotypic and clinical characteristics between NPM1 mutated acute myeloid leukemia (AML) (NPM1m(+)AML) and unmutated AML(NPM1m(-)AML) not otherwise characterized (NOS) under similar FAB subtypes constituent ratio.

METHODSImmunophenotyping and NPM1 gene mutation type-A, B and D and other leukemic related fusion genes were detected by multiparameter flow cytometry and real time RT-PCR or PCR, respectively. 104 AML patients with NPM1m(+)AML and performed immunophenotyping assay were included, 97 with NPM1m(-)AML.

RESULTSThere were significant difference between the two groups at presentation in terms of sex, white blood count(WBC), platelet counts (PLT), blast ratio, normal karyotype ratio, WT1 expression level, FLT3-ITD mutation positive rate and remission rate of first course of induction therapy (P < 0.05). On the immunophenotype, the expression of early differentiation antigens (CD34, HLA-DR, CD117, CD38), lymphocytic antigens (CD7, CD4, CD19, CD2), myeloid and monocytic differentiation-associated antigens (CD13, CD14, CD15) were lower, and that of CD33 as well as CD123 were higher in NPM1m(+)AML patients. Among them, only CD34, HLA-DR, CD7, and CD4 positive cases were significantly lower in NPM1m(+)AML group than in NPM1m(-)AML group (P < 0.05), the rest of them had significant difference in the number of positive cells (P < 0.05). Above features were further analyzed between the M1/M2 and M4/M5 subgroups. M1/M2 cases retained the women prominent and had a higher WT1 expression level (P < 0.05). The expression of monocytic differentiation-associated antigens including HLA-DR and lymphocytic antigens were higher and that of CD117 were lower in M4/M5 subtype (P < 0.05). Among them, the positive rates of HLA-DR, CD64, CD11b, CD10, CD15, and CD4 were significantly higher in M4/M5 than in M1/M2 in NPM1m(+)AML group (P < 0.05).

CONCLUSIONThe most clinical characteristics in NPM1m(+)AML patients are consistent with reports, but some immunophenotype are different to the previous reports under similar FAB subtypes constituent ratio. The major immunophenotypic features of NPM1m(+)AML patients are lower expression of progenitor, myeloid and lymphoid lineage antigens. Monocytic differentiation-associated antigens are only higher expression in M4/M5 cases when comparison with M1/M2 cases within NPM1m(+)AML group.

Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Child ; Child, Preschool ; Female ; HLA-DR Antigens ; immunology ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; immunology ; Male ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; Young Adult

OBJECTIVETo study the interaction between Toll-like receptor (TLR) 4 and myeloid differentiation protein-2 (MD-2) in living cells using fluorescence resonance energy transfer (FRET) technology.

METHODSThe coding sequences of TLR4 and MD-2 (without the signal peptide sequence) were amplified by PCR and cloned into enhanced cyan fluorescence protein (CFP) and enhanced yellow fluorescence protein (YFP) expression vectors carrying TLR4 signal peptides (pECFP-C1-SP and pEYFP-C1-SP). HEK293 cells were transfected respectively or together with the reconstructed plasmids verified by enzyme digestion and sequence analysis, and the expression and sublocalization of these fluorescence proteins in the cells were observed using fluorescence microscope. FRET in the cells coexpressing CFP-TLR4 and YFP-MD-2 was detected using routine and acceptor photobleaching method.

RESULTSThe reconstructed plasmids were expressed in HEK293 cells. The cyan or yellow fluorescence was located in the cytoplasm, mainly around the nucleus in the cells transfected with pECFP/TLR4 or pEYFP/MD-2, and both the cyan and yellow fluorescence located mainly in the membrane and occasional in the cytoplasm of cells cotransfected with pECFP/TLR4 and pEYFP/MD-2. Routine or acceptor photobleaching detected FRET phenomena in cells coexpressing CFP-TLR4 and YFP-MD-2, suggesting direct interaction between TLR4 and MD-2.

CONCLUSIONThis study provides direct evidence of the interaction between TLR4 and MD-2 in living cells.

Cell Line ; Fluorescence Resonance Energy Transfer ; methods ; Humans ; Luminescent Proteins ; genetics ; metabolism ; Lymphocyte Antigen 96 ; genetics ; metabolism ; Plasmids ; genetics ; Protein Binding ; Recombinant Fusion Proteins ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Transfection

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.

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.

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 (x2=50.624,P=0.000).The Prevalence rates related to resistance of NNRTIs and NRTIs were also significantly different ( x2＝48.771,P=0.000 and x2=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％,x2=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 thanA and B (x2=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 (x2=14.213,P=0.001 ) in the three areas.Two PIs mutations were detected in 257 patients:M461/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.