[The influence of T lymphocyte activation on HIV-1 susceptibility of Han Chinese].

OBJECTIVE: To explore the influence of T lymphocyte activation on HIV-1 susceptibility of Han Chinese. METHODS: In 2008, 37 HIV-1 highly exposed persistently seronegative individuals (ESNs) and 101 healthy controls were screened from Shenzhen. Flow cytometer was used to assay the expression difference of HIV-1 infection related co-receptor, the difference between the two groups were analyzed by Mann-Whitney U statistics methods. RESULTS: T cell HLA-DR(+) CD4 T cells and HLA-DR(+) expression of ESNs (12.64 (5.94 - 21.90), 21.12 (10.74 - 30.21)) were all significantly lower than that of healthy controls (22.52 (7.91 - 58.60), 32.28 (14.72 - 67.82)) (P values all < 0.05). T cell CD45RA-RO(+), CCR5(+)CD4 expression of ESNs (58.68 (49.06 - 72.44), 21.93 (15.84 - 25.89)) were all significantly higher than that of healthy controls (53.17 (42.63 - 63.21), 16.14 (11.94 - 21.98)) (P values all < 0.05). T cell CXCR4(+)CD4 T cells expression of ESNs (93.67 (92.17 - 94.96)) was significantly lower than that of healthy controls (95.16 (92.99 - 96.77)) (P values all < 0.05). Healthy controls and ESNs could be divided into low expression group and high expression group according to HLA-DR(+)CD8 T cells bimodal distribution. A total of 89.2% (33/37) ESNs fell into HLA-DR + CD8 low expression group, and 58.4% (59/101) of the healthy controls located in low expression group (P < 0.05). CONCLUSION: To Han Chinese, the low activation status of T lymphocyte has significant correlation with HIV-1 low susceptibility.

Three fatal cases of pandemic 2009 influenza A virus infection in Shenzhen are associated with cytokine storm.

China had taken strict measures for pandemic 2009 H1N1 infection with enhanced surveillance and hospital isolation since April 2009. In Shenzhen, over 1200 confirmed cases of H1N1 infection were identified. Three young patients died of severe pneumonia. Among them, two boys developed neurological complications. Cytokine storm seemed an important cause.

[Molecular epidemiology on vpr gene of HIV-1 strains].

OBJECTIVE: To understand the rule of vpr gene variance of HIV-1 strains. METHODS: RT-PCR was used to amplify vpr gene of HIV-1 strains in Shenzhen. PCR products were sequenced and used for gene phylogenetic analysis and the 32 - 46 amino acids of Vpr protein were compared. The difference of 77 amino acid polymorphism distribution between domestic region and foreign region was analyzed. RESULTS: 01_AE was the major HIV-1 subtype in Shenzhen. The gene distance among subtype B was larger than in other subtypes. 77-amino acid of Vpr protein had three polymorphism forms as Arginin, Glutamine and Histidine, with Glutamine as the wild form. There were no significant differences in the three amino acid distributions between HIV-1 strains from domestic region and foreign region. CONCLUSION: vpr genes of different HIV-1 strains belonged to 01_AE subtype. There was polymorphism seen in the vpr gene which was consistent with both domestic and international HIV-1 strains.

[Molecular epidemiological survey on HIV-1 among the injecting drug users population in Shenzhen].

OBJECTIVE: To study the prevalent status of human immunodeficiency virus-1 (HIV-1) subtypes in IDU (injecting drug users) population in Shenzhen and to study their source of infection in order to predict the epidemic trend and evolution. METHODS: 166 HIV-1 positive plasma from the IDUs was collected from 1996 to 2008. HIV-1 env genes were amplified by nested-PCR from RNA. The C2-V3 regions (450 bp) of HIV-1 env were sequenced for analyses. Phylogenetic analyses were performed on the nucleotide sequence data. RESULTS: Among 166 samples, there were 6 HIV-1 strains including CRF01_AE, CRF08_BC, CRF07_BC 3 circulating recombinant forms (CRFs) and B', C, A1 3 subtypes. Data from the genotype analyses showed that 65.06% (108/166) were CRF01_AE, 19.88% (33/166) were CRF07_BC, 6.02% (10/166) were CRF08_BC, 7.23% (9/166) were subtype B', 0.60% (1/166) were subtype C and 1.20% (2/166) were subtype A1. Phylogenetic tree analysis showed that some of HIV-1 clusters defined in CRF01_AE, CRF07_BC and subtype B' in different time groups. Significant increase of gene distance in CRF01_AE and CRF07_BC strains in the three different periods. CONCLUSION: CRF01_AE and CRF07_BC were the major epidemic CRF strains among the IDU population in Shenzhen while the subtype B', C, A1 and CRF08_BC were also circulating in IDU population in this region. The variation of all different subtypes was increasing through these years.

[Analysis of the tandem-repeat polymorphisms in DC-SIGNR alleles among drug users population with or without HIV/HCV infection].

OBJECTIVE: To study the distribution of DC-SIGN/DC-SIGNR alleles among drug user (DUs) populations with or without HIV/HCV infection in Shenzhen, and to evaluate the role of these alleles in the construction of genetic resistance to HIV or HCV and screen out the anti-HIV/HCV gene in Shenzhen. METHODS: All 500 DU blood samples were collected from Shenzhen Detoxification Center, including 313 from injected drug users (IDUs). All samples were screened for HIV and HCV antibody by means of ELISA. The genomic DNA were extracted and amplified by PCR. The neck domain repeat regions of DC-SIGN/DC-SIGNR were sequenced directly from the PCR products to confirm the amplification for some samples and all positive PCR products were analyzed by agarose gel electrophoresis. RESULTS: Of 500 samples, 97 were found HIV positive, all of which were IDUs and HCV positive. The total positive rate of HCV among all HIV negative DU was 57.57% (232/403), and it was 63.89% (138/216) among IDUs; in comparing to the 50.26% (94/187) of DUs with other manners there showed significant difference (chi(2) = 7.61, P = 0.0058). Among HIV + DUs, there was a higher proportion of patient with the DC-SIGNR 5/6 and 5/8 (Fisher's exact, P = 0.043 and P = 0.034) with statistical significance; there was no statistically significant difference between HCV + and HCV-DUs and no significant difference between IDUs and other DUs for the DC-SIGNR polymorphism. CONCLUSION: The results might indicate that DC-SIGN/DC-SIGNR polymorphism might not influence the susceptibility to HCV. Genotype 5/6 might probably have a relation with HIV infection, but still need further investigation for the low frequency.

[Human immunodeficiency virus type 1 strains epidemic in Shenzhen].

OBJECTIVE: To study the epidemic status of human immunodeficiency virus type 1 (HIV-1) subtypes in Shenzhen and to study their transmission source and routes. METHODS: HIV-1 env and gag genes were amplified by nested PCR from uncultured peripheral blood mononuclear cells (PBMCs) obtained from 122 HIV-1 carriers confirmed in Shenzhen. The C2-V3 region (about 450 bp) of HIV-1 env and P17/ P24 region were sequenced. RESULTS: Among 122 samples, 6 HIV-1 strains including 3 circulating recombinant forms (CRFs) of CRF01_AE, CRF08_BC, CRF07_BC and 3 subtypes of B', B, C were found in Shenzhen, and the percentages were 45.1% (55/122) for CRF01_AE, 31.1% (38/122) for CRF08_BC, 6.6% (8/122) for CRF07_BC, 14.8% (18/122) for B' subtype, 1.6% (2/122) for B subtype, and 0.8% (1/122) for C subtype. The intragroup genetic distances were (4.455 +/- 1.478)%, (2.997 +/- 1.345)%, (4.380 +/- 2.024)%, (5.186 +/- 2.487)%, and (4.869 +/- 2.638)%, respectively. In comparison with the sequence of respective international strains 01AE. TH. 90. CM240, 97CNGX-9F, CN. 97. C54A, B. US. 83. JRFL, and RLA2, the genetic distances were (5. 228 +/- 0.823)%, (3.634 +/- 1.073)%, (4.233 +/- 1.119)%, (4.950 +/- 2.564)%, and (5.795 +/- 2.198)%, respectively. The major subtypes found in injection drug users (IDUs) were CRF07_BC, CRF08_BC, and CRF01_AE strains. CRF01_AE and B' strains were epidemic mainly in sexual workers. CONCLUSION: There are 3 HIV-1 subtypes (B', B, C) and 3 CRFs (CRF01_AE, CRF08_BC, CRF07_BC) epidemics in Shenzhen. The predominant subtypes varies among different transmission routes. While CRF01_AE is predominant among sexual workers, CRF08_BC and CRF01_AE are major subtypes among IDU population.

[Effect of CCR5delta32, CCR5m303, CCR2-64I and SDF1-3'A gene polymorphism on the prognosis of Chinese HIV-1 carriers].

OBJECTIVE: To study how CCR5delta32, CCR5m303, CCR2-64I, SDF1-3'A gene polymorphisms affect the prognosis of Chinese HIV-1 carrier. METHODS: Epidemiologic survey was done to the HIV-1 carriers who were found in Shenzhen area. PCR/RFLP technology was applied to analyze CCR5delta32, CCR5m303, CCR2-64I, SDF1-3'A gene polymorphisms of the HIV-1 carriers. The plasma virus load and CD4+ cell counting was assayed. The incubation period of some carriers was estimated. SPSS11.0 software was used to analyze the data. RESULTS: No persons with CCR5delta32 and CCR5m303 mutation genotype were found from 189 HIV-1 carriers. SDF1-3'A allele frequency was 26.14% and CCR2-64I allele 19.82%. The carriers were divided into high virus load group (virus load < 20,000 copies/ml) and low virus load group (virus load > or =20,000 copies/ml). It was found by one-way ANOVA analysis on the logarithm of virus load that there was no significant difference between CCR2-64I wild genotype and cross bred genotype (P=0.272). One-way ANOVA analysis on delitescence of some carriers showed that there was not significant difference between CCR2-64I wild genotype and cross bred genotype (P=0.662). One-way ANOVA analysis on the logarithm of virus load showed that there was significant difference among SDF1-3'A wild genotype, cross bred genotype and pure mutation genotype (P=0.001). CONCLUSION: CCR2-64I gene mutation may not significantly affect virus load of Chinese HIV-1 carriers, nor it affect the incubation period of HIV-1 carriers. SDF1-3'A gene mutation can decrease virus load, but it may not prolong the incubation period of HIV-1 carriers.

[Effect of urotensin II on secretion of adrenomedullin from human vascular endothelial cells].

OBJECTIVE: To study the effect of human urotensin II (HU II) on secretion of adrenomedullin (ADM) from human vascular endothelial cells (HVEC) and its mechanism. METHODS: In cultured HVEC, different concentrations of HUII were used to stimulate the ADM secretion from HVEC, and the inhibitors of different signal transduction pathway were used to investigate their effects on ADM secretion. The contents of ADM in medium were determined by radio immunoassay. RESULTS: HUII stimulated secretion of ADM from HVEC in a time-dependent and concentration-dependent manner. The contents of ADM in the experiment groups were changed compared with that in control group (P < 0.05). The increase of ADM could be inhibited by inhibitor of extracellular signal-regulated protein kinase (PD(98059)), inhibitor of P38 kinase (SB(202190)), inhibitor of calmodulin (W(7)) and inhibitor of Ca(2+) (nicardipine) (P < 0.05). The inhibition ratio in those groups was 68%, 78%, 24% and 25% respectively. But the inhibitor of Calcineurin (CaN) and inhibitor of protein kinase C (H(7)) had no influence on the secretion of ADM from HVEC (P > 0.05). CONCLUSION: The stimulated effect of HUII on the ADM secretion from HVEC may be mediated by Ca(2+), ERKs, CaM-PK and P38 signal transduction pathways.

[Observation of short- and mid-term clinical outcome of percutaneous coronary intervention for acute myocardial infarction with heart failure or cardiogenic shock].

OBJECTIVE: To observe the short- and mid-term effects of percutaneous coronary intervention (PCI) in patients with ST-segment elevation acute myocardial infarction (AMI) complicated by heart failure and/or cardiogenic shock . METHODS: Altogether 90 patients with AMI were recruited, of whom 58 were treated by PCI, 20 by thrombolytic therapy, and the other received general treatment without reperfusion therapy. The length of hospital stay, major adverse cardiac events (MACE) and left ventricular ejection fraction (LVEF) were compared between PCI and thrombolysis groups. The relationship between the patency time of the infarct-related artery (IRA), thrombolysis in myocardial infarction (TIMI) grade after PCI and prognosis were analyzed in PCI group. RESULTS: The patency rate of IRA was significantly improved in patients receiving PCI therapy in comparison by those with thrombolytic therapy (98.3% vs 65.0%, P<0.01), and the LVEF was also higher in PCI group with lower mortality (6.9% vs 25.0%, P<0.05) during in-hospital and follow-up period. CONCLUSION: PCI can be a more effective therapy than thrombolytic therapy in the treatment of ST-segment elevation AMI accompanied with heart failure and/or cardiogenic shock.

[Mechanism of urotensin II-stimulated adrenomedullin secretion in human vascular endothelial cells].

OBJECTIVE: To study the mechanism of urotensin II(U II)-stimulated adrenomedullin secretion in human vascular endothelial cells. METHODS: In cultured human vascular endothelial cells (HEVCs), different concentrations of U II was used to stimulate the secretion of Adm, and different inhibitors were used to study the changes in the secretion after block of different signal transduction pathways. The contents of Adm in the medium were detected with radioimmunoassay. RESULTS: U II-stimulated Adm secretion in the HEVCs in a time- and concentration-dependent manner. Adm contents of the treatment groups were comparable with that of the control group (P<0.05 ), and the secretion of Adm could be inhibited by the inhibitor of extracellular signal-regulated protein kinases (PD098059), p38 kinase inhibitor (SB202190), calmodulin inhibitor (W7) and Ca(2+) inhibitor (nicardipine)(P<0.05), but calcineurin inhibitor and protein kinase C inhibitor (H7) had no such effect (P>0.05). CONCLUSION: Ca(2+), MAPK, CaM-PK and p38 signal transduction pathways may play major roles in U II-stimulated secretion of Adm in HVECs.