2006年 · 第119卷第23期

One of the major advances and few real breakthroughs in HIV/AIDS research is the discovery of anti-retroviral drugs and highly active anti-retroviral therapy (HAART)。

Abstract：Background This study transferred a recombinant gene encoding human insulin like growth factor-1 (hIGF-1)into modified primary skeletal myoblasts with a retroviral vector (pLgXSN) and determined whether the hIGF-1 promoted growth of skeletal muscle in rat.Methods hIGF-lcDNA was amplified in vitro from normal human liver cells by using RT-PCR and cloned into plasmid vector pLgXSN. The recombinant vector pLghIGF-1SN and control vector pLgGFPSN were transfected into packaging cell PT67 and G418 was used to select positive colony. Myoblasts were infected with a high titre viral supernatant and transduction efficiency was evaluated as GFP expression. The expression of hIGF-1 mRNA in myoblasts was investigated by immunocytochemistry and RT-PCR. MTT assays detected the growth of myoblasts in vitro. Myoblasts transduced with pLghIGF-1SN were injected into hind limb muscles of 10-12 week male SD rats. Formed tissues were harvested 4 weeks later. Myocyte diameter, mean weight of hind limb and body were measured to evaluate the skeletal muscle growth.Results Recombinant retroviral plasmid vector pLghIGF-1SN was constructed successfully. The titre of the packaged recombinant retrovirus was 1 × 106 cfu/ml. The transfection rate of PT67 cells reached 100% after G418 screening. hIGF-1 expression was positive in myoblast-IGF-1. The proliferation rate of myoblast-IGF-1 in vitro was higher than GFP-myoblast or myoblast (P＜ 0.05). The mean weights of hind limb and body of rats injected myoblast-IGF-1 were higher than those of the rats injected with myoblast-GFP or myoblast (P＜ 0.05). Myocyte diameter had a significant increase in IGF-1 group compared to GFP group and myoblast group (P＜ 0.05).Conclusions The transfection of the human IGF- 1 gene mediated by a retroviral vector can promote the growth of skeletal muscle in rats. Genetically modified primary skeletal myoblasts provide a possibly effective approach to treat some skeletal muscle diseases。

Abstract：Background Asthma is a chronic airway disease with inflammation characterized by physiological changes(airway hyper-responsiveness, AHR) and pathological changes (inflammatory cells infiltration and mucus production). Eosinophils play a key role in the allergic inflammation. But the causative relationship between eosinophils and airway inflammation is hard to prove. One of the reasons is lack of activation marker of murine eosinophils. We investigated the expression of CD69 on murine eosinophils in vitro, the relationship between the expression of CD69 on eosinophils from peripheral blood and bronchoalveolar lavage fluid and on airway inflammation in asthmatic mice.Methods Eosinophils from peripheral blood of IL-5 transgenic mice (NJ.1638) were purified. Mice were divided into five groups: wild type mice sensitized and challenged with saline (WS group), wild type mice sensitized and challenged with ovalbumin (WO group), IL-5-/- mice sensitized and challenged with saline and transferred with purified eosinophils (ISE group), IL-5-/- mice sensitized and challenged with OVA and transferred with purified eosinophils (IOE group), IL-5-/- mice sensitized and challenged with OVA and transferred with purified eosinophils, pretreated with anti CD4 monoclonal antibody (IOE+antiCD4mAb group).IL-5-/- mice were sensitized with OVA at day 0 and day 14, then challenged with OVA aerosol. On days 24, 25, 26and 27 purified eosinophils were transferred intratracheally to IL-5-/- mice. On day 28, blood and BALF were collected and CD69 expression on eosinophils measured by flowcytometry.Results Purified eosinophils did not express CD69. But eosinophils cultured with PMA+MA, IFN- γ, IL-5 or GM-CSF expressed CD69 strongly. Eosinophils from blood of WO, WS group did not express CD69 at all. The numbers of eosinophils in BALF of WO group, IOE group, ISE group and IOE+antiCD4mAb group were significantly higher than in mice of WS group which did not have eosinophils at all. CD69 expression on eosinophils in BALF of IOE and WO groups was strong. Eosinophils in BALF of ISE and IOE+antiCDmAb groups did not expre ss CD69. The mucus production result was similar to CD69 expression. There were eosinophils infiltration in lung slides of all groups except WS group.Conclusion Activation in airway of eosinophils could directly lead to airway inflammation。

Abstract：Background Improvement of clinical symptoms following hyperbaric oxygen (HBO) treatment of neuropsychiatric disorders arising from traumatic brain injury was proved by our previous study. This study was aim to obtain the evidence of other changes.Methods Three hundred and ten patients with neuropsychiatric disorders arising from traumatic brain injury were treated twice with hyperbaric oxygen. Cerebral single photon emissions computed tomography (SPECT)images and computed tomography scans (CT) before and after hyperbaric oxygen treatment, were compared.Results Before treatment, the proportion of abnormal cerebral changes detected by SPECT was 81.3% but only 15.2% by CT. After HBO treatment, 70.3% of SPECT scans showed no abnormalities and these patients were clinically improved. Treatment improved regional cerebral blood flow.Conclusion SPECT was much more sensitive than CT in the diagnosis of neuropsychiatric disorders following hyperbaric oxygen treatment of neuropsychiatric disorders arising from traumatic brain injury。

Abstract：Background This study was aim to explore the characteristics of phenotypic resistance of resistant strains of HIV type-1 (HIV-1) subtype B and to compare the concordance between the phenotypic resistance and genotypic resistance. Methods The genotypic resistance assay for the HIV-1 clinical isolates was performed. One isolate without resistance mutation was chosen as a drug-sensitive reference strain and seven subtype B isolates with resistance mutations were phenotypically tested. Fifty percent inhibitory concentrations (IC50) between resistant and sensitive viruses were compared. The resistance extent was determined by the folds of the increased IC50. The concordance between the phenotypic resistance and genotypic resistance was also analyzed.Results IC50 of resistant isolates were 0.0006-0.1300 μmol/L for zidovudine (AZT), 0.0016-0.0390 μmol/L for lamivudine (3TC), 0.0104-0.4234 μmol/L for nevirapine (NVP), and 0.0163-0.1142 μmol/L for indinavir (IDV), respectively. Genotypic and phenotypic resistance assays indicated that the resistant strains were intermediately and highly resistant to nucleotide analog reverse transcriptase inhibitors and non-nucleotide analog reverse transcriptase inhibitors. The phenotypic assay was consistent with the genotypic assay. For measuring the potential resistance, the genotypic assay was more sensitive than the phenotypic. In evaluating the resistance to protease inhibitors, these two assays were discrepant.Conclusions Both the phenotypic and genotypic assays indicate that the resistant viruses exist in HIV-infected patients in China who have received treatment. Phenotypic and genotypic assays have high concordance, and the genotypic assay could replace the phenotypic assay to predict the HIV-1 resistance。

Abstract：Background At the end of 2005, 650 000 people lived with human immunodeficiency virus type-1 (HIV-1) in (HAART) supported by the "China CARES" program but the immune responses of HAART were seldom reported. This study investigated the effect of HAART on the activation and coreceptor expression of T lymphocytes in Chinese HIV/AIDS patients and evaluated its effect on immune reconstitution.Methods Seventeen HIV/AIDS patients were enrolled and three-color-flow cytometry was used to detect the activation of HLA-DR CD38 and the coreceptor CCR5, CXCR4 expression on T lymphocytes in whole blood samples taken from the patients before and after 3- or 6-month HAART.Results The activation percents of CD4+, CD8+ T lymphocytes were significantly higher before therapy than the normal controls (HLA-DR/CD4: 40.47± 18.85 vs 11.54±4.10; CD38/CD4: 81.34± 10.86 vs 53.34± 11.44;HLA-DR/CD8:63.94±12.71 vs 25.67±9.18; CD38/CD8:86.56±11.41 vs 58.84±6.16, all P＜0.01). After 6-month combined antiretroviral treatment, the activation of T lymphocytes in HIV/AIDS patients was significantly decreased (HLA-DR/CD4:28.31 ± 13.48; CD38/CD4:69.88 ± 12.64; HLA-DR/CD8: 46.56±18.64;CD38/CD8: 70.17± 14.54, all P＜0.01 compared with the pre-treatment values). Before the treatment, CCR5 expression on CD8+ T lymphocytes was up-regulated while CXCR4 expression on CD8+ T lymphocytes downregulated in HIV/AIDS patients compared with the normal controls (CD8/CCR5:70.9 1± 10.03 vs 52.70 ±7.68; CD8/CXCR4: 24.14± 11.08 vs 50.05± 11.68, all P＜0.01). After 6-month HAART, CCR5 expression on CD8+ T lymphocytes significantly decreased (56.35±12.96, P＜0.01), while CXCR4 expression on CD8+ T lymphocytes increased (36.95±9.96, P＜0.05) compared with the pre-treatment and the normal controls. A significant statistical relationship was observed between the expression of activation markers, CCR5 and the CD4+ T lymphocyte counts after HAART (P＜0.05).Conclusions Reduced activation of T lymphocytes and a normalization of coreceptor expression were observed in Chinese HIV/AIDS patients after HAART. Immunity can be restored in HIV/AIDS patients receiving HAART。

Abstract：Background Man who has sex with man (MSM) is one of the high risk groups for spreading HIV/AIDS. It was reported that the most prevalent human immunodeficiency virus type 1 (HIV-1) strain among MSM is subtype B; however, T cell immunity remains unknown across the HIV-1 B genome in this population.Methods Using Elispot assay with synthetic peptides spanning the sequence of HIV-1 consensus B,HIV-1-specific cytotoxic T-cell lymphocyte responses were quantified among 3 treated and 19 untreated HIV-1 infected MSM from Beijing, China. Cross-sectional association between viral loads and cellular immune responses were analyzed.Results Peptide pools corresponding to each HIV-1 protein were used for Env, Gag, Pol, Nef, Tat/Rev, Vpr/Vpu and Vif. The results showed that the magnitude of T cell responses in the 3 treated HIV+ MSM group [median,770 spot forming cells (SFCs) per 106 peripheral blood mononuclear cells (PBMCs)] might be significantly lower than that in the 19 untreated HIV+ MSM group (median, 6175 SFCs per 106 PBMCs). Nef, Gag and Pol are the most frequently targeted HIV-1 antigens; and 16 subjects (73%) were identified with vigorous T cell immunity against each of these three proteins. The overall magnitude of T cell immunity closely related to its breadth (r=-0.72, P＜0.05) and was inversely but weakly associated with viral loads (r=-0.15). Further analysis showed that both Gag (r=-0.24) and Pol specific T cells (r=-0.12) contributed to this inverse association whereas Nef specific T cells showed no association with viral loads.Conclusions The magnitude of HIV-1 specific T cells is inversely but weakly associated with viral loads among MSM; HIV-specific T cell responses against conservative sequences (Gag and Poi) are the main contributors to this association among Chinese HIV+ MSM. These findings have important implications for vaccine design。

Abstract：Background Few studies have examined the properties of human immunodeficiency virus type 1 (HIV-1) epitope-specific cytotoxic T lymphocyte (CTL) responses in children. To address this issue, we characterized epitope-specific CTL responses and analyzed the determinants that may affect CTL responses before and after highly active antiretroviral therapy (HAART) in children with HIV-1 infection.Methods A total of 22 HIV-1-infected children and 23 uninfected healthy children as control were enrolled in the study. Circulating CD4 T cells and HIV-1 RNA load in plasma were routinely measured. Peripheral HIV-1-specific CTL frequency and HIV-1 epitope-specific, interferon-γ (IFN-γ)-producing T lymphocytes were measured using tetramer staining and enzyme-linked immunospot (ELISPOT) assay, respectively.Circulating dendritic cell (DC) subsets were monitored with FACS analysis.Results More than 80% of the children with HIV-1 infection exhibited a positive HIV-1-epitope-specific CTL response at baseline, but HIV-specific CTLs and IFN-γ-producing lymphocytes decreased in patients who responded to HAART in comparison with non-responders and HAART-naive children. The duration of virus suppression resulted from HAART was inversely correlated with CTL frequency. While in HAART-naive children, HIV-1-specific CTL frequency was positively correlated with myeloid DC (mDC) frequency,although the cause and effect relationship between the DCs and CTLs remains unknown.Conclusions HIV-1-epitope-specific CTL responses are dependent on antigenic stimulation. The impaired DC subsets in blood might result in a defect in DC-mediated T cell responses. These findings may provide insight into understanding the factors and related mechanisms that influence the outcome of HIV-1 carriers to HAART or future antiviral therapies。

Abstract：Background Unregulated commercial blood/plasma collection among farmers occurred between 1992 and 1995 in central China and caused the second major epidemic of human immunodeficiency virus type 1 (HIV-1)infection in China. It is important to characterize HIV-1-infected former blood donors and to study characteristics associated with disease progression for future clinical intervention and vaccine development.Methods A cross-sectional study was performed on HIV-1-infected former blood donors (FBDs) and age-matched HIV-seronegative local residents. Demographic, epidemiologic, clinical and key laboratory data were collected from all study participants. Both unadjusted and adjusted multivariate linear regressions were employed to analyze the association of the decrease of CD4+ T-cell counts with other characteristics.Results Two hundred and ninety-four HIV-1-infected FBDs and 59 age-matched HIV-seronegative local residents were enrolled in this study. The unregulated blood/plasma collection occurred more than a decade (10.8- 12.8 years) ago, which caused the rapid spread of HIV-1 infection and the high prevalence of co-infection with hepatitis C virus (HCV, 89.5%); hepatitis B virus (HBV) co-infection was observed in only 11 HIV+participants (3.7%). Deterioration in both clinical manifestation and laboratory parameters and increase of viral loads were observed in parallel with the decrease of CD4+ T-cell counts. The decrease of total lymphocyte counts (P＜0.001)and hemoglobin levels (P＜0.001) and the appearance of dermatosis (P=0.03) were observed in parallel with the decrease of CD4+ T-cell counts whereas viral loads (P＜0.001) and CD8+ T-cell counts (P=0.01) were inversely associated with CD4+ T-cell counts.Conclusions Co-infection with HCV but not HBV is highly prevalent among HIV-1-infected FBDs. CD4+ T-cell counts is a reliable indicator for disease progression among FBDs. Total lymphocyte counts, hemoglobin level and appearance of dermatosis were positively, whereas CD8+ T-cell counts and viral loads were inversely associated with the decreased CD4+ T-cell counts。

Pediatric cardiac surgery in China has made great progress since the first successful open heart operation for correction of ventricular septal defect (VSD) with an artificial heart-lung machine in 1958.1 In recent years, the state has paid much attention to both the basic research and the primary,secondary and tertiary prevention of congenital heart disease.…… Biography：Correspondence to:Dr. LIU Ying-long, Pediatric Cardiac Center, Department of Surgery, Cardiovascular Institute and Fuwai Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100037, China ( Tel: 86-10-88398188.Fax: 86-10-68332747. Email: pumcwu@yahoo.com.cn)

Since the initiation of China's nationwide free antiretroviral therapy (ART) in 2002, the availability of highly active antiretroviral therapy (HAART) in China has increased dramatically。

Cytochrome P4502D6 (CYP2D6) is an important cytochrome P450 enzyme which catalyzes the metabolism of more than 80 drugs.1 Many of these drugs have a narrow therapeutic index and the combined use of substrates may result in significant pharmacokinetic interactions, leading to substrate accumulation and deleterious drug effects。

Posttraumatic foreign bodies in the heart or great vessels is rare, which may cause cardiac tamponade, bleeding, shock, infection, embolism,arrhythmia, valve dysfunction, etc.1-3 The foreign bodies can be removed by surgery or percutaneous intervention.4 In this report we reviewed our experience in managing posttraumatic foreign bodies in 13 patients at our institution from 1992 to 2002。

To the Editor: The recent case report of primary cardiac hemangioendothelioma1 prompted me to write this letter-to-the-editor for 5 reasons。

The Fourth Asian Pacific Congress on Thrombosis and Haemostasis (APCTH) was held from September 21 to 23, 2006 in Suzhou, China and organized by the Asian Pacific Society on Thrombosis and Haemostasis and the Chinese Society of Haematology with professor RUAN Chang-geng as the chairperson.