Role of C6ORF120, an N-glycosylated protein, is implicated in apoptosis of CD4⁺ T lymphocytes.

BACKGROUND: Although CD4(+) T cell apoptosis and CD8(+) T cell responses have been extensively studied during HIV infection, how apoptosis signals being initiated in CD4(+) T cells still need to be elucidated. The present study was designed to characterize the function-unknown gene, C6orf120, and elucidates its primary role in tunicamycin-induced CD4(+) T apoptosis. METHODS: The C6orf120 coding sequence was amplified from peripheral blood mononuclear cells (PBMCs) total RNA of AIDS patients. The DNA fragment was inserted into the pET-32a expression system, transformed into Escherichia coli, and preparation of C6ORF120 recombinant protein. The magnetic cell separation technology was used to prepare primary CD4(+) T cells and CD8(+) T cells. The primary T cells were cultured at 1 × 10(6) cells/ml, treated with 0, 0.1, 1, 10, 100, and 200 ng/ml of C6orf120 recombinant protein for 48 hours, then harvested for cell cycle and apoptosis analysis. Tunicamycin (0.5 µmol/L) was used to induce endoplasmic reticulum stress in Jurkat cells. The biomarker 78 KDa glucose-regulated protein (GRp78) and growth arrest and DNA damage (GADD) were used to evaluate endoplasmic reticulum stress of Jurkat cells. RESULTS: We prepared C6ORF120 recombinant protein and its polyclonal antibody. Immunohistochemical analysis showed that C6orf120 mainly expressed in hepatocytes and cells in germinal center of lymph node. At concentration of 0.1, 1, 10, 100, and 200 ng/ml, C6orf120 recombinant protein could induce apoptosis of Jurkat cells and primary CD4(+) T cells, and promoting G2 phase of its cell cycle. Western blotting analysis showed that C6ORF120 recombinant protein increased the expression of GRp78 and GADD in Jurkat cells in vitro. CONCLUSION: Our results suggested that C6ORF120 could induce apoptosis of CD4(+) T cells, at least in part, mediated with endoplasmic reticulum stress.

[The analysis of the clinical features between survivors and non-survivors with the severe form of new influenza A (H1N1) viral infection].

OBJECTIVE: To understand the clinical features of critically ill patients with pandemic 2009 influenza A (H1N1) and investigate the risk factors associated with death cases. METHODS: The clinical features of 55 critically ill patients with pandemic 2009 influenza A (H1N1) viral infection hospitalized at Beijing Ditan Hospital from October 3 to December 15, 2009 were retrospectively analyzed, and a comparative analysis was performed on the manifestations of the survival and the death groups of patients. RESULTS: There were 31 males and 24 females. The age ranged from 10 months to 84 year old, and the mean (SD) was 38 (20) year old. The critically ill cases were more in patients under age 65 (48/55), with obesity (33/49), with underlying diseases (26/49), and pregnancy (6/24). Both the survivors and non-survivors of patients had high fever, cough, sputum (some sputum with blood), dyspnea, räles of both lungs fields, and all further developed severe pneumonia. The patients also showed respiratory failure (54/55) and ARDS (26/55). All of them received oseltamivir therapy, and 38 patients received mechanical ventilation and 30 were given steroid therapy. Secondary infection occurred in 27 cases, and ventilator-associated pneumonia happened in 10 patients. In the early stage of onset, C-reactive protein (CRP) increased [(131 ± 130) mg/L] and low counts of T lymphocytes were present [CD(4)(+), CD(8)(+) T was (217 ± 139)/µl and (162 ± 82)/µl]. With the progress of disease, the non-survival cases had persistently increased CRP and the counts of T lymphocytes did not recover, while the secondary fungal infection was significantly higher than in the survivor cases (P < 0.05). By using BMI, underlying diseases, ARDS, the day of Oseltamivir initiated, steroid therapy, following bacterial and fungal infection as variables through logistic regression analysis, it was shown that higher BMI and following fungal infection were associated with higher fatal risks (OR was 6.512, 19.631 respectively, both of P value was low than 0.05). There was no death case who received oseltamivir treatment within 48 hours of onset of disease. CONCLUSIONS: Critical illness in pandemic 2009 influenza A (H1N1) was associated with patients under age 65, with obesity, underlying diseases, and pregnancy. Persistently increased CRP and lower counts of T lymphocytes were associated with unfavorable prognosis. The patients with higher BMI and secondary fungal infection had higher fatal risks. Oseltamivir treatments at early stage would probably reduce mortality.

Clinical features of the initial cases of 2009 pandemic influenza A (H1N1) virus infection in China.

BACKGROUND: The first case of 2009 pandemic influenza A (H1N1) virus infection in China was documented on May 10. Subsequently, persons with suspected cases of infection and contacts of those with suspected infection were tested. Persons in whom infection was confirmed were hospitalized and quarantined, and some of them were closely observed for the purpose of investigating the nature and duration of the disease. METHODS: During May and June 2009, we observed 426 persons infected with the 2009 pandemic influenza A (H1N1) virus who were quarantined in 61 hospitals in 20 provinces. Real-time reverse-transcriptase-polymerase-chain-reaction (RT-PCR) testing was used to confirm infection, the clinical features of the disease were closely monitored, and 254 patients were treated with oseltamivir within 48 hours after the onset of disease. RESULTS: The mean age of the 426 patients was 23.4 years, and 53.8% were male. The diagnosis was made at ports of entry (in 32.9% of the patients), during quarantine (20.2%), and in the hospital (46.9%). The median incubation period of the virus was 2 days (range, 1 to 7). The most common symptoms were fever (in 67.4% of the patients) and cough (69.5%). The incidence of diarrhea was 2.8%, and the incidence of nausea and vomiting was 1.9%. Lymphopenia, which was common in both adults (68.1%) and children (92.3%), typically occurred on day 2 (range, 1 to 3) and resolved by day 7 (range, 6 to 9). Hypokalemia was observed in 25.4% of the patients. Duration of fever was typically 3 days (range, 1 to 11). The median length of time during which patients had positive real-time RT-PCR test results was 6 days (range, 1 to 17). Independent risk factors for prolonged real-time RT-PCR positivity included an age of less than 14 years, male sex, and a delay from the onset of symptoms to treatment with oseltamivir of more than 48 hours. CONCLUSIONS: Surveillance of the 2009 H1N1 virus in China shows that the majority of those infected have a mild illness. The typical period during which the virus can be detected with the use of real-time RT-PCR is 6 days (whether or not fever is present). The duration of infection may be shortened if oseltamivir is administered.

[Effects of traditional Chinese medicine on CD4 + T cell counts and HIV viral loads during structured treatment interruption in highly active antiretroviral therapy].

OBJECTIVE: To explore the impacts of traditional Chinese medicine (TCM) on CD4 + T cell counts and human immunodeficiency virus (HIV) viral loads during the course of structured treatment interruption (STI) in highly active antiretroviral therapy (HAART). METHODS: Nineteen HIV/ADIS patients were treated for 14 months as follows: initiated with zidovudine/lamivudine + efavirdine for 6 months, then discontinued the therapy and treated with TCM instead for 2 months. HAART was then reinitiated for another 3 months, and then discontinued and replaced with TCM for another 3 months. The changes of CD4 + T cell counts and HIV viral loads were measured. RESULTS: During the first STI of HAART, 43.8% of patients had no viral rebounds one month later, and 62.6% had stable or increased immune functions; 18.8% had no viral rebounds two months later, and 43.8% had stable or increased immune functions. Changes of viral loads were not significantly different between these two months (P = 0.097), while CD4 + T cell counts significantly decreased two months later compared with one month later (P = 0.043). During the second STI of HAART, 33.3% of patients had no viral rebounds one month later, and 64.3% had stable or increased immune functions; 13.3% had no viral rebounds 3 months later and 46.6% had stable or increased immune functions. Changes of viral loads had significant difference (P = 0. 017), while CD4 + T cell counts at month 12 elevated significantly compared with the baseline (P = 0.014). CONCLUSIONS: TCM can suppress the viral rebounds during STI-HAART, maintain immune functions. However, this effect may decrease along with the prolongation of STI-HAART.

Application of Molecular Adsorbents Recirculating System to remove NO and cytokines in severe liver failure patients with multiple organ dysfunction syndrome.

BACKGROUND: Molecular Adsorbents Recirculating System (MARS) is a new promising artificial liver support therapy, the aim of this study was to assess the effectiveness of MARS to remove nitrous oxide (NO) and cytokines in severe liver failure patients with multiple organ dysfunction syndrome (MODS). METHODS: Sixty single MARS treatments were performed with length of 6-24 h on 24 severe liver failure patients (18 males/6 females) with MODS. RESULTS: The MARS therapy was associated with a significant removal of NO and certain cytokines such as TNF-alpha, IL-6, IL-8, and INF-gamma, together with marked reduction of other non-water-soluble albumin bound toxins and water-soluble toxins, these were associated with a improvement of the patients' clinical conditions including hepatic encephalopathy, deranged hemodynamic situation and as well as renal and respiratory function, thus resulted into marked decrease of Sequential Organ Failure Assessment (SOFA) score and improved outcome: nine patients were able to be discharged from the hospital or bridged to successful liver transplantation, the overall survival of 24 patients was 37.5%. CONCLUSION: We can confirm the positive therapeutic impact and safety to use MARS on liver failure patients with MODS associated with elevated levels of NO and cytokines.

[The treatment of molecular adsorbents recirculating system artificial liver in severe liver failure patients with multiple organ dysfunction syndrome].

OBJECTIVE: To evaluate the effectiveness and mechanisms of molecular adsorbents recirculating system (MARS) treatment in severe liver failure patients with multiple organ dysfunction syndrome (MODS). METHODS: 60 single MARS treatments were performed for 6 - 24 hours on 24 severe liver failure patients with MODS. RESULTS: MARS therapy was associated with marked reduction of albumin bound toxins and water soluble toxins, together with a significant removal of NO and certain cytokines, such as TNF-alpha, IL-6, IL-8, and INF-gamma. These were associated with a improvement of the patients' clinical conditions including hepatic encephalopathy, deranged hemodynamic situation, as well as renal and respiratory function, thus resulted into marked decrease of sequential organ failure assessment (SOFA) score (from 9.72+-1.89 to 6.98+-2.34), and improving outcome: 9 patients were able to be discharged from the hospital or bridged to successful liver transplantation. The overall survival rate of 24 patients was 37.5%. CONCLUSIONS: There is positive therapeutic impact and safety to use MARS on liver failure patients with MODS. The effectiveness of MARS is correlated with reducing the levels of NO and cytokines, except for completely removing of accumulated toxins in liver failure patients.