TOSO interacts with SYK and enhances BCR pathway activation in chronic lymphocytic leukemia.

BACKGROUND: TOSO, also named Fas inhibitory molecule 3 (FAIM3), has recently been identified as an immunoglobulin M (IgM) Fc receptor (FcµR). Previous studies have shown that TOSO is specifically over-expressed in chronic lymphocytic leukemia (CLL). However, the functions of TOSO in CLL remain unknown. The B-cell receptor (BCR) signaling pathway has been reported to be constitutively activated in CLL. Here, we aimed to investigate the functions of TOSO in the BCR signaling pathway and the pathogenesis of CLL. METHODS: We over-expressed TOSO in B-cell lymphoma cell lines (Granta-519 and Z138) by lentiviral transduction and knocked down TOSO by siRNA in primary CLL cells. The over-expression and knockdown of TOSO were confirmed at the RNA level by polymerase chain reaction and protein level by Western blotting. Co-immunoprecipitation with TOSO antibody followed by liquid chromatography coupled with tandem mass spectrometry (IP/LCMS) was used to identify TOSO interacting proteins. Western blotting was performed to detect the activation status of BCR signaling pathways as well as B-cell lymphoma 2 (BCL-2). Flow cytometry was used to examine the apoptosis of TOSO-over-expressing B lymphoma cell lines and TOSO-down-regulated CLL cells via the staining of Annexin V and 7-AAD. One-way analyses of variance were used for intergroup comparisons, while independent samples t tests were used for two-sample comparisons. RESULTS: From IP/LCMS, we identified spleen tyrosine kinase (SYK) as a crucial candidate of TOSO-interacting protein and confirmed it by co-immunoprecipitation. After stimulation with anti-IgM, TOSO over-expression increased the phosphorylation of SYK, and subsequently activated the BCR signaling pathway, which could be reversed by a SYK inhibitor. TOSO knockdown in primary CLL cells resulted in reduced SYK phosphorylation as well as attenuated BCR signaling pathway. The apoptosis rates of the Granta-519 and Z138 cells expressing TOSO were (8.46 ±â€Š2.90)% and (4.20 ±â€Š1.21)%, respectively, significantly lower than the rates of the control groups, which were (25.20 ±â€Š4.60)% and (19.72 ±â€Š1.10)%, respectively (P < 0.05 for both). The apoptosis rate was reduced after knocking down TOSO in the primary CLL cells. In addition, we also found that TOSO down-regulation in primary cells from CLL patients led to decreased expression of BCL-2 as well as lower apoptosis, and vice versa in the cell line. CONCLUSIONS: TOSO might be involved in the pathogenesis of CLL by interacting with SYK, enhancing the BCR signaling pathway, and inducing apoptosis resistance.

Effects of air pollutants on occurrences of influenza-like illness and laboratory-confirmed influenza in Hefei, China.

Accumulating evidence suggests that air pollution is a risk factor for adverse respiratory and cardiovascular health outcomes. However, the different impacts of exposure to air pollutants on influenza virus activity and influenza-like illness (ILI) have not been well documented in epidemiological studies. We examined the association between air pollutants of particular matters < 2.5 µm (PM2.5), particular matters < 10 µm (PM10), nitrogen dioxide (NO2), sulfur dioxide (SO2), and influenza occurrences in Hefei, China, from December 2013 to December 2015 by generalized Poisson additive regression models. The result suggested that PM2.5 and PM10 had similar effects on clinical ILI and influenza incidence. PM10 was negatively associated with clinical ILI (relative risk (RR) 0.980, 95% confidence interval (CI) 0.974-0.987), while PM2.5 were positively associated with clinical ILI (RR 1.040; 95% CI 1.032-1.049). RRs for the laboratory-confirmed cases of influenza were 0.813 (95% CI, 0.755-0.875) for PM10 and 1.216 (95% CI, 1.134-1.304) for PM2.5. Nevertheless, the impacts of SO2 and NO2 on ILI and influenza were distinct. SO2 had significant influence on laboratory-confirmed influenza and had no significant linear relationship with ILI. NO2 was negatively correlated with influenza but had no obvious effect on clinical ILI cases. The present study contributes novel evidence on understanding of the effects of various air pollutants on influenza activities, and these findings can be useful and important for the development of influenza surveillance and early warning systems.

Seasonal pattern of influenza activity in a subtropical city, China, 2010-2015.

Influenza seasonality study is critical for policy-makers to choose an optimal time for influenza vaccination campaign, especially for subtropical regions where influenza seasonality and periodicity are unclear. In this study, we explored the seasonality and periodicity of influenza in Hefei, China during 2010 to 2015 using five proxies originated from three data sources of clinical surveillance of influenza-like illness (ILI), laboratory surveillance of influenza and death registration of pneumonia and influenza. We combined both wavelets analysis and de-linear-trend regression with Fourier harmonic terms to estimate seasonal characteristics of epidemic phase, peak time, amplitude, ratio of dominant seasonality. We found both annual cycle of influenza epidemics peaking in December-February and semi-annual cycle peaking in December-February and June-July in subtropical city Hefei, China. Compared to proxies developed by ILI and death registration data separately, influenza proxies incorporated laboratory surveillance data performed better seasonality and periodicity, especially in semi-annual periodicity in Hefei. Proxy of ILI consultation rate showed more timeliness peak than other proxies, and could be useful in developing the early warning model for influenza epidemics. Our study suggests to integrate clinical and laboratory surveillance of influenza for future influenza seasonality studies in subtropical regions.

Excess mortality associated with influenza after the 2009 H1N1 pandemic in a subtropical city in China, 2010-2015.

OBJECTIVES: The aim of this study was to use a quasi-Poisson regression model to estimate the mortality burden associated with influenza type/subtypes in a subtropical city in China, for the years 2010-2015. METHODS: Quasi-Poisson models were fitted separately to weekly numbers of deaths from various causes. The exploratory variables were products of weekly proportions of specimens positive for influenza type/subtypes and weekly influenza-like illness consultation rates to represent influenza activity. Adjustments were made for long-term and seasonal trends, absolute humidity, and population size as confounding factors in the models. Excess deaths associated with influenza were regarded as the measurement for disease burden of influenza. RESULTS: The excess mortality for all-cause death associated with influenza was 9.9 per 100000 population in Hefei, with influenza A(H3N2) virus having the highest excess mortality rate, followed by influenza A(H1N1) virus and influenza B virus. Following the 2009 H1N1 pandemic, the highest excess mortality rate associated with influenza for different causes was consistently found in the year 2014, with the excess mortality rate for all-cause death reaching 17.47 per 100000 population. The sex differences in influenza-associated mortality were not statistically significant (p>0.05). CONCLUSIONS: The mortality burden of influenza has been substantial in Hefei since the 2009 influenza pandemic, while the evidence on sex differences in mortality burden is limited. The severity profile of influenza type/subtypes in China needs to be assessed and confirmed in more cities in future studies.

Selective estrogen receptor modulator BC-1 activates antioxidant signaling pathway in vitro via formation of reactive metabolites.

AIM: Benzothiophene compounds are selective estrogen receptor modulators (SERMs), which are recently found to activate antioxidant signaling. In this study the molecular mechanisms of antioxidant signaling activation by benzothiophene compound BC-1 were investigated. METHODS: HepG2 cells were stably transfected with antioxidant response element (ARE)-luciferase reporter (HepG2-ARE cells). The expression of nuclear factor erythroid 2-related factor 2 (Nrf2) in HepG2-ARE cells was suppressed using siRNA. The metabolites of BC-1 in rat liver microsome incubation were analyzed using LC-UV and LC-MS. RESULTS: Addition of BC-1 (5 µmol/L) in HepG2-ARE cells resulted in a 17-fold increase of ARE-luciferase activity. Pretreatment with the estrogen receptor agonist E2 (5 µmol/L) or antagonist ICI 182,780 (5 µmol/L) did not affect BC-1-induced ARE-luciferase activity. However, transfection of the cells with anti-Nrf2 siRNA suppressed this effect by 79%. Addition of BC-1 in rat microsome incubation resulted in formation of di-quinone methides and o-quinones, followed by formation of GSH conjugates. BC-1 analogues with hydrogen (BC-2) or fluorine (BC-3) at the 4' position did not form the di-quinone methides. Both BC-2 and BC-3 showed comparable estrogenic activity with BC-1, but did not induce ARE-luciferase activity in HepG2-ARE cells. CONCLUSION: Benzothiophene compound BC-1 activates ARE signaling via reactive metabolite formation that is independent of estrogen receptors.

[Study on the score criteria of severe hand, foot and mouth disease cases].

OBJECTIVE: To explore the score criteria of severe hand, foot and mouth disease (HFMD) cases and to provide evidence for unified criteria and treatment on severe HFMD cases. METHODS: All severe cases and partial mild cases reported by two designated hospitals of HFMD in Fuyang during March to June, 2008 were scored by the methods of criteria constructed in advance. ROC curve was adopted to evaluate the score criteria and the gold standard was defined according to ICU, intubation and clinical outcomes, etc. Sensitivity, specificity and Youden's index were used to determine the division scores on critical, severe and mild cases. RESULTS: 97% of the cases (34 cases) were scored less than 6 points. 88% of cases (24 cases) who were intubated or mechanical ventilated had the scores of 6 points or higher. 79% of deaths (11 cases) were scored 10 points or higher. The area of receiver operation characteristic (ROC) curve was 0.90 (95%CI: 0.83 - 0.98) between severe and mild cases and the area of ROC curve was 0.95 (95%CI: 0.92 - 0.98) between critical and severe, mild cases. When comprehensively considering the sensitivity and specificity, severe cases were best judged when score was 4 points (sensitivity, specificity and Youden's index were 0.94, 0.68 and 0.62 respectively). When score was 6 points, critical cases were judged very well (sensitivity, specificity and Youden's index were 0.92, 0.84 and 0.76 respectively). CONCLUSION: Score criteria could be quantified to determine the degree of seriousness and with high-value for diagnosis on HFMD.

[Study on a fatal pregnant woman died from by avian influenza (H5N1)].

OBJECTIVE: To ascertain the causation of a pregnant woman with undefined pneumonia reported from the People's Hospital of Tongling city in Anhui province on November 2005. METHODS: Epidemiological and clinical information of the case was collected from the keypersons close to the case and referring to the medical record. A medical observation was carried out on the close contacts of the case and sick or dead poultry. Tracheal aspirates being collected were tested by both RT-PCR and real-time PCR to detect viral nucleic acids of A/H5N1, and were inoculated into special pathogen free (SPF) embryonated hens' eggs. RESULTS: The pregnant woman was found to have been contacted with the sick/dead poultry directly on the 4th day before onset of illness. All the 122 close contacts were healthy after a 10-day medical observation. The major clinical features of the case were viral pneumonia with rapidly developed leukopenia and lymphopenia. The progress to acute respiratory distress syndrome and multiple organ dysfunction syndromes was found at clinical presentation. HA and NA gene of A/H5N1 virus were positive. The 8 gene fragments of A/Anhui/1/2005 (H5N1) isolated from the tracheal aspirates had not carried genes from a human virus through reassortment, and the receptor-binding site of the hemagglutinin was polybasic cleavage site. CONCLUSION: This was the first documented case of H5N1 infection in pregnant woman. The immunotolerant state of pregnancy might have predisposed to the fatal outcome of the patient.