Tak1 is Required for the Survival of Hematopoietic Cells and Hepatocytes in Mice

Transforming growth factor beta-activated kinase 1 (TAK1), a member of the MAPKKK family, is a key mediator of proinflammatory and stress signals. Activation of TAK1 by proinflammatory cytokines and T and B cell receptors induces the nuclear localization of nuclear factor kappaB (NF-kappaB) and the activation of c-Jun N-terminal kinase (JNK)/AP1 and P38, which play important roles in mediating inflammation, immune responses, T and B cell activation, and epithelial cell survival. Here, we report that TAK1 is critical for the survival of both hematopoietic cells and hepatocytes. Deletion of TAK1 results in bone marrow (BM) and liver failure in mice due to the massive apoptotic death of hematopoietic cells and hepatocytes. Hematopoietic stem cells and progenitors were among those hematopoietic cells affected by TAK1 deletion-induced cell death. This apoptotic cell death is autonomous, as demonstrated by reciprocal BM transplantation. Deletion of TAK1 resulted in the inactivation of both JNK and NF-kappaB signaling, as well as the down-regulation of expression of prosurvival genes

Serological analysis of allergic components of house dust mite provides more insight in epidemiological characteristics and clinical symptom development in North China

BackgroundHouse dust mite (HDM) is the most common airborne source causing complex allergy symptoms. There are geographic differences in the allergen molecule sensitization profiles. Serological testing with allergen components may provide more clues for diagnosis and clinical management.ObjectiveThis study aims to investigate the sensitization profile of eight HDM allergen components in a large number of patients enrolled in the clinic and to analyze the relation of gender, age, and clinical symptoms in North China.MethodsThe 548 serum samples of HDM-allergic patients (ImmunoCAP® d1 or d2 IgE ≥0.35) were collected in Beijing City and divided in four different age groups and three allergic symptoms. The specific IgE of HDM allergenic components, Der p 1/Der f 1, Der p 2/Der f 2, Der p 7, Der p 10, Der p 21, and Der p 23, was measured using the micro-arrayed allergen test kit developed by Hangzhou Zheda Dixun Biological Gene Engineering Co., Ltd. The new system was validated by comparing to single-component Der p 1, Der p 2, and Der p 23 tests by ImmunoCAP in 39 sera. The epidemiological study of these IgE profiles and the relation to age and clinical phenotypes were analyzed.ResultsA greater proportion of male patients was in the younger age groups, while more female patients were in the adult groups. Both the sIgE levels and the positive rates (approximately 60%) against Der p 1/Der f 1 and Der p 2/Der f 2 were higher than for the Der p 7, Der p 10, and Der p 21 components (below 25%). The Der f 1 and Der p 2 positive rates were higher in 2–12-year-old children. The Der p 2 and Der f 2 IgE levels and positive rates were higher in the allergic rhinitis group. The positive rates of Der p 10 increased significantly with age. Der p 21 is relevant in allergic dermatitis symptom, while Der p 23 contributes to asthma development.ConclusionHDM groups 1 and 2 were the major sensitizing allergens, with group 2 being the most important component relevant to respiratory symptoms in North China. The Der p 10 sensitization tends to increase with age. Der p 21 and Der p 23 might be associated with the development of allergic skin disease and asthma, respectively. Multiple allergen sensitizations increased the risk of allergic asthma

Identification of RegIV as a Novel GLI1 Target Gene in Human Pancreatic Cancer

GLI1 is the key transcriptional factor in the Hedgehog signaling pathway in pancreatic cancer. RegIV is associated with regeneration, and cell growth, survival, adhesion and resistance to apoptosis. We aimed to study RegIV expression in pancreatic cancer and its relationship to GLI1.GLI1 and RegIV expression were evaluated in tumor tissue and adjacent normal tissues of pancreatic cancer patients and 5 pancreatic cancer cell lines by qRT-PCR, Western blot, and immunohistochemistry (IHC), and the correlation between them. The GLI1-shRNA lentiviral vector was constructed and transfected into PANC-1, and lentiviral vector containing the GLI1 expression sequence was constructed and transfected into BxPC-3. GLI1 and RegIV expression were evaluated by qRT-PCR and Western blot. Finally we demonstrated RegIV to be the target of GLI1 by chromatin immunoprecipitation (CHIP) and electrophoretic mobility shift assays (EMSA).The results of IHC and qRT-PCR showed that RegIV and GLI1 expression was higher in pancreatic cancer tissues versus adjacent normal tissues (p<0.001). RegIV expression correlated with GLI1 expression in these tissues (R = 0.795, p<0.0001). These results were verified for protein (R = 0.939, p = 0.018) and mRNA expression (R = 0.959, p = 0.011) in 5 pancreatic cancer cell lines. RegIV mRNA and protein expression was decreased (94.7±0.3%, 84.1±0.5%; respectively) when GLI1 was knocked down (82.1±3.2%, 76.7±2.2%; respectively) by the RNAi technique. GLI1 overexpression in mRNA and protein level (924.5±5.3%, 362.1±3.5%; respectively) induced RegIV overexpression (729.1±4.3%, 339.0±3.7%; respectively). Moreover, CHIP and EMSA assays showed GLI1 protein bound to RegIV promotor regions (GATCATCCA) in pancreatic cancer cells.GLI1 promotes RegIV transcription by binding to the RegIV gene promoter in pancreatic cancer

Association of serum chemokine ligand 21 levels with asthma control in adults

OBJECTIVES: The chemokine ligand (CCL) 21 regulates the maturation, migration, and function of dendritic cells, and has been implicated in the pathogenesis of asthma. This study aimed to investigate the association between serum CCL21 levels and asthma control. METHODS: The serum levels of CCL21 and other inflammatory cytokines were analyzed in patients with asthma (n=44) and healthy controls (n=35) by enzyme-linked immunosorbent assay. IgE levels and eosinophil counts were determined by turbidimetric inhibition immunoassay and fully automatic blood analysis, respectively. The Asthma Control Test (ACT) questionnaire was used, and spirometry and fractional exhaled nitric oxide (FENO) measurements were performed. A multiple unpaired Student’s t-test was performed to analyze the differences in CCL21 and interleukin levels between patients with asthma and healthy controls. The correlation of CCL21 levels with disease severity was evaluated using the Pearson’s rank correlation test. RESULTS: Serum CCL21 levels were lower in patients with asthma (254.78±95.66 pg/mL) than in healthy controls (382.95±87.77 pg/mL) (po0.001). Patients with asthma had significantly higher levels of IL-1b (19.74±16.77 vs. 2.63±5.22 pg/mL), IL-6 (7.55±8.65 vs. 2.37±2.47 pg/mL), and tumor necrosis factor-a (12.70±12.03 vs. 4.82± 3.97 pg/mL) compared with the controls. CCL21 levels were positively correlated with the ACT score (rs=0.1653, p=0.0062), forced expiratory volume in 1s (FEV1)/forced vital capacity (rs=0.3607, po0.0001), and FEV1 (rs=0.2753, p=0.0003), and negatively correlated with FENO (rs=0.1060, p=0.0310). CCL21 levels were negatively correlated with serum IgE levels (rs=0.1114, p=0.0268) and eosinophil counts (rs=0.3476, po0.0001). CONCLUSIONS: Serum CCL21 levels may be a new biomarker for assessing asthma control

Spatial and temporal trends in total organic carbon (TOC), black carbon (BC), and total nitrogen (TN) and their relationships under different planting patterns in a restored coastal mangrove wetland: case study in Fujian, China

This study aims to determine the sediment changes and the trends in TOC, BC and TN before and after restoration of the mangrove wetland in the Jinjiang Estuary and to determine the effect of the wetland restoration process on the biogeochemical cycle of carbon and nitrogen. The results suggest that the sediments were mainly silt-sized. Among different sites with different types of plants and vegetation densities, the adsorptive ability of N in the plots in plantations of Kandelia obovata, Avicennia marina and Acanthus ilicifolius was the highest. The TOC content differed (p < 0.05) with the density of the plot and significantly differed (p < 0.01) with the mangrove species at the densities of 0.5 × 1 m and 0.5 × 0.5 m. There was a positive relationship between the TOC and TN and the TOC and carbon-nitrogen ratio (C/N) (P < 0.05)

BM was collected from and mice on days 4, 6, and 8 after the first polyI:C injection

After lysis of RBCs, BM-nucleated cells were stained with cell surface markers and analyzed by flow cytometry for HSCs and CPs. Progressive loss of HSCs and CPs in mouse BM is shown by lineage marker and Sca1 c-kit staining (B–D). Day 8 BM cells were used as a control (A). (E) Progressive reduction of colony-forming ability of BM cells after deletion. (F) BM cells were collected from and mice 16 h after a single polyI:C injection, and apoptosis was checked by annexin V staining. *, P < 0.05; **, P < 0.01.<p><b>Copyright information:</b></p><p>Taken from "TAK1 is required for the survival of hematopoietic cells and hepatocytes in mice"</p><p></p><p>The Journal of Experimental Medicine 2008;205(7):1611-1619.</p><p>Published online 7 Jul 2008</p><p>PMCID:PMC2442639.</p><p></p

BM cells were collected into lyse/fix buffer immediately after the mice were killed to fix the BM cells and lyse the RBCs simultaneously

BM-nucleated cells were washed three times with PBS containing 5% FBS to completely remove all traces of the lyse/fix buffer. The cells were stained with cell surface markers as indicated and then permeabilized using permeabilization buffer for TAK1 and p-TAK1 staining. (A–C) High level of TAK1 protein expression in c-kit hematopoietic cells and Gr1 myeloid cells, but low expression in B220 B cells. (D–F) TAK activity (shown here by p-TAK1 levels) is higher in c-kit hematopoietic cells (including B220c-kit B cell progenitors, red arrow in F). (G and H) No obvious difference in expression in LSK-HSC/Ps and LK-CPs, but increased TAK1 activity in LK-CPs compared with LSK-HSC/Ps. (I) is expressed in all lineages of hematopoietic cells, including HSC/Ps, as shown by quantitative RT-PCR. BM cells from knockout mice (4 d after mutation induction) were used as negative controls. Compared with differentiated cells, including Gr1 myeloid cells, B220 B cells, Ter119-nucleated erythoid cells, and CD3 T cells, RNA expression was significantly higher in LK-CPs and B cell progenitors (B220c-kit). Both * and ** are P < 0.01. *, compared with BM cells; **, compared with mature hematopoietic cells, including Gr1, B220, and CD3 cells.<p><b>Copyright information:</b></p><p>Taken from "TAK1 is required for the survival of hematopoietic cells and hepatocytes in mice"</p><p></p><p>The Journal of Experimental Medicine 2008;205(7):1611-1619.</p><p>Published online 7 Jul 2008</p><p>PMCID:PMC2442639.</p><p></p