Investigation of lncRNA-mRNA co-expression network in ETV6-RUNX1-positive pediatric B-cell acute lymphoblastic leukemia.

ETV6/RUNX1 gene fusion is the most common chromosomal translocation abnormality occurred in pediatric B-cell acute lymphoblastic leukemia (B-ALL). Compared with ETV6-RUNX1-negative patients, ETV6-RUNX1-positive patients possess more improved treatment strategies but higher risk to relapse. In this research, the potential gene interaction networks were constructed intending for elucidating the pathogenesis of B-ALL. We performed the weighted gene co-expression network analysis (WGCNA) to assess the involvement of lncRNA-mRNA pairs in B-ALL patients consisting of 24 ETV6-RUNX1-positive patients and 18 ETV6-RUNX1-negative patients and found a module that was significantly associated with positive/negative trait. Gene Ontology analysis showed that mRNAs in this module were enriched in the positive regulation of MAPK cascade, positive regulation of JNK cascade, and myeloid cell differentiation pathway. To further investigate the relationship between lncRNAs and mRNAs in this significant module, we constructed the lncRNA-mRNA co-expression network. 3 lncRNAs (RP11-170J3.2, RP11-135F9.1 and RP1-151B14.9) were found at the core of the lncRNA-mRNA co-expression network, which had the most co-expression connections with mRNAs. And several related mRNAs (ACTN1, TNFRSF21 and NLRP3) had a significant correlation with the patient survival prediction. Our findings may explicate the pathogenesis of B-ALL, and the disease-associated genes could provide clues to find novel biomarkers for prognosis.

Programmed cell death 4 and BCR-ABL fusion gene expression are negatively correlated in chronic myeloid leukemia.

Programmed cell death 4 (PDCD4) is a tumor suppressor that inhibits carcinogenesis, tumor progression and invasion by preventing gene transcription and translation. Downregulation of PDCD4 expression has been identified in multiple types of human cancer, however, to date, the function of PDCD4 in leukemia has not been investigated. In the present study, PDCD4 mRNA and protein expression was investigated in 50 patients exhibiting various phases of chronic myeloid leukemia (CML) and 20 healthy individuals by reverse transcription-quantitative polymerase chain reaction and western blot analysis. PDCD4 expression and cell proliferation was also investigated following treatment with the tyrosine kinase inhibitor, imatinib, in K562 cells. The results demonstrated that PDCD4 mRNA and protein expression was decreased in all CML samples when compared with healthy controls, who expressed high levels of PDCD4 mRNA and protein. No significant differences in PDCD4 expression were identified between chronic phase, accelerated phase and blast phase CML patients. In addition, PDCD4 expression was negatively correlated with BCR-ABL gene expression (r=-0.6716; P<0.001). Furthermore, K562 cells treated with imatinib exhibited significantly enhanced PDCD4 expression. These results indicate that downregulation of PDCD4 expression may exhibit a critical function in the progression and malignant proliferation of human CML.

[Performance of an Intermittent Aeration Membrane Bioreactor for Removal of Veterinary Antibiotics from Piggery Wastewater].

The removal performance of 11 veterinary antibiotics in piggery wastewater and their accumulation in the activated sludge were studied in an intermittent aeration membrane bioreactor (IAMBR) at different COD/TN ratios and organic loads. The results showed that both antibiotics and chemical oxygen demand (COD) were efficiently removed at hydraulic retention time (HRT) of no less than five days and COD/TN ratio of 2.1, the removal rate was (79.1 ± 0.7)% for total antibiotics and was (88.4 ± 1.4)% for COD. As HRT was shortened to three days, the removal rate of COD was little changed but the removal rate of total antibiotics was significantly decreased. As COD/TN decreased from 2.1 to 0.7, the removal rate of total antibiotics was little changed, but the COD removal rate was significantly decreased. Antibiotics of tetracycline and quinolone species kept accumulating in the sludge at long SRT, and the accumulation amount was decreased with shorter SRT. The concentration and composition of antibiotic in the sludge were influenced by the raw wastewater.

[Method for Simultaneous Determination of 11 Veterinary Antibiotics in Piggery Wastewater and Sludge and Its Application in Biological Treatment].

In order to determine eleven commonly used veterinary antibiotics (including four tetracyclines, two sulfonamides, three quinolones and two macrolides) in piggery wastewater and activated sludge in the Yangtze River Delta region, the conditions of solid phase extraction and high performance liquid chromatography-tandem mass spectrometry were optimized. The recovery rate and relative standard deviations of the method were confirmed as 73% - 105.2%, 3.1% - 10.2% for piggery wastewater (n = 3) and 57.4% - 104.6%, 1.9% - 10.9% (n = 3) respectively for the activated sludge. Removal of antibiotics was then studied in a membrane bioreactor. The results showed that antibiotics of both tetracycline and sulfonamide species took a large portion in the wastewater, while tetracycline species were the dominant in the sludge. Tetracycline species in the wastewater were removed by 85.2%, mainly through biodegradation (51.9%) and secondly by sludge adsorption (33.2%). By comparison, sulfonamide species was removed by 95.8%, almost all through biodegradation while little by sludge adsorption. Flask tests suggested that the accumulated antibiotics in the sludge give no significant influence on the microbial removal of organics and ammonium.

[A study on the veterinary antibiotics contamination in groundwater of Jiaxing].

Liquid chromatography and tandem mass (LC-MS/MS) followed with solid-phase extraction (SPE) was developed for simultaneously determining four classes (tetracyclines, quinolones, macrolides and sulfonamides) of ten commonly used veterinary antibiotics in groundwater of Jiaxing city, an important pig breeding base in the Yangtze River Delta region. Samples were taken from 10 typical rural river sections and 21 main urban river sections. Results revealed severe pollution existed in the rural river environment. The total concentration of ten antibiotics was as high as 65.6-467.0 ng x L(-1), among which tetracyclines and sulfonamides respectively ranged in 40.8-253.0 ng x L(-1) and undetected (nd)- 165.0 ng x L(-1), macrolides and quinolones respectively ranged in 3.1-14.68 ng x L(-1) and nd-14.54 ng x L(-1). By comparison, the pollution level in urban rivers was much lower. The total concentration of ten antibiotics ranged in 20.1 ng x L(-1) to 61.2 ng x L(-1), among with tetracyclines varied from undetected to 44.0 ng x L(-1), while sulfonamides, macrolides and quinolones were respectively below 2.7 ng x L(-1), 6.3 ng x L(-1) and 21.6 ng x L(-1).

[Role of dendritic cells and their Toll-like receptor 4 in the pathogenesis of idiopathic thrombocytopenic purpura].

OBJECTIVE: To study the surface antigen of the dendritic cells (DC) and their Toll-like receptor 4 (TLR4) expression in patients with idiopathic thrombocytopenic purpura (ITP), and to explore their role in ITP pathogenesis. METHODS: The peripheral blood mononuclear cells isolated from complete remission patients (CR), non-complete remission patients (n-CR) and normal controls were stimulated by rhGM-CSF and rhIL-4. The surface antigen of the DC was analyzed by flow cytometry. The level of IL-12p70 in the supernatant was detected by enzyme linked immunosorbent assay. The expression of TLR4 mRNA of DC was detected by real time PCR. RESULTS: In the 21 CR ITP patients, the expression of both CD80 and CD86 in DC was significantly increased compared with that in normal controls \[(51.60 ± 13.47)% vs (36.03 ± 15.43)%, (61.50 ± 15.93)% vs (40.28 ± 11.49)%, respectively\] (P < 0.01). The expression of CD80 and CD86 in n-CR group was also significantly increased \[(53.29 ± 19.49)% and (62.91 ± 18.43)%, respectively\] (P < 0.01). After HD-DXM treatment, both CD80 and CD86 in CR patients were decreased (P < 0.01). There was no difference between the DXM treatment patients and the normal controls. In n-CR group, there was no difference in CD80 and CD86 expression before and after DXM therapy \[(52.30 ± 20.98% and (49.79 ± 20.28)%, respectively\] (P > 0.05). CD80 was still higher than normal (P < 0.05), while CD86 was not changed. The level of IL-12p70 in CR ITP patients before treatment was significantly higher \[(67.52 ± 14.43) pg/ml\] than that of the controls \[(39.78 ± 10.03) pg/ml\](P < 0.01), and after treatment, was significantly decreased to (43.90 ± 8.49) pg/ml, being no difference from that in control. In n-CR group, IL-12p70 was lower after treatment \[(48.45 ± 9.68) pg/ml\] than that before treatment \[(65.35 ± 12.52) pg/ml\] (P < 0.01), but still higher than that in control (P < 0.05). The TLR4 mRNA level in DCs of CR ITP patients before treatment were significantly higher 0.69 ± 0.17 than that of controls (0.31 ± 0.09) (P < 0.01) and after treatment, was reduced to 0.35 ± 0.11, being no difference from that in control. In n-CR group, TLR4 mRNA was decreased from 0.65 ± 0.09 to 0.52 ± 0.21 after treatment (P < 0.01), but still higher than normal (P < 0.01). CONCLUSION: DC may play an important role in ITP by their Toll-like receptor and cytokine secretion.

Dexamethasone inhibits immunoreactivity of dendritic cells in patients with chronic idiopathic thrombocytopenic purpura.

The objective of this study was to investigate the possible effects of dexamethasone treatment on the immunoreactivity of dendritic cells in patients with chronic idiopathic thrombocytopenic purpura (ITP). Thirty-six newly diagnosed patients with chronic ITP received an oral high dose of dexamethasone (HD-DXM) at single daily doses of 40 mg for 4 consecutive days. The CD14 leukocytes isolated from the 21 remission patients and 10 normal controls were stimulated by recombinant human granulocyte-macrophage colony-stimulating factor and rhIL-4. The surface antigens of the dendritic cells were analyzed by flow cytometry and the level of IL-12p70 in the supernatant was detected by enzyme-linked immunosorbent assay. In ITP patients, the expression of both CD80 and CD86 in dendritic cells were significantly increased compared with those of the normal controls (51.60 +/- 13.47 vs. 36.03 +/- 15.43%, 61.50 +/- 15.93 vs. 40.28 +/- 11.49%, respectively; P < 0.05). After HD-DXM treatment, both CD80 and CD86 were decreased to levels comparable to normal controls (P > 0.05). The level of IL-12p70 in ITP patients was significantly higher (67.52 +/- 14.43 pg/ml) than the controls (39.78 +/- 10.03 pg/ml, P < 0.05). After treatment, IL-12p70 was reduced to 43.90 +/- 8.49 pg/ml with no significant differences between ITP group and control (P > 0.05). Dendritic cells and their cytokine secretion play important roles in ITP, and DXM may achieve its therapeutic effect on ITP by inhibiting immune responses through suppressing the function of dendritic cells.