Molecular Mechanism of the Cell Death Induced by the Histone Deacetylase Pan Inhibitor LBH589 (Panobinostat) in Wilms Tumor Cells.

BACKGROUND: Wilms tumor (WT) is an embryonic kidney cancer, for which histone acetylation might be a therapeutic target. LBH589, a novel targeted agent, suppresses histone deacetylases in many tumors. This study investigated the antitumor activity of LBH589 in SK-NEP-1 and G401 cells. METHODS: SK-NEP-1 and G401 cell growth was assessed by CCK-8 and in nude mice experiments. Annexin V/propidium iodide staining followed by flow cytometry detected apoptosis in cell culture. Gene expressions of LBH589-treated tumor cells were analyzed using an Arraystar Human LncRNA Array. The Multi Experiment View cluster software analyzed the expression data. Differentially expressed genes from the cluster analyses were imported into the Ingenuity Pathway Analysis tool. RESULTS: LBH589 inhibited cell proliferation of SK-NEP-1 and G401 cells in a dose-dependent manner. Annexin V, TUNEL and Hochest 33342 staining analysis showed that LBH589-treated cells showed more apoptotic features compared with the control. LBH589 treatment inhibited the growth of SK-NEP-1 xenograft tumors in nude mice. Arraystar Human LncRNA Array analysis of genes and lncRNAs regulated by LBH589 identified 6653 mRNAs and 8135 lncRNAs in LBH589-treated SK-NEP-1 cells. The most enriched gene ontology terms were those involved in nucleosome assembly. KEGG pathway analysis identified cell cycle proteins, including CCNA2, CCNB2, CCND1, CCND2, CDK4, CDKN1B and HDAC2, etc. Ingenuity Pathway Analysis identified important upstream molecules: HIST2H3C, HIST1H4A, HIST1A, HIST1C, HIST1D, histone H1, histone H3, RPRM, HSP70 and MYC. CONCLUSIONS: LBH589 treatment caused apoptosis and inhibition of cell proliferation of SK-NEP-1and G401 cells. LBH589 had a significant effect and few side effects on SK-NEP-1 xenograft tumors. Expression profiling, and GO, KEGG and IPA analyses identified new targets and a new "network" of genes responding to LBH589 treatment in SK-NEP-1 cells. RPRM, HSP70 and MYC may be important regulators during LBH589 treatment. Our results provide new clues to the proapoptotic mechanism of LBH589.

C3 glomerulonephritis in 7 children / 中华实用儿科临床杂志

Objective To analyze the clinical characteristics of 7 pediatric patients with C3 glomerulonephritis (C3-GN).Methods The clinical manifestations,pathological features,therapies,prognosis of patients from Jun.2006 to Nov.2011 were analyzed retrospectively.Results All of the patients were presented as acute nephritic syndrome,and 4 patients with macroscopic hematuria,other 3 cases with severe proteinuria.Five patients with eyelid edema.All the patients showed decreased level of serum complement C3,while serum complement C4 was normal.Investigations showed elevation of 24 h urine protein,5 patients with elevated antistreptolysin O titers.Three patients with renal dysfunction.Isolated C3 deposition in mesangial and endothelial areas(+ +-+ + +) was confirmed by immunofluorescence.Light microscope revealed membrane proliferative glomerulonephritis and mesangial proliferative glomerulonephritis.Electron microscope showed swelling and hyperplasia of endothelial cells without electron-dense deposition or podocyte foot fusion.Based on conventional treatment,administration of immunosuppressant was performed in 3 patients with severe pathological changes.After a follow-up of 2 months to 5 years,the prognosis seems to be benign.Conclusions Children with C3-GN are usually presented as acute nephritic syndrome,characterized by isolate C3 deposition in immunofluorescence.Electron microscope showed lesion of endothelial cells,while no electron-dense deposits in mesangial and endothelial areas.The mechanism may be associated with dysregulation of alternative complement pathway,and seems had a good short-term prognosis.

Inhibition of the ecto-beta subunit of F1F0-ATPase inhibits proliferation and induces apoptosis in acute myeloid leukemia cell lines.

BACKGROUND: Leukemia, a heterogeneous clonal disorder of hematopoietic progenitor cells, presents a world-wide health problem, especially in childhood. F1F0 ATPase, an inner mitochondrial enzyme, is expressed on the plasma membrane of tumor cells, and its inhibition induces both anti-angiogenic and anti-tumorigenic activity. METHODS: Monoclonal Antibody (McAb) against ATPase was produced by polyethylene glycol-mediated fusions and screened by ELISA. Proliferation, cell cycle and apoptosis of cells were analyzed when the surface ATPase of cells was blockaded with McAb. RESULTS: We detected cell-membrane expression of the F1F0 ATPase ß subunit on 0.1% to 56% of the 11 cell lines derived from leukemia, including acute myeloid leukemia (AML). We produced a monoclonal antibody, McAb7E10, which recognizes both the native and recombinant ATPase ß subunit, with a dissociation constant (KD) of 3.26E-10. We demonstrate that McAb7E10 binds to ATPase at the cell surface, where it is able to inhibit ATP synthesis. McAb7E10 significantly inhibited proliferation of AML cell lines in vitro: the relative inhibitory rates of 50 µg/mL McAb7E10 treated MV4-11and HL-60 cells were 69.6% and 81.9% respectively. Cell cycle analysis indicated that McAb7E10 significantly induced apoptosis in MV4-11 and HL-60 cells: the relative rates of apoptosis in 5, 10 and 50ug/mL McAb7E10 treated MV4-11 cells was 3.6 ± 0.83%, 8.4 ± 1.69% and 17.3 ± 2.56% compared to 1.5% ± 0.85% in mouse IgG treated cells (p < 0.01). The relative rate of apoptosis in 5, 10 and 50ug/mL McAb7E10 treated HL-60 cells was 5.5 ± 2.37%, 11.3 ± 3.62% and 19.9 ± 3.31% compared to 1.56% ± 0.97% in mouse IgG treated cells (p < 0.01). Annexin V staining demonstrated that the relative apoptotic rates in 50 µg/mL McAb7E10 treated MV4-11 and HL-60 cells were 50.5% ± 7.04% and 32.9% ± 4.52%, respectively, significantly higher than IgG control antibody treated cells were 21.9% ± 3.11% and 15.3% ± 3.95%, p < 0.01. CONCLUSIONS: These findings indicate that ectopic expression of ATPase ß subunit may be a tumor-associated antigen in hematological malignancies. The F1F0 ATPase ß subunit provides a potential target for immunotherapy in AML and hematological malignancies.

Three cases of eosinophilichyperplastic lymphogranuloma in children's parotid area / 华西口腔医学杂志

From March 2009 to October 2009, three pediatric patients with parotid tumor were cured. Preoperative physical examination showed regional swelling in parotid area, the surface skin was in moderate reddish purple, the border was vague, and the swelling was inactive. The patients' IgE were significantly increased. B ultrasound examination demonstrated the focus was an isoecho with ringlike dark band around, which was concluded as bull's-eye sign. Magnetic resonance imaging (MRI) examination indicated a cystic mass between the skin and parotid. Preoperative diagnosis was eosinophilichyperplastic lymphogranuloma (Kimura's disease) and the granuloma was excised by operation. Pathological examination revealed the capillary vessel hyperplasia in local tissue with a plenty of eosinophils and lymphocytes infiltrating. The disease was confirmed. Although the disease is rare, the diagnosis still could be made by preoperative physical examination, laboratory and imaging examinations.

Correlations of Immunopathologic Types with Clinical Manifestations and Histological Classifications in Children with Henoch Schonlein Purpura Nephritis / 实用儿科临床杂志

0.05).Compared with other immunopathologic types,IgA plus IgG plus IgM deposition type had higher proportion of histological grade Ⅲb-Ⅵ(P

Clinical and laboratory studies on childhood acute leukemia with 11q23 abnormalities / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the interrelations among morphology, immunology, cytogenetics and clinical outcome in childhood acute leukemia with 11q23 abnormalities.</p><p><b>METHODS</b>Eighteen patients with 11q23 abnormalities, from 320 childhood acute leukemia patients, were retrospectively analysed for cell morphology, flow cytometry, immunophenotyping, R-banding karyotype as well as clinical features and prognosis. Twenty cases of childhood AL with normal karyotype during the same period were used as control.</p><p><b>RESULTS</b>The incidence of 11q23 abnormalities in our childhood acute leukemia patients was 5.63% including 14 acute lymphoblastic leukemia (ALL) and 4 acute myeloid leukemia (AML). Of 16 cases immunophenotypically tested, 13 expressed lymphoid antigens and 3 CD(34) and other myeloid antigens. Karyotype analysis disclosed the following abnormalities: t(4; 11)(q21; q23) in 6 cases, t(10; 11)(p13; q23) in 3, t(11; 19)(q23; p13) in one and del(11)(q23) in 6. The complete remission rate for these patients with 11q23 abnormalities was comparable to that of the control (72.2% vs 80.0%, P > 0.05), while the mortality rate in the former was significantly higher than that in the latter (61.1% vs 25.0%, P < 0.05).</p><p><b>CONCLUSIONS</b>11q23 abnormalities were mainly seen in childhood ALL and acute monocytic leukemia with unique prognostic features.</p>