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1.
Blood ; 135(1): 41-55, 2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31697823

RESUMO

To study the mechanisms of relapse in acute lymphoblastic leukemia (ALL), we performed whole-genome sequencing of 103 diagnosis-relapse-germline trios and ultra-deep sequencing of 208 serial samples in 16 patients. Relapse-specific somatic alterations were enriched in 12 genes (NR3C1, NR3C2, TP53, NT5C2, FPGS, CREBBP, MSH2, MSH6, PMS2, WHSC1, PRPS1, and PRPS2) involved in drug response. Their prevalence was 17% in very early relapse (<9 months from diagnosis), 65% in early relapse (9-36 months), and 32% in late relapse (>36 months) groups. Convergent evolution, in which multiple subclones harbor mutations in the same drug resistance gene, was observed in 6 relapses and confirmed by single-cell sequencing in 1 case. Mathematical modeling and mutational signature analysis indicated that early relapse resistance acquisition was frequently a 2-step process in which a persistent clone survived initial therapy and later acquired bona fide resistance mutations during therapy. In contrast, very early relapses arose from preexisting resistant clone(s). Two novel relapse-specific mutational signatures, one of which was caused by thiopurine treatment based on in vitro drug exposure experiments, were identified in early and late relapses but were absent from 2540 pan-cancer diagnosis samples and 129 non-ALL relapses. The novel signatures were detected in 27% of relapsed ALLs and were responsible for 46% of acquired resistance mutations in NT5C2, PRPS1, NR3C1, and TP53. These results suggest that chemotherapy-induced drug resistance mutations facilitate a subset of pediatric ALL relapses.

2.
Eur J Pediatr ; 2019 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-31807902

RESUMO

Juvenile myelomonocytic leukemia (JMML) is a heterogeneous childhood leukemia. The management of patients with JMML requires accurate assessment of genetic and clinical features to help in patient risk stratification. This study aimed to investigate the association between genomic alterations and prognosis in children with JMML. Genomic DNA was extracted from a total of 93 patients with JMML for targeted sequencing. Univariable and multivariable analysis were used to evaluate the correlation between gene mutations and prognosis of the patients. Patients with PTPN11 mutation exhibited significantly lower event-free survival (EFS) compared with non-PTPN11 mutations (P = 0.005). Patients without or with one somatic alteration at diagnosis showed significantly better prognosis in comparison with those with more than two alterations (P = 0.009). PTPN11 mutation with additional alterations showed significantly the poorest outcome in comparison with those with only one non-PTPN11 mutation, only one PTPN11 mutation, and combined mutations without PTPN11, respectively (P < 0.0001).Conclusion: Both PTPN11 mutation and the number of somatic alterations detected at diagnosis are likely to be the major determinant of outcome in JMML. The subgroup of patients with PTPN11 mutation showed the shortest survival which was even worsened when a secondary mutation was present.

3.
Leukemia ; 33(10): 2365-2378, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-30940905

RESUMO

Bone marrow (BM) niche responds to chemotherapy-induced cytokines secreted from acute lymphoblastic leukemia (ALL) cells and protects the residual cells from chemotherapeutics in vivo. However, the underlying molecular mechanisms for the induction of cytokines by chemotherapy remain unknown. Here, we found that chemotherapeutic drugs (e.g., Ara-C, DNR, 6-MP) induced the expression of niche-protecting cytokines (GDF15, CCL3 and CCL4) in both ALL cell lines and primary cells in vitro. The ATM and NF-κB pathways were activated after chemotherapy treatment, and the pharmacological or genetic inhibition of these pathways significantly reversed the cytokine upregulation. Besides, chemotherapy-induced NF-κB activation was dependent on ATM-TRAF6 signaling, and NF-κB transcription factor p65 directly regulated the cytokines expression. Furthermore, we found that both pharmacological and genetic perturbation of ATM and p65 significantly decreased the residual ALL cells after Ara-C treatment in ALL xenograft mouse models. Together, these results demonstrated that ATM-dependent NF-κB activation mediated the cytokines induction by chemotherapy and ALL resistance to chemotherapeutics. Inhibition of ATM-dependent NF-κB pathway can sensitize ALL to chemotherapeutics, providing a new strategy to eradicate residual chemo-resistant ALL cells.

4.
Exp Hematol ; 66: 32-41.e8, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30031030

RESUMO

Fanconi anemia (FA) is a rare recessive disease characterized by progressive bone marrow failure, congenital abnormalities, and increased incidence of cancers. To date, mutations in 22 genes can cause FA or an FA-like phenotype. In China, in addition to clinical information, FA diagnosis primarily relies on genetic sequencing because the chromosome breakage test is rarely performed. Here, we employed multiple genetic diagnostic tools (DNA sequencing, multiplex ligation-dependent probe amplification, and chromosome microarray) and a variant-based functional assay platform to investigate the genetic cause in 25 Chinese suspected FA patients. A total of 45 distinct candidate variants were detected in six FA genes (FA-A, FA-B, FA-C, FA-D2, FA-G, and FA-J), of which 36 were novel. Eight missense variants and one indel variant were unable to restore FANCD2 mono-ubiquitination and mitomycin C resistance in a panel of FA indicator cell lines, indicating that these mutations are deleterious. Three missense variants (FANCA-L424V, FANCC-E273K, and FANCG-A153G) were harmless. Finally, 23 patients were molecularly diagnosed with FA, consistent with their clinical phenotype. In the FA-A subgroup, large deletions accounted for 14% of the disease-causing variants. We have established a comprehensive molecular diagnostic workflow for Chinese FA patients that can substitute for standard FA cytogenetic analysis.


Assuntos
Proteína do Grupo de Complementação A da Anemia de Fanconi/genética , Proteína do Grupo de Complementação C da Anemia de Fanconi/genética , Proteína do Grupo de Complementação D2 da Anemia de Fanconi/genética , Proteína do Grupo de Complementação G da Anemia de Fanconi/genética , Anemia de Fanconi/genética , Mutação , Sequência de Bases , Criança , Pré-Escolar , China , Éxons , Anemia de Fanconi/diagnóstico , Anemia de Fanconi/etnologia , Anemia de Fanconi/patologia , Feminino , Expressão Gênica , Humanos , Lactente , Íntrons , Masculino , Reação em Cadeia da Polimerase Multiplex , Análise de Sequência de DNA , Sequenciamento Completo do Exoma
5.
Pediatr Blood Cancer ; 65(10): e27266, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29943896

RESUMO

BACKGROUND: Mixed-lineage leukemia (MLL) with multifarious partner genes leads to aggressive leukemia with dismal outcomes. METHODS: Using panel-based targeted sequencing, we examined 90 cases with MLL-rearranged (MLL-r) childhood acute leukemia, including 55 with acute lymphoblastic leukemia (ALL) and 35 with acute myeloid leukemia (AML). RESULTS: MLL breakpoints and complete rearrangements were identified. A total of 37.8% (34/90) of patients displayed a single direct MLL fusion gene, 15.6% (14/90) carried a single reciprocal fusion, and 27.8% (25/90) had both reciprocal MLL fusion alleles. The remaining 17 MLL-r cases exhibited complex translocations with homozygous disruptions on chromosome 11 or two breakpoints on the same MLL allele with a deletion of functional regions. A total of 77 patients (45 ALL and 32 AML) received chemotherapy with a median follow-up of 2.5 years. Unexpectedly, we identified children with reciprocal MLL fusions who exhibited relatively favorable outcomes compared with those in children with complex translocations or a single direct MLL fusion allele (66.1% vs. 24.6% and 27.6%, P = 0.001). Reciprocal MLL fusion may be functionally rescued by a partially truncated MLL protein. CONCLUSION: Comprehensive MLL-r analysis by targeted next-generation sequencing can provide detailed molecular information and is helpful for precise stratified treatment and clinical prognosis determination.


Assuntos
Histona-Lisina N-Metiltransferase/genética , Leucemia Mieloide Aguda/genética , Proteína de Leucina Linfoide-Mieloide/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Criança , Pré-Escolar , Feminino , Rearranjo Gênico , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Lactente , Estimativa de Kaplan-Meier , Leucemia Mieloide Aguda/mortalidade , Leucemia Mieloide Aguda/patologia , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/mortalidade , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Prognóstico , Intervalo Livre de Progressão , Translocação Genética
6.
J Pediatr Hematol Oncol ; 40(5): e299-e304, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29702541

RESUMO

Fanconi anemia (FA) is a rare autosomal recessive or X-linked disorder with highly variable clinical manifestations and an incidence of ∼1 to 5 in 1 million births. To date, 15 bona fide FA genes have been reported to be responsible for the known FA complementation groups and the FANCA gene accounts for almost 60%. In the present study, we report a special Chinese family, which has 2 children with classic FA characteristics. Via 2-step analysis of the whole-exome sequencing data and verification using multiplex ligation-dependent probe amplification test, one child was found to have a novel compound heterozygous mutation of a splicing variant (c.1471-1G>A) and a large intragenic deletion (exons 23-30 del) of the FANCA gene. The other child had the same splicing variant and another novel large deletion (exons 1-18 del) in the FANCA gene. Clone sequencing showed the c.1471-1G>A variant generate an altered transcript with 1 cryptic splice site in intron 15, resulting in a premature termination codon (p.Val490HisfsX6). This study not only shows the complexity of FA molecular diagnosis via comprehensively studying the FA pathogenic genes and the mutational spectrum, but also has significant reference value for the future molecular diagnosis of FA.


Assuntos
Família , Proteína do Grupo de Complementação A da Anemia de Fanconi/genética , Anemia de Fanconi/diagnóstico , Anemia de Fanconi/genética , Mutação , Sítios de Splice de RNA , Grupo com Ancestrais do Continente Asiático , Criança , China , Códon de Terminação , Humanos , Masculino
7.
Ann Hematol ; 96(8): 1389-1397, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28623394

RESUMO

Hematopoietic stem cell transplantation (HSCT) using an optimized conditioning regimen is essential for the long-term survival of patients with inherited bone marrow failure syndromes (IBMFS). We report HSCT in 24 children with Fanconi anemia (FA, n = 12), Diamond-Blackfan anemia (DBA, n = 7), and dyskeratosis congenita (DC, n = 5) from a single HSCT center. The graft source was peripheral blood stem cells (n = 19) or cord blood stem cells (n = 5). FA and DC patients received reduced-intensity conditioning, while DBA patients had myeloablative conditioning. The median numbers of infused mononuclear cells and CD34+ cells were 14.20 × 108/kg and 4.3 × 106/kg, respectively. The median time for neutrophil and platelet recovery was 12 and 18 days, respectively. Complete donor engraftment was achieved in 23 of 24 patients. There was one primary graft failure. During a median follow-up of 27.5 months (range, 2-130 months), the overall survival in all patients was 95.8%. The incidence of grade II-III acute graft versus host disease (GvHD) and chronic GvHD was 29.2% and 16.7%, respectively. We conclude that HSCT can be a curative option for patients with IBMFS. Modification of the conditioning regimen based on the type of disease may lead to encouraging long-term outcomes.


Assuntos
Anemia Aplástica/terapia , Doenças da Medula Óssea/terapia , Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Transplante de Células-Tronco Hematopoéticas/métodos , Hemoglobinúria Paroxística/terapia , Transplante de Células-Tronco de Sangue Periférico/métodos , Adolescente , Anemia de Diamond-Blackfan/terapia , Criança , Pré-Escolar , Transplante de Células-Tronco de Sangue do Cordão Umbilical/efeitos adversos , Seleção do Doador , Disceratose Congênita/terapia , Anemia de Fanconi/terapia , Feminino , Doença Enxerto-Hospedeiro/diagnóstico , Doença Enxerto-Hospedeiro/etiologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Hematúria/diagnóstico , Hematúria/etiologia , Humanos , Lactente , Estimativa de Kaplan-Meier , Masculino , Transplante de Células-Tronco de Sangue Periférico/efeitos adversos , Condicionamento Pré-Transplante
8.
Anal Chem ; 87(19): 9810-6, 2015 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-26356097

RESUMO

(13)C NMR spectroscopic integration employing short relaxation delays and a 30° pulse width was evaluated as a quantitative tool for analyzing the components of polysorbate 80. (13)C NMR analysis revealed that commercial polysorbate 80 formulations are a complex oligomeric mixture of sorbitan polyethoxylate esters and other intermediates, such as isosorbide polyethoxylate esters and poly(ethylene glycol) (PEG) esters. This novel approach facilitates the quantification of the component ratios. In this study, the ratios of the three major oligomers in polysorbate 80 were measured and the PEG series was found to be the major component of commercial polysorbate 80. The degree of polymerization of -CH2CH2O- groups and the ratio of free to bonded -CH2CH2O- end groups, which correlate with the hydrophilic/hydrophobic nature of the polymer, were analyzed, and were suggested to be key factors for assessing the likelihood of adverse biological reactions to polysorbate 80. The (13)C NMR data suggest that the feed ratio of raw materials and reaction conditions in the production of polysorbate 80 are not well controlled. Our results demonstrate that (13)C NMR is a universal, powerful tool for polysorbate analysis. Such analysis is crucial for the synthesis of a high-quality product, and is difficult to obtain by other methods.


Assuntos
Espectroscopia de Ressonância Magnética/métodos , Polissorbatos/química , Ésteres/análise , Isossorbida/análise , Ácido Oleico/análise , Polietilenoglicóis/análise , Polimerização
9.
Nat Med ; 21(6): 563-71, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25962120

RESUMO

Relapse is the leading cause of mortality in children with acute lymphoblastic leukemia (ALL). Among chemotherapeutics, thiopurines are key drugs in ALL combination therapy. Using whole-exome sequencing, we identified relapse-specific mutations in the phosphoribosyl pyrophosphate synthetase 1 gene (PRPS1), which encodes a rate-limiting purine biosynthesis enzyme, in 24/358 (6.7%) relapsed childhood B cell ALL (B-ALL) cases. All individuals who harbored PRPS1 mutations relapsed early during treatment, and mutated ALL clones expanded exponentially before clinical relapse. Our functional analyses of PRPS1 mutants uncovered a new chemotherapy-resistance mechanism involving reduced feedback inhibition of de novo purine biosynthesis and competitive inhibition of thiopurine activation. Notably, the de novo purine synthesis inhibitor lometrexol effectively abrogated PRPS1 mutant-driven drug resistance. These results highlight the importance of constitutive activation of the de novo purine synthesis pathway in thiopurine resistance, and they offer therapeutic strategies for the treatment of relapsed and thiopurine-resistant ALL.


Assuntos
Retroalimentação Fisiológica/efeitos dos fármacos , Leucemia de Células B/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Ribose-Fosfato Pirofosfoquinase/genética , Adolescente , Criança , Pré-Escolar , Exoma/genética , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Lactente , Leucemia de Células B/tratamento farmacológico , Leucemia de Células B/patologia , Masculino , Mercaptopurina/administração & dosagem , Mutação , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Purinas/biossíntese , Recidiva , Ribose-Fosfato Pirofosfoquinase/química , Tetra-Hidrofolatos/administração & dosagem
10.
Zhonghua Er Ke Za Zhi ; 53(11): 817-23, 2015 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-26758319

RESUMO

OBJECTIVE: To enrich our national database with data of rare diseases by analyzing molecular diagnosis and hematopoietic stem cell transplantation (HSCT) in children with inherited bone marrow failure syndromes (IBMFS). METHOD: Next-generation sequencing (NGS)-based genetic diagnosis panel was applied for the clinical diagnosis and management of IBMFS. Retrospective analysis was performed on clinical and genetic data of 17 consecutive children who received HSCT over a long time interval (November. 2005-June 2015). RESULT: Three patients were diagnosed only by clinical manifestation before 2012. After that NGS-based genetic diagnosis panel was used to identify IBMFS-related genes in 12/14.IBMFS patients (except two Diamond-Blackfan anemia (DBA) patients). Two Fanconi anemia (FA) patients were confirmed to be new variations through family-genotype-analysis and 3 families accepted prenatal diagnosis to avoid birth of affected fetuses. Seventeen IBMFS patients (10 FA,5 DBA and 2 dyskeratosis congenital (DKC)) were treated with HSCT from matched sibling donors (n=2), matched unrelated donors (n=8) or mismatched unrelated donors (n=7). The source of stem cells for transplantation included peripheral blood (n=12) and cord blood (n=5). With regard to the conditioning regimens, FA and DKC patients received fludarabine-based reduced intensity conditioning, while DBA patients received classical busulfan-based myeloablative conditioning. Median age at the time of HSCT was 36 months (7-156 months). The number of infused mononuclear cells and CD34⁺ cells was (10.6 ± 6.7) × 108 and (5.9 ± 7.0) × 106 per kilogram of recipient body weight, respectively. The median number of days to neutrophil recovery was 13 days after HSCT (range: 10-19 days). Platelet recovery was faster in the PBSCT group than in the CBT group ((16.3 ± 6.0) days vs. (30.0 ± 17.1) days,t=-2.487,P=0.026). During a median follow-up of 17 months (range: 2-114 months), except one FA patient who was transplanted with HLA-matched unrelated cord blood (CB) died from pneumonia and heart failure because of engraftment failure, other 16 children are alive after the successful HSCT. The failure-free survival rate of the patients three years after HSCT was 94%. CONCLUSION: NGS-based molecular diagnosis technology and effective HSCT have significantly facilitated the treatment of children with IBMFS in our country, and our national database about this rare disease is to be further exploited.


Assuntos
Transplante de Células-Tronco Hematopoéticas , Hemoglobinúria Paroxística/diagnóstico , Hemoglobinúria Paroxística/terapia , Anemia Aplástica , Anemia de Diamond-Blackfan/terapia , Doenças da Medula Óssea , Criança , Disceratose Congênita/terapia , Anemia de Fanconi/terapia , Sangue Fetal , Hemoglobinúria Paroxística/genética , Humanos , Estudos Retrospectivos , Irmãos , Taxa de Sobrevida , Condicionamento Pré-Transplante , Doadores não Relacionados , Vidarabina/análogos & derivados , Vidarabina/uso terapêutico
11.
Inflammation ; 38(2): 800-11, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25227280

RESUMO

Macrophage polarization is a dynamic and integral process of tissue inflammation and remodeling. Here we demonstrate an important role of Aurora kinase A in the regulation of inflammatory M1 macrophage polarization. We found that there was an elevated expression of Aurora-A in M1 macrophages and inhibition of Aurora-A by small molecules or specific siRNA selectively led to the suppression of M1 polarization, sparing over the M2 macrophage differentiation. At the molecular level, we found that the effects of Aurora-A in M1 macrophages were mediated through the down-regulation of NF-κB pathway and subsequent IRF5 expression. In an autoimmune disease model, experimental autoimmune encephalitis (EAE), treatment with Aurora kinase inhibitor blocked the disease development and shifted the macrophage phenotype from inflammatory M1 to anti-inflammatory M2. Thus, this study reveals a novel function of Aurora-A in controlling the polarization of macrophages, and modification of Aurora-A activity may lead to a new therapeutic approach for chronic inflammatory diseases.


Assuntos
Aurora Quinase A/fisiologia , Polaridade Celular/fisiologia , Encefalomielite Autoimune Experimental/enzimologia , Macrófagos/enzimologia , Inibidores de Proteínas Quinases/farmacologia , Animais , Polaridade Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Encefalomielite Autoimune Experimental/tratamento farmacológico , Humanos , Macrófagos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Piperazinas/farmacologia , Piperazinas/uso terapêutico , Inibidores de Proteínas Quinases/uso terapêutico
13.
Oncotarget ; 5(21): 10732-44, 2014 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-25313141

RESUMO

B cell acute lymphoblastic leukemia (B-ALL) is the most common hematological malignancy diagnosed in children, and blockade of the abnormally activated PI3Kδ displayed promising outcomes in B cell acute or chronic leukemias, but the mechanisms are not well understood. Here we report a novel PI3Kδ selective inhibitor X-370, which displays distinct binding mode with p110δ and blocks constitutively active or stimulus-induced PI3Kδ signaling. X-370 significantly inhibited survival of human B cell leukemia cells in vitro, with associated induction of G1 phase arrest and apoptosis. X-370 abrogated both Akt and Erk1/2 signaling via blockade of PDK1 binding to and/or phosphorylation of MEK1/2. Forced expression of a constitutively active MEK1 attenuated the antiproliferative activity of X-370. X-370 preferentially inhibited the survival of primary pediatric B-ALL cells displaying PI3Kδ-dependent Erk1/2 phosphorylation, while combined inhibition of PI3Kδ and MEK1/2 displayed enhanced activity. We conclude that PI3Kδ inhibition led to abrogation of both Akt and Erk1/2 signaling via a novel PI3K-PDK1/MEK1/2-Erk1/2 signaling cascade, which contributed to its efficacy against B-ALL. These findings support the rationale for clinical testing of PI3Kδ inhibitors in pediatric B-ALL and provide insights needed to optimize the therapeutic strategy.


Assuntos
Benzimidazóis/farmacologia , MAP Quinase Quinase 1/antagonistas & inibidores , MAP Quinase Quinase 2/antagonistas & inibidores , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Leucemia-Linfoma Linfoblástico de Células Precursoras B/tratamento farmacológico , Inibidores de Proteínas Quinases/farmacologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Purinas/farmacologia , Apoptose/efeitos dos fármacos , Benzimidazóis/química , Western Blotting , Proliferação de Células/efeitos dos fármacos , Desenho de Drogas , Humanos , Imunoprecipitação , MAP Quinase Quinase 1/metabolismo , MAP Quinase Quinase 2/metabolismo , Estrutura Molecular , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Leucemia-Linfoma Linfoblástico de Células Precursoras B/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patologia , Inibidores de Proteínas Quinases/química , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Purinas/química , Transdução de Sinais/efeitos dos fármacos , Células Tumorais Cultivadas
14.
Clin Transplant ; 28(11): 1225-33, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25123053

RESUMO

Graft failure and survival are the major problems for patients with aplastic anemia undergoing hematopoietic stem cell transplantation (HSCT). Previous studies showed that anti-HLA antibodies negatively impact engraftment in HSCT. This retrospective study of 51 pediatric patients with acquired aplastic anemia who underwent allogeneic HSCT at a single institution between 2006 and 2012 investigated the influence of anti-HLA antibodies on the outcome of HSCT. Serum samples collected before HSCT were tested for the presence of anti-HLA antibodies. Pre-existing anti-HLA antibodies were detected in 54.9% (28/51) of patients, among whom 39.2% (20/51) had anti-HLA class I antibodies. Anti-HLA antibodies were associated with worse five-yr survival (78.6% vs. 100%, p = 0.021) and higher treatment-related mortality (21.4% vs. 0%, p = 0.028) compared with antibody-negative patients. Anti-HLA class I antibody-positive patients had poorer five-yr survival (75.0%) than anti-HLA class I&II antibody-positive and antibody-negative patients (87.5% and 100.0%, respectively, p = 0.039). Presence of anti-HLA class I antibodies (p = 0.024) and older age (10 yr or more; p = 0.027) significantly increased the risk of post-HSCT mortality. Pre-existing anti-HLA antibodies negatively affect the outcome of HSCT in pediatric patients with aplastic anemia. Routine testing for anti-HLA antibodies concurrent with efficient treatment should be conducted prior to HSCT.


Assuntos
Anemia Aplástica/sangue , Anemia Aplástica/terapia , Anticorpos/sangue , Antígenos HLA/imunologia , Transplante de Células-Tronco Hematopoéticas , Adolescente , Anemia Aplástica/mortalidade , Criança , Pré-Escolar , Feminino , Teste de Histocompatibilidade , Humanos , Masculino , Estudos Retrospectivos , Taxa de Sobrevida , Resultado do Tratamento
15.
Cell Immunol ; 290(1): 138-44, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24978614

RESUMO

Inducible regulatory T-cells (iTReg) can be generated from CD4(+)Foxp3(-) naïve conventional T-cells by a combination of TGF-ß and T-cell receptor (TCR) signaling. It is of enormous clinical importance to identify agents that can promote the generation and differentiation of functional iTreg cells. We have established a phenotypic screening platform to identify new compounds that can promote the TGFß-mediated iTreg differentiation. We have found Kenpaullone, a potent CDK1, CDK2 and CDK5 inhibitor, as new enhancer for iTreg cell differentiation. Kenpaullone promotes iTreg cell differentiation through increased and prolonged transcription of foxp3 gene by enhancing TGFß-Smad3 signaling pathway. Thus, we have demonstrated that CDK2 is the biological target of Kenpaullone and proven that CDK2 is a novel negative regulator of iTreg cell differentiation.


Assuntos
Quinase 2 Dependente de Ciclina/antagonistas & inibidores , Proteína Smad3/imunologia , Linfócitos T Reguladores/imunologia , Fator de Crescimento Transformador beta/imunologia , Animais , Apoptose , Benzazepinas/farmacologia , Proteína Quinase CDC2/antagonistas & inibidores , Proteína Quinase CDC2/biossíntese , Proteína Quinase CDC2/genética , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/imunologia , Proliferação de Células , Células Cultivadas , Quinase 2 Dependente de Ciclina/biossíntese , Quinase 2 Dependente de Ciclina/genética , Quinase 5 Dependente de Ciclina/antagonistas & inibidores , Quinase 5 Dependente de Ciclina/genética , Quinase 3 da Glicogênio Sintase/antagonistas & inibidores , Glicogênio Sintase Quinase 3 beta , Indóis/farmacologia , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Interferência de RNA , RNA Interferente Pequeno , Receptores de Antígenos de Linfócitos T/imunologia , Transdução de Sinais/imunologia , Transcrição Genética/efeitos dos fármacos
16.
Se Pu ; 32(2): 151-6, 2014 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-24822449

RESUMO

An analytical method using ultra-high performance liquid chromatography (UPLC) was developed for qualitative and quantitative analysis of 25 illegally added drugs in diet health foods. The diet food samples were extracted using 40 mL methanol by sonication. After centrifugation, the supernatants were separated on a Waters HSS T3 column with gradient elution at a flow rate of 0.3 mL/min, coupling with diode array detection (DAD) in wavelength range from 200 nm to 400 nm. The binary mobile phase was acetonitrile and 10 mmol/L ammonium acetate solution (containing 0.1% formic acid). The correlation coefficient of standard curve for each drug in linearity range was not less than 0. 997, as well as the recoveries of all the drugs in diet health foods were 70.7%-104% with the relative standard deviations (RSDs) of 0.132%-5.03% at three spiked levels. Seventeen diet food samples were tested, in which phenolphthalein was found in three samples and emodin was found in one sample. The method is specific, easy, quick, and suitable for confirmation of the 25 illegally added drugs in diet health foods.


Assuntos
Cromatografia Líquida de Alta Pressão , Resíduos de Drogas/análise , Contaminação de Alimentos/análise , Alimentos Orgânicos/análise , Emodina/análise , Fenolftaleína/análise
17.
Zhongguo Dang Dai Er Ke Za Zhi ; 15(7): 509-13, 2013 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-23866268

RESUMO

OBJECTIVE: To study the expression of zinc finger protein X-linked (ZFX) in bone marrow mononuclear cells (BMMCs) of children with B lineage acute lymphoblastic leukemia (B-ALL) and its relationship with prognosis. METHODS: The expression of ZFX in human leukemia cell lines (REH, HL-60, NB(4) and K562) was measured by Western blot. ZFX gene was cloned by PCR from one patient and DNA sequencing technology was used to confirm it. Real-time PCR was used for detecting ZFX mRNA expression in the BMMCs of 82 children with newly-diagnosed B-ALL, 24 children with complete remission (CR) after induction therapy and 64 control children (fracture or congenital heart disease patients). According to the presence of bone marrow or central nervous system relapse during a follow-up of 3 years, the patients were identified as having a good or poor prognosis. Their ZFX mRNA levels in BMMCs at diagnosis were compared. RESULTS: ZFX protein was expressed in human leukemia cell lines REH, HL-60, NB(4) and K562. ZFX mRNA expression was significantly higher in the newly-diagnosed ALL group than in the control group (P < 0.01). ZFX mRNA expression in the ALL CR group was significantly reduced compared with the newly-diagnosed ALL group (P < 0.01). Children with a poor prognosis had significantly higher ZFX mRNA levels at diagnosis than those with a good prognosis (P < 0.05). CONCLUSIONS: ZFX is over-expressed in children with B-ALL and its levels are higher in those with a poor prognosis than those with a good prognosis, which suggests that ZFX is important in the prognosis evaluation of B-ALL.


Assuntos
Fatores de Transcrição Kruppel-Like/fisiologia , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patologia , Adolescente , Linhagem Celular Tumoral , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Fatores de Transcrição Kruppel-Like/análise , Fatores de Transcrição Kruppel-Like/genética , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras B/metabolismo , Prognóstico , Reação em Cadeia da Polimerase em Tempo Real
18.
Guang Pu Xue Yu Guang Pu Fen Xi ; 31(9): 2462-6, 2011 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-22097849

RESUMO

In the present study, based on the leaf-level hyperspectral data of BaiMu, LeiZhu and WuHuanZi, the authors come up with two solutions through the theory of statistics; the first one is that optimal discriminating band between tree species is extracted by mean interval confidence, the other one is that tree species is discriminated by the Manhattan distance and the Min Max interval similarity. The research results showed that (1) the optimal discriminating bands between BaiMu and LeiZhu are around 350-446, 497-527, 553-1 330, 1 355-2 400 and 2 436-2 500 nm; the optimal discriminating bands between BaiMu and WuHuanZi are around 434-555, 580-1 903, 1 914-2 089, 2 172-2 457 and 2 475-2 500 nm; the optimal discriminating bands between LeiZhu and WuHuanZi are around 434-555, 580-1 903, 1 914-2 089, 2 172-2 457 and 2 475-2 500 nm; and this result is helpful for us to find maximum difference to identifying tree species respectively. (2) In these optimal discriminating bands, we find that the Manhattan distance between the same species is far less than the different species; but the Min-Max interval similarity between the same species is far more than the different species, so this result could help us to discriminate and identify different types of tree species effectively.


Assuntos
Análise Espectral , Árvores/classificação , Intervalos de Confiança , Folhas de Planta
19.
Guang Pu Xue Yu Guang Pu Fen Xi ; 31(11): 3010-3, 2011 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-22242506

RESUMO

In the present study, based on the leaf-level hyperspectral data of MaoZhu, LeiZhu and XiaoShunZhu, We come up with two solutions to discrimination through the theory of non-parametric test and pattern recognition; the first one is that optimal discriminating band between bambusoideae species is extracted by Mann-Whitney non-parametric test, the other is that bambusoideae species is discriminated by the support vector machine. The research results showed that (1) the optimal discriminating band between MaoZhu and LeiZhu is around 503-655, 689-732, 757-1 000, 1 038-1 084, 1 238-1 311, 1 404-1 591, 1682-1 800, 1 856-1 904, and 1 923-2 500 nm; the optimal discriminating band between MaoZhu and XiaoShunZhu is around 350-386, 731-1 430, 1 584-1 687, and 1 796-1 873 nm; the optimal discriminating band between LeiZhu and XiaoShunZhu is around 355-356, 498-662, 689-745, and 1 344-2 500 nm; and it can eliminate 30.0%, 57.7%, and 35.8% of the invalid distinction between bands by Mann-Whitney non-parametric test method. (2) In these optimal discriminating bands, we found that the accuracy of bambusoideae discrimination is 98.4%, 93.5%, and 95.1%, the generalization accuracy is 93.3%, 90.0%, and 86.7% by sequential minimal optimization algorithm. It indicates that this method is valid for selecting feature band and discriminating bambusoideae species.


Assuntos
Bambusa/classificação , Análise Espectral , Máquina de Vetores de Suporte , Algoritmos , Folhas de Planta , Estatísticas não Paramétricas
20.
Guang Pu Xue Yu Guang Pu Fen Xi ; 30(8): 2157-60, 2010 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-20939328

RESUMO

During Raman spectroscopy analysis, the organic molecules and contaminations will obscure or swamp Raman signals. The present study starts from Raman spectra of prednisone acetate tablets and glibenclamide tables, which are acquired from the BWTek i-Raman spectrometer. The background is corrected by R package baselineWavelet. Then principle component analysis and random forests are used to perform clustering analysis. Through analyzing the Raman spectra of two medicines, the accurate and validity of this background-correction algorithm is checked and the influences of fluorescence background on Raman spectra clustering analysis is discussed. Thus, it is concluded that it is important to correct fluorescence background for further analysis, and an effective background correction solution is provided for clustering or other analysis.


Assuntos
Glibureto/análise , Prednisona/análise , Análise Espectral Raman , Algoritmos , Análise por Conglomerados , Fluorescência , Análise de Componente Principal , Soluções , Comprimidos
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