Thrombocytosis is a frequently occured disease in pediatrics.It can be classified into primary thrombocythemia and reactive thrombocytosis.The former can be divided into familial thrombocythemia and essential thrombocythemia.Essential thrombocythemia,chronic myeloproliferative disorders,is very rare in children,but may be associated with thromboembolic and hemorrhagic complications.Reactive thrombocytosis is very common and is due to a variety of conditions,such as acute and chronic infections,bleeding,iron deficiency and so on.Treatment of reactive thrombocytosis should be directed to the primary disease.This review briefly describes thrombocytosis etiology,pathogenesis,prognosis,and treatment of research progress.

Acute mveloid leukemia(AML) is a heterogeneous group of leukemias that arise from clonal transformation of hematopoietic precursors.This review summarizes classification of pediatric AML,more precise risk-group stratification,ongoing phase Ⅲ studies and minimal residual diseasse monitoring.In addition,we discuss the opportunities for innovative chemotherapy drugs in refractory AML and relapsed AML,such as gemtuzumab ozogamicin,liposomal daunorubicin,clofarabine,cladribine and lmatinib.Finally,the roles of allogeneic hematopoietic stem cell transplantation in pediatric AML are also discussed.

Objective To prepare butenafine hydrochloride plastics,investigate the prescription composition and make a quality control standard for the preparation.Methods Film-forming time and appearance quality as the evaluation index,the quality control standard of butenafine hydrochloride according to the Chinese pharmacopoeia two section ( 2010 edition ) was made.ResuIts The prescription of butenafine hydrochloride plastics was identified as:1%butenafine hydrochloride(w/w),10%glycerol(w/w),3%carbomer 971PNF(w/w),0.1% ethyl p-hydroxybenzoate(w/w),moderate anhydrous sodium sulfite(pH adjusting agent) and 95% ethanol (solution).The preparation was colorless,transparent and viscous semi-solid with pH4.5.A content determination method of butenafine hydrochloride with HPLC was established and the result was stable and reliable .ConcIusion The butenafine hydrochloride has several advantages such as preparation simply , stable property,application convenience and quality control.It is a potential preparation to develop.

Objective To analysis the clinical characteristics and the long-term effect of children with acute lymphoblastic leukemia (ALL).Methods From 2005 to 2010,80 newly diagnosed ALL children were enrolled and treated with protocol based on ALL-BFM2002.The five-years overall survival (OS)and event-free survival(EFS) were analyzed by the method of Kaplan-Meier.Results For the 80 patients,male to female ratio is 1.22∶1.The median age was 4.3 years.33 were in standard risk(41.2％),37 were in medium risk(46.3％),and 10 were in high risk(12.5％).22 had white blood cell count ≥20 x 109/L(27.5％).three patients with BCR-ABL translocation(3.8％),one patient with MLL gene rearrangement(1.3％),17 patients with TEL-AML translocation (21.3％).During induction therapy,79 patients (98.8 ％) achieved complete remission(CR).The five-years OS and EFS were (85.9 ± 4.0) ％ and (79.2 ± 4.7) ％ respectively.The five-years EFS:SR group (86.6 ± 6.4) ％,IR group (81.1 ± 6.4) ％,HR group (48.0 ± 16.4) ％.The difference among risk groups was statistically significant(x2 =7.03,P ＜0.05).12 patients relapsed(15.0％),the median time from diagnosis to relapse was 23.5 months.11 patients died (13.8 ％).Conclusion According to stratification by risk factors and risk-adapted therapy,the quality of ALL children's life had improved.

Objective To investigate the effects of tongue cancer resistance-associated protein 1 (TCRP1) in proliferation of chronic myeloid leukemia cells (CML),and explore the new thoughts of pathogenesis of CML.Methods The expression of TCRP1 was detected in the peripheral blood mononuclear cells (PBMC) of CML with real-time quantitative polymerase chain reaction (PCR) and Western blot.After the expression of TCRP1 was interfered in K562 cells,the proliferation of cells was detected by 3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and soft agar colony forming assay,and the expression of protein kinase B (AKT) and its phosphorylation were tested by Western blot.Results In PBMC of CML patients,the mRNA and protein levels of TCRP1 were significantly higher than those of normal controls.The results of MTS assay and soft agar colony forming assay showed that the proliferation of K562 cells was significantly decreased after the expression of TCRP1 was interfered.After knockdown of TCRP1 in K562 cells,the phosphorylation of AKT was significantly decreased while the expression of total AKT did not change.Conclusions The expression of TCRP1 was increased in CML cells.High expression of TCRP1 might contribute to proliferation of K562 cells via the phosphorylation of AKT.

Engraftment syndrome (ES) is a clinical syndrome that occurs after hematopoietic stem cell transplantation and during the recovery process of neutrophils.The main clinical manifestations include non-infectious fever, rash, capillary leakage and non-cardiogenic pulmonary edema, which may be similar with many early complications after transplantation.Therefore, it is sometimes difficult to be diagnosed and differentiated among different kinds of complications.Typical ES is self-limiting and has good response to steroids.However, patients with ES may result in encephalopathy and multi-organ failure if it is untreated without notice.In this review, we discussed the pathophysiological mechanisms, clinical manifestations, diagnosis and differential diagnosis, risk factors, treatment and prognosis of ES, aiming to provide guidance and reference for clinicians.

Objective To investigate the changes of CD4 +CD25highFoxp3 +regulatory T (Treg) cells and their significance in immune escape of childhood B-cell acute lymphocytic leukemia ( B-ALL ) . Methods Forty-two children with B-ALL and twenty-eight age-matched healthy children were enrolled in this study.Flow cytometry analysis was performed to evaluate the proportion of CD 4 +CD25high Foxp3 +Treg cells as well as CD4 +CD25high ICOS+Foxp3 +and CD4 +CD25high ICOS-Foxp3 +subsets in peripheral blood samples.The expression of associated molecules including IL-10, TGF-β, IL-35, TGF-βRII, ICOS and CD28 at protein level were also measured by flow cytometry analysis .The transcription level of Smad3/4, TIEG1 and Itch by CD4 +T cells were determined by quantitative real-time PCR.The concentration of TGF-βin plasma was detected by enzyme-linked immunosorbent assay.Results (1)The proportion of CD4 +CD25highFoxp3 +Treg cells in children with B-ALL were significantly higher than those of health subjects (P<0.05).The proportion of both ICOS +Foxp3 +and ICOS -Foxp3 +subsets were increased in comparison with those of control group (P<0.05), while the ratio of ICOS +Foxp3 +to ICOS-Foxp3 +was decreased (0.73 ±0.21 vs 1.87 ±0.59, P<0.05).(2) The expression of Foxp3, TGF-β, IL-10 and IL-35 by ICOS+Foxp3 +Treg cells and the expression of membrane bound TGF-βby ICOS -Foxp3 +Treg cells were significantly increased in children with B-ALL (P<0.05).However, the expression of Foxp3 by ICOS -Foxp3 +Treg cells showed no significant difference between the two groups (P>0.05).(3)The concentra-tion of TGF-βin plasma from children with B-ALL were higher than those from control group [ ( 25 .83 ± 12.65) ng/ml vs (8.59 ±5.73) ng/ml, P<0.05].The expression of TGF-βRII and its associated mole-cules (Smad3/4, TIEG1 and Itch) by CD4 +T cells were significantly up-regulated.Moreover, an increased expression of ICOS and CD28 by CD4 +CD25highFoxp3 +Treg cells were also observed in children with B-ALL (P<0.05).Conclusion The hyper-activity of TGF-β, ICOS and CD28 signaling might be closely associ-ated with the increased proportion of CD4 +CD25high Foxp3 +Treg cells and the imbalance of its subsets in children with B-ALL.

PURPOSE: The role of systemic sensitization in the pathophysiology of chronic rhinosinusitis with nasal polyps (CRSwNP) remains elusive. This study sought to characterize the pattern of cytokines in polyp tissues from atopic and nonatopic patients with CRSwNP. METHODS: Atopic and nonatopic polyp and normal tissues were collected from 70 CRSwNP patients and 26 control subjects, respectively. The distribution of inflammatory cells (eosinophils, neutrophils, mast cells, etc.) were examined using immunohistochemistry, the mRNA levels of the transcription factors GATA-3, T-bet, RORc, and FOXP3 were determined using quantitative real-time polymerase chain reaction. The levels of inflammatory mediators (IFN-gamma, IL-5, IL-17A, etc.) in tissue homogenates were measured using enzyme-linked immunosorbent assay (ELISA). Moreover, the levels of inflammatory mediators in the supernatant of anti-IgE stimulated polyp tissues were measured using ELISA. RESULTS: Atopic CRSwNP patients were characterized by increased eosinophil accumulation, enhanced eosinophilic inflammation (elevated IL-5, ECP, and total IgE), and significantly increased GATA-3 mRNA levels (P<0.05), whereas both atopic and non-atopic CRSwNP patients showed decreased FOXP3 mRNA expression (P<0.05). After addition of anti-IgE stimulation, atopic CRSwNP patients produced more IL-5, IL-2, IL-10, IL-17A, and PGD2 in the supernatant of stimulated polyp tissues than nonatopic CRSwNP patients did. CONCLUSIONS: Atopic and nonatopic CRSwNP patients may possess the patterns of inflammatory response in polyp tissues.
China* ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; Humans ; Immunoglobulin E ; Immunohistochemistry ; Inflammation ; Interleukin-10 ; Interleukin-17 ; Interleukin-2 ; Interleukin-5 ; Mast Cells ; Nasal Polyps* ; Neutrophils ; Polyps ; Prostaglandin D2 ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Transcription Factors

Objective:To investigate the effects of Notch1 signaling on histone acetylation of Foxp3 gene and its roles in regulating regulatory T (Treg) cells in children with acute B-cell precursor lymphoblastic leukemia (BCP-ALL).Methods:Blood samples were collected form 38 children with BCP-ALL before treatment and 15 age-matched healthy children (control group). Flow cytometry was performed to detect the proportion of peripheral blood CD4 + CD25 hiFoxp3 + Treg cells and the expression of Foxp3, cytotoxic lymphocyte antigen 4 (CTLA4), glucocorticoid-induced tumor necrosis factor receptor (GITR), CD39 and Notch1 at protein level. Histone 4 acetylation (H4Ac) at Foxp3 gene promoter and the binding abilities of Foxp3 gene promoter to NICD1 and p300 in CD4 + T cells were measured by chromatin immunoprecipitation. Quantitative real-time PCR was performed to detect the expression of Foxp3, presenilin 1 (PSEN1), mastermind-like transcriptional coactivator 1 (MAML1), SKI-interacting protein (SKIP), F-box and WD40 domain protein 7 (FBXW7), glycogen synthase kinase-3 beta (GSK3β) and IKAROS at mRNA level in CD4 + T cells. The concentrations of TGF-β and IL-10 in plasma were evaluated by ELISA. Results:(1) The proportion of peripheral blood CD4 + CD25 hiFoxp3 + Treg cells, the expression of differentiation- and function-associated molecules (Foxp3, CTLA4, GITR and CD39) and the concentrations of TGF-β and IL-10 in plasma were higher in the BCP-ALL group than in the control group ( P<0.05). (2) In children with acute BCP-ALL, H4Ac at Foxp3 promoter and the binding abilities of Foxp3 gene promoter to NICD1 and p300 were significantly increased as compared with those in control group( P<0.05). The binding abilities of Foxp3 gene promoter to NICD1 and p300 were positively correlated with the expression of Foxp3 at mRNA level ( r=0.58 and 0.46, both P<0.05). After competitive inhibition, the three aforementioned indexes in the acute BCP-ALL group were significantly lower than those in untreated group ( P<0.05); the binding ability of Foxp3 gene promoter to NICD1 in the control group was also significantly lower than that in untreated control group ( P<0.05), but no statistical differences in the other two indexes were found between the control groups with or without treatment ( P>0.05). ⑶ Compared with the control group, the expression of Notch1, PSEN1, MAML1 and SKIP in CD4 + T cells were elevated significantly ( P<0.05), while the transcription level of negative regulatory factor FBXW7 was decreased remarkably in children with acute BCP-ALL ( P<0.05). No statistical differences in the expression of GSK3β or IKAROS were found between the two groups ( P>0.05). Conclusions:Overactivation of Notch1 signaling caused by low expression of FBXW7 might be the key factor resulting in histone 4 hyperacetylation at foxp3 gene promoter and Treg cell dysfunction in children with acute BCP-ALL.

OBJECTIVE: To investigate the genetic etiology, clinical diagnosis and treatment of a child with pancytopenia, failure to thrive and pulmonary infection. METHODS: Peripheral blood samples of the child and her parents were collected. Genomic DNA was extracted. Genetic variants associated with hematological diseases were detected by high-throughput sequencing. RESULTS: Three variants of TCN2 gene were found, one of which located in exon 5 upstream(c.581-8A>T), the parents has carried this variant; one in exon 6 (c.924_927del), the variant was originated from the mother; one in exon 7 (c.973C>T), the variant has ocurred de novo. The variants pathogenic analysis combined with clinical manifestation, pancytopenia, the increase in methylmalonic acid level and increased homocysteine, the child was diagnosed with transcobalaminIIdeficiency. The patient presented with respiratory infection, which was confirmed to be pneumocystosis by lung radioscopy and pathogenic high-throughput sequencing of broncho-alveolar lavage fluid. The patient presented with acute respiratory distress syndrome during the treatment with intramuscular injection of vitamin B CONCLUSION We reported a case of Chinese child with TCNII deficiency due to novel gene variant, and analyzed the pathogenicity of the three variants. The treatment of TCNII deficiency with cobalamin should be individualized.
Amino Acid Metabolism, Inborn Errors ; Child ; Female ; Genetic Testing ; Humans ; Rare Diseases ; Transcobalamins/genetics* ; Vitamin B 12