The Factors Affecting Relapse in Pediatric B-cell Acute Lymphoblastic Leukemia Patients without Prognostic Fusion Genes Following Up for 10 years / 中国实验血液学杂志

OBJECTIVE@#To analyze the efficacy of children with B-cell acute lymphoblastic leukemia (B-ALL) without prognostic fusion genes treated by CCLG-ALL 2008, and investigate the related factors affecting the recurrence of the patients.@*METHODS@#B-ALL patients without prognostic fusion genes treated by the protocol of CCLG-ALL 2008 in our hospital from March 2008 to December 2012 were retrospectively analyzed. Follow-up time was ended in August 31, 2019. The median follow-up time was 92 months (range 0-136 months). Kaplan-Meier was used to detect the RFS, and COX multivariate regression analysis was employed to identify the independent factors affecting the recurrence of the patients.@*RESULTS@#There were 140 males and 99 females enrolled in this study. The ratio of male to female was 1.41∶1. The median age was 4.4 years old and the median number of WBC at initial stage was 4.98×109/L. There were 77 cases relapsed during the observation while 162 without relapsed, 16 cases lost to follow-up and 72 cases died. The recurrence and mortality rate was 32.22% and 30.1%, respectively, in which 45 cases died of recurrence (62.5% of the total deaths). Univariate analysis showed that the age≥6 years old, WBC >100×109/L, the bone marrow blasts on day 15≥25%, the bone marrow minimal residual disease (MRD) at week 12 >10-4, and the higher risk were the main factors affecting the recurrence of the patients (P<0.05). Multivariate COX regression analysis showed that age≥6 years old, WBC >100×109/L, bone marrow MRD >10-4 at the 12th week were the independent risk factors affecting recurrence of the patients.@*CONCLUSION@#Age, initial WBC, and bone marrow MRD at the 12th week were correlated with recurrence in children with B-ALL without prognostic fusion genes, which can be used as prognostic indices of recurrence risk in clinical.

Safety and Efficacy of VDMP Re-induction Regimen in Chinese Children with Relapsed Acute Lymphoblastic Leukemia-A Report from Jiangsu Childhood Hematology Group / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the therapeutic safety and efficacy of VDMP re-induction regimen in Chinese children with relapsed acute lymphoblastic leukemia (ALL).</p><p><b>METHODS</b>Forty-one patients with relapsed ALL were prospectively enrolled in this study. All the patients were distributed in 3 children's hospitals and treated with VDMP regimen as the first re-induction chemotherapy. Therapeutic efficacy and side-effects were analyzed.</p><p><b>RESULTS</b>The ratio of male to female was 27:14. The median age was 7.9 (2.2-15.4) years old. Patients relapsed at very early, early, and late stage were 7 cases, 11 cases, and 23 cases, respectively.The immunophenotype analysis showed that 38 cases were B-ALL, and 3 cases were T-ALL. All patients suffered from grade 4 of neutropenia and forty(97.6%) cases got infection, of them one case died. Thirty-nine(95.1%) cases had nonhematologic adverse event at least one organ involved grade 3 in 38 out of 41 cases, the VDMP therapy was completed, 34(89.5%) cases achieved a complete remission (CR), 1 case achieved partial remission(PR), and 3 cases didn't get remission. Follow-up data of 38 cases with completing VDMP chemotherapy were obtained, only one case was lost. Among 37 cases available for evaluation, 16 cases received allo-hematopoietic stem cell transplantation(allo-HSCT) after chemotherapy, and 13 patients survived, while 21 cases did not receive allo-HSCT(treated with chemotherapy only), and 8 patients survived.The overall survival rate of allo-HSCT group was significantly higher than that of those treated with chemotherapy only(P<0.05).</p><p><b>CONCLUSION</b>VDMP re-induction regimen is effective and well tolerable for pafients in the treated children with relapsed ALL. After remission, allo-HSCT is recommended with the aim of long survival.</p>

Clinical Features and Prognostic Factors of Children with Acute Lymphoblastic Leukemia in High-Risk Group / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical features and prognostic factors of pediatric acute lymphoblastic leukemia (ALL) in high-risk (HR) group.</p><p><b>METHODS</b>A total of 421 children with ALL in the Children's Hospital of Soochow University from August 2008 to March 2013 were diagnosed and treated according to the Chinese Children Leukemia Group (CCLG)-2008 Protocol. Among different risk-groups, 148 cases were stratified into the low-risk group and 191 cases were included in the moderate-risk group. Eight-two patients of the high-risk group were analyzed retrospectively for their clinical features, 5-year event-free survival (EFS) rate and overall survival (OS) rate.</p><p><b>RESULTS</b>The median follow-up times of 82 patients were 64 months(3.0-76.3 months), 55 patient achieved complete remission(CR) after 1 cycle of induction chemotherapy(CR rate 67.1%), 25 patients relapsed(30.5%) mainly in very early and early relapse phases, significantly different from the low-risk group (P=0.013), 27 pateitns died(32.9%). The 5-year pEFS and pOS were 57.20% and 58.5%, respectively. Phor BCR/ABLand MRD>10on the 33rd day in the high-risk group were 2 main factors influencing EFS and OS according to single factor analysis. Phor BCR/ABLwas an independent prognostic factor, however, the MRD value on the 33rd day was not statistically significant differente by virtue of COX regression analysis.</p><p><b>CONCLUSION</b>The clinical feature of children with ALL in high risk group display low induction CR rate, high recurrence rate and the lower 5-year pEFS. Phor BCR/ABLis regarded as an independent factor of poor prognosis.</p>

Efficacy Analysis of Allogeneic Hematopoietic Stem Cell Transplantation for Children with Severe Aplastic Anemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children with severe aplastic anemia (SAA).</p><p><b>METHODS</b>A total of 11 children with SAA were treated with HLA matched siblings (n = 7), umbilical cord blood (n = 2) and haploidentical HSCT (n = 2).</p><p><b>RESULTS</b>Among 11 children patients, 10 patients achieved engraftment, but 3 children patients experienced secondary graft failure, after donor lymphocyte infusions (DLI), they achieved engraftment again. One patient received cord blood transplantation and experienced primary graft rejection, but acquired autologous recovery. The median time for neutrophils to reach over 0.5 × 10(9)/L was 14 days (10-19 days) in the 9 children received bone marrow or bone marrow and peripheral blood allo-HSCT, while the median time for platelets to reach over 20 × 10(9)/L was 17 days (8-42 days). For the patient received double cord blood transplantation, the time of neutrophile and platelet level recovery was 16 days and 41 days, respectively.</p><p><b>CONCLUSION</b>If HLA-matched sibling donor is available, allo-HSCT can be recommended as the first line of treatment for children with SAA. It is feasible for children with SAA to receive allo-HSCT from selective donor, including cord blood and haploidentical HSCT. Donor lymphocyte infusions can improve engraftment.</p>

Expression of platelet membrane glycoproteins in pediatric idiopathic thrombocytopenic purpura and its clinical significance / 中华实用儿科临床杂志

Objective To explore the relationship between the expression of platelet membrane glycoprotein in pediatric idiopathic thrombocytopenic purpura (ITP) and its clinical significance.Methods A modified monoclonal antibody immobilization of platelet antigen (MAIPA) method was used to detect the positive expression rates of 4 platelet membrane glycoproteins (GP Ⅰ b/Ⅸ,GP Ⅰ b,GP Ⅲ a,and GP Ⅰ b) in 80 pediatric patients with ITP.The correlation was explored between the GP positive rate and the clinical efficacy in pediatric ITP.Trying to observe the correlationship between the GP positive rate of pediatric ITP in the total,the different gender,the acute and chronic and the treatment response rate in pediatric ITP respectively.Results There was a significant difference in curative rate statistically between the GP positive group and the GP negative group(x2 =8.535,P ＜ 0.01) in 80 pediatric ITP patients,but no statistic difference in curative rate existed between the 36 female and 44 male(x2 =0.013,P ＞0.05).Markedly statistic difference was found in the female(x2 =4.433,P ＜ 0.05),the same to the male (x2 =4.156,P ＜ 0.05).Meanwhile,there was an extremely statistic difference between 67 acute and 13 chronic patients(x2 =23.513,P ＜ 0.001).Apparently statistic difference also occurred in the acute (x2 =4.157,P ＜ 0.05),but not in the chronic cases (x2 =0.410,P ＞ 0.05).Conclusions The clinical response rate is significantly correlated with the GP positive rate in pediatric ITP,but not correlated with gender.The GP positive rate can reflect the disease status of pediatric ITP to a certain extent and be used as an indicator for judging the efficacy and monitor prognosis of pediatric ITP.

The expression and clinical significance of miR-203 in pediatric acute leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the methylation, expression and clinical significance of miR-203 in pediatric acute leukemia.</p><p><b>METHODS</b>The methylation status of miR-203 promoter CpG islands was detected with methylation-specific polymerase chain reaction. The expression of miR-203 was detected by Taqman real- time quantitative polymerase chain reaction. And the clinical significance of miR-203 in pediatric acute leukemia (ALL) was also analyzed.</p><p><b>RESULTS</b>The promoter of miR-203 was unmethylated in all of 31 pediatric acute lymphoblastic leukemia, all of 15 pediatric acute myeloid leukemia (AML) and all of 23 controls. The relative expression levels of miR-203 in controls, pediatric acute leukemia, ALL and AML were 16.93±6.31, 48.97±10.38, 55.88±12.91, 24.28±9.10 respectively. The results indicated that miR-203 was significantly up- regulated in pediatric acute leukemia (P=0.011) and ALL (P=0.009), not in pediatric AML (P=0.514) compared with control. The expression of miR-203 was significantly related with the gender, immunophenotype, chromosome, fusion gene, BCR-ABL, SIL-TAL1 and prednisone experiment in pediatric ALL and the gender, chromosome, fusion gene, SIL-TAL1 in pediatric acute leukemia (P<0.05). And in risk stratification pairwise comparisons, the expression of miR-203 in the medium-risk and high-risk groups appeared significantly different (P=0.022).</p><p><b>CONCLUSION</b>miR-203 may not be regulated with methylation mechanism in pediatric acute leukemia. miR- 203 may be a protooncogene involved in the formation of pediatric acute leukemia and ALL. Further analyses indicated that high expression of miR-203 may be associated with poor prognosis of pediatric ALL and acute leukemia.</p>

Clinical significance of glucocorticoid induction test in Chinese childhood acute lymphoblastic leukemia / 中华儿科杂志

<p><b>OBJECTIVE</b>Acute lymphoblastic leukemia (ALL) is the most common childhood cancer, while glucocorticoid (GC) is a critical component in multi-agent chemotherapy protocols currently used for the treatment of ALL. The purpose of this study was to investigate the relationship between the glucocorticoid induction test and the clinical features and the prognosis of Chinese childhood ALL.</p><p><b>METHOD</b>The study recruited 309 hospitalized patients (187 male and 122 female) with childhood ALL, the sex, age, initial WBC count, immunophenotype, chromosome and gene expression were recorded. After diagnosis, all patients received GC induction test for 7 days. Then they were divided into prednisone good response (PGR) group and prednisone poor response (PPR) group according to the peripheral lymphoblast count on D8. Early responses to chemotherapy and treatment outcomes of the patients in the two groups were also analyzed.</p><p><b>RESULT</b>Of the 309 patients, 263 belonged to PGR group and 46 belonged to PPR group. Initial WBC count was higher in PPR group than in PGR group (86.30×10(9)/L vs. 30.97×10(9)/L, P < 0.01) . B lineage ALL showed more sensitive to GC than T-ALL (86.6% vs. 60%, P < 0.05). Different initial-risk-group's sensitivity to GC differed from one another (high-risk:51.4%, medium-risk: 82.7%, standard risk: 93.7%, P < 0.0125). There was no significant difference between two groups in chromosomal karyotypes (P > 0.05). BCR-ABL positive ALL showed lower sensitivity to GC (P < 0.05) , while MLL, TEL-AML1, E2A-PBX1 positive rates in two groups were of no statistical significance (P > 0.05). Bone marrow was reviewed on D15 and D33, and the CR rates in PGR group were significantly higher than that in PPR group (D15: 60.5% vs. 32.6%, D33: 94.6% vs. 73.3%, P < 0.01) ; Minimal residual disease (MRD) levels were examined on D33, W12, and both were much lower in PGR group (D33: P < 0.01, W12: P < 0.05). Of the PGR group 215 patients (81.7%) remained continuously in complete remission (CCR) while only 28 cases (60.9%) in PPR group did so. The CCR rate was much higher in PGR group than that in PPR group (P < 0.01).</p><p><b>CONCLUSION</b>Closely related to clinical features and the outcomes of treatment, GC induction test is also an important prognostic factor in Chinese childhood ALL.</p>

The effect of the adverse events with thiopurine S-methyltransferase gene mutation on outcome of childhood acute lymphoblastic leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate thiopurine S-methyltransferase (TPMT) activity and gene promoter polymorphism to probe its significance of individual chemotherapy in acute lymphoblastic leukemia (ALL) children.</p><p><b>METHODS</b>HPLC method was carried out to determine TPMT activity (n=100), which activity at newly diagnosed. At the same time determination of TPMT activity in healthy children (n=180), these children come from the health care clinic. Using online primer3 software design primers, PCR products were purified. To sequence TPMT gene of the patients with clinical events(n=30). According to the method to analysis of correlation between TPMT activity and toxicity.</p><p><b>RESULTS</b>The average TPMT activities were (31.72±10.31) nmol·g⁻¹Hb·h⁻¹ and (30.70±9.67) nmol·g⁻¹Hb·h⁻¹ in ALL and healthy groups respectively, without gender differences of TPMT activities (P=0.45) in both groups. The TPMT activity with clinical events in newly diagnosed ALL patients (n=30) was (24.07±11.43) nmol·g⁻¹Hb·h⁻¹. There are significant differences of TPMT activities between severe bone marrow suppression [(20.96±7.24) nmol·g⁻¹Hb·h⁻¹] and ALL patients with clinical events groups (P<0.05). The TPMT activity of (40.46±8.18) nmol·g⁻¹Hb·h⁻¹ in recurrence children was also significantly different (P<0.05). TPMT activity in severe liver toxicity group was not significantly different (P=0. 930). Of TPMT gene sequencing in ALL patients with clinical events, only 3 children were heterozygosity mutations of TPMT*3C, while others homozygous genotype. There were significant differences of TPMT activities between heterozygosity genotype [(11.99±1.32) nmol·g⁻¹Hb·h⁻¹] and homozygous genotype groups [(24.95±11.32) nmol·g⁻¹Hb·h⁻¹] (P<0.05). There were five kinds of variations at the vicinity of the promoter region of -100 of tandem repeats (VNTR) polymorphism（*V3/*V3、*V3/*V4、*V4/*V4、*V5/*V5、*V4/*V6）without significant differences of TPMT activities among five kinds (P=0.186).</p><p><b>CONCLUSION</b>TPMT activity was related to the gene polymorphism. TPMT activity determination had prognostic value and guided individualized treatment.</p>

Relationship between genetic polymorphism of multidrug resistance 1 gene and the risk of childhood acute lymphocytic leukemia / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the relationship between genetic polymorphism in exon 12 C1236T, exon 21 G2677T/A and exon 26 C3435T of the multidrug resistance 1 (MDR1) gene and the risk of childhood acute lymphocytic leukemia (ALL).</p><p><b>METHOD</b>A total of 176 patients with ALL and a cohort of 170 matched healthy subjects were included. SNaPshot SNP typing was used to determine the genotypes of MDR1 C1236T, G2677T/A, C3435T. Based on the clinical data, the relationship between genetic polymorphism of MDR1 and the risk of childhood ALL was analyzed.</p><p><b>RESULT</b>There was significant difference in the distribution of genotype of MDR1 C3435T between the group of controls and cases. The mutant homozygous TT genotype was found to be associated with occurrence of ALL (P = 0.000; OR = 4.504). The data show evidence of pairwise linkage disequilibrium between the three common SNPs (C1236T-G2677T/A-C3435T). The haplotypes of TTT, TGC, CGC and CAC were predominant. The haplotype CGT distributed significantly differently between the groups of controls and cases (P = 0.034). The frequency of the haplotype TTT/TTT in the high risk group was higher than the other groups (P = 0.037).</p><p><b>CONCLUSION</b>The present findings suggest that 3435C→T polymorphism in MDR1 gene may be a genetic susceptibility factor for ALL. The haplotype of MDR1 (C1236T-G2677T/A-C3435T) could be the clinical parameter at diagnosis.</p>

Clinical significance of dynamic monitoring the minimal residual disease in childhood B-lineage acute lymphoblastic leukemia by multiparameter flow cytometry / 中国实验血液学杂志

This study was aimed to explore the clinical significance of monitoring level of minimal residual disease (MRD) at different time point in B-lineage childhood acute lymphoblastic leukemia (B-ALL). Two hundred and six children with B-ALL were enrolled in this study from Augest 2008 to September 2011 in our hospital. MRD levels were detected by flow cytometry at day 15, 33 and week 12 after initial chemotherapy. The event-free survival (EFS) for patients based on MRD levels measured at different stages of chemotherapy were compared by Kaplan Meier analyses. The results showed that out of 206 cases 196 cases achieved complete remission (CR) after induction therapy (CR rate 95.1%), the 1- and 3-year EFS rate were (92.7 ± 1.8)% and (78.7 ± 3.7)%, respectively, and the 3-year EFS rate was (85.6 ± 4.9)% in standard risk group, (82.1 ± 5.8)% in intermediate risk group and (58.1 ± 9.2)% in high risk group, there was significant statistical difference between above mentioned 3 groups (P < 0.001). The MRD analysis at different time points showed that the higher the MRD level, the lower the 3-year EFS rate of children with ALL, in which the 3-year EFS rate of MRD ≥ 10(-2) at day 15, MRD ≥ 10(-3) at day 33 and MRD ≥ 10(-3) at week 12 were significantly lower. The MRD ≥ 10(-3) at week 12 was proven to be an independent predictor by multivariate Cox proportional-hazards regression model. The 3-year EFS rate for patients with MRD < 10(-3) and MRD ≥ 10(-3) at week 12 were (86.3 ± 4.1)% vs (55.8 ± 9.1)% (P < 0.05); 8 relapsed among 98 cases with negative MRD (MRD < 10(-4)) at day 33, 19 relapsed among 108 cases with positive MRD at day 33 between the two groups for recurrence rate has significant difference (P < 0.05). It is concluded that dynamically monitoring MRD by multi-parameter flow cytometry can precisely evaluate treatment response, judge treatment outcome and predict relapse in childhood B-ALL. The MRD 10(-2) at day 15, MRD 10(-3) at day 33 and MRD 10(-3) at week 12 should be considered as the best cut-off. MRD ≥ 10(-3) at week 12 was proven to be an independent factor of poor prognosis.

Analysis of clinical features and prognostic significance of childhood T-lineage acute lymphoblastic leukemia / 中国实验血液学杂志

This study was aimed to explore the clinical features and prognosis outcome of childhood T-cell acute lymphoblastic leukemia (T-ALL). The clinical data of 38 cases of newly diagnosed T-ALL from Jan 2005 to Aug 2010 were analyzed retrospectively, and 78 cases of B-ALL with intermediate and high risk were collected as control group, then the sensitive rate of patients to prednisone pretreatment, complete remission (CR) rate at day 33 after induction chemotherapy, relapse rate and 3-year event-free survival (EFS) were compared between T-ALL and B-ALL children. The results showed that no significant statistic difference were found in distribution of age, infiltration of liver, spleen and lymph nodes as well as central nervous system disease, chromosome abnormality, expression level of fusion gene and so on between T-ALL and B-ALL groups (p > 0.05), but there were significant differences in sex and number of cases with WBC count ≥ 50 × 10(9)/L between them (p < 0.05). The sensitive rate of T-ALL and B-ALL patients to prednisone pretreatment was 51.9% and 89.3% respectively (p < 0.05). The ratio failed to achieve CR at day 33 after induction chemotherapy was 15.4% and 8.1% in the two groups (p > 0.05). The relapse rate of T-ALL and B-ALL cases was 30.8% (8/26) and 14.9% (11/74) respectively (p > 0.05). The time from CR to relapse was (9.78 ± 3.48) month and (21.28 ± 14.32) month (p < 0.05). The 3 year EFS of T-ALL cases with intermediate and high risk was (37.5 ± 17.1)% and (22.2 ± 9.8)%, while 3 year EFS of B-ALL cases was (66.7 ± 7)% and (51.7 ± 9.3)% respectively (p < 0.05) according to Kaplan-Meier survival curve. It is concluded that as compared with B-ALL cases, the male ratio and initial WBC count are higher, moreover the early response to prednisone pretreatment and 3 year EFS are poor in T-ALL cases, the prognosis outcome is poor also.

Preliminary study of gene expression profile associated with risk classification of childhood patients with acute lymphoblastic leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore genes associated with risk classification of childhood acute lymphoblastic leukemia (ALL) by gene chip technology.</p><p><b>METHODS</b>Group A and B were both composed of three newly diagnosed ALL cases with standard risk. After re-evaluation, group B was relegated to high-risk. The control group was composed of three idiopathic thrombocytopenic purpura (ITP) patients. The gene expression profiles of group A and B were studied by Illumina Human-6 Beadchip. Eighty-two ALL patients were selected as the experimental group and 21 with normal bone marrow as control group for real-time quantitative RT-PCR (RQ-PCR).</p><p><b>RESULTS</b>(1) There were 19 genes expressed differently between group B and A, including 14 up-regulated as ABCC4 and BCL11A, 5 down-regulated genes as TOP2A. (2) ABCC4 and BCL11A were validated by RQ-PCR and their expression level was higher in the high risk group than in the standard risk group (P < 0.05). The gene expression level in the group A and B was higher than that in the normal control group (P < 0.01). TOP2A was also validated by RQ-PCR and its expression level in the high risk group was lower than that in the standard risk group (P < 0.05). The gene expression level in the groups A and B was lower than that in the normal control group and the difference was statistically significance (P < 0.01). (3) There was a significant difference in the expression level of ABCC4 between the remission and unremission patients (P < 0.05). There was no significant difference in the expression level of BCL11A between different clinical indicators (P > 0.05). There was significant difference in the expression level of TOP2A between remission and prednisone good responder groups (P < 0.05).</p><p><b>CONCLUSIONS</b>Fourteen genes studied were involved in the pathogenesis and drug resistance mechanism in childhood ALL patients. Investigation of gene expression profile will be helpful for predicting drug resistance, prognosis, early intervention and target therapy in childhood ALL.</p>

Relationship between pharmacokinetics and efficacy and toxicity of daunorubicin in children with acute leukemia / 中华儿科杂志

<p><b>OBJECTIVE</b>To study relationship between daunorubicin (DNR) pharmacokinetics and efficacy and toxicity in children with acute leukemia.</p><p><b>METHODS</b>(1) The concentration of DNR in plasma of children with acute leukemia was determined by high performance liquid chromatography (HPLC)-fluorescence detection method. Plasma was sampled frequently from the start of the infusion till the end of 24 h. DNR pharmacokinetics was studied by determination of the concentrations. (2) Efficacy and toxicity were monitored in each period after chemotherapy. Laboratory studies included examination of bone marrow, white blood cell count, electrocardiogram, echocardiogram, myocardial enzymogram, the liver and kidney function.</p><p><b>RESULTS</b>(1) DNR was eliminated from plasma in a two-compartment manner. The maximum concentration was seen 1 - 3 h after infusion. Cmax was 63.50 microg/L. Tmax was 1.45 h. The concentration decreased quickly to a low level of about 11.52 microg/L from the end of 2 hours infusion. There was a large inter-individual difference in pharmacokinetic parameters of DNR. The difference of CL was 9-fold, AUC was 8-fold, Cmax was 5-fold. (2) CL of male patients [57.99 L/(h.m(2))] was significantly lower than that of female patients [93.71 L/(h.m(2))] (P < 0.05). Tmax of children older than 6 years was 1.1 h, and that of children younger than 6 years was 1.8 h (P < 0.05); Cmax of children older than 6 years was 90.77 microg/L, younger than 6 years was 57.44 microg/L (P < 0.05).</p><p><b>CONCLUSION</b>(1) There is a large inter-individual difference in pharmacokinetic parameters of DNR in children. It may predict individual variety of efficacy and toxicity. Therapeutic drug monitoring is important. (2) Male patients and children older than 6 years had a higher bioavailability and lower metabolism, toxicity may easily occur in such children, therefore they may need lower dose.</p>

Expression of CYP3A5 mRNA in children with acute leukemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>The cytochrome P450 subfamily IIIA5 (CYP3A5) gene is responsible for the metabolism of many clinically used anticancer agents. So far the studies on CYP3A5 gene has only been focused on the leukemia cell lines. This study examined the polymorphism of CYP3A5 and tried to find the possible relationship between CYP3A5 gene expression and treatment outcome or prognosis in children with acute leukemia.</p><p><b>METHODS</b>The genotype distribution of CYP3A5-6986A/G gene polymorphism was detected with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 66 children with newly diagnosed acute leukemia (AL) and 22 control individuals. Quantitative real-time RT-PCR was used to examine wt-CYP3A5 and SV1-CYP3A5 mRNA levels in the bone marrow.</p><p><b>RESULTS</b>Three genotypes of CYP3A5-6986A/G polymorphisms were found: CYP3A5*1/*1, CYP3A5*1/*3 and CYP3A5*3/*3. There were significant differences in the wt-CYP3A5 mRNA expression among the AL patients with different genotypes (p<0.05). In patients with acute lymphocytic leukaemia (ALL), the complete remission (CR) rate in the group with a low expression of wt-CYP3A5 mRNA was significantly higher than that in the group with a high expression (p<0.05). A dynamic monitoring for wt-CYP3A5 mRNA expression was performed in two cases of ALL. The expression increased before ALL relapse compared with that in CR in a patient, while in the other patient, the expression was kept in a low level and the patient remained in CR CONCLUSIONS: wt-CYP3A5 mRNA expression was associated with the treatment outcome and prognosis in children with AL. Dynamic monitoring for wt-CYP3A5 mRNA expression in the bone marrow may be useful in the evaluation of the disease severity in childhood acute leukemia.</p>

Influences of interferon-alpha on expression of Th cytokines and CCR7 in dendritic cells from patients with chronic myeloid leukemia in vitro / 中国实验血液学杂志

This study was aimed to investigate the influences of interferonalpha (IFN-alpha) on expressions of CCR7, interleukin10 (IL-10) and IL-12p70 in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to stem cell factor (SCF), granulocyte-macrophage colony stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-alpha) and IL-4, IFN-alpha was added to the serum-free medium of DCs. After culture for 10-14 days, phenotypes and function of CML-DCs were evaluated respectively by flow cytometry and methyl thiazolyl tetrazolium (MTT) assay. Chromosome of DCs was analyzed by displaying G banding assay. The concentrations of IL-10 and IL-12P70 in supernatants were evaluated by enzyme-linked immunosorbent assay (ELISA). The results showed that the expressions of CD40, CD83, CD86 and CCR7 and the OD value in allogeneic mixed-lymphocyte reaction (MLR) in group with IFN-alpha (300 U/ml) were twice as high as those in group without IFN-alpha. The percentage of Ph1 positive cells and concentrations of IL-10 and IL-12 P70 were reduced in group with IFN-alpha. It is concluded that the defective phenotypes and functions of CML-DCs can be recruited partly by IFN-alpha. The mechanism may lie in the facts that expression of CCR7 and co-stimulatory molecules is promoted and the inhibitory effect of IL-10 on CML-DCs is relieved partly through the regulation of IFN-alpha.

Influence of interferon alpha on expression of Fas and Fas ligand in dendritic cells from patients with chronic myeloid leukemia / 中国实验血液学杂志

The study was aimed to investigate the influence of interferon alpha (IFN-alpha) on the expressions of Fas and Fas ligand (FasL) in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to adding stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor alpha (TNF-alpha) and interleukin 4 (IL-4), the IFN-alpha was added to the serum-free medium for DCs. After culturing for 10 - 14 days, cell phenotype and percentage of Ph(1) chromosome were detected by different methods. The expression of Fas or FasL on CML-DCs and cell cycle of DCs labeled with propidium iodine (PI) were measured by flow cytometry. The concentration of sFas in supernatants was analyzed by enzyme-linked immunosorbent assay (ELISA). The results indicated that the expression of co-stimulatory molecules were improved significantly while the percentages of Ph(1) positive cells decreased. The level of Fas on cells was up-regulated and the concentration of sFas decreased. However, the expression of FasL was negative. The ratio of apoptosis rose gradually while the concentration of IFN-alpha increased. It is concluded that IFN-alpha can accelerate the apoptosis of Ph(1) positive cells through Fas/FasL pathway, so the number of Ph(1) negative cells increases relatively.

Classical and molecular cytogenetic abnormalities in 124 pediatric patients with acute lymphoblastic leukemia / 中华儿科杂志

<p><b>OBJECTIVE</b>In childhood acute lymphoblastic leukemia (ALL), cytogenetics plays an important role in diagnosis, allocation of treatment and prognosis. On the basis of the conventional cytogenetic analysis, molecular methods have improved pediatric hematologists/oncologist's ability to accurately and rapidly perform risk-stratification on patients with childhood ALL during the last few years. The aim of the present study was to assess the demography of cytogenetic abnormalities in childhood ALL.</p><p><b>METHOD</b>The study subjects consisted of 124 newly diagnosed ALL patients younger than 16 years of age, who were diagnosed at the Department of Pediatric Hematology/Oncology, Soochow University Children's Hospital. The diagnosis and FAB subtypes of ALL was determined by Wright-Giemsa-stained bone marrow smears and cytochemical staining. Immunophenotyping of the bone marrow samples was performed by flow cytometry. Multiplex polymerase chain reaction (Multiplex PCR) analysis was performed to detect the 29 most common leukemia translocations for routine molecular diagnostic hematopathology practice, and complement the information gained from conventional cytogenetic analysis.</p><p><b>RESULTS</b>Cytogenetic analysis was successful in 112 of 124 children with ALL. Sixty-eight (60%) of them had clonal chromosomal abnormalities. Numerical imbalances consisted of hyperdiploid (> 47 chromosomes, 36 cases), hypodiploid (< 46 chromosomes, 14 cases), pseudodiploidy (18 cases). Chromosomal translocations were observed in 13 patients by conventional cytogenetic analysis. Three cases were found positive for 4; 11 translocation, 3 cases for 9; 22 translocation, 1 case for 1; 19 translocation and 6 cases for other rare translocations. Multiplex-PCR analysis detected 116 of the 124 ALL patients. Thirteen cases of TEL-AML1, 10 cases of rearrangement in the MLL gene, 4 cases of E2A-PBX1, 4 cases of E2A-HLF, 3 cases of BCR-ABL, 2 cases of TLS-ERG, 32 cases of HOX11 were detected by Multiplex PCR in B-lineage leukemias. SIL-TAL1 had been found in 4 of 7 of T-lineage leukemias.</p><p><b>CONCLUSIONS</b>Sixty-eight cases of ALL showed chromosomal aberrations. Multiplex PCR positivity was detected in 59 (50%) of the 116 ALL patients studied. Multiplex PCR combined with chromosomal analysis uncovered chromosomal abnormalities in 95 of 124 (77%) of ALL patients and supplemented each other in detecting chromosomal abnormalities.</p>

Clinical Features of Acute Leukemia with Positive Mixed Lineage Leukemia Fusion Gene in Children / 实用儿科临床杂志

Objective To analyze the clinical features of acute leukemia(AL) with positive mixed lineage leukemia(MLL)fusion gene in children,and explore their treatment protocols,prognosis factors,and so on.Methods Clinical features,treatment protocols,and prognosis factors were studied retrospectively among 51 AL patients with MLL fusion gene.MLL fusion gene was detected by morphology immunology,cytogenetics,molecul arbiology and reverse transcrption polymerase chain reaction(RT-PCR).Results Fifty-one AL patients with MLL fusion gene positive,included 37 cases of acute lymphoblastic leukemia(ALL) and 14 cases of acute myelocytic leukemia(AML).Forty-two patients exhibited abnormal clonal chromosome 11.MLL fusion gene rearrangements and MLL fusion gene partial tandem duplication were found among 36 cases and 15 cases,respectively.Thirty-two cases who received regular chemotherapy were followed up.Twenty-four cases including 19 cases of ALL and 5 cases of AML had achieved complete remission(CR).Six cases including 5 cases of ALL and 1 cases of AML had achieved more than 2 years CR.Sixteen cases were alive update including 12 cases of ALL and 4 cases of AML.Ten cases of positive MLL fusion gene were turning negative.Up to now,6 cases relapsed and 6 cases were dead.Conclusions The incidence of AL children with positive MLL fusion gene is low.It has some features,such as,high replapse rate and poor prognosis.A few patients sensitive to chemotherapies can achieve CR.They live with constant negative MLL fusion gene.

Selective depletion of donor alloreactive T cells by using immuno-magnetic cell sorting / 中国实验血液学杂志

This study was aimed to explore a new method of alleviating graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation through selective elimination of human alloreactive T cells expressing either CD25(+) or CD69(+) by immuno-magnetic cell sorting (MACS). Healthy donor peripheral blood mononuclear cells were cocultivated with bone marrow mononuclear cells from HLA-nonidentical leukemia recipient with remission in one-way mixed lymphocyte culture (OWMLC). After 3 days, both CD25(+) and CD69(+) lymphocytes were removed by MACS. The depleted donor fraction and untreated donor cells were then rechallenged in a secondary mixed lymphocyte culture (MLC) with the original stimulator cells or a third party to assess relative alloreactivity. The results showed that 50% inhibition of the secondary MLC was observed in the depleted donor fraction. Alloreactivity against unrelated third-party cells was largely preserved. It is concluded that this method reduces alloreactivity while retaining reactivity against a third party target in vitro.

Therapeutic effectiveness of CCLG-97 protocol on standard-risk childhood acute lymphoblastic leukemia / 中华儿科杂志

<p><b>OBJECTIVE</b>With the improvement of the diagnosis and treatment, the complete remission (CR) rate and the survival rate of childhood acute lymphoblastic leukemia have been increased in the recent 10 years. The objective of this study was to analyze the outcomes of 119 standard-risk childhood acute lymphoblastic leukemia (SR-ALL) patients, and explore how to improve the survival rate in ALL.</p><p><b>METHODS</b>A total of 119 patients aged 14 months to 15 years were diagnosed as SR-ALL according to the Suggestion of Diagnosis And Treatment for Childhood Acute Leukemia-1993. Among them, seventy-nine were boys and 40 were girls. All of the patients were treated with the CCLG-97 protocol and were followed up for a period of 20 approximately 78 months.</p><p><b>RESULTS</b>The complete remission rate reached 97.4% in four-week induction. Twenty-one patients were out of follow-up, comprising 63%, 14%, 10%, 8% and 5% of all subjects in 1998, 1999, 2000, 2001 and 2002, respectively. The overall survival rates were 93.3%, 90.2%, 88.0%, 85.0%, 85.0% and 85.0% in 1 year, 2 years, 3 years, 4 years and 5 years, respectively. Relapses occurred in 13 patients (13.8%). Among 9 isolated hematologic relapses, 5 patients (56%) were given irregular therapy, 2 did not reach CR within 4 weeks and relapsed 2 years later, 2 accepted regular therapy, 1 was of hypodiploidy and 1 T-ALL. Isolated central nervous system (CNS) relapse occurred in 4 patients (4.3%). Fifteen patients (12.6%) died, 5 of whom (4.2%) died of complications.</p><p><b>CONCLUSION</b>Reinforcing administration and regular therapy are important to improve the long-term survival rate in childhood ALL. The clinical classification should be adjusted with the improvement of diagnostic methods. CCLG-97 protocol decreased the rate of the relapses in SR-ALL and didn't increase the rate of therapy-related death. High-dose methotrexate should be used in therapy and its dosage, usage and individualized therapeutic regimen should be further studied.</p>