Kinetics of MDSC in Patients Treated Steroids-Ruxolitinib as the First Line Therapy for aGVHD / 中国实验血液学杂志

OBJECTIVE@#To analyze the kinetic characteristics of lymphocyte subsets and myeloid-derived suppressor cell (MDSC) in patients who newly diagnosed intermediate- to high-risk aGVHD and treated with steroids-ruxolitinib as the first line therapy from a single-arm, open clinical trial (NCT04061876).@*METHODS@#We prospectively observed the efficacy of 23 patients having intermediate- to high-risk aGVHD and treated with steroids-ruxolitinib as the first line therapy. The kinetic characteristics of lymphocyte subsets and MDSC were monitored, and then we compared them in steroids-ruxolitinib group (n=23), free-aGVHD group (n=20) and steroids group (n=23).@*RESULTS@#Of the 23 patients, the CR rate was 78.26% (18/23) on day 28 after first-line treatment with steroids-ruxolitinib. On day 28 after treatment, patients had lower level of CD4+CD29+ T cells (P=0.08) than that of pre-treatment, whereas levels of other lymphocyte subsets in this study were higher than that of pre-treatment; CD4+CD29+ T cells in CR patients decreased, compared with refractory aGVHD patients. On day 28 of treatment, CD8+CD28- T cells (P=0.03) significantly increased in patients with aGVHD than that in patients without aGVHD, so did CD8+CD28- T / CD8+CD28+ T cell ratio (P=0.03). Compared with patients without aGVHD, patients with aGVHD had lower level of G-MDSC, especially on day 14 after allo-HSCT (P=0.04). Compared with pre-treatment, M-MDSC was higher in CR patients on day 3 and 7 post-treatment (P3=0.01, P7=0.03), e-MDSC was higher on day 28 post-treatment (P=0.01). Moreover, compared with CR patients, M-MDSC was lower in refractory aGVHD patients on day 3 post-treatment (P=0.01) and e-MDSC was lower on day 28 post-treatment (P=0.01). Compared with steroids group, MDSC in steroids-ruxolitinib group was higher, with the most significant difference in M-MDSC (P3=0.0351; P7=0.0142; P14=0.0369).@*CONCLUSION@#We found that patients newly diagnosed intermediate- to high-risk aGVHD receiving first-line therapy with steroids-ruxolitinib achieved high response rate. Moreover, the novel first-line therapy has a small impact on the immune reconstitution of patients after allo-HSCT. Elevated MDSC might predict a better response in aGVHD patients receiving this novel first-line therapy. M-MDSC responded earlier to steroids-ruxolitinib than e-MDSC, G-MDSC.

Characteristics of N6-methyladenosine modification patterns in t(8;21) acute myeloid leukemia / 南方医科大学学报

OBJECTIVE@#To investigate the relationship between AML1-ETO (AE) fusion gene and intracellular N6-methyladenosine (m6A) modification pattern in t(8;21) acute myeloid leukemia (AML).@*METHODS@#RNA m6A sequencing was performed in SKNO-1 and AE knockdown SKNO-1 (SKNO-1 siAE) cells using RNA-protein co-immunoprecipitation and high-throughput sequencing (methylated RNA immunoprecipitation sequencing, MeRIP-Seq) to analyze the changes in m6A modification of the entire transcriptome. Transcriptome sequencing (RNA-seq) was performed using high-throughput sequencing. The differentially modified mRNAs were further functionally annotated by Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. The changes in m6A-related enzyme expressions were detected using real-time PCR.@*RESULTS@#A total of 26 441 genes were identified in AE knockdown AML cells and AE-expressing cells, containing 72 036 m6A peaks. AE knockdown caused a reduction of the number of intracellular m6A peaks from 37 042 to 34 994, among which 1278 m6A peaks were significantly elevated and 1225 were significantly decreased; 1316 genes with newly emerged m6A modification were detected and 1830 genes lost m6A modification after AE knockdown. The differential peaks were mainly enriched in pathways involving cancer and human T-lymphocytic leukemia virus I. RNA-seq results showed that 2483 genes were up-regulated and 3913 genes were down-regulated after AE knockdown. The combined analysis of MeRIP-Seq and RNA-Seq results revealed relatively high expression levels of m6A-modified genes as compared with the genes without m6A modification (SKNO-1: 0.6116±1.263 vs 2.010±1.655, P < 0.0001; SKNO-1 siAE: 0.5528±1.257 vs 2.067±1.686, P < 0.0001). The m6A modified genes located in the 3'UTR or 5 'UTR had significantly higher expression levels than those located in exonic regions (SKNO-1: 2.177± 1.633 vs 1.333 ± 1.470 vs 2.449 ± 1.651, P < 0.0001; SKNO-1 siAE: 2.304 ± 1.671 vs 1.336 ± 1.522 vs 2.394 ± 1.649, P < 0.05). Analysis of RNA-seq data identified 3 m6A-related enzymes that showed significantly elevated mRNA expression after AE knockdown, namely WTAP, METTL14, and ALKBH5 (P < 0.05), but the results of real-time PCR showed that the expressions of WTAP and ALKBH5 were significantly increased while the expression of METTL14 was lowered after AE knockdown (P < 0.05).@*CONCLUSION@#AE knockdown results in differential expressions of m6A-associated enzymes, suggesting that the AE fusion gene regulates the expression of one or more m6A-associated enzymes to control cellular methylation levels.

Value of SQLE Expression in Prognostic Evaluation of Patients with Acute Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To analyze the relationship between the expression level of SQLE and the prognosis of patients with acute myeloid leukemia (AML) through large sample data.@*METHODS@#The data of genome, transcriptome, gene chip expression, and clinical information were statistically analyzed in multiple cohorts of AML patients with large samples.@*RESULTS@#It was found that the expression level of SQLE gene in tumor cells of AML patients was significantly higher than that of healthy controls (P=0.001). In the three AML corhort, the SQLE high expression group showed a worse therapeutic outcome (OS, P=0.009, P=0.0001, P=0.006; EFS, P=0.005, P=0.001). The unvariate and multivariate survival prognosis analysis indicated that the high expression of SQLE suggests lower event-free survival rate (EFS, HR=1.551, P<0.05) and overall survival rate (OS, HR=1.484, P<0.05). At the same time, it was also found that among different risk subgroups, the expression of SQLE in high risk group was higher (P<0.001, P=0.01), while the patients with high SQLE expression, who received allogeneic HSCT, had longer overall survival time (P=0.006).@*CONCLUSION@#The up-regulation SQLE expression suggests a poor prognosis for the patients with AML.

Increased risk of cardio-cerebrovascular disease after hematopoietic cell transplantation in patients with previous history / 中华医学杂志(英文版)

BACKGROUND@#The impacts of previous cardio-cerebrovascular disease (pre-CCVD) on the outcomes of hematopoietic cell transplantation (HCT) are not well described. Patients with pre-CCVD may often be poor candidates for HCT. This study aimed to investigate the impact of pre-CCVD on transplant outcomes.@*METHODS@#A retrospective study was conducted between patients with and without pre-CCVD who consecutively received allogeneic or autologous HCT between November 2013 and January 2020 with a matching of age and disease status. The cardiovascular complications and HCT outcomes of the two groups were evaluated and compared. The primary endpoints were post-transplant cardio-cerebrovascular disease (post-CCVD) and non-relapse mortality (NRM). We used a multivariable Cox proportional hazard model and the Fine-Gray competing risk regressions for analyses to estimate the hazard ratios (HRs).@*RESULTS@#The outcomes of 23 HCT recipients with pre-CCVD were compared with those of 107 patients in the control group. No significant differences were noted in terms of engraftment, overall survival (OS) (67.00% vs. 67.90%, P = 0.983), or relapse (29.78% vs. 28.26%, P = 0.561) between the pre-CCVD group and the control group. The cumulative incidences of 2-year NRM were similar between patients with pre-CCVD and the controls (14.68% vs. 17.08%, P = 0.670). However, pre-CCVD was associated with an increased incidence of post-CCVD (HR: 12.50, 95% confidence interval [CI]: 3.88-40.30, P < 0.001), which was an independent risk factor for increased NRM (HR: 10.29, 95% CI: 3.84-27.62, P < 0.001) and inferior OS (HR: 10.29, 95% CI: 3.84-27.62, P < 0.001).@*CONCLUSIONS@#These findings suggest that the existence of pre-CCVD before transplantation might not result in increased mortality directly but superpose the toxicity of the transplantation procedure, leading to a risk of post-CCVD. Post-CCVD was a powerful predictor for high NRM and inferior OS. Further risk stratification of pre-CCVD is needed to reduce NRM in various transplantation settings.

Clinical Characteristics and Prognosis of U2AF1 Mutation in Patients with Acute Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the incidence, clinical features of U2AF1 gene mutation in patients with acute myeloid leukemia(AML) and its effect of prognosis.@*METHODS@#A total of 161 patients with AML were enrolled. The second-generation sequencing method was used to detect U2AF1 gene mutation, and the relationship between U2AF1 mutation and clinical features, prognosis was analyzed.@*RESULTS@#The mutation rate of U2AF1 gene in 161 AML patients was 3.73%. The counts of peripheral blood leukocytes and platelets in the U2AF1 gene mutation group were lower than those in the wild type group. The complete response rate of U2AF1 gene mutation group was 66.67%, while that in wild type group was 55.48%, which shows no significant difference between the two groups (P=0.70). The median EFS of wild type group and the mutant group was not reached and reached to 133 days, respectively (P=0.03), while the medium OS in two groups was not reached and reached to 210 days (P=0.01).@*CONCLUSION@#The AML patients with U2AF1 mutation positive have a poor prognosis as compared with the wild type group, which may be a poor prognostic factor for acute myeloid leukemia.

Single Center Analysis of Pathogenic Characteristics of Lung Candida Infection and Other Pathogenic Bacteria Infection in the Patients with Hemotological Maliglancies in the Tropical Region of Hainan / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the infection characteristics of sputum and venous blood pathogen in the patients with hematological malignancies and respiratory symptom in the Hematology Department in tropical region and to investigate its drug-resistance so as to provide etiological evidence for clinical treatment.</p><p><b>METHODS</b>The pathogens and their drug-resistance of 2466 samples in the patients with hematological malignancies and respiratory symptom in the Department were analyzed retrospectively from January 2013 to November 2017. The samples were collected from sputum and venous blood.</p><p><b>RESULTS</b>The sputum sample culture in patients with hematologic diseases showed that 224 strains were isolated, out of them 98 strains (43.75%) were fungi mainly candida albicans (41 strains); and then 88 Gram-negative strains (39.28%), among them the main pathogenic bacteria were Escherichia coli(22 strains) and klebsiella Klebsiella pneumoniae(12 strains); and then 38 Gram-positive strains (16.96%), among them the main pathogeni-bacteria were Enteroccocus (14 strains) and Gram-positive bacilli (14 strains). The blood samples culture of patients with hematologic diseases showed that 61 strains were isolated, out of them the isolated rate of Gram-positive bactetia was higherst, which accounted for 55.74%(34/61), mainly including staphylococcus lominis (19 strains); and the isolated rate of Gram-negative bacteria was 44.26% (27/61), among them main pathogenic bacteria was Klebsiella pneumoniae (12 strains). The resistance test of pathogenic bacteria to drugs showed that the resistant rate of Gram-negative bacteria to tigecycline, imipenem and atl-962 duenner was lowest, while the Gram-positive pathogenic bacteria such as Enterococcus, Gram-positive bacilli and Staphylococcus aureus were sensitive to vancomycin, tigecycline and linezolid was high.</p><p><b>CONCLUSION</b>the patients in hematology department are infected easily in the hospital in tropical region. The main pathogens are fungal strains in the respiratory tract of patients with hematological malignancy according to the sputum culture results. The clinician in tropical regions should choose suitable antibiotics for anti-infective therapy, which is different from the situation in North China or other northern areas.</p>

Efficacy and Safety of Unmanipulated Haploidentical Related Donor Allogeneic Peripheral Blood Stem Cell Transplantation in Patients with Relapsed/Refractory Acute Myeloid Leukemia / 中华医学杂志(英文版)

<p><b>Background</b>Studies of haploidentical-related donor (HRD) stem cell transplantation using a combination of peripheral blood stem cells (PBSCs) and bone marrow as the graft have reported encouraging results for patients with hematological diseases. However, few studies specifically reported transplantation of only PBSCs from HRDs among patients with relapsed or refractory acute myeloid leukemia (AML). Here, the long-term outcomes and side effects of unmanipulated HRD PBSC transplantation (HRD-PBSCT) for relapsed/refractory AML were analyzed.</p><p><b>Methods</b>We performed a retrospective analysis of the outcomes in relapsed/refractory AML patients who underwent PBSCT from HRDs (n = 36).</p><p><b>Results</b>Thirty-one (86.1%) patients in the HRD-PBSCT group achieved platelet recovery. The cumulative incidence of acute graft-versus-host disease (aGVHD) in the HRD-PBSCT group was 40.00%, and the cumulative incidence of grades 2-4 aGVHD in this group was 13.33%. A total of 13 patients in the HRD-PBSCT group had recurrent disease at a median of 183 days after transplantation (range: 10-1700 days), reaching cumulative incidences of relapse of 50.28% at 5 years. On multivariate analysis, donor age and patient age >40 years were independent risk factors for inferior disease-free survival or overall survival (P < 0.05). The results of the present study demonstrate rapid and complete neutrophil engraftment, a low incidence of grade 2-4 aGVHD, and promising survival rates in patients after HRD-PBSCT. Thus, granulocyte colony-stimulating factor-primed PBSCs may be a reliable graft source in unmanipulated HRD-HSCT under myeloablative conditioning when no matched sibling donor is available.</p><p><b>Conclusions</b>Our results support the feasibility, effectiveness, and tolerability of PBSCs as a graft source in unmanipulated HRD transplantation under myeloablative conditioning in patients with leukemia.</p>

Clinical Features and Prognosis of t(8;21) AML Patients in China: A Multicenter Retrospective Study / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To summarize the clinical characteristics of peripheral blood, immune phenotypes, fusion genes and cytogenetics of patients with t(8;21) acute myeloid leukemia(AML) through the retrospective analysis of 586 patients with t(8;21) AML from 15 blood disease research centers in Northern area of China.</p><p><b>METHODS</b>The factors affecting prognosis of patients with t(8;21) AML were investigated by using univariate and multivariate COX regression.</p><p><b>RESULTS</b>The immune type of t(8;21) AML patients was mainly with HLA-DR, CD117, CD34, MPO, CD38, CD13and CD33(>95%), part of them with CD19and CD56; the most common accompanied mutation of t(8;21) AML patients was C-KIT mutation (37.8%); in addition to t(8;21) ectopic, the most common chromosomal abnormality was sex chromosome deletions (38.9%). The univariate analysis revealed a significant survival superiority of OS and PFS in t(8;21) AML patients of WBC≤3.5×10/L without C-KIT mutation, the newly diagnosed ones achieved HSCT(P<0.05), only survival superiority on OS in t(8;21) AML patients with extramedullary infiltration and CD19 positive; the results of multivariate analysis showed a significant survival superiority on OS and PFS in t(8;21) AML patients with WBC≤3.5×10/L(P<0.05).</p><p><b>CONCLUSION</b>The clinical features of t(8;21) AML patients in China are similar to those in other countries, WBC≤3.5×10/L is a good prognostic factor while the C-KIT mutation is a poor one in t(8;21) AML patients.</p>

Analysis of Clinical Characteristics in 10 Patietns with T Large Granular Lymphocytic Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the clinical manifestations and laboratory features of patients with T large granular lymphocytic leukemia (T-LGLL), so as to improve the understanding of this disease.</p><p><b>METHODS</b>The clinical data of 10 patients with T-LGLL in General Hospital of Chinese PLA from October 2015 to March 2010 were analyzed retrospectively.</p><p><b>RESULTS</b>Their median age at diagnosis was 51 years old. 9/10 (90%) patients showed symptoms of anemia, with a median Hb level of 82.5 g/L, 5/10 (50%) patients combined with autoimmune disorders and with a median Hb level of 77 g/L. 7/10 (70%) patients had splenomegaly, 2/10 (20%) patients had complex karyotype, 2/10 (20%) patients had gene mutations, the median age of 4 patients with complex karyotype and gene mutation was 49 years old, all of them suffered from splenomegaly. The immunophenotype of 6/10 patients was CD3+ CD4- CD8+ and that of 2/10 patients (20%) was CD3+ CD4- CD8-, that of another 2/10 (20%) was CD3+ CD4+ CD8-, the clinical features between different types of immunization were not statistically different.</p><p><b>CONCLUSION</b>T-LGLL patients often are old men, combined with anemia and splenomegaly, often associated with autoimmune diseases; the patients with complex karyotype and gene mutation are younger and they are more with hepatosplenomegaly; the guide role of different immunotypes for clinical strategy is no significant.</p>

Construction of SIRT1 promoter expression vector and its activity analysis as well as influence of AML1-ETO on transcriptional regulation of SIRT1 gene / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was aimed to analyze the expression and regulation mechanism of SIRT1 in AML1-ETO positive leukemia to find the core promoter.</p><p><b>METHODS</b>The real-time RT-PCR was used to detect the expression of SIRT1 in AML1-ETO positive leukemia cell line and clinical samples of leukemia patients, a SIRT1 promoter-luciferase reporter vector was constructed and the promoter activity was evaluated in the 293T cell line. A series of possible core promoter fragments of the SIRT1 5'-untranslated region were amplified by PCR, the PCR products were cloned into XhoI/HindIII-digested pGL3-Basic reporter vector, the poly-cationic compound SuperFect reporter vector complexes were transfected into 293T cells.The dual-luciferase Reporter Assay System was used to quantitate the reporter vector luciferase activity.</p><p><b>RESULTS</b>The six kinds of promoter fragment of SIRT1 gene were successfully constructed and cloned into the pGL3-Basic reporter vector, which was authenticated by XhoI/HindIII co-digestion and DNA sequencing. The luciferase activity of the promoter construct was significantly higher than that of the pGL3-Basic promoter in 293T cells. The luciferase report gene assay was also used to detect the regulation of AML1-ETO on the transcription activity of SIRT1 promoter. The results showed that the expression level of SIRT1 increased with the increase mens of AML1-ETO, the promoter of SIRT1 could be bound by AML1-ETO.</p><p><b>CONCLUSION</b>The SIRT1 promoter-luciferase reporter vector is successfully constructed, the transfection system used in this study can effectively transfer gene in 293T cells. The SIRT1 core promoter possesses higher activity in 293T cells and can promote significantly expression of luciferase reporter gene in 293T cells. The transcription regulation of AML1-ETO on SIRT1 is carried out via promoting its promoter activity.</p>

Advances in the Therapy of t(8;21) Acute Myeloid Leukemia / 中国实验血液学杂志

The t(8;21)(q22;q22) translocation is the most common chromosomal abnormalities in AML, and the chromosomal translocation forms AML1-ETO. The t(8;21) AML is a heterogeneity disease. It is unclear for how to treat the relapsed or refractory AML. Recently, the clinical trials and pathogenesis have made great progress. This article summarizes the current clinical trials and recruiting t(8;21) AML clinical trials and researches that related to treatment are as followed: epigenetics, JAK/STAT signaling, steroid, Chinese traditional medicine, and interferon. With the progress of pathogenesis researches, more and more treatments will translate into clinical trials, which can provide more optional choice for relapsed or refractory t(8;21) AML. In this article the AML1-ETO structure and t(8;21) AML pathogenesis, the clinical researches of t(8;21) AML treatment and basic researches of t(8;21) AML treatment are summarized.

Outcome and prognosis in acute myeloid leukemia patients treated with related peripheral blood hematopoietic stem cell transplantation / 中国实验血液学杂志

This study was purposed to evaluate the outcome of acute myeloid leukemia patients treated with related peripheral blood hematopoietic stem cell transplantation (PBHSCT) and analyse the potential prognostic factors. A total of 64 acute myeloid leukemia patients treated with related peripheral allo-HSCT from march 2008 to august 2012 in our hospital were enrolled in the analysis. All the patients received either HLA-matched related or mismatched related donor mobilized peripheral blood stem cells. All the patients were followed up and evaluated for overall survival (OS), leukemia-free survival (LFS) and relapse rate (RR) , and the potential prognostic factors well analyzed. The results showed that the 3 year OS , LFS and RR were 61.9%, 52% and 39.1% respectively. Univarite analysis demonstrated that the disease status before transplantation (P < 0.01) , donor type (P < 0.01), white blood cell count at initial diagnosis (P < 0.05) are related with outcome, and severe aGVHD has some influence on the outcome (P > 0.05) . Multivariate analysis indicated the status of disease before transplantation, donor type, severe aGVHD are the most important prognostic factors. It is concluded that the related PBHSCT is effective treatment method for AML patients, recurrence is the main reason for the failure after transplantation, disease status before transplantation, donor type, and severe aGVHD are independent prognostic factors.

Effect of autologous peripheral blood stem cell transplantation in 13 patients with AML1/ETO (+) acute myeloid leukemia / 中国实验血液学杂志

This study was purposed to analyse the clinical efficacy of autologous peripheral blood stem cell transplantation (APBSCT) in 13 Patients with AML1/ETO (+) acute myeloid leukemia, and to evaluate the role of quantitative detecting the AML1/ETO gene in treatment of AML patients. A total of 13 patients with AML1/ETO (+) acute myeloid leukemia treated with APBSCT from August 2007 to November 2012 were retrospectively analyzed. The median follow-up time was 26 (7.8-75.8)months. Kaplan-Meier analysis was used to calculate the overall survival (OS), leukemia-free survival (LFS) and cumulative relapse rate (RR). Log rank method was used to perform univariate analysis. The results showed that the 3 year-OS, LFS, and RR were (70.5 ± 15.3)%, (51.3 ± 16.7)%, 48.7%, respectively. The AML1/ETO expression level in 4 cases out of 5 relapsed patients was quantified during and after therapy, and the result showed that AML1/ETO expression level significantly increased before morphological relapse. In univariate analysis, there was no statistic significance in terms of age, sex, count of white blood cells at diagnosis, interval from diagnosis to transplantation, count of MNC for infusion. It is concluded that APBSCT has good therapeutic effect on AML1/ETO (+) AML, and regular quantitative monitoring of AML1/ETO expression level can predict early recurrence. Allogeneic hematopoietic stem cell transplantation after relapse may contribute to obtain opportunity to achieve the long-term survival for intermediate and high risk patients.

Analysis of influence factors on G-CSF-mobilized hematopoietic stem cells of 191 healthy donors in a mono center / 中国实验血液学杂志

This study was aimed to investigate the factors influencing mobilization efficiency of peripheral hematopoietic stem cells with granulocyte colony stimulating factor (G-CSF) and their impact on healthy donors. 181 donors were mobilized subcutaneously with G-CSF at 5 - 10 µg/(kg·d), and 10 donors were mobilized with G-CSF at 3.3 - 4.9 µg/(kg·d), once 12 h, for 4 - 5 d. Peripheral blood mononuclear cell (MNC) and CD34(+) cell counts were analyzed by flow cytometry. Mobilization-related side effects were also monitored. The results showed that white blood cell counts increased by 6 times averaged after mobilization (P < 0.01). The platelet count obviously decreased (P < 0.01), while the hemoglobin level did not show significant difference. No significant differences were observed in MNC and CD34(+) cell counts between those subjects harvested on the 4th and 5th day after mobilization. Male donors were superior to female ones in cell harvest (P < 0.01). Donor body weight played positive role in cell yield, while impact of age on harvest was not remarkable. Neither MNC nor CD34(+) cell count showed a linear relationship with G-CSF dose. Only slight side effects were observed on the donors in this study. It is concluded that mobilization with G-CSF is sufficient in healthy donors without remarkable side effects.

Protective effect of ademetionine 1, 4-butanedisulfonate on liver injury caused by chemotherapeutic agents / 中国实验血液学杂志

The aim of this study was to observe the protective effect of ademetionine 1, 4-butanedisulfonate on liver injury caused by chemotherapy in patients with leukemia. The clinical data of protective effect were analyzed retrospectively from January 2010 to April 2012. A total of 62 acute leukemia patients were divided into A group (27 cases) and B group (35 cases), the polyene phosphatidyl choline combined with ademetionine or combined with compound glycyrrhizin were given in A and B group, respectively. The changes of liver function were observed after 2 weeks, 5 patients in B group suffered from acute liver injury were treated by ademetionine as rescue therapy. Liver function was compared before and after treatment. The results showed that ALT and AST levels were significantly reduced in A group (P < 0.05), none of the patients (0/27) suffered from acute liver injury, but 14.29% (5/35) patients in B group suffered from acute liver injury, and liver function could be recovered by substitution treatment of ademetionine (the median time is 8 days, 5-14 days). It is concluded that the protective and therapeutic effect of ademetionine against liver injury caused by chemotherapy in patients with leukemia is better than that of compound glycyrrhizin.

Prognostic significance of detecting MLL-AF9 fusion gene expression in patients with acute myeloid leukemia by real-time fluorescence quantitative PCR / 中国实验血液学杂志

<p><b>UNLABELLED</b>This study was aimed to explore the value of detecting the expression levels of MLL-AF9 (mixed lineage leukemia, MLL) fusion gene during the treatment of acute myeloid leukemia (AML) by real-time fluorescence quantitative PCR (RQ-PCR), and to evaluate its prognostic significance in monitoring minimal residual disease (MRD). The expression levels of 11 patients with MLL-AF9 fusion gene positive were detected precisely by RQ-PCR during the treatment in order to analyze the correlation of detection results with clinical manifestations. The results showed that the expression levels of MLL-AF9 fusion gene in patients at initial diagnosis were 1.3%-55.28%.</p><p><b>RESULTS</b>obtained from 5 patients who received chemotherapy alone during the interval between first and second courses of chemotherapy indicated that 2 patients with <0.1% of MLL-AF9 fusion gene expression levels all achieved hematologic complete remission and survived, while the remaining 3 patients with ≥ 0.1% of MLL-AF9 fusion gene did not achieve hematologic complete remission and only 1 case survived. Moreover, results obtained from 6 transplant patients within a month before the transplantation suggested that 4 of them with < 0.1% of MLL-AF9 fusion gene expression levels survived without relapses, while the remaining 2 patients with ≥ 0.1% of MLL-AF9 fusion gene expression levels relapsed and died. Besides, MLL-AF9 fusion gene expression levels were ≥ 0.1% within one month before the morphological relapse of bone marrow in 2 recurrent patients. It is concluded that the detecting the expression level of MLL-AF9 fusion gene by RQ-PCR is an effective and accurate method to quantify and monitor the MRD level of MLL-AF9 gene positive AML patients and may be used for early detecting molecular relapse of AML.</p>

Analysis of clinical characteristics and prognosis of 13 cases of acute erythroleukemia / 中国实验血液学杂志

The aim of this study was to investigate the clinical characteristics and prognosis of acute erythroleukemia (AEL, AML-M6). The clinical features and results of morphologic, immunophenotypic, cytogenetic and molecular biologic detections were retrospectively analyzed in 13 cases of AEL from 305 acute leukemia patients hospitalized between October 2007 and October 2012. The results showed that the expression of erythroid and non-erythroid cells increased at the same time. The myeloid antigens mainly expressed CD13/CD33/CD117/CD34, while the erythroid antigens expressed Gly and CD71. The karyotypic detection indicated that there were 1 case with normal karyotype, 3 cases with simple karyotypic abnormality and 2 cases with complex karyotypic abnormality, the other cases were not detected. The molecular biological detection found that the poor prognosis gene existed in 5 cases [38.5% (5/13)], including 3 cases with MLL-MLL fusion gene, 1 case with MLL mutation, and 1 cases with NRAS gene mutation, the abnormal genes were not detected in remainder 8 cases. After chemotherapy with decitabine, the complete remission (CR) rate achieved 53.5% (7/13), partial remission (DR) rate achieved 15.4% (2/13). Finally, 8 patients received allo-HSCT, the median overall survival (OS) was 20.7 months, 3 year survival rate was 79%, 3 year disease-free survival rate was 78%. It is concluded that the acute erythroleukemia is a rare subtype of AML, which is transformed from MDS and has harmful genes and poor prognosis. Allo-HSCT and treatment with decitabine may enhance the survival rate of AEL.

Clinical analysis of acute myeloid leukemia with CBFB-MYH11-positive / 中国实验血液学杂志

The study was aimed to investigate the clinical characteristics of acute myeloid leukemia (AML) with CBFB-MYH11 gene. The clinical data of 12 cases were analyzed retrospectively, including age, clinical characteristics, immunophenotype, treatment protocols and efficacy as well as the prognosis. The results indicated that 12 patients with CBFB-MYH11 were detected in 293 AML patients. The median age of the 12 patients was 32.5 (21 - 57) years old. According to French-American-British (FAB) classification, 66.7% (8/12) patients was diagnosed as M4Eo and 33.3% patients was diagnosed as M4. At new diagnosis, the median WBC count was 19.8×10(9)/L (2.46 - 164.30×10(9)/L). The WBC count > 100×10(9) was found in 16.7% patients (2/12). The complete remission (CR) rate after 1 and 2 cycles of induction chemotherapy were 83.3% and 16.7% respectively, so the total CR rate was 100%. Estimated 5-year relapse-free survival (RFS) and overall survival (OS) were 80% and 83%, respectively. It is concluded that patients with CBFB-MYH11 are usually M4Eo and M4. Patients with this fusion gene are often associated with high frequency of CD33, CD34, CD117, HLA-DR, CD15, CD64 and CD14 expression. Patients with CBFB-MYH11 have a tendency of higher CR rate, longer RFS and OS.

Clinical efficacy of decitabine plus improved CAG chemotherapy and haplo-identical donor peripheral lymphocyte infusion regimen on elderly patients with high risk myelodysplastic syndrome and acute myeloid leukemia / 中国实验血液学杂志

This study was aimed to observe the clinical efficacy and adverse effects of decitabine plus improved CAG chemotherapy and haploid-identical donor peripheral lymphocyte infusion regimen on elderly patients with myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Five elderly patients with MDS and AML were treated with decitabine plus improved CAG chemotherapy and donor peripheral lymphocyte infusion regimen. Examinations on liver and renal function, electrocardiogram and bone marrow analysis were performed before and after treatment, and adverse effects were observed. The results indicated that after a course of treatment by decitabine plus improved CAG chemotherapy and haplo-identical donor peripheral lymphocyte infusion regimen, the total effective rate was 100%, and 4 patients (80%) achieved complete remission, 1 patient achieved partial remission. The dominant clinical adverse effect was bone marrow depression, the median time of neutrophil>0.5×10(9)/L and platelet>20×10(9)/L was 15 d and 16 d respectively for patients without previous MDS. It is concluded that decitabine plus improved CAG chemotherapy and haploid-identical donor peripheral lymphocyte infusion regimen may be effective with less adverse effects for elderly primary AML and high risk MDS patients, it is a promising therapeutic methods and worthy to deeply study.

Silence mechanism of WT1 gene in leukemic cell line U937 / 中国实验血液学杂志

This study was aimed to investigate the methylation status of WT1 gene in leukemia cell lines and its relation with expression of WT1 gene. The WT1 gene was silenced by DNA methylation or histone deacetylation, and the expression of WT1 gene was induced by using HDAC inhibitor and/or demethylation agent of DNA. Some leukemia cell lines (U937, HL-60, K562, KG1) were detected by RT-PCR, MS-PCR, restriction analysis, and DNA sequencing. U937 leukemic cells without WT1 mRNA expression were incubated with HDAC inhibitor Trichostatin A (TSA) and/or demethylation agent decitabine. The results showed that the U937 cells did not express WT1 gene, but HL-60, K562 and KG1 cells highly expressed WT1 gene; WT1 gene was unmethylated in HL-60 cells, but methylated in K562 and U937 cells. WT1 expression could be reactivated by co-incubation with TSA and decitabine, but not was observed by using single drug. It is concluded that WT1 promoter is methylated in some leukemia cells, however, the methylation can not affect its expression. DNA methylation and deacetylation of histones are synergistic to inhibit the expression of WT1 in leukemic U937 cells, the combination of TSA with decitabine can induce expression of WT1 gene.