Effect of Age on the Time of Neutropenia after Initial Induction Therapy for Patients with Newly Diagnosed Acute Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of age on the time of neutropenia after initial induction therapy for newly diagnosed acute myeloid leukemia (AML) patients.@*METHODS@#Data of 18-65 years old AML patients treated in our hospital from Junuary 2015 to July 2020 were retrospectively analyzed. The clinical characteristics, time of neutropenia after initial induction treatment, early responses, and related influencing factors for the time of neutropenia were analyzed and compared between 18-40 years old group and 41-65 years old group.@*RESULTS@#There were 112 patients enrolled in this study, including 66 (58.9%) males, and their median age was 46 years old. Compared with 18-40 years old group, the incidence of FLT3-ITD gene mutation increased (P=0.039) but core binding factor (CBF) decreased (P=0.003) significantly in 41-65 years old group. The incidence of neutropenia was 97.3%, and the average time was (18.70±1.192) days. The time of neutropenia was (21.43±1.736) days in 41-65 years old group, which was longer than (14.91±1.356) days in 18-40 years old group (P=0.006). The time of neutropenia in CBF positive group was shorter than that in negative group (P=0.012), as well as in patients with remission (CR+CRi) (≤ 2 courses) than those with non-remission (NR) (P=0.024), while in high-risk group was longer than that in low-risk group (P=0.040). Multivariate analysis showed that age, FLT3-ITD gene mutation positive, and non-remission (NR) after two courses of treatment were independent risk factors for the time of neutropenia.@*CONCLUSION@#In non-elderly patients with newly diagnosed AML, age is an influencing factor for the time of neutropenia. Key words　　;

PX-12 Promoting Apoptosis of Multiple Myeloma Cell Line H929 Induced by Bortezomib / 中国实验血液学杂志

OBJECTIVE@#To study the effect of PX-12 on apoptosis of multiple myeloma (MM) cell line induced by bortezomib.@*METHODS@#MM cell line H929 cells were divided into PX-12 group, bortezomib group, combination group, and control group. 5.0 μmol/L PX-12, 20 nmol/L bortezomib, combination of the two drugs, and DMSO were given to the above mentioned group, respectively. After culture for 24, 48, and 72 hours, the changes of cell viability were observed, the MM cell activity was detected by MTT method, and the cell cycle distribution and apoptosis of each group was detected by flow cytometry. The intracellular ROS level was measured by H@*RESULTS@#MTT assay showed that after culture for 72 hours, the activity of H929 cells in PX-12 group (P<0.05) and bortezomib group (P<0.01) was significantly lower than that in the control group, while that in the combination group was decreased most significantly (P<0.01). After culture for 48 hours, cells in G1 phase in PX-12 group was decreased to 40%, while cells in S phase and G@*CONCLUSION@#PX-12 can increase the apoptosis of MM cell line H929 induced by bortezomib, which may be caused by increasing of ROS level.

CLAG Regimen Composed of Continuous Intravenous Infusion of Cladribine in the Treatment of Refractory/Relapsed Acute Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To study the efficacy and safety of continuous intravenous infusion of 2-Chlorodeoxyadenosine (2-CdA) combined with high-dose cytarabine (Ara-C) and granulocyte colony-stimulating factor (G-CSF) (CLAG regiem) in the treatment of relapsed/refractory acute myeloid leukemia (AML).@*METHODS@#Fifteen patients with refractory/relapsed AML hospitalized in 5 medical units such as Department of Hematology, the Affiliated Tumor Hospital of Zhengzhou University and received one course of CLAG regimen from June 2014 to August 2019 were analyzed retrospectively (specifically: cladribine 5 mg/M@*RESULTS@#Among the 15 patients with refractory/relapsed AML, 9 males and 6 females, the median age was 35 (13-63) years old. FAB classification: 1 case of M@*CONCLUSION@#The CLAG regimen consisting of continuous intravenous infusion of cladribine shows high CR in the treatment of AML patients, but the duration of CR is short, myelosuppression is sever, so that infection control is the key. Allogeneic hematopoietic stem cells transplantation should be performed as soon as possible after CR.

Synergistic Mechanism of Interferon alpha-1b, Interleukin-2 and Thalidomide for Immune Regulation in Patients with Acute Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To explore the synergistic immunomodulatory mechanism of interferon alpha-1b, interleukin-2 and thalidomide (ITI) regimen on patients with acute myeloid leukemia (AML).@*METHODS@#Sixty eight untreated de novo or relapsed or refractory or maintenance therapy patients with AML admitted in the Affiliated Cancer Hospital of Zhengzhou University and the other 11 medical units from March 2016 to May 2019 were treated with ITI regimen. Peripheral blood specimen per patient was collected into EDTA-K3 anticoagulation vacuum tube before the administration of ITI and 3 months after the treatment; peripheral blood lymphocyte subsets and perforin and Granzyme B expression were analyzed by using flow cytometry; the levels of VEGF, IFN-γ, TNF-α and IL-6 in the plasma were detected by using a cytometric bead array. Thirty-five healthy subjects from the hospital physical examination centre were selected as normal controls.@*RESULTS@#The ratio of CD4@*CONCLUSION@#The ITI regimen can raise the ratio of CD4

Clinical Analysis of Gene Mutation in Adult Patients with B-ALL and Its Influence on Clinical Prognosis / 中国实验血液学杂志

OBJECTIVE@#To investigate the gene mutation in adult patients with B-ALL and its influence on clinical prognosis.@*METHODS@#Clinical data of 226 adult patients with B-ALL were retrospectively analyzed in the period from August 2011 to February 2018. The incidence of gene mutation in all patients were detected, and the influence of mutation gene on clinical prognosis were estimated. Cox regression model were used to evaluate the independent prognostic factors.@*RESULTS@#208 (92.04%) of 226 patients showed gene mutations, and the median mutation number was 2 (0-8). Among them, 54 cases (23.89%) showed 14 or more mutations. The top five mutation types of all patients were SF1, FAT1, MPL, PTPNII and N-RAS respectively. The median OS and median RFS times of 226 patients were 27.0 (5.5-84.0) months and 22.5 (0-81.0) months respectively. The OS and RFS times of Ph@*CONCLUSION@#Gene mutations are common in all adult B-ALL patients, and the clinical prognosis of patients with JAK and epigenetics-related signaling pathway mutations is worsen, while the WBC level closely relates to the clinical prognosis of the patients.

Expression of Ki-67 in Adult and Children Acute Lymphoblastic Leukemia and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the relationship of Ki-67 level with clinical features, immunophenotype, gene mutation, curative efficacy and prognosis in patients with acute lymphoblastic leukemia(ALL).</p><p><b>METHODS</b>Flow cytometry gated at CD45/SSC was used to detect the expression of Ki-67, and the correlation of Ki-67 expression with clinical manifestation, laboratorial indexes, curative efficacy and prognosis was analysed.</p><p><b>RESULTS</b>Ki-67 expression level increased in ALL patients, the median expression rate was 29.22%, there was significant difference as compared with the healthy control (P<0.01). In adult ALL, the median expression rate of Ki-67 in the high-risk group was 31.49%, and the difference was statistically significant as compared with the low-risk group (P<0.05). In children ALL, the median expression rate of Ki-67 in high-risk group was 42.28%, and the difference was statistically significant (P<0.05). The results of unvariate analysis showed that the age, WBC count at newly diagnosed and extramedullary invasion were adverse factors affecting OS and DFS; the results of multivariate analysis showed that age and extramedullary invasion were independent risk factors for OS and DFS in patients.</p><p><b>CONCLUSION</b>Age≥14 years old, intramedullary invasion are the poor factors for prognosis; the Ki-67 level is not an independent factor for the prognosis of patients.</p>

Clinical Efficacy of EPOCH±R Followed by DICE±R Regimen for Primary Breast Diffuse Large B-Cell Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical efficacy and safety of EPOCH±R followed by DICE±R regimen for primary breast diffuse large B-cell lymphoma.</p><p><b>METHODS</b>Forty-three patients with primary breast diffuse large B-cell lymphoma were admitted in our hosptial from January 2000 to April 2016. Among them 24 patients were treated with CHOP±R regimen, 19 patients were treated with EPOCH±R followed by DICE±R regimen. The clinical efficacy, survival rate and adverse effects were observed and compared between them.</p><p><b>RESULTS</b>The complete rate in EPOCH±R followed by DICE±R regimen group was higher than that in the CHOP±R group (84.2% vs 70.8%), and the relapsed rate was lower in EPOCH±R followed by DICE±R regimen group than that in the CHOP±R group (6.25% vs 35.3%). Progression-free survival (PFS) and overall survival (OS) rates of 5 years after diagnosis in the EPOCH±R followed by DICE±R group were significantly higher as compared with that in CHOP±R group (PFS, 75% vs 47.4%, P=0.035; OS, 73.3% vs 45.2%, P= 0.043). Treatment-related hematologic adverse events were more serious in the EPOCH±R followed by DICE±R group(63.2% vs 25%). However, these adverse events were controlled and no treatment-related deaths were observed. Multivariate analysis showed that age (P=0.008; 95% CI, 0.026 to 0.579), radiotherapy (P=0.045; 95% CI, 1.028 to 14.719) and LDH level (P=0.007; 95% CI, 0.017 to 0.531) were independent prognostic factors for 5 year overall survival.</p><p><b>CONCLUSION</b>EPOCH±R followed by DICE±R regimen is an effective and safe treatment regimen for PB-DLBCL. Prognostic factors for survival are age, LDH level and radiotherapy.</p>

Effect of RNA Interference-silenced TAK1 on Kasumi-1 cell Proliferation Inhibition Induced by AsOand Its Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of transforming growth factor-β activated kinase-1(TAK1) gene silenced by RNA interference on proliferation inhibition of Kasumi-1 cells induced by AsOand its mechanism.</p><p><b>METHODS</b>The experiments were divided into 4 groups, including control group(Kasumi-1 cells treated with non-specific siRNA), TAK1 specific siRNA treated group (Kasumi 1 treated with TAK specific siRNA), AsOtreated group (Kasumi 1 cells treated with AsO) and combined treated group (Kasumi 1 cells treated with TAK1 specific siRNA plus AsO). The proliferation inhibition rate of Kasami 1 cells was detected by CCK-8 method, the apoptotic rate of cells was detected by flow eytometry, the expressions of TAK1, phosphorylated c-Jun N-terminal kinase(p-JNK) and apoptosis-related proteins were detected by Western blot.</p><p><b>RESULTS</b>AsOcould inhibit Kasumi-1 cell proliferation in a dose-dependent manner between 0.5 to 20 µmol/L with ICof (3.79±0.36) µmol/L at 24 h, and also inhibit Kasumi-1 cell proliferation in a dose-dependent manner between 0.5 to 10 µmol/L with ICof (2.38±0.17) µmol/L at 48 h, but then the inhibitory effect reached plateau. After treating Kasumi-1 cells with TAK1 siRNA and 3.5 µmol/L AsOfor 24 h, the proliferation inhibition rate was (10.86±1.64)% and (49.80±2.19)%, meanwhile the apoptosis rate was (8.47±0.75)% and (24.78±2.14)%, all significantly higher than those in control group (P<0.05, P<0.01). The proliferation inhibition rate and apoptosis rate of the combined treated group were significantly higher than that in control and single treated groups (P<0.05, P<0.01), TAK1 silencing and 3.5 µmol/L of AsOcould decrease the expression of TAK1, p-JNK, c-Fos, c-Jun and BCL-2 in different degrees, and increase the expression levels of BAX and the activated (cleaved) caspase-3, 9 with statistically significant differences as compared with control group (P< 0.05). When Kasumi-1 cells were treated with TAK1 specific siRNA plus AsOfor 24 h, protein expression levels were all significantly greater than that in the single-treated groups (P< 0.05).</p><p><b>CONCLUSION</b>TAK1 silencing and AsOcan separately and synergistically inhibit Kasumi-1 cell proliferation which probably relates with the inducing apoptosis via the JNK and mitochondrial pathway. Meanwhile, TAK1 silencing enhances the inhibitory effect of AsOon Kasumi-1 cell proliferation.</p>

Rituximab and Dexamethasone Combined with Cyclophosphamide for Treatment of Relapsed and Refractory Immune Thrombocytopenia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the efficiency and safety of rituximab and dexamethasone combined with cyclophosphamide for treating patients with relapsed and refractory immune thrombocytopenia (ITP).</p><p><b>METHODS</b>Twelve patients with relapsed and refractory immune thrombocytopenia were prospectively enrolled in this study, and received rituximab 375 mg/m(2) once a week for 4 weeks, dexamethasone 40 mg once a day for consecutive 4 days, and cyclophosphamide 500 mg/m(2) biweekly for 2 weeks. The levels of IFN-r and IL-4 in peripheral blood of patients were measured by enzyme-linked immunosorbent assay (ELISA), and the percentages of Breg, Treg and Th17 cells were detected by flow cytometry before and after treatment. Efficiency was evaluated according to platelet counts, and side effects were observed.</p><p><b>RESULTS</b>Six out of 12 patients reached to complete remission and 4 patients reached to partial remission, with the total response rate 83.33%. The platelet counts [(115.42 ± 76.60) × 10(9)/L] after treatment were significantly higher than that before treatment [(115.42 ± 76.60) × 10(9)/L] (P < 0.001). The ratio of IFN-r/ IL4 after treatment (5.89 ± 2.30) was very significantly lower than that before treatment (7.00 ± 2.73) (P = 0.002). The percentage of Breg cells after treatment [(21.27 ± 4.28)%] were much significantly higher than that before treatment [(15.48 ± 1.67)%] (P < 0.001). The ratio of Treg/Th17 after treatment (3.07 ± 1.50) was significantly higher than that before treatment (0.98 ± 0.45) (P < 0.001). Infusion reaction was observed in 1 patient, secondary hypertension and hyperglycemia were in 1 patient, and pneumonia in 2 patients.</p><p><b>CONCLUSION</b>Rituximab and dexamethasone combined with cyclophosphamide can improve the outcomes of patients with relapsed and refractory immune thrombocytopenia patients and they were well tolerated, its mechanism may be related with the balance between T cell sunsets and Treg cells.</p>

Expression of TRAF-6,TAK1 and TGF-β mRNA in peripheral blood mononuclear cells of patients with diffuse large B cell lymphoma before and after chemotherapy / 中国实验血液学杂志

This study was purposed to detect the expression levels of TRAF6, TAK1 and TGF-β mRNA in peripheral blood mononuclear cell (PBMNC) of patients with diffuse large B cell lymphoma (DLBCL) before and after chemotherapy, and to explore the effect of chemotherapy on the activity of TRAF6/TAK1 signal pathway. The expression levels of TRAF-6, TAK1 and TGF-β mRNA in PBMNC of 38 patients with DLBCL were detected by using the quantitative real time PCR before treatment or after two cycles of chemotherapy, 12 healthy people were served as the control. The results showed that the expression levels of TRAF-6, TAK1 and TGF-β mRNA in PBMNC of DLBCL patients' were higher than those in healthy people. Before treatment, the expression levels of TRAF-6 and TAK1 mRNA had no significant difference as compared with healthy people (P > 0.05); after chemotherapy, the expression levels of these two genes significantly increased, and the differences both had statistically significant as compared with healthy people (P < 0.05); meanwhile the increased expression levels of these two genes after chemotherapy had statistically significant difference as compared with levels before treatment (P < 0.05) , and those expression levels were positively correlated. While the expression level of TGF-β mRNA decreased after chemotherapy as compared with level before treatment, and the differences had statistically significantse(P < 0.05). It is concluded that the activity of TRF6/TAK1 signal pathways in PBMNC of DLBCL patients' significantly increases after chemotherapy, while the expression level of TGF-β mRNA after chemotherapy is abviously lower than level before treatment.

Effects of As2O3 in combination with TPA on K562 cells / 中国实验血液学杂志

This study was aimed to investigate the effects of arsenic trioxide (As2O3) combined with TPA on cell cycle, cell differentiation and apoptosis of K562 cell line, and their possible mechanisms. K562 cells were treated with 200 nmol/L TPA, 2 µmol/L As2O3 alone and 200 nmol/L TPA combined with 2 µmol/L As2O3. The proliferative inhibition rates were determined with CCK-8. Annexin V and agarose gel electrophoresis were adopted to detect apoptosis. Colony formation test was used to determine the colony-formation efficiency. Flow cytometry was used to detect the cell differentiation and cell cycle changes. Western blot was employed to detect the expression of P38 and p-P38 proteins. The results showed that combination treatment had synergistic effects on the proliferative inhibition and apoptosis, which were much higher than those treated alone. As2O3 could decrease the colony formation ability of K562 cells. The cells treated with both TPA and As2O3 expressed far more CD11b antigens compared with cells exposed to As2O3 alone. K562 cells treated with TPA were arrested in G1 phase compared with the control group, As2O3 increased the percentage of K562 cells in the G2 phase. The combination treatment increased the expression of p-P38 of K562 cells compared with the cells exposed As2O3 alone. It is concluded that TPA can enhance the effect of As2O3 on inducing apoptosis and adjusting cell cycle , which will expect to provide a new therapeutic program.

Clinical study of COAD-B regimen in treatment of patients with relapsed/refractory non-Hodgkin lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy, adverse events and long-term survival of cyclophosphamide, vindesine, cytarabine, dexamethasone and bleomycin (COAD-B) regimen for relapsed and refractory non-Hodgkin lymphoma (NHL).</p><p><b>METHODS</b>Eighty six patients diagnosed with relapsed or refractory NHL were included in our study from January 2007 to January 2013. The chemotherapy regimen was COAD-B, the therapeutic efficacy was evaluated every 2 courses. Once the stable disease (SD) or progress of the disease (PD) achieved, the patients would switch to other second-line regimens.</p><p><b>RESULTS</b>The overall response rate (ORR) was 67.4%, median remission duration was 13 months (3-51 months); 1-,2- and 4-year overall survival (OS) rates were 75.4%, 56.8% and 40.0%, respectively; 1-, 2- and 4-year progression-free survival (PFS) rates were 50.3%, 39.4% and 27.5%, respectively. The main adverse reaction of patients was myelosuppression. The response to chemotherapy and long- term survival of the relapsed patients were significantly better than that of the refractory ones, and the difference had statistical significance.</p><p><b>CONCLUSION</b>COAD-B could be the salvage regimen for relapsed and refractory NHL.</p>

Effect of TAK1 gene silencing on the apoptosis of Kasumi-1 cells induced by arsenic trioxide / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the effect of transforming growth factor-β activated kinase-1 (TAK1) gene silencing on the proliferation and apoptosis of Kasumi-1 cells induced by arsenic trioxide (As₂O₃).</p><p><b>METHODS</b>Acute myeloid leukemia with t(8;21) cell line Kasumi-1 cells were treated with As₂O₃ or in combination with TAK1 siRNA interference technology. The experiment was divided into four groups: Kasumi-1 cells without any treatment, TAK1 specific siRNA transfection alone, Kasumi-1 cells treated with different concentration of As₂O₃, TAK1siRNA transfection combined with As₂O₃. CCK-8 was used to detect the cell viability. The expression of phosphorylated c-Jun N-terminal kinase (P-JNK) was determined by Western Blot. Cell apoptosis and growth were examined by morphological and colony formation assay.</p><p><b>RESULTS</b>After Kasumi-1 cells were treated with As₂O₃, the rate of cell inhibition was concentration-dependent, and the 50% inhibitory concentration was 3.5 μmol/L. The highest expression level of P-JNK appeared in 30 minutes after cells were treated with As₂O₃. The apoptosis rates of Kasumi-1 cells without any treatment, TAK1 siRNA interference alone group, As₂O₃ alone group and the combined group were (5.02 ± 1.13)%, (6.18 ± 0.28)%, (48.33 ± 2.70)% and (86.07 ± 2.21)%; colony formation rates were (73.83 ± 2.78)%, (76.03 ± 1.46)%, (55.07 ± 1.50)% and (22.20 ± 1.15)%; apoptosis rate of TAK1 siRNA group and the untreated group has no significant difference (P = 0.052); colony formation rate between TAk1 siRNA group and the untreated group has no significant difference (P = 0.179), but the difference in other groups was significant (P = 0.000).</p><p><b>CONCLUSION</b>Silencing the expression of TAK1 can enhance the anti-proliferative and pro-apoptotic effect of As₂O₃ on Kasumi-1 cells, and its mechanism may be through the TAK1 downstream JNK signal pathway.</p>

Inhibitory effect of PD98059 on MAPK signaling pathway in acute lymphocytic leukemia cells / 中国实验血液学杂志

This study was purposed to investigate the effect of blocking Ras/Erk signaling pathway on expression of important transcription factor c-fos, c-jun and TAK1 gene in primary acute lymphocytic leukemia (ALL) cells. The best effective concentration and effect time of PD98059 were screened; the expression levels of c-fos, c-jun and TAK1 in primary cultured cells of normal persons, primary cultured ALL cells and primary cultured ALL cells treated by PD98059 were detected by SYBR GreenI real-time quantitative-PCR. The results showed that before treatment by PD98059 the expression levels of c-fos and TAK1 mRNA were significantly up-regulated in primary cultured ALL cells as compared with primary cultured cells of normal persons (P = 0.014 and P = 0.017 respectively). After treatment by PD98059, the expression levels of c-fos, c-jun mRNA decreased in all 7 serum samples, while expression of TAK1 was down-regulated in 5 samples, and up-regulated in 2 samples. After treatment with PD98059, there was no statistical difference of c-fos, c-jun and TAK1 expression levels in primary cultured ALL cells and primary cultured normal cells. It is concluded that the c-fos and TAK1 activity of primary cultured ALL cells increases, and blocking the Ras/Erk signaling pathway of ALL cells can lead to obvious decrease of important transcription factors c-fos, c-jun, TAK1 genes expression.

Investigation on biological characteristics of CD133+ cancer stem cells in human laryngeal carcinoma Hep-2 cell line / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the biology characters of CD133+ cancer stem cells from Hep-2 cell line.</p><p><b>METHODS</b>The Flow cytometry was applied to purify CD133+ cells from Hep-2 cell line. The sorted CD133+ cells were cultured in RPMI1640. The ability of migration, invasion and clonality of CD133+ cells were performed to characterize the properties of the cells. CD133+ cells and CD133- control cells were treated with paclitaxel and exposed respectively to X-rays emitted by linear accelerator with a dose of 10 Gy. The surviving rates and growth inhibition ratio of cells in two groups were detected with MTT assay to observe the resistance to irradiation and chemotherapy in CD133+ cells.</p><p><b>RESULTS</b>The percentage of CD133+ cells in the unsorted and the sorted cells were 3.1% +/- 0.2% and 90.2% +/- 5.5%, respectively. CD133+ cells showed the higher proliferation and colony ability than CD133- cells. The numbers of CD133+ and CD133- cells that passed the membrane of Transwell chamber were 526 +/- 39 and 220 +/- 20 respectively (t = 22.08, P < 0.001). The colony forming units of three passages were 30.0% +/- 4.7%, 32.2% + 3.6%, 32.7% + 3.4% in CD133 cells and 15.2% +/- 2.2%, 12.0% + 2.5%, 13.8% +/- 3.3% in CD133- cells. There were statistic difference between two groups (t = 8.99, t = 14.66, t = 12.69, P < 0.01). At 24, 48, and 72 hours of treatment with paclitaxel, the cell surviving rates of CD133+ cells were 90.1% +/- 5.9%, 85.1% +/- 7.1% and 70.3% +/- 6.4% and lower than those of CD133- cells, respectively (t = 5.24, t = 8.18, t = 8.14, P < 0.01) . After radiotherapy, growth inhibition ratio of CD133+ and CD133- cells were 30.0% +/- 7.1% and 55.0% +/- 6.3% (t = 8.30, P < 0.01).</p><p><b>CONCLUSIONS</b>CD133+ cells exist in a small proportion in Hep-2 cell line and they show the properties of cancer stem cells, with the resistance to irradiation and chemotherapy.</p>

Detection of NPM1, FLT3 and C-KIT mutations in acute myeloid leukemia and their prognostic analysis / 中国实验血液学杂志

This study was aimed to evaluate the frequencies and prognostic significance of the nucleophosmin 1 (NPM1) mutation, the fms-like tyrosine kinase 3 (FLT3) mutation and c-KIT mutation in acute myeloid leukemia (AML) and to explore their relevance to clinical characteristics, cytogenetics and survival. Genomic DNA from 78 newly diagnosed AML from August 2010 to October 2012 was screened by PCR and sequencing or capillary electrophoresis (CE) for NPM1, FLT3 and c-KIT mutations. The results showed that the incidence of NPM1 mutation was 14.1% in AML patients and 26.7% in normal karyotype AML patients. NPM1 mutant cases were significantly associated with old age (P < 0.05), high peripheral white cell count and platelet counts (P < 0.05) and low expression of CD34 (P < 0.05), but no statistic difference was found in sex, percentage of bone marrow blasts, Hb, expression of CD117 and HLA-DR, complete remission rate, overall survival and relapse rate (P > 0.05). The prevalences of FLT3-ITD and FLT3-TKD mutations were 11.5% (9/78) and 3.8% (3/78) respectively, and no one patient has both of the two mutations. Patients with FLT3-ITD mutation had higher white blood cell counts and percentage of in bone marrow blasts (P < 0.05), and lower overall survival (P < 0.05), more relative to normal karyotype (P < 0.05), while no statistic difference was found in sex, age, platelet count, Hb level, complete remission rate and relapse rate (P > 0.05). No statistic analysis was performed due to the cases of less FLT3-TKD mutation. C-KIT mutation accounts for 7.7% (6/78). Patients with C-KIT mutation had a higher percentage in abnormal karyotype (P < 0.05), and higher relapse rate (P < 0.05), and lower overall survival, whereas no statistic difference was found in sex, age, percentage of bone marrow blasts, peripheral blood cell count, complete remission rate (P > 0.05). It is concluded that the detection of NPM1, FLT3 and C-KIT mutations may contribute to guiding treatment and evaluating prognosis of patients with AML.

Clinical significance of dynamic monitoring of thymic recent output function in different stages of treatment in patients with diffuse large B-cell lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>To detect the changes of naive T cell level of thymic recent output at different stages of treatment in patients with diffuse large B-cell lymphoma (DLBCL), thereby to evaluate the relationship of thymic recent output function with prognosis and the impact of chemotherapy on the potential of immunological recovery.</p><p><b>METHODS</b>The levels of T-cell receptor rearrangement excision circles (TREC) in DNA of peripheral blood mononuclear cells (PBMNC) from 30 DLBCL patients were monitored before, during, until 3 months and 6 months after chemotherapy by real-time PCR (TaqMan), and TREC-level was detected according to the number of CD3 positive(CD3(+)) cells. Twelve normal individuals who matched in age were served as controls.</p><p><b>RESULTS</b>There was a dramatic reduction of TREC values in all DLBCL patients among which TREC values in germinal center B-cell-like-DLBCL (GCB-DLBCL) were higher than those in non-GCB-DLBCL, as compared with TREC values of normal individual in peripheral blood. The mean values of TREC were 0.91 ± 0.15/1000 PBMNCs and (1.22 ± 0.69)/1000 CD3(+) cells in GCB-DLBCL, (0.43 ± 0.29)/1000 PBMNCs and (0.64 ± 0.44)/1000 CD3(+) cells in non-GCB-DLBCL before chemotherapy. TREC values were significantly associated with lower international prognostic index (IPI) grade (r = -0.441, P = 0.015). TREC-level in DLBCL patients was further decreased after chemotherapy, and reached to the lowest level after the 6th cycle of chemotherapy, and during the corresponding period, the mean values of TREC were (0.63 ± 0.34)/1000 PBMNCs and (0.89 ± 0.65)/1000 CD3(+)cells in GCB-DLBCL, (0.19 ± 0.11)/1000 PBMNCs and (0.27 ± 0.25)/1000 CD3(+) cells in non-GCB-DLBCL. TREC-level began to rise obviously 3 months after the last cycle of chemotherapy in most of the DLBCL patients, and came close to normal level in five cases of patients 6 months after the last cycle of chemotherapy.</p><p><b>CONCLUSIONS</b>Thymic recent output function was impaired severely in DLBCL patients. There was an important relationship between thymic recent output function before chemotherapy and prognosis, and chemotherapy had influenced the potential of immunological recovery.</p>

Clinical significance of IDH1 and IDH2 mutations in patients with acute myeloid leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To assess the frequencies and prognostic significance of the isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2) mutations in acute myeloid leukemia (AML) and to explore their relevance to clinical, cytogenetic and molecular feature.</p><p><b>METHODS</b>Genomic DNA from 96 newly diagnosed AML patients from Sep. 2009 to Jan. 2011 was screened by RT-PCR and sequencing for IDH1 and 1DH2 mutation.</p><p><b>RESULTS</b>The prevalence of IDH1 (p. P127 and p. I130) and IDH2 mutations (p. R140) was 14.6% (14/ 96) and 2.17% (2/96) respectively. The IDH1 mutations of p. P127 and p. I130 were not reported so far in literature. Of 14 IDH1 mutation patients, 10 were with normal karyotype and the differences had statistical significance (P=0.021). Two patients with IDH2 mutation were also with normal karyotype. IDH2 mutations were in older patients at diagnosis. Patients with IDH mutation had higher white blood cell counts, lower platelet counts, expression of HLA-DR, CD34, CD33 and CD13, lower remission rate and higher relapse rate.</p><p><b>CONCLUSION</b>IDH mutation is recurring genetic change in AML and indicates poor prognosis.</p>

Hyperthermia increases the sensitivity of RPMI 8226 cells to adriamycin / 中国实验血液学杂志

This study was purposed to explore the effect of hyperthermia on sensitivity of multiple myeloma cells RPMI 8226 to adriamycin (ADM) and its mechanism. The working concentration of ADM against RPMI 8226 cells was defined by MTT assay. RPMI 8226 cells were divided into 4 groups: control group, hyperthermia (42°C) group, chemotherapy (ADM) group and combination group (42°C + ADM), the survival rate of RPMI 8226 cells in 4 groups was detected by trypan blue exclusion, the inhibitory effect of hyperthermia on proliferation of RPMI 8226 cells was detected by MTT assay, the cell cycle distribution, apoptosis rate of cells, intracellular ADM concentration and P-gp expression level were measured by flow cytometry. The 1/4 IC(50) of ADM was defined as the working concentration in the experiment. The results indicated that the hyperthermia promoted the entering the cells from in G(0)/G(1) phase into S and G(2)/M phases, the expression of P-gp protein on cells in hyperthermia and combination groups was down-regulated, the intracellular ADM concentration in combination group obviously increased. It is concluded that the hyperthermia combined with ADM obviously enhance the inhibitory effect on proliferation of RPMI 8226 cells. The hyperthermia increases the sensitivity of RPMI 8226 cells to chemotherapy through down-regulating the expression of P-gp protein on cells and increasing the intracellular ADM concentration.

Expression of NF-kappaB mRNA in acute myeloid leukemia / 中国实验血液学杂志

This study was aimed to investigate the expression level of NF-kappaB mRNA in acute myeloid leukemia (AML) using real time fluorescence quantitative polymerase chain reduction (qPCR) detection and to explore the effect of NF-kappaB mRNA in pathogenesis of AML. The fresh bone marrow was collected from 60 newly diagnosed patients with AML, the total RNA was extracted by means of RTIzoL, the cDNA was synthesized, the expression of NF-kappaB mRNA was detected by qPCR using GAPDH as internal reference. 12 normal healthy persons were selected as controls. The results showed that the expression of NF-kappaB mRNA in patients with AML was higher than that in normal healthy persons with significant difference (p<0.05), the expression of NF-kappaB mRNA in patients with AML-M4 and -M5 were higher than that in patients with AML-M1, -M2 and -M3. It is concluded that the expression of NF-kappaB mRNA is higher in patients with AML. The up-regulation of NF-kappaB expression in patients with AML may play an important role in pathogenesis of AML.