Efficacy of Different Doses of Daunorubicin Induced Chemotherapy in Patients with Newly Diagnosed Primary Acute Myeloid Leukemia Under 65 Years Old / 中国实验血液学杂志

OBJECTIVE@#To compare the efficacy and safety of different doses of daunorubicin combined with a standard dose of cytarabine as induction chemotherapy in newly diagnosed primary acute myeloid leukemia (AML) patients.@*METHODS@#The clinical data and outcome were retrospectively analyzed in 86 newly diagnosed primary AML patients who were under 65 years old and treated with daunorubicin combined with cytarabine (DA regimen) at West China Hospital of Sichuan University from January 2017 to June 2019. Patients were divided into 2 groups based on the dose of daunorubicin they received, 35 cases in the escalated-dose group ［75 mg/(m@*RESULTS@#Median follow-up time of all the patients was 15 months. The CR rate and MRD@*CONCLUSION@#The escalated dose of daunorubicin can induce higher complete remission rate, deeper remission and longer duration of remission without increasing adverse events in newly diagnosed primary AML patients.

Bendamustine treatment of Chinese patients with relapsed indolent non-Hodgkin lymphoma: a multicenter, open-label, single-arm, phase 3 study / 中华医学杂志(英文版)

BACKGROUND@#Bendamustine was approved in China on May 26th, 2019 by the National Medical Product Administration for the treatment of indolent B-cell non-Hodgkin lymphoma (NHL). The current study was the registration trial and the first reported evaluation of the efficacy, safety, and pharmacokinetics of bendamustine in Chinese adult patients with indolent B-cell NHL following relapse after chemotherapy and rituximab treatment.@*METHODS@#This was a prospective, multicenter, open-label, single-arm, phase 3 study (NCT01596621; C18083/3076) with a 2-year follow-up period. Eligible patients received bendamustine hydrochloride 120 mg/m2 infused intravenously on days 1 and 2 of each 21-day treatment cycle for at least six planned cycles (and up to eight cycles). The primary endpoint was the overall response rate (ORR); and secondary endpoints were duration of response (DoR), progression-free survival (PFS), safety, and pharmacokinetics. Patients were classified according to their best overall response after initiation of therapy. Proportions of patients in each response category (complete response [CR], partial response [PR], stable disease, or progressive disease) were summarized along with a two-sided binomial exact 95% confidence intervals (CIs) for the ORR.@*RESULTS@#A total of 102 patients were enrolled from 20 centers between August 6th, 2012, and June 18th, 2015. At the time of the primary analysis, the ORR was 73% (95% CI: 63%-81%) per Independent Review Committee (IRC) including 19% CR and 54% PR. With the follow-up period, the median DoR was 16.2 months by IRC and 13.4 months by investigator assessment; the median PFS was 18.6 months and 15.3 months, respectively. The most common non-hematologic adverse events (AEs) were gastrointestinal toxicity, pyrexia, and rash. Grade 3/4 neutropenia was reported in 76% of patients. Serious AEs were reported in 29 patients and five patients died during the study. Pharmacokinetic analysis indicated that the characteristics of bendamustine and its metabolites M3 and M4 were generally consistent with those reported for other ethnicities.@*CONCLUSION@#Bendamustine is an active and effective therapy in Chinese patients with relapsed, indolent B-cell NHL, with a comparable risk/benefit relationship to that reported in North American patients.@*CLINICAL TRIAL REGISTRATION@#ClinicalTrials.gov, No. NCT01596621; https://clinicaltrials.gov/ct2/show/NCT01596621.

Mutation analysis of FLT3-ITD gene in acute myeloid leukemia / 中国病理生理杂志

AIM:To investigate the mutation of FLT3-ITD gene in the patients with newly diagnosed acute myeloid leukemia (AML). METHODS:From March 1, 2015 to June 1, 2017, 207 patients with AML admitted to de-partment of hematology, sichuan provincial people′s hospital were enrolled in this study. The bone marrow samples were collected from the patients. PCR was used to detect the mutation of FLT3-ITD gene. After the corresponding chromosome was obtained by R-banding, the cells were made into strips and banding. Twenty karyotypes with relatively cleavage were automatically selected from each specimen to complete karyotyping. By analysis of the clinical data and following-up the prognosis, the FLT3-ITD gene mutation in diagnostic and evaluative values for AML were performed. RESULTS:FLT3-ITD gene mutation was found in 42 cases of 207 AML patients, the positive rate was 20. 29 % . FLT3-ITD positive patients showed 3 bands. FLT3-ITD gene mutation in 42 patients with positive results showed that FLT3-ITD gene mutations in turn met the end to end, and insert a number of nucleotides, but all the mutations were in-frame mutations. According to the FAB and WHO standard, in 42 cases of FLT3-ITD positive positive patients, M0 accounted for 0.00% , M1 accounted for 2.38% (1/42), M2 accounted for 23.81% (10/42), M3 accounted for 0.00% , M4 accounted for 2.38% (1/42), M5 accounted for 69.05% (29/42), M6 accounted for 0.00% , M7 accounted for 2.38% (1/42). The white blood cell ( WBC) level and complete response (CR) rate in FLT3-ITD positive patients were lower than those in FLT3-ITD negative patients (P<0.05). CONCLUSION:The WBC level and CR rate, which are lower in FLT3-ITD positive patients than those in negative patients, are the clinical risk factors. It will be helpful to determine the prognosis evaluation for AML pa-tients.

Imbalance of Th17 and Treg cells in peripheral blood from multiple myeloma patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the ratio of Th17 cells and CD4⁺CD25⁺Foxp3⁺ regulatory T (Treg) cells in peripheral blood from patients with multiple myeloma (MM) and explore its pathological effects.</p><p><b>METHODS</b>70 MM patients were divided into three groups: newly diagnosed group (n=30), plateau stage group (n=23) and relapsed/refractory group (n=17). The controls consisted of 20 healthy donors. The frequencies of Th17 and Treg cells were detected by flow cytometry.</p><p><b>RESULTS</b>Compared with controls [(0.72±0.33)%] and plateau stage group [(0.74±0.29)%], frequencies of Th17 cells were higher in newly diagnosed group [(1.62±0.65)%] and relapsed/refractory group [(1.45±0.51)%], respectively (P<0.05). Compared with controls [(2.33±0.90)%] and plateau stage group [(1.69±0.70)%], frequencies of Treg cells were significantly lower in newly diagnosed group [(0.55±0.23)%] and relapsed/refractory group [(0.82±0.54)%], respectively (P<0.05). The ratios of Th17/Treg in newly diagnosed group and relapsed/refractory group were higher than those in controls (P<0.05). There were no differences of the frequencies of CD3⁺CD4⁺ T cells and Th17 cells between plateau stage group and controls. The frequencies of Treg cells were significantly lower in plateau stage group than that in controls (P<0.05), and the ratio of Th17/Treg was significantly higher in plateau stage group than that in controls (P<0.05).</p><p><b>CONCLUSION</b>The remarkable abnormality of T cells subsets was reduction of CD4⁺ T cells in MM. Higher frequency of Th17 and lower ratio of Treg could lead to imbalance of Th17/Treg, which may play a critical role in the pathogenesis of MM.</p>

COX-2 inhibitor celecoxib can suppress the proliferation of FLT3-ITD positive acute myeloid leukemia cells with prominent down regulation of MEK/MCL-1 expression in vitro / 中国实验血液学杂志

The purpose of this study was to investigate the effects of Celecoxib on the proliferation of the FLT3-ITD positive and negative acute myeloid leukemia cells and its mechanism. The proliferation inhibition effect of Celecoxib with different doses on the FLT3-ITD positive cells MV4-11 and the FLT3-ITD negative K562 cells was detected by CCK-8 method, the cell apoptosis was determined by flow cytometry, and the MEK, Mcl-1, pAKT expression was tested by Western blot. The results showed that Celecoxib inhibited the proliferation of both MV4-11 and K562 cells, but the IC50 for MV4-11 was (29.14 ± 2.4) µmol/L, which was significantly lower than that of K562 cells (39.84 ± 1.0) µmol/L (P < 0.05); The induced apoptosis rate of Celecoxib at 20-80 µmol/L on MV4-11 was not observed, but there was apparent influence on K562 at the same concentration. Western blot showed that Celecoxib down-regulated the expression of MEK and Mcl-1 but did not change the expression of pAKT obviously on MV4-11 cells, while the expression of Mcl-1 was reduced a little, but no obvious change were found in the expression of MEK and pAKT on K562 cells. It is concluded that the Celecoxib can inhibit the proliferation of FLT3-ITD positive AML cells distinctly, and the potential mechanism may be related to the inhibition of the MEK/Mcl-1 signaling pathway.

Individualized leukemia cell-population profiles in common B-cell acute lymphoblastic leukemia patients / 癌症

Immunophenotype is critical for diagnosing common B-cell acute lymphoblastic leukemia (common ALL) and detecting minimal residual disease. We developed a protocol to explore the immunophenotypic profiles of common ALL based on the expression levels of the antigens associated with B lymphoid development, including IL-7Rα (CD127), cytoplasmic CD79a (cCD79a), CD19, VpreB (CD179a), and sIgM, which are successive and essential for progression of B cells along their developmental pathway. Analysis of the immunophenotypes of 48 common ALL cases showed that the immunophenotypic patterns were highly heterogeneous, with the leukemic cell population differing from case to case. Through the comprehensive analysis of immunophenotypic patterns, the profiles of patient-specific composite leukemia cell populations could provide detailed information helpful for the diagnosis, therapeutic monitoring, and individualized therapies for common ALL.

Ratio balance of Th17 and Treg cells in peripheral blood of patients with chronic lymphocytic leukemia / 中国实验血液学杂志

This study was purposed to investigate the ratio of Th17 cells and CD4(+)CD25(+)Foxp3(+) regulatory T (Treg) cells in peripheral blood of patients with chronic lymphocytic leukemia (CLL) and to explore their roles in the pathogenesis and clinical diagnosis. Based on the number of peripheral lymphocytes and treatment condition, the CLL patients were divided into 2 groups: untreated group (n = 30) and remission group (n = 15), the healthy control group (n = 20) was set up as well. The frequencies of Th17 and Treg cells of all cases were detected by flow cytometry (FCM). The results showed that frequencies of CD3(+)CD4(+)T cells and Th17 cells were significantly higher in untreated group than that in healthy control group (P < 0.05), the frequencies of CD3(+)CD8(+)T cells and Treg cells were significantly lower in untreated group than that in healthy control group (P < 0.05), the ratio of Th17/Treg was significantly higher in untreated group than that in healthy control group (P < 0.05). The frequencies of Th17 were not statistically different between remission and healthy control groups, the frequencies of Treg cells were significantly lower in remission group than that in healthy control group (P < 0.05), the ratio of Th17/Treg was significantly higher in remission group than that in healthy control group (P < 0.05), frequencies of Th17 cells were markedly lower in remission group than that in untreated group (P < 0.05). It is concluded that Th17/Treg imbalance exists in patients with CLL, which may play a key role in pathogenesis and development of CLL.

Early detection of MRD in peripheral blood after induction chemotherapy of newly diagnosed patients with AML and its correlation with curative effects / 中国实验血液学杂志

The purpose of this study was to detect the minimal residual disease (MRD) in peripheral blood of newly diagnosed patients with acute myeloid leukemia (AML) on day 8 of induction chemotherapy and analyze the correlation between day 8 MRD (D8RD) and therapeutic effectiveness. 29 adult patients (13 males and 16 females, aged 16 - 75 years, median 41 years) with AML diagnosed and treated in West China Hospital from September 2009 to June 2010 were analyzed and followed up in the study. The leukemia-associated aberrant immunophenotype (LAIP) of all the patients were detected by multiparameter flow cytometry (FCM) before therapy. The level of MRD in the peripheral blood at day 8 of induction chemotherapy was detected by FCM based on the LAIP. The overall survival curve was drawn by calculation using Kaplan-Meier method using, and the comparison between different groups was carried out by Log-rank test. The results indicated that after first course therapy, the levels of peripheral D8RD in 7 out of 29 AML cases were lower than 0.01% (negative group), and that in another 22 cases were higher than 0.01% (0.08% - 55%, positive group). The sex, age, WBC, LDH, percentage of bone marrow blasts at diagnosis in these groups were not statistically different. 6 cases achieved CR (86%) in D8RD negative group, and also 6 cases achieved CR (27%) in D8RD positive group, CR rate in D8RD negative group was higher than in D8RD positive group (86% vs 27%, P < 0.05). The median follow-up of 29 cases lasted for 15 months; the 1-year overall survival rate of D8RD negative and D8RD positive groups was 100% and 39.4%, respectively (P < 0.01). It is concluded that MRD level in peripheral blood at day 8 of induction chemotherapy is an early index to predict clinical efficacy of induction therapy in AML.

Influence of immunosuppressive therapy on expression of TNF-α/IFN-γ in cytoplasm of peripheral blood lymphocytes of patients with aplastic anemia / 中国实验血液学杂志

The purpose of this study was to investigate the influence of immunosuppressive therapy on the expression of TNF-α/IFN-γ in cytoplasm of peripheral blood lymphocytes of patients with aplastic anemia (AA). The expression of TNF-α and IFN-γ in cytoplasm of peripheral CD3(+) lymphocytes were measured by flow cytometry in 25 cases of de novo AA patients and 20 cases of AA after immunosuppressive therapy. The results showed that the positive rates of CD3(+)/TNF-α(+) and CD3(+)/IFN-γ(+) in de novo AA patients were (5.97 ± 6.78)% and (15.20 ± 11.28)% respectively, and (1.56 ± 0.87)% and (1.76 ± 0.87)% in normal controls respectively. There was significant difference between de novo AA patients and normal controls (p < 0.05). The positive rates of CD3(+)/TNF-α(+) and CD3(+)/IFN-γ(+) in immunosuppressive therapy group were (1.67 ± 1.26)% and (4.35 ± 4.33)% respectively. The difference between immunosuppressive therapy group and de novo AA group was statistically significant (p < 0.05). It is concluded that the levels of intracellular TNF-α and IFN-γ in AA patients are higher than those in normal controls. Immunosuppressive therapy significantly reduces the expression of intracellular TNF-α and IFN-γ. Its relationship with the clinical treatment is worth further observing.

Immunophenotype characteristics and prognosis of acute leukemia patients with cross expressing lymphoid and myeloid lineage associated antigens / 中国实验血液学杂志

The aim of study was to investigate the immunophenotype characteristics and prognosis of acute leukemia patients with cross-expressing lymphoid and myeloid lineage-associated antigens. The immunophenotypes of leukemic cells were examined by using flow cytometry. All patients were classified into several groups according to FAB subtypes and immunophenotyping. The cross-expressed antigens analyzed for AML included CD2, CD7, CD19, CD56 and other co-expressed lymphoid antigens. The myeloid antigens analyzed for ALL included CD13 and co-expressed CD13/CD33. ALL and AML patients without expression of cross-expressing antigens were selected as control. Complete remission (CR) ratio and relapse-free survival (RFS) of patients in all groups were compared. The results indicated that among 161 patients analyzed, 91 cases of AML with cross-expressing lymphoid and myeloid antigens included that 24 cases of AML expressed lymphoid surface marker-CD7, namely CD7(+) AML, 14 cases of AML only expressed lymphoid surface marker-CD19, namely CD19(+) AML, 8 cases of AML expressed lymphoid surface marker-CD2 (including CD2/CD19 co-expressed), namely CD2(+) AML, 10 cases of AML expressed lymphoid surface marker-CD56 (including CD56/CD19 or CD56/CD2 co-expressed), namely CD56(+) AML, 16 cases of AML expressed two or more lymphoid surface markers, namely Ly ≥ 2(+) AML, 9 cases of ALL expressed myeloid surface markers CD13, namely CD13(+) ALL, 10 cases of ALL expressed myeloid surface markers CD13 and CD33, namely CD13/CD33(+) ALL. 29 cases of ALL did not expressed myeloid surface markers, namely My(-) ALL, and 41 case of AML did not expressed lymphoid surface markers, namely Ly(-) AML. CR ratio and RFS of Ly ≥ 2(+) AML patients were lower than those of Ly(-) AML patients. RFS of CD56(+) AML patients was lower, but CR ratio had no significant difference, when compared with Ly(-) AML patients. CR ratio and RFS of other AML patients with cross-expressing antigens had no significant difference when compared with Ly(-) AML patients. CR ratio and RFS of CD13(+) ALL and CD13/CD33(+) ALL patients had no significant difference when compared with My(-) ALL patients. It is concluded that the importance of cross-expressing antigens for prognosis of patients should be analyzed concretely. CD56(+) AML and Ly ≥ 2(+) AML have bad prognosis, while other cross-expressed lymphoid and myeloid lineage-associated antigens have no impact on prognosis of acute leukemia patients.

Expression of B7-H1 molecule on human bone marrow mesenchymal stem cells and its effects on T lymphocyte proliferation / 中国实验血液学杂志

The mechanisms of human bone marrow mesenchymal stem cells (hBMMSCs)-mediated immunomodulation are still not be completely clarified. In order to investigate the expression of B7-H1 on hBMMSCs and to explore whether B7-H1 mediated signaling pathway (B7-H1/PD-1) involves in the mechanisms of hBMMSCs-mediated immunomodulation, the hBMMSCs were isolated, cultured and identified, the B7-H1 expression on hBMMSCs was detected by flow cytometry, RT-PCR, and Western blot. The inhibitory effect of hBMMSCs on proliferation of T lymphocytes was observed in mixed lymphocyte culture, and then the functional anti-B7-H1 monoclonal antibody (mcAb) was used to block B7-H1, the proliferation of T lymphocytes was detected by using CCK-8. The results indicated that hBMMSCs highly expressed B7-H1 molecule, hBMMSCs effectively inhibited the proliferation of T lymphocytes with a dose-dependent manner, and the inhibitory proliferation of T lymphocytes by hBMMSCs could be partially restored when the anti-B7-H1 mAb was used to block the B7-H1, the inhibitory rate of T lymphocyte proliferation decreased from 64.1% to 38.75%. It is concluded that B7-H1 highly expresses on hBMMSCs, the B7-H1 mediated signaling pathway (B7-H1/PD-1) involves in the mechanisms for hBMMSCs-mediated immunomodulation.

FLT3-ITD detection of free DNA in plasma from 235 patients with acute myeloid leukemia and its clinical significance / 中国实验血液学杂志

This study was purposed to evaluate the clinical significance of FLT3-ITD of free DNA in plasma from patients with AML. Free DNA in plasma of 235 patients with AML were extracted and identified by globin gene. FLT3 was amplified by PCR and compared with detected results of leukemic cellular DNA (BM or PB). The results indicated that out of total 235 patients, globin gene in plasma free DNA was successfully amplified from 190 cases. In 188 newly diagnosed, replaced and refractory cases, 35 cases showed ITD mutation (19%). And they also showed ITD mutation in leukemic cellular DNA. But in 47 patients in remission, 2 patients with FLT3-ITD mutation of free DNA in plasma had no mutation in cellular DNA, but got relapse early. Compared with patients of FLT3-wt, patients with FLT3-ITD mutation had increased WBC count and expression rate of CD7, CD56 and decreased CR rate. It is concluded that leukemic-specific DNA in plasma can be detected in AML patients and consistent with detected results of leukemic cellular DNA. Furthermore, the free DNA in plasma is more sensitive for MRD monitoring in remitted patients. FLT3-ITD detection plays an important role in evaluation of prognosis and molecular target therapy for AML patients.

The clinical significance of Ig heavy chain and TCR gamma gene rearrangement detected in free DNA in plasma in patients with non-Hodgkin lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the clinical significance of IgH and TCR gamma gene rearrangement in plasma free DNA in patients with non-Hodgkin Lymphoma (NHL).</p><p><b>METHODS</b>Plasma free DNA in 74 patients with NHL were extracted and identified by Globin gene. IgH (FR3A/VLJH), TCR gamma (TVG/TJX) clonal rearrangements were amplified by PCR and compared with results of mononuclear cell DNA and pathological biopsy sample DNA.</p><p><b>RESULTS</b>Plasma free DNAs were successfully obtained from 58 cases (35 B-NHL and 23 T-NHL) of newly diagnostic, refractory and relapsed NHL out of total 74 patients (78.4%), but not found in the rest 16 patients in remission. Of 35 B-NHL cases, 31 showed IgH rearrangement (88.6%), and none with TCR gamma rearrangement; of 23 T-NHL cases, 8 showed TCR gamma rearrangement (34.8%), and 2 with IgH gene rearrangement synchronously. In comparison with the results of IgH and TCR gamma gene rearrangement in biopsy samples in 30 B-NHL cases, 26 cases in plasma free DNA (86.7%) and 24 in biopsy samples (80%) were positive (P > 0.05). In 20 T-NHL patients, 7 cases in plasma cell-free DNA (35%) and 6 cases in biopsy samples (30%) were positive (P >0.05).</p><p><b>CONCLUSIONS</b>Tumor-derived DNA could be detected in plasma from underlying cancer patients. For NHL patients, detecting IgH and TCR gamma gene rearrangement in plasma free DNA has the same clinical significance as in biopsy samples.</p>

Establishment and evaluation of hematology review criteria for action following automated complete blood counts and leukocytes differential / 中华检验医学杂志

Objective To establish and evaluate criteria applied to review of complete blood counts (CBC)and differential results from automated hematology analyzers.Methods Temporary criteria were established by using alarm system of XE-2100 automated hematology analyzer and by consulting the 41 suggested rides of international consensus group.2 795 out-and in-patient samples were run as clinical samples.Stained blood films were prepared and manual differential with smear review were performed on all samples.Statistical analysis was done for each temporary rule and instrument flag which indicated abnormal cell quantity and morphology.Results Of all rules,instrument flags of ‘Immature Gran/Left Shift?’, ‘ Atypical Lympho?’and‘NRBC(nucleated red blood cell)?’showed most frequent false positive and false negative instrument flag.Evaluation on rnles about cell quantity change showed false positive and false negative rates were both low.Results of morphology evaluation showed that true positive rate was 17.44%, false positive rate was 15.82%,true negative rate was 63.49%,false negative rate was 3.25%.‘ Atypical Lymphocyte?’,‘Immature Gran?’and‘blast?’were the most frequent false positive flags.According to those results and clinicians opinions,our hematology review criteria for action following automated CBC and leukocytes differential was established.Conclusions The hematology review criteria have high true positive rate and low false negative rate.To clinical hematology laboratory using automated hematology analyzer,new criteria can reduce work load,bring lower false negative rate and higher work efficiency.