Oral arsenic plus imatinib versus imatinib solely for newly diagnosed chronic myeloid leukemia: a randomized phase 3 trial with 5-year outcomes.

PURPOSE: The synergistic effects of combining arsenic compounds with imatinib against chronic myeloid leukemia (CML) have been established using in vitro data. We conducted a clinical trial to compare the efficacy of the arsenic realgar-indigo naturalis formula (RIF) plus imatinib with that of imatinib monotherapy in patients with newly diagnosed chronic phase CML (CP-CML). METHODS: In this multicenter, randomized, double-blind, phase 3 trial, 191 outpatients with newly diagnosed CP-CML were randomly assigned to receive oral RIF plus imatinib (n = 96) or placebo plus imatinib (n = 95). The primary end point was the major molecular response (MMR) at 6 months. Secondary end points include molecular response 4 (MR4), molecular response 4.5 (MR4.5), progression-free survival (PFS), overall survival (OS), and adverse events. RESULTS: The median follow-up duration was 51 months. Due to the COVID-19 pandemic, the recruitment to this study had to be terminated early, on May 28, 2020. The rates of MMR had no significant statistical difference between combination and imatinib arms at 6 months and any other time during the trial. MR4 rates were similar in both arms. However, the 12-month cumulative rates of MR4.5 in the combination and imatinib arms were 20.8% and 10.5%, respectively (p = 0.043). In core treatment since the 2-year analysis, the frequency of MR4.5 was 55.6% in the combination arm and 38.6% in the imatinib arm (p = 0.063). PFS and OS were similar at five years. The safety profiles were similar and serious adverse events were uncommon in both groups. CONCLUSION: The results of imatinib plus RIF as a first-line treatment of CP-CML compared with imatinib might be more effective for achieving a deeper molecular response (Chinadrugtrials number, CTR20170221).

Current Treatment Patterns of Aplastic Anemia in China: A Prospective Cohort Registry Study.

Aplastic anemia (AA) is a hematologic disease characterized by pancytopenia and hypocellular bone marrow, potentially leading to chronic anemia, hemorrhage, and infection. The China Aplastic Anemia Committee and British Committee for Standards in Haematology guidelines recommend hematopoietic stem-cell transplantation (HSCT) or immunosuppressive therapy (IST) comprising antithymocyte globulin (ATG) with cyclosporine (CsA) as initial treatment for AA patients. With limited epidemiological data on the clinical management of AA in Asia, a prospective cohort registry study involving 22 AA treatment centers in China was conducted to describe the disease characteristics of newly diagnosed AA patients and investigate real-world treatment patterns and patient outcomes. Of 340 AA patients, 72.9, 12.6, and 3.5% were receiving IST, traditional Chinese medicine, and HSCT, respectively, at baseline; only 22.2% of IST-treated patients received guideline-recommended ATG with CsA initially. Almost all patients received supportive care (95.6%) as blood transfusion (97.8%), antibiotics (63.7%), and/or hematopoietic growth factors (58.2%). Overall, 64.8% achieved a partial or complete response, and 0.9% experienced relapse. No new safety concerns were identified; serious adverse events were largely unrelated to the treatment regimen. These results demonstrate the need to identify and minimize treatment barriers to standardize and align AA management in China with treatment guideline recommendations and further improve patient outcomes.

[Clinical Efficacy of Second-generation Tyrosine Kinase Inhibitor in Treating Chronic Myeloid Leukemia].

OBJECTIVE: To evaluate the efficacy of second-generation tyrosine kinase inhibitor (TKI) in treating chronic myeloid leukemia (CML). METHODS: A total of 97 patients with CML were enrolled. The patients were treated with TKI and monitored with complete blood count, cytogenetic and molecular indicators during the course of therapy. Survival analysis was performed to evaluate its clinical efficacy. RESULTS: The treatment achieved 97.9% complete hematologic response (CHR), 63.9% major cytogenetic response (MCyR), 60.0% complete cytogenetic response (CCyR) and 44.3% major molecular response (MMR) rates. Apart from CHR, better effects were shown in those indicators during chronic phase compared with progressive phase (P < 0.05). The 1-year, 2-year, 3-year and 5-year overall survival (OS) rate was (90.6 ± 3.0)%, (80.1 ± 4.5)%, (77.5 ± 5.0)% and (64.6 ± 9.3)%, respectively, compared with an event-free survival (EFS) rate of (81.1 ± 4.0)%, (64.4 ± 5.3)%, (56.4 ± 6.0)% and (46.2 ± 8.2)%, respectively. The patients had a 1-year, 2-year, 3-year and 5-year progession-free survival (PFS) rate of (87.4 ± 3.4)%, (73.2 ± 4.9)%, (68.9 ± 5.5)% and (57.4 ± 8.7)%, respectively. A difference between chronic phase (better results) and progressive phase (P < 0.05) was also found in survival indicators. The first-line TKI therapy had 100% CHR, 95% MCyR, 95% CCyR and 70% MMR, compared with 97.3% CHR, 56.8% MCyR, 48.6% CCyR and 36.5% MMR for the second-line TKI therapy. Apart from CHR, the first-line therapy produced better results than the seond-line therapy (P < 0.05). CONCLUSION: CML patients in chronic phase and first-line use of TKI have better outcomes.

Standardized fluorescence in situ hybridization testing based on an appropriate panel of probes more effectively identifies common cytogenetic abnormalities in myelodysplastic syndromes than conventional cytogenetic analysis: a multicenter prospective study of 2302 patients in China.

In an attempt to establish the advantages of fluorescence in situ hybridization (FISH) studies over conventional cytogenetic (CC) analysis, a total of 2302 de novo MDS patients from 31 Chinese institutions were prospectively selected in the present study for both CC and standardized FISH analysis for +8, -7/7q-, -5/5q-, 20q- and-Y chromosomal abnormalities. CC analysis was successful in 94.0% of the patients; of these patients, 35.9% of the cases were abnormal. FISH analysis was successful in all 2302 patients and detected at least one type of common cytogenetic abnormality in 42.7% of the cases. The incidences of +8, -7/7q-, -5/5q-, 20q- and-Y chromosomal abnormalities by FISH were 4.1% to 8.7% higher than those by CC. FISH identified abnormalities in 23.6% of the patients exhibiting normal CC results and revealed that 20.7% of the patients with adequate normal metaphases (≥20) had abnormal clones. FISH identified cytogenetic abnormalities in 50.4% of the patients with failed CC analysis. In summary, our multicenter studies emphasised and confirmed the importance of applying standardized FISH testing based on an appropriate panel of probes to detect common cytogenetic abnormalities in Chinese de novo MDS patients, particularly those with normal or failed CC results.

[Efficacy of tyrosine kinase inhibitor in treatment of chronic myeloid leukemia patients].

OBJECTIVE: To evaluate the effects of tyrosine kinase inhibitors (TKI) in the treatment of chronic myeloid leukemia. METHODS: There were total 655 cases of chronic myeloid leukemia treated in one single-institution enrolled in this study. The dosage of TKI Imatinib was 400 mg/d for chronic phase, 600 mg/d for accelerated and blast phase respectively. Complete blood count, cytogenetic and molecular studies were regularly monitored during the course of therapy. The therapeutic effect was evaluated and the survival analysis was performed. RESULTS: The total complete hematologic response (CHR), major cytogenetic response (MCyR), complete cytogenetic response (CCyR) and major molecular response (MMR) rates were 92.1%, 75.8%, 73.1% and 47.9% respectively. 1-year, 3-year, 5-year and 10-year overall survival (OS) rates were (96.3 +/- 0.8)%, (86.3 +/-1.8)%, (79.0 +/- 2.4)% and (66.5 +/- 4.8)% respectively. 1 year, 3-year, 5-year and 10-year event-free survival (EFS) rates were (92.2 +/- 1.1)%, (77.9 +/- 2.1)%, (67.9 +/- 6.8)% and (35.8 +/- 6.0)% respectively. The proportion of the patients in chronic phase achieving CHR, MCyR, CCyR and MMR were 98.7%, 82.5%, 79.4% and 52.4% respectivly. Compared with chronic phase patients, the efficacy of IM in the treatment of accelerated phase and blast phase patients was significantly lower. The effect of TKI in early chronic phase was better than that in late chronic phase. Early molecular response was associated with a better 5-year EFS, but not OS. CONCLUSION: CML patients in chronic phase treated with TKI have a better outcome. The earlier TKI be used, the better the prognosis and efficacy be achieved.

Effect of Antiviral Prophylaxis Strategy for Chemotherapy-Associated Hepatitis B Reactivation in Non-Hodgkin's Lymphoma Patients with Hepatitis B Virus Infection: A Retrospective Cohort Study.

Recent data indicates that nucleoside/nucleotide analogue (NUC) is effective in preventing and controlling hepatitis B virus (HBV) reactivation in HBV-carrying cancer patients who undergo chemotherapy, but the ideal antiviral agent and optimal application protocol still needs to be determined. Meanwhile, it is uncertain whether those with past HBV infection require antiviral prophylaxis during chemotherapy. This report retrospectively analyzed non-Hodgkin's lymphoma (NHL) patients seen from January, 2004 to June, 2009 in West China Hospital. We found that the prevalence of chronic HBV infection in our NHL patients was 20.7 % while that of past HBV infection was 21.05 %. Compared with the high rate (25.6 %) of HBV reactivation in patients with chronic HBV infection, none of those with past HBV infection in fact had occult HBV infection thus none experienced reactivation. Of the 82 patients with chronic HBV infection who received chemotherapy, antiviral prophylaxis could significantly reduce the incidence of HBV reactivation (5.0 vs. 45.2 % in the control group) and the incidence of liver function damage (32.5 vs. 73.8 % in the control group). The results of the current study confirmed previous reports that prophylactic NUCs administration can effectively prevent HBV reactivation and significantly reduce the incidence of HBV reactivation especially for patients receiving rituximab-containing regimens. Due to the fact that none of individuals who had past HBV infection developed HBV reactivation reported in our study, antiviral prophylaxis may not be required for patients with past HBV infection. Close observation of alanine aminotransferase and HBV-DNA contributes to early diagnosis and timely treatment of HBV reactivation.

[Effects of imatinib mesylate on the levels of endocrine hormones].

OBJECTIVE: To measure the levels of hormones in chronic myelogenous leukemia (CML) patients receiving imatinib mesylate (IM) and evaluate the effects of IM on endocrine system. METHODS: 69 patients with CML while taking IM were enrolled and a total of 86 peripheral blood samples were detected. The levels of total triiodothyronine (TT3), total tetraiodothyronine (TT4), thyroid stimulating hormone (TSH), testosterone, progesterone, estradiol (E2), plasma total cortisol (PTC) at 8:00-10:00 am measured. Concentration of hormones in different groups were measured to evaluate the effects of IM on endocrine system and relationships with its administration duration, plasma concentration and clinical symptoms. RESULTS: (1) Of the 7 types of hormones, an elevation of TSH level was found in 14 patients (20.3%), a decrease of TT3 and testosterone in 8 patients (11.6%) and 8 males (18.6%), respectively. (2) A significant decline of TT3 and testosterone was observed in all patients divided by different administration duration. Negative correlation was seen between TT3 level and duration of administration (r=-0.273, P=0.010), which was also found for testosterone (r=-0.302, P=0.025). (3) There was no correlation between serum levels of the seven hormones and concentration of IM. CONCLUSION: IM affect the levels of thyroid and sex hormones in some patients with clinical manifestations: a decrease of TT3, testosterone and testosterone, an increase of TSH, which have relationship with the duration of administration.

[Early detection of MRD in peripheral blood after induction chemotherapy of newly diagnosed patients with AML and its correlation with curative effects].

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.

Flow cytometric immunophenotyping is of great value to diagnosis of natural killer cell neoplasms involving bone marrow and peripheral blood.

Natural killer (NK) cell neoplasms are unusual disorders. In this study we compared results of flow cytometric immunophenotype (FCI) with cytomorphology, histopathology and clinical findings in a series of patients with NK cell neoplasms with peripheral blood and/or bone marrow involvement, and the FCI of neoplastic and normal NK cells were compared. Retrospective data and specimens (bone marrow aspiration or peripheral blood) from 71 cases of NK cell neoplasms were obtained. All patients have been demonstrated laboratory and clinical features consistent with NK cell neoplasms, and the subtypes were determined by integrated clinical estimation. Routine 4-color flow cytometry (FCM) using a NK/T cell related antibody panels was performed. NK cell neoplasms were divided into two major subtypes by FCI, namely malignant NK cell lymphoma, including extranodal nasal type NK cell lymphoma (ENKL, 11 cases) and aggressive NK cell lymphoma/leukemia (ANKL, 43 cases), and relative indolent chronic lymphoproliferative disorder of NK cell (CLPD-NK, 17 cases). The former exhibited stronger CD56-expressing, larger forward scatter (FSC) and more usually CD7- and CD16-missing. FCI of CLPD-NK was similar to normal NK cells, but CD56-expressing was abnormal, which was negative in five cases and partially or dimly expressed in eight cases. Cytomorphologic abnormal cells were found on bone marrow slides of 4 cases of ENKL and 30 cases of ANKL. Eight cases of ENKL were positive in bone marrow biopsies, and other three cases were negative. In 32 cases of ANKL which bone marrow biopsies were applied, 21 cases were positive in the first biopsies. Lymphocytosis was found only in six cases of CLPD-NK by cytomorphology, and biopsy pathology was not much useful for diagnosing CLPD-NK. These results suggest that FCM analysis of bone marrow and peripheral blood was superior to cytomorphology, bone marrow biopsy, and immunohistochemistry in sensitivity and early diagnosis for ANKL, stage III/IV ENKL and CLPD-NK. FCI could not only define abnormal NK cells but also determine the malignant classification. It is beneficial for clinical management and further study of NK cell neoplasms.

[Immunophenotype characteristics and prognosis of acute leukemia patients with cross expressing lymphoid and myeloid lineage associated antigens].

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.

[Flow cytometric analysis of cerebrospinal fluid in the diagnosis of central nervous system leukemia].

OBJECTIVE: To evaluate the value of flow cytometric immunophenotyping of cerebrospinal fluid (CSF) cells in the diagnosis of central nervous system leukemia. METHODS: Ninety two CSF samples were analyzed with 4-color flow cytometry. Antibody panles CD19/CD34/CD3/CD45, CD117/CD34/CD5/CD45, CD7/CD34/ CD19/CD45, CD7/CD3/HLA-DR/CD45, CD20/CD10/CD3/CD45, and anti-g/anti-lambda/CD19/CD45 were used in determining cell composition and detecting abnormal cells. The results of flow cytometry were compared with conventional cell count and morphology. Flow cytometry analysis was repeated for five samples 48 hours after the initial test. RESULTS: Abnormal cells were found in 33 out of the 92 (35.9%) samples. Among the 59 samples taken from patients with lymphocyte neoplasm, CD19 + blast cells were found in the CSF in 13 patients with B-cell lymphoblastic leukemia; CD7+ blast cells were found in 4 T-ALL cases; and monoclonal CD19+ cells were found in 6 other types of lymphoma cases. In the 32 patients with clinically diagnosed myeloid leukemia, CD117+ myeloid cells were found in the CSF of 7 patients and B cell blast cells were found in 2 CML cases. The abnormal cells in the CSF detected by immunophenotyping decreased significantly 48 hours after the initial test. Abnormal cells were detected in 25 samples (27.2%) by morphology, less than those detected by immunophenotyping. The cell concentrations of the eight samples in which abnormal cells were only detected by flow cytometry were lower than 10 X 10(6)/L. The immunophenotyping results of two ALL patients were still positive when morphologic results had become negative after chemotherapy. CONCLUSION: Flow cytometric analysis of CSF may be helpful in the diagnosis of meningeal leukemia. It has higher positive rate and better accuracy than cytomorphology and cell count.

[Expression of hOCT1 in patients with chronic myelogeneous leukemia treated by imatinib mesylate].

OBJECTIVE: To measure the expression of hOCT1 gene in patients with Chronic Myelogeneous Leukemia (CML) and to explore its role in the progress of the disease and responding to Imatinib Mesylate (IM) treatment. METHODS: Sixty three peripheral blood or bone marrow samples were taken from 30 patients with CML (20 in chronic phase, 10 in advanced phase). The samples were divided into two groups: responding (optimal and suboptimal) and non-responding according to effectiveness of the IM treatment. The mRNA levels of hOCT1 gene were detected with RT-PCR (SQ-PCR), 3, 6, 9, 12 and 15 months after the IM therapy. The associations between hOCT1 gene levels and clinical presentations, laboratory indicators and cytogenetic findings were analysed. RESULTS: No significant difference in the expression levels of hOCT1 gene was found before and after the IM treatment (0.5110+/-0.1629 vs 0.5207+/-0.1909, P=0.5840). No significant difference in the expression levels of hOCT1 genes was found between the patients in chronic phase and the patients in advanced phase before the IM treatment (0.5525+/-0.1985 vs 0.4490+/-0.1717, P=0.1090). The levels of hOCT1 did not have significant changes 3, 6, 9, 12 and 15 months after the IM treatment (P=0.3412). No significant difference in the expression levels of hOCT1 genes was found between the 15 patients with optimal and suboptimal responding to IM and the 5 patients with no responding to IM (0.5820+/-0.1460 vs 0.4640+/-0.1781, P=0.127). Although the hOCT1 levels of the 16 chronic patients increased after IM treatment compared to the baseline (0.5207+/-0.1909 vs 0.5110+/-0.1629, P=0.001), there was no significant correlation between the increase of hOCT1 and the decrease of BCR-ABL (P=0.821). CONCLUSION: hOCT1 has no association with the stage and course of CML, nor with the effectiveness of IM therapy.

[Influence of hOCT1 polymorphism on imatinib mesylate effectiveness in chronic myelogenous leukemia patients].

OBJECTIVE: To explore the correlation between hOCT1 polymorphism and imatinib mesylate (IM) effectiveness in chronic myelogenous leukemia(CML) patients, and to provide for the clinical individual personalized therapy. METHODS: Fifty-three CML and 23 non-CML patients were enrolled in this study. Blood or bone marrow samples were collected. Amplification refractory mutation system (ARMS)-polymerase chain reaction was used to amplify the polymorphisms gene segment of hOCT1-P283L, R287G and M408V and their frequencies were statistically analysed. With clinical outcomes, the correlation between hOCT1 polymorphism and IM effectiveness in CML was analyzed. RESULTS: (1) For 74 Han Chinese, the allele frequencies of hOCT1-P283L, R287G and M408V were 39.86%, 29.05% and 45.27%, respectively. (2) The genotypes of hOCT1-P283L, R287G and M408V in 2 Tibetan Chinese were CC, CC, AG and CC, CG, AG, respectively. (3) In the CML patients with IM optimal response, the frequencies of 283T and 287G allele were predominant (P<0.05). No significant difference was found in the frequency distribution of hOCT1-M408V genotype and allele among the 3 different response groups (P>0.05). CONCLUSION: (1) Three single nucleotide polymorphisms (cSNP) P283L, R287G and M408V were found in the hOCT1 gene from 76 Chinese. (2) hOCT1 gene polymorphism is associated with the long-term molecular response of CML patients received IM therapy, indicating that the polymorphisms of hOCT1-283T, 287G may be good predictors for IM response. (3) There is no correlation between the polymorphisms of hOCT1-P283L, R287G, M408V and secondary IM resistance in CML patients.

[MMP-9 expression profile in inflammatory cells of Mip-1alpha knockout mice and Mip-1alpha receptor knockout mice].

OBJECTIVE: To explore the MMP-9 expression profile in peritoneal inflammatory macrophages and granulocytes in Mip-1alpha-, CCR1- and CCR5-deficient mice. METHODS: In sodium thioglycolate-induced murine peritonitis models, peritoneal macrophages and granulocytes were harvested, identified and purified from WT mice and Mip-1alpha-, CCR1-, CCR5-deficient mice. The RT-PCR was applied to evaluate the expression of MMP-9 in macrophages and granulocytes of different group of mice. RESULTS: The expressions of MMP-9 of macrophages in Mip-1alpha-, CCR1-, CCR5-deficient mice were significantly lower than that of WT mice (P<0.05); MMP-9 expression of granulocytes in Mip-1alpha-, CCR5-deficient mice were also significantly lower than that of WT mice (P<0.05), while the MMP-9 expression of granulocytes in CCR1-deficient mice was significant higher than that of WT mice. CONCLUSION: Deletion of Mip-1alpha and CCR5 could reduce the MMP-9 expression in both macrophages and granulocytes, while deletion of CCR1 could reduce MMP-9 expression in macrophages but increase MMP-9 expression in granulocytes.

[Overview of chronic myelogenous leukemia and its current diagnosis and treatment patterns in 15 hospitals in China.].

OBJECTIVE: To explore demographic characteristics, current diagnosis and treatment patterns of chronic myelogenous leukemia (CML) patients in China. METHODS: Data of hospitalized CML patients in 2005 whole year and outpatient information (July 1 through September 30, 2006) from 15 hospitals throughout China were analyzed. RESULTS: A total of 1824 CML cases were analyzed, including 722 inpatients and 1102 outpatients. The male/female ratio was 1.78:1. The median age at diagnosis was 40.02 (2.45 - 83.29) years old, 90.41% of the patients were diagnosed at chronic phase. Proportion of accelerated phase or blast crisis patients increased to 21.66% during study period. 93.20% of the patients received blood routine and bone marrow morphologic examination at diagnosis and in monitoring; 70.29% were performed cytogenetic analysis and 51.54% performed molecular measurement in addition. The most common therapy for CML treatment was hydroxycarbamide. The proportion of patients treated with imatinib and interferon was 37.45% and 25.55%, respectively. Of 722 inpatients, 164 (22.72%) received hemotopoietic stem cell transplantation (HSCT). The proportions of accelerated phase and blast crisis patients treated with imatinib were 48.28% and 48.42%, respectively, being significantly higher than that of chronic phase patients (35.9%) (P < 0.05). The mean imatinib dosage administered in the three phases patients did not differ significantly. Imatinib resistance rates were 6.87% and 16.28% for outpatient and inpatient, respectively. In the outpatient group, the primary resistance to imatinib occurred comparably to the secondary resistance (68.75%), while primary resistance was predominant in inpatient group (65.71%). The intolerance rates of imatinib for outpatient and inpatient were 3.21%, 11.63%, respectively. The majority of patients treated with imatinb were not monitored in time: 63.38% patients evaluated hematologic response after 3 months of treatment, proportions of patients received cytogenetic examination after 6 months and 12 months of treatment were 41.41% and 27.35%, respectively. Mean cost for HSCT was 213 092 +/- 125 890 RMB. CONCLUSIONS: CML in China tends to afflict younger population than in Western countries. Most patients were diagnosed in the chronic phase. Due to restriction of financial support, only one third of CML patients were treated with imatinib, and the majority of the treated were not monitored in time. Clinicians should pay attention to resistance and intolerance to imatinib treatment in accelerated phase or blast crisis patients.

[Role of gammadeltaT cells in pathogenesis of acquired pure red cell aplastic anemia].

This study was purposed to investigate the changes in quantum and function of gammadelta T cell subsets, and to explore its significance in pathogenesis of acquired pure red cell aplastic anemia (A-PRCA). Eleven patients were diagnosed as A-PRCA based on bone marrow smear and biopsy, and were treated with cyclosporine A and glucosidorum tripterygll totorum. The flow cytometry technique was used for analyses of T cells subsets and gammadelta T cells. Furthermore, peripheral mononuclear cells (MNC) isolated from A-PRCA patients were cultured in RPMI 1640 medium (10(5) cells/ml) containing 10% FCS, phytohemagglutinin (PHA, 10 microg/ml), and recombinant human interleukin-2 (rIL-2, 50 U/ml) for two weeks, then gammadelta T cells were isolated with the TCRgammadelta Microbead Kit from cultured cells. The collected gammadelta T cells were incubated with normal control bone marrow MNC in RPMI 1640 medium (37 degrees C, 5% CO2 atmosphere) for CFU-E, CFU-GM, and BFU-E colony assay. The result showed that compared with the control group, CD3(+), CD8(+) cells increased significantly in the patient group (P < 0.05), the CD4(+)/CD8(+) ratio decreased and reversed, and gammadelta T cells were significantly increased in patient group (P < 0.05). After treatment with cyclosporine A, 9 out of 11 patients got good response, and CD3(+), CD8(+) cells in the responding patient decreased, the ratio of CD4(+)/CD8(+) returned to normal, and gammadelta T cells also decreased to normal range. Moreover, in vitro culture, the gammadelta T cells isolated from A-PRCA patients showed an inhibiting action to CFU-E and BFU-E but not to CFU-GM in a dose-dependent manner. It is concluded that gammadelta T cells increase in A-PRCA patients, and decrease in parallel to normal range with significant improvement of anemia symptoms after immune suppressive therapy. The gammadelta T cells isolated from A-PRCA patients showed an inhibiting action to CFU-E and BFU-E but not to CFU-GM in vitro culture, suggesting that gammadelta T cells may bring an impact on the research of A-PRCA pathogenesis. Cyclosporine A demonstrated better therapeutic effect on A-PRCA patients.

[Establishment of an imatinib resistance cell line K562R and its resistant principia].

OBJECTIVE: To establish an imatinib resistance cell line and to study its resistant principia. METHODS: K562 cells were cultured in imatinib at gradually increased concentrations to generate their resistance cell line. MTT assay, RT-PCR, flow cytometry and HPLC were used to clarify the possible mechanisms of the resistance. RESULTS: (1) Imatinib resistance cell line K562R was successfully induced by continuous culture in the presence of gradually increasing doses of imatinib up to 5 micromol/L. K562R cells were maintained in the media containing 5 micromol/L imatinib. (2) Proliferation data showed that cell growth of K562R was not inhibited in 5 micromol/ L imatinib, whereas the parental sensitive cell was significantly inhibited by up to 2.0 micromol/L imatinib. (3) The IC50 of K562R was about 7.5 micromol/L which was ten times higher than that of the parental cell. (4) HPLC revealed that the intracellular imatinib concentration of K562R was strikingly lower than that of the parental cells (up to 27. 8-fold). By flow cytometry, P-gp was not detected on K562R cell, indicating that low intracellular imatinib concentration may not be due to P-gp mediated efflux. (5) Sequence analysis of the 374 bp ABL kinase domain showed no mutation in K562R cell. CONCLUSION: An imatinib resistance cell line K562R has been successfully established. Low intracellular imatinib concentration is a key link in the chain of cell resistance.

[Effect of arsenic trioxide on bone marrow stromal cells of patients with multiple myeloma].

To explore the effect of arsenic trioxide (As2O3) on growth and secretion of interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) of bone marrow stroma cells (BMSC) from the patients with multiple myeloma (MM). Specimens of bone marrow aspiration from MM patients were used to establish BMSC cultures. BMSC and human MM cell line CZ-1 were cultured together or alone in the absence or presence of As2O3 at various concentrations (1-20.0 micromol/L). Cell growth inhibition was assessed by MTT assay, cytokines in the culture supernatants were measured with ELISA. The results showed that As2O3 had cytostatic effect on CZ-1 with fifty percent growth inhibition (IC50) for 48 hours at 2.3 micromol/L. As2O3 did not inhibit the growth of BMSC. High levels of IL-6 and VEGF have been found in the culture supernatants of BMSC from MM patients. Cytokine production of BMSC treated with As2O3 significantly decreased as compared with controls (P < 0.05). Excitingly, even the increased cytokine production triggered by adhesion of MM cell and BMSC was also inhibited by As2O3. It is concluded that As2O3 has no inhibitory effect on cell growth of BMSC, but inhibit the production of IL-6 and VEGF by BMSC.

[Immune regulatory effect of human bone marrow mesenchymal stem cells on T lymphocyte].

To investigate the immune regulatory effects of human bone marrow mesenchymal stem cells on alloantigen T lymphocyte in vitro, human MSCs were isolated and expanded from bone marrow cells, and identified with cell morphology, and the phenotypes were assessed by immunohistochemistry and flow cytometry. As the stimulation factor of T lymphocytes proliferation, either PHA or dendritic cells isolated from cord blood were cocultured with CD2(+) T lymphocytes from peripheral blood mononuclear cells by magnetic beads with or without MSC in 96-well plats for seven days. T cell proliferation was assessed by [(3)H]-thymidine incorporation using a liquid scintillation counter. T cell subsets, Th1, Th2, Tc1 and Tc2 were analyzed by flow cytometry after co-culture of CD2(+) T cells with MSCs for 10 days. The results showed that a significant decrease of CD2(+) T cell proliferation was evident when MSC were added back to T cells stimulated by DC or PHA, and an increase of Th2 and Tc2 subsets were observed after co-culture of MSC with T lymphocytes. It is suggested that allogeneic MSC can suppress T cell proliferation in vitro and the cause of that was partly depend on interaction of cells and the alteration of T cell subsets.