Decitabine combined with arsenious acid in the treatment of patients with higher-risk myelodysplastic syndromes and chronic myelomonocytic leukemia / 中国实用内科杂志

OBJECTIVE: To evaluate the clinical efficacy of decitabine combined with arsenious acid in the treatment of patients with higher-risk myelodysplastic syndromes(MDS) and chronic myelomonocytic leukemia(CMML). METHODS: Totally 39 patients with MDS and 8 patients with CMML received the treatment of decitabine and arsenious acid from April 2016 to December 2018. Decitabine [20 mg/(m~2·d)] and arsenious acid [0.15 mg/(m~2·d)] were administered intravenously for 5 consecutive days every 4-6 weeks. Patients who achieved complete or partial remission entered into the consolidation cycle. Efficacy and influencing factor were analyzed. RESULTS: Clinical response were observed in 31 patients after a median of 2 courses(ranging 1-12) of treatment. The overall response rate(ORR) was 66.0%. The median duration of response was 16 weeks(ranging 2-52 weeks). There were 8 cases(17.0%) of complete remission(CR), 10 cases(21.3%) of partial remission(PR),12 cases(25.5%) of hematological improvement(HI), 1 case(2.1%) of marrow complete remission(mCR), 8 cases(17.0%) of stable disease(SD), and 1 case(2.1%) of progressive disease(PD). By next generation sequencing, 25 genes mutated with 70 times in 33 cases. The mutation frequency of epigenetic regulators(57.6%) was higher than splicing factors(33.5%), transcription factors and kinase signaling(54.5%),and TP53(21.2%)(P<0.01). There was no significant difference in response rates among these patients(47.4%, 54.5%, 50.0% and85.7%, P=0.977). Gene mutation frequency(VAF) of patients who responded to the regimen declined significantly(16.67% vs. 10.26%,P=0.014). CONCLUSION: Decitabine combined with arsenious acid has significant effect in the treatment of patients with higher-risk MDS and CMML and is well-tolerated. Gene mutation test results by next generation sequencing might be related to clinical response.

Effect of miR-214 on Fludarabine Resistance in Chronic Lymphocytic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate effect and mechanism of miR-214 in fludarabine resistance of chronic lympho-cytic leukemia (CLL).@*METHODS@#A total of 10 patients with CLL resistante to fludarabine (Flu) and 10 healthy persons admitted to Hematology Department of our hospital in August 2014 - July 2018 were selected. Expression level of miR-214 in mononuclear cells in patients with CLL and healthy persons were determined by RT-PCR. Primary CLL cells from patients with CLL were divided into normal control group (control group), negative control group (miR-214-NC group) and viral transinfection group (miR-214-ASO group). After 24 h-transfection, CLL cells were cultured with different con-centration of Flu for 48 h, then the cell proliferation and apoptosis were detected, and the levels of down-stream genes and proteins releted with PTEN and PI3K/AKT signialing pathway were determined.@*RESULTS@#The expression level of miR-214 in mononuclear cells of CLL patients significantly increased in comparison with healthy persons(P<0.05); the expression level of miR-214 in miR-214-ASO group significantly decreased (P<0.05); Absorbance in control group at Flu concentration of 3, 10 and 30 μmol/L was significantly decreased (P<0.05). Apoptosis rate in miR-214-ASO group at Flu concentration of 10 mmol/L significantly increased (P<0.05). At Flu concentration of 10 mmol/L, mRNA levels PTEN and BAD in miR-214-ASO group significantly increased (P<0.05), but mRNA levels of MDM2 and NF-κB significantly decreased (P<0.05). At Flu concentration of 10 mmol/L, protein levels of PTEN and p-BAD in miR-214-ASO group significantly increased (P<0.05), but protein levels of MDM2 and NF-κB significantly decreased (P<0.05).@*CONCLUSION@#Inhibition of miR-214 can enhance the sensitivity of drug-resistant CLL cells to fludarabine, which may be raleted with the promotion of cell apotosis and regulation of down-stream molecules expression of PTEN/AKT signaling pathway.

Influence of Chromosome Abnormality on Therapeutic Efficacy and Prognosis of Newly Diagnosed Multiple Myeloma Treated with Bortezomib / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the influence of chromosome abnormality on therapeutic efficacy and prognosis of patients with newly diagnosed multiple myeloma(MM) treated with bortezomib.</p><p><b>METHODS</b>The clinical data of 152 patients with newly diagnosed MM were collected from January 2008 to December 2011. All patients received bortezo-mib-based chemotherapy and the therapeutic efficacy were investigated for 4 cycles later. The R banding and DNA probe were used to analyze the chromosome and gene (RB1 deletion, D13S319 deletion, P53 deletion, IgH rearrangement and 1q21 amplification) of chromosome specimens. Moreover, the therapeutic efficacy and long-term survival data were analyzed among the patients with different types of chromosomal abnormality. The Kaplan-Meier was applied to analyze survival, and COX risk proportional model was used for multivariate analysis.</p><p><b>RESULTS</b>Among 152 patients with MM, there were 47 cases(30.92%) of abnormal karyotype, 43 cases(28.29%) of abnormal RB1,49 cases (32.24%) of abnormal D13S319, 30 cases (19.74%) of abnormal P53, 58 cases (38.16%) of abnormal IgH and 33 cases (21.71%) of abnormal chromosome 1q21. All the patients were evaluable for the therapeutic efficacy, including 24 CR, 54 nCR, 21 PR, 14 MR and 39 PD with response rate of 74.34% and remission rate of 50.66%. Compared with normal controls, the response and remission rate were lower than that in the patients with abnormal karyotype of D13S319, P53 or IgH, and remission rate was lower in the patients with RB1 or 1q21 (P<0.05). All the patients were followd-up (median: 52.0 months, range: 22-72 months), but median overall survival(OS) was not yet reached at the end of the follow-up. The median OS was in the patients with different chromosome versus the normal subjects (P<0.05). The chromosome abnormality was found to affect the prognosis of MM by COX multivariate analysis. In regard to the normal subjects, the risk for poor prognosis increased by 1.177, 2.639, 6.552, 3.124, 2.045 and 7.264 fold in the patients with abnormal Karyotype of RB1, D13S319, P53, IgH and 1q21, respectively.</p><p><b>CONCLUSION</b>The abnormality of chromosome can influence the efficacy and prognosis of newly diagnosed MM patients treated with bortezomib. The detection of chromosomal abnormalities has a certain reference value for the treatment of primary MM.</p>

Effect of Pin1 inhibitor juglone on proliferation, migration and angiogenic ability of breast cancer cell line MCF7Adr / 华中科技大学学报（医学）（英德文版）

This study aimed to evaluate the effects of Pin1 inhibitor Juglone on proliferation, migration and the angiogenic ability of breast cancer cell line MCF7Adr. MCF7Adr cells were cultured and separately treated with Pin1 inhibitor Juglone (treatment group) and DMEM without drug (control group). The cell cycle was examined by flow cytometry. Cell migration was measured by wound-healing assay. Cyclin E protein content was detected by Western blotting. The angiogenesis factor vascular endothelial growth factor (VEGF) in cell media was determined by enzyme linked immunosorbent assay. The results showed that the percentage of cells in G2/M phase in treatment group was significantly higher than that in control group (25.5% vs. 10.1%, P<0.05), and that in G0/G1 phase and S stage in treatment group was significantly lower than that in control group (40.5% vs. 48.2%, and 33.7% vs. 41.7%, P<0.05). Cyclin E protein content in treatment group was significantly lower than that in control group (39.2 ± 7.4 vs. 100 ± 23.1, P<0.05). (A0-A24)/A0 value in treatment group was significantly lower than that in control group (23.9 ± 3.8 vs. 100 ± 14.4, P<0.05). VEGF-A, -B, and -C contents in cell media of treatment group were significantly lower than those in control group (P<0.05). It was suggested that Pin1 inhibitor Juglone can effectively inhibit the proliferation, migration and the angiogenic ability of MCF7Adr cells, and can be used as an alternative drug therapy for breast cancer.

Polymorphisms of FcγRIIA, FcγRIIIA and FcγRIIB in patients with immune thrombocytopenia and their clinical significance / 中国实验血液学杂志

This study was aimed to investigate the correlation of FcγR polymorphisms with the susceptibility, severity and efficacy of immunotherapy for patients with immune thrombocytopenia (ITP). PCR and DNA sequencing were used to determine the polymorphisms of FcγRIIA, FcγRIIIA and FcγRIIB in 44 ITP patients, and in 97 healthy control subjects. The results indicated that FcγRIIIA-158V/F polymorphisms between patients and controls were statistically significantly different (P = 0.015); among FcγRIIIA genotypes, the frequency of 158V/V homotype was higher in ITP (P = 0.005). However, the FcγRIIA-131H/R or FcγRIIB-232T/I polymorphisms were not significantly different between patients and controls; there were no correlation of FcγRIIA, FcγRIIIA and FcγRIIB genotype frequencies with the platelet counts or the courses of ITP; among the 38 ITP patients who received treatments, the complete response (CR) rate was 42% (16/38), and partial response (PR) rate was 34% (13/38). The therapeutic response was significantly different between FcγRIIIA-158V/V homotype and 158F/V heterotype (P = 0.034). The CR of patients with 158V/V homotype was obviously lower than that of patients with 158F/V, but the frequencies of FcγRIIA and FcγRIIB genotypes not correlated with the responsiveness to treatment. The CR rate of 6 patients treated with rituximab was 67%, and PR rate was 17%. The overall response rate was as high as 84%, the adverse reactions were not observed. It is concluded that the polymorphism of FcγRIIIA-158V/F, but not FcγRIIA-131H/R or FcγRIIB-232T/I, correlates with the patient susceptibility and therapeutic response of ITP.

Autoimmune hemolytic anemia associated with B-cell chronic lymphoproliferative disorders / 中国实验血液学杂志

This study was purpose to investigate the clinical characteristics of B-cell chronic lymphoproliferative disorders (B-CLPD) complicated by autoimmune hemolytic anemia (AIHA) so as to improve the understanding of this disease. The clinical characteristics, laboratory data, therapy and outcome of 14 patients suffering from B-CLPD complicated by AIHA were retrospectively analyzed in Wuxi People Hospital and the First Affiliated Hospital of Nanjing Medical University from 2000 to 2012. The results showed that 9 cases of the 14 patients were patients with chronic lymphocytic leukemia (CLL), 5 cases were patients with lymphoma, at time of hemolysis the median level of hemoglobin was 61 (33 - 84)g/L, the median ratio of reticulocytes was 12.0 (3.1 - 35.0)%, the positive rate of Coombs test was 100%. 1 case received corticosteroid alone, 5 cases were treated with chemotherapy combined with corticosteroid, 8 cases were treated with immunochemotherapy rituximab combined with corticosteroid. Overall response rate was 100%, in which CR was 78.6% (11/14), PR was 21.4% (3/14). The follow-up for these patients were performed to now, 35.7% (5/14) patients relapsed with hemolysis again, but they showed therapeutic response to treatment with above-mentioned therapy. From patients treated with rituximab alone, only 1 patient relapsed. Among 14 patients, 6 cases died, 1 case was lost, the other cases are still alive. It is concluded that the AIHA is the commonest complication of B-CLPD, it can be observed at different stages of B-CLPD, the treatment with corticosteroids can give well therapeutic effect for these patients, but the long time CR is lower, the rituximab has been confirmed to be effective for B-CLPD complicated by AIHA.

Research advances on correlation of ARID5B gene with childhood acute lymphoblastic leukemia - review / 中国实验血液学杂志

Childhood acute lymphoblastic leukemia (C-ALL) is the most common pediatric cancer. Although its etiology remains poorly understood, the hypothesis of ALL correlated with a genetic basis was examined through association studies based on candidate genes. Recently, two independent large-scale genome-wide association studies reported that the five single nucleotide polymorphisms (rs7073837; rs10821936; rs10994982; rs7089424; rs10740055) in the gene AT rich interactive domain 5B (ARID5B) at 10q21.2, were associated with the high incidence risk of C-ALL, especially with hyperdiploid lymphoblastic leukemia. Variations in these single nucleotide polymorphisms influence the risk of specific disease subtypes, and also possess race- and sex-differences in leukemia incidence. Further elucidation of the mechanisms through which ARID5B variants are involved in C-ALL not only has a great diagnostic value, but also a guidance for the clinical therapy, ultimately improving the prognosis of disease. Therefore, the related studies of ARID5B with C-ALL were summarized briefly in this review.

Isocitrate dehydrogenase gene mutations in acute myeloid leukemia / 中国实验血液学杂志

The purpose of this study was to identify point mutation of the isocitrate dehydrogenase gene (IDH1 and IDH2) in patients with acute myeloid leukemia(AML) and its clinical significance. 90 de novo AML patients were selected for this study, the genomic DNA was served as template, the exon4 of IDH1 and IDH2 were amplified respectively. The IDH mutation was detected by using directly sequencing method for PCR product. The results indicated that among 90 de novo AML patients, 4 patients (4.4%) showed the IDH1 gene mutation positive, and 7(7.8%) patients showed IDH2 gene mutation positive. None was found harboring both mutations, the overall rate of mutation positive of them was 12.2%. In the AML patients with IDH gene mutation positive, the rate of normal karyotype was 72.7%, which was significantly higher than that in abnormality karyotype. The CR rate in mutation positive patients was 72.7%, which seemed as if higher than that in mutation negative patients, but without statistical significance. The mutation disappeared when the patients gained CR, and reappeared in same loci after relapse occurred. It is concluded that the IDH gene point mutation appears in normal karyotype patients, especially in patients combined with NPM1 gene mutation. The IDH gene mutation may be an important target for therapy and evaluating clinical prognosis of patients with normal karyotype.

Mutation of tet2 gene and malignant blood disease / 中国实验血液学杂志

Tet2 (the 2nd member of tet oncogene family) is a newly discovered antioncogene on the chromosome 4q24 of the patient with malignant myeloma, which has a potential for functional deletion. Recent studies demonstrated that tet2 mutation was found in polycythemia vera (PV), essential thrombocythemia (ET), myelofibrosis, systematic mastocytosis (SM), and myelodysplastic syndrome (MDS). However, a great number of perspective researches are still needed for exploring the role of tet2 in the pathogenesis of malignant blood diseases. In this review, the relation of tet2 mutation with myeloproliferative neoplasm, systemic mastocytosis, myelodysplastic syndrome, acute myeloid leukemia and other malignant blood diseases are summarized.

Expression of the CDH13 gene and BCR/ABL fusion gene in chronic myeloid leukemia patients and their relationship / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the expression of the CDH13 gene and BCR/ABL fusion gene in chronic myeloid leukemia(CML) patients at different stages and explore their relationship.</p><p><b>METHODS</b>RNA was isolated from peripheral blood in 30 healthy adults, 22 primary CML patients and 25 blastic crisis of CML patients. We examined the expression of the CDH13 gene and BCR/ABL fusion gene using EVA Green real-time reverse transcriptase-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The expression of the BCR/ABL fusion gene in the patients with blastic crisis CML was 4.72 folds as that of the patients with primary CML. The expression of the CDH13 gene in the primary and blastic crisis CML patients was significantly reduced to 36% and 25% of that in healthy controls, respectively. Moreover, negative correlation was found between the expressions of these two genes.</p><p><b>CONCLUSION</b>The study showed that the reduction of the CDH13 expression at different clinical stage of CML may account for the defective cell adhesion in CML, and the expression of the CDH13 gene was probably down-regulated by the BCR/ABL fusion gene.</p>

Correlations of JAK2V617F point mutation with clinical and laboratory features in patients with polycythemia vera / 中国实验血液学杂志

To evaluate JAK2V617F point mutation in patients with polycythemia vera (PV) and its clinical significance, the point mutation was detected by allele specific polymerase chain reaction (AS-PCR), and the clinical and laboratory features of 50 PV patients with JAK2V617F positive and negative mutations were analyzed and compared each other. The results showed that among 50 patients, 31 patients (62.0%) had JAK2V617F point mutation; 12 patients (24.0%) showed thrombosis and microvascular disturbances; 3 patients had chromosome karyotype abnormalities. As compared with negative mutation group, the age and leukocyte count in patients with JAK2V617F point mutation were older (57.5 +/- 10.0 vs 45.6 +/- 14.9, p < 0.05) and higher (16.2 +/- 6.7 vs 9.0 +/- 5.2, p < 0.05) respectively. It is concluded that the frequency of the JAK2V617F point mutation is 62.0% in PV patients, the age and leukocyte count of patients with JAK2V617F point mutation are older and higher respectively than those in negative mutation group.

Study of trisomy 22 and inversion 16 in acute myeloid leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the value of trisomy 22 ( +22) in the diagnosis of inv(16) acute myeloid leukemia (AML).</p><p><b>METHODS</b>Interphase fluorescence in situ hybridization (FISH) was performed in 18 AML patients with +22. The probe was two-color break apart probe for CBFbeta with SpectrumRed on the centromeric side and SpectrumGreen on the telomeric side. The FISH results were compared with that of R-banding conventional cytogenetics (CC). Multiplex FISH (M-FISH) was used to analyze the relationship of +22 and inv(16).</p><p><b>RESULTS</b>CC revealed inv(16) in none of the 18 AML, with +22, but FISH revealed inv (16) in 11 of them and del( 16) (q22) in one. As CC results, 9 of the 11 cases were sole +22, one complicated with trisomy 8, and one del(16) (q22). Four patients with +22 and inv(16) were analyzed by M-FISH and revealed +22 only.</p><p><b>CONCLUSION</b>+22 can be regarded as an important marker for the diagnosis of inv(16) AML.</p>

Study on molecular cytogenetic abnormalities in multiple myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the molecular cytogenetic abnormalities in multiple myeloma (MM).</p><p><b>METHODS</b>Bone marrow plasma cells from 23 previously untreated MM patients were purified by CD138 McAb magnetic cell sorting system, and a panel of probes for interphase fluorescence in situ hybridization were used to detect the 13q14 deletion, p53 deletion and IgH gene translocation in the sorted MM cells.</p><p><b>RESULTS</b>Among 23 MM patients, 13q14 deletion was observed in 10 (43.5%) cases, with the positive rate of 13q14 deleted cells ranged from 79% to 96%; 14q32 translocation was observed in 11 (47.8%) cases; 13q14 deletion and 14q32 translocation were simultaneously observed in 7 (30.4%) cases; and p53 deletion was observed in none of the 23 cases.</p><p><b>CONCLUSION</b>The frequency of 13q14 deletion and IgH gene translocation in multiple myeloma are high; and the relationship between 13q14 deletion, IgH gene translocation and prognosis is worth further investigating.</p>

Multiplex fluorescence in situ hybridization for detecting complex chromosomal aberrations in chronic myeloid leukemia in blast crisis / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the value of multiplex fluorescence in situ hybridization (M-FISH) for the detection of complex chromosomal abnormalities (CCA) of chronic myeloid leukemia in blast crisis (CML-BC).</p><p><b>METHODS</b>M-FISH was used to study 26 cases of CML-BC with CCA assayed by conventional cytogenetics (CC).</p><p><b>RESULTS</b>Sixty-nine kinds of structural rearrangements were detected by M-FISH besides typical t (9;22) translocation, among them only 10 were balanced ones and 59 unbalanced ones including 1 insertion, 6 deletions, 52 translocations and derivative chromosomes. In addition, 23 numerical abnormalities were detected. All chromosomes were involved in CML-BC, and chromosomes 17, 2, 8, 16 involvements were the most frequent. M-FISH failed to find out the abnormal clone in 1 case, discovered CCA clones that were missed CC in 6 cases. Clarified 16 kinds of aberrations which could not be identified CC and corrected 5 aberrations made wrong description by CC. Thirty-five kinds of translocations were found by M-FISH which were missed by CC. The aberrations of der (9) t (16; 6; 9; 22) and der (18) t (16; 18; 19) we found were reported in the literature for the first time.</p><p><b>CONCLUSIONS</b>M-FISH can refine CCA in CML-BC, find out or correct the missed or misidentified abnormalities by CC. The frequent secondary chromosomal abnormalities in CML-BC with CCA are different from that in CML.</p>

Molecular genetics in chronic myeloid leukemia with variant Ph translocation / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the value of fluorescence in situ hybridization (FISH) and multiplex fluorescence in situ hybridization (M-FISH) techniques in the detection of genetic changes in chronic myeloid leukemia (CML) with variant Philadelphia translocation (vPh).</p><p><b>METHODS</b>Cytogenetic preparations from 10 CML patients with vPh confirmed by R banding were assayed with dual color dual fusion FISH technique. If only one fusion signal was detected in interphase cells, metaphase cells were observed to determine if there were derivative chromosome 9[der (9)] deletions. Meanwhile, the same cytogenetic preparations were assayed with M-FISH technique.</p><p><b>RESULTS</b>Of the 10 CML patients with vPh, 5 were detected with der (9) deletions by FISH technique. M-FISH technique revealed that besides the chromosome 22, chromosomes 1, 3, 5, 6, 8, 10, 11 and 17 were also involved in the vPh. M-FISH technique also detected the abnormalities which were not found with conventional cytogenetics (CC), including two never reported abnormalities.</p><p><b>CONCLUSION</b>The combination of CC, FISH and M-FISH technique could refine the genetic diagnosis of CML with vPh.</p>

Detection of circulating and bone marrow myeloma cells in patients with multiple myeloma and its clinical significance / 中国实验血液学杂志

This study was aimed to investigate the correlation between circulating myeloma cells (CMC) and bone marrow myeloma cells (MMC) in patients with multiple myeloma (MM) and its clinical significance. Four-color flow cytometry was used to detect the percentage of CMC and MMC in 55 patients with MM. Other prognosis-associated factors such as beta(2) microglobulin (beta(2)-MG), serum albumin (Alb), chromosomal abnormalities and renal function were simultaneously analyzed. The patients were divided into four groups: group A, in which CMC and MMC were simultaneously detected; group B, in which only MMC were detected; group C, in which only CMC were detected; group D, in which no myeloma cells were detected in peripheral blood or bone marrow. The results showed that the concentrations of beta(2)-MG and creatinine were significantly increased and Alb markedly decreased in group A as compared with other groups. Statistical differences existed in the above-mentioned factors between patients with myeloma cells detected and not detected. The percentages of CMC or MMC in newly diagnosed, refractory and relapsed patients were apparently higher than those in patients with partial and complete remission, respectively. CMC were strikingly correlated with MMC. It is concluded that the percentages of CMC and MMC not only imply tumor load in MM patients, but also predict the progression of MM, respectively for patients with MM, in those patients CMC and MMC were simultaneously detected.

Molecular cytogenetic characteristics of chronic lymphocytic leukemia / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the molecular cytogenetic characteristics in patients with chronic lymphocytic leukemia (CLL).</p><p><b>METHODS</b>Interphase fluorescence in situ hybridization (FISH) was used to detect trisomy 12, deletion of 13q14 and 17p13 in 60 patients with CLL.</p><p><b>RESULTS</b>Out of the 60 patients, 41 (68.3%) had at least one kind of molecular cytogenetic aberrations. Two (3.3%) had two kinds of abnormalities. Trisomy 12 was found in 12 (20.0%) cases, 13q14 deletion in 24 (40.0%) cases and 17p13 deletion in 5 (11.7%) cases. The number of trisomy 12 cells ranged from 4.0% to 34.0%, 13q14 deletion ranged from 22.0% to 93.0% and 17p13 deletion ranged from 6.0% to 68.0%. There was no significant difference among each Binet stages.</p><p><b>CONCLUSION</b>FISH is a more rapid, accurate and sensitive technique in analysis of chromosome aberrations in CLL. FISH may provide accurate information of molecular cytogenetics for CLL.</p>