Gene Mutation in Acute Lymphoblastic Leukemia by DNA Sequencing / 中国实验血液学杂志

OBJECTIVE@#To analyze the characteristics of gene mutation in adult ALL and its clinical significance.@*METHODS@#Clinical data of 134 primary adult ALL patients and DNA sequencing results of 16 kinds of gene mutation were collected. The characteristic of gene mutation and clinical significances were statistically analyzed.@*RESULTS@#In 31 cases of 134 ALL cases (23.13%) the gene mutations were detected as follows: 19 cases of 114 B-ALL cases (16.67%), 11 cases of 19 T-ALL cases (57.89%) and 1 case of T/B-ALL. The incidence of T-ALL gene mutation was significantly higher than that of B-ALL (χ@*CONCLUSION@#There may be multiple gene mutations in adult ALL patients. IL7R and NOTCH1 are the most common gene mutations and NOTCH1 mutation may indicate poor prognosis. Detection of gene mutations is helpful to understand the pathogenesis of ALL and evaluate the prognosis of adult ALL patients.

Therapeutic observation of cranial suture acupuncture in treating depression / 针灸推拿医学（英文版）

Objective:To observe the clinical efficacy and safety of cranial suture acupuncture plus paroxetine in treating depression,and to discuss the action mechanism of this acupuncture method.Methods:One hundred depression patients were allocated to an observation group and a control group according to the random number table,with 50 cases in each group.The control group was intervened by oral administration of paroxetine tablets,20 mg each time,once a day for successive 6 weeks;the observation group was additionally given cranial suture acupuncture,once a day for 6 weeks.They were scored by Hamilton depression scale-17 (HAMD-17) before the treatment and respectively after 1-week,2-week,4-week and 6-week treatment.The clinical efficacy and safety were also observed.Results:After 6-week treatment,the total effective rate was 94.0％ in the observation group versus 78.0％ in the control group,and the between-group difference was statistically significant (P＜0.01).The HAMD-17 scores respectively after 1-week,2-week,4-week and 6-week treatment were significantly lower than the score before the treatment in the observation group (all P＜0.05);the HAMD-17 scores respectively after 2-week,4-week and 6-week treatment were significantly different from the score before the treatment in the control group (all P＜0.05).There were significant differences in the HAMD-17 score between the two groups respectively after 4-week and 6-week treatment (both P＜0.05).Conclusion:Cranial suture acupuncture plus paroxetine can ease the symptoms of depression,with faster onset and more significant therapeutic efficacy compared with paroxetine alone.

Expression of CD146 in Adult and Children's Acute B Cell Lymphoblastic Leukemia and Its Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the differences of CD146 expression in adult and children's acute B cell lymphoblastic leukemia(B-ALL), and its relation with clinical features, molecular biological and cytogenctic claracteristics.</p><p><b>METHODS</b>The expression of CD146 in bone marrow samples from adult and children's B-ALL patients were detected by flow cytometry (FCM) and the relation of CD146 abnormal high expression with the patients' clinical features, molecular biological and cytogenetical characteristics, as well as other antigens were analyzed.</p><p><b>RESULTS</b>The abnormal high expression rates of CD146 in adult and children's B-ALL patients were 29.17% and 9.09% respectively, showing that the expression rate of CD146 in adult patients was higher than that in children's patients(P<0.05). In adult B-ALL, CD146 was positively related with CD64 and CD117, while in children's B-ALL CD146 was positively related with CD71 and CD58 (P<0.05). After 1 course of standardized chemotherapy, the complete remission rates in adult and children's B-ALL patients with abnormal high expression of CD146 both were low as compared with adult and children's B-ALL without abnormal high expression of CD146 (P<0.05).</p><p><b>CONCLUSION</b>The expression rate of CD146 in adult B-ALL is higher than that in children's B-ALL. The CD146 positively relates with poor prognostic antigens, the CD146 may be one poor prognosis marker.</p>

Expression and Clinical Significance of N-cadherin in Bone Marrow Leukemic Cells Derived from Patients with Acute Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of N-cadherin in bone marrow leukemic cells derived from acute leukemia patients and its clinical significances.</p><p><b>METHODS</b>A total of 113 patients with acute leukemia were enrolled in this study. Flow cytometry was employed to detect the expression of N-Cadherin in bone marrow leukemic cells from acute leukemia patients and the relationships between the N-cadherin expression and the clinical characteristics of patients with acute leukemia were analyzed.</p><p><b>RESULTS</b>The expression of N-Cadherin in bone marrow leukemic cells deriveted from patients with acute leukemia was variable with 0%-99.7%. For adult AML patients, the positive rate of CD34 in N-cadheringroup was significantly higher than that in N-cadheringroup(67.39% vs 33.33%)(P=0.013), while the differences of total CR rate and rate of CR after 1 cycle of induction treatment were not significant between these 2 groups(P>0.05). As to ALL patients, N-cadheringroup had significant lower WBC count (21.31±7.07 vs 51.10±23.69)(P=0.008) and lower percentage of peripheral blood blast (43.22±5.75% vs 66.45±5.65%)(P=0.015). The CR rate after 1 cycle of induction treatment and rate of overall CR were lower and the relapse rate was higher in N-cadherinALL group than those in N-cadherinALL group, but the differences were not significant (P>0.05). For childhood ALL, the positive rate of CD33 in N-cadheringroup was significantly higher than that in N-cadheringroup(47.62% vs 0%)(P=0.012). The relapse rate was higher in N-cadheringroup than that in N-cadheringroup (30.00% vs 0%)(P=0.115). The median survival time, 3-year overall OS rate and 3-year relapse-free survival rate in N-cadheringroups of adult AML, non-M3 AML, ALL and chidhood ALL paients were superior to N-cadheringroups, but the differences were not significant.</p><p><b>CONCLUSION</b>The expression of N-cadherin in bone marrow leukemic cells relates to some clinical features of patients with acute leukemia and to some extent has inferior effect on survival of patients with acute leukemia.</p>

Expression of N-Cadherin in Patients with Multiple Myeloma and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of N-Cadherin in the patients with multiple myeloma (MM) and to explore its clinical significance.</p><p><b>METHODS</b>A total of 64 patients with multiple myeloma were enrolled in this study. The expression of N-Cadherin in bone marrow CD38⁺/CD138⁺ cells from multiple myeloma patients was detected by flow cytometry. The relationship between N-Cadherin expression and clinical prognostic factors was analyzed.</p><p><b>RESULTS</b>Among 64 cases of MM, the expression of N-Cadherin in 17 patients (26.56%) was high (> 20%), while that in 47 cases (73.44%) was low (< 20%); The differences of N-Cadherin expression in disease staging and classification, known prognostic factors, myeloma cell antigen expression and bone damage between patients with high and low N-Cadherin expression were not statistically different; the difference N-Cadherin expression in genetic abnormalities such as D13S319 deletion, RB1 deletion and IGH gene rearrangement between above-methioned two groups was not significant. The 1q21 amplification rate in the group with high expression of N-Cadherin was enhanced significently; the overall survival (OS) times of patients with abnormally high and low expression levels of N-Cadherin were 26.7 months and 55.5 months respectively, and the difference was statistically significant (P < 0.05).</p><p><b>CONCLUSION</b>The high expression of N-Cadherin in multiple myeloma may be one of the indicator for poor prognosis of MM, which may be related with 1q21 amplification.</p>

Aplastic anemia with macrocytic anemia: a study based on long-term follow-up / 中华血液学杂志

<p><b>OBJECTIVE</b>To elucidate the clinical features, response rate, prognosis and clonal evolution of aplastic anemia (AA) with macrocytic anemia (mAA).</p><p><b>METHODS</b>The clinical features at initial diagnosis and data in follow up of mAA hospitalized from January 2000 to October 2011 were analyzed retrospectively.</p><p><b>RESULTS</b>(1) Of 153/568 (26.9%) cases of mAA at initial diagnosis, 114(74.5%)were non-severe AA (NSAA), 39(25.5%)severe AA (SAA) and 0 very severe AA (VSAA), while the proportion was 16.2%, 45.2%, and 38.6% in 376 normocytic anemia AA (nAA), and the difference is statistically significant(χ(2) = 181.390; P = 0.000). The median age of mAA was significantly higher than that of nAA \[30(4 - 70)years vs 19 (3 - 68) years, P = 0.001\]. (2) There were no statistical difference in hemoglobin, absolute neutrophil count (ANC), platelet count (PLT), response rate after 6 months treatment and overall survival (OS) between mAA and nAA grouped in SAA and NSAA respectively. In SAA, the reticulocyte count (Ret) of mAA was significantly higher than that of nAA \[23.90(2.99 - 61.00)×10(9)/L vs 13.1(0 - 70.60)×10(9)/L, P = 0.000\] and the proportion of erythroid cells in bone marrow of mAA was also higher \[23.5 (0 - 58) vs 14.5 (0 - 65), P = 0.043\], while they did not differ significantly in NSAA. (3) The proportion of AA with PNH clones or abnormal cytogenetics did not differ significantly in mAA and nAA groups before treatment. The incidences of AA evolved to PNH in mAA and nAA was not statistically significant (7/153 vs 9/376, χ(2) = 1.099, P = 0.294) and so was the incidence of evolution to MDS/AML(3/153 vs 13/376, χ(2) = 0.399, P = 0.528).</p><p><b>CONCLUSION</b>In presented with macrocytic anemia at initial diagnosis of AA, higher proportion of NSAA, elderly age, higher Ret and proportion of erythroid cells are features, but being no statistical difference in the response rate, OS, and proportion of clonal evolution.</p>

Effect of proteasome inhibitor bortezomib on proliferation, apoptosis and XIAP expression in K562 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of proteasome inhibitor bortezomib on proliferation, apoptosis of K562 cells and the expression of XIAP.</p><p><b>METHODS</b>K562 cells were treated with bortezomib at different concentration. Cell proliferation was analyzed by WST-1 assay, cell apoptosis by flow cytometry and TUNEL, XIAP mRNA expression from 5 - 100 nmol/L by RT-PCR, and XIAP protein expression by SP immunohistochemistry.</p><p><b>RESULTS</b>K562 cells were treated with bortezomib at different concentrations for 24 h respectively, the cells growth was significantly inhibited with inhibition rates from (13. 6 ± 0. 2)% to (81. 4 ± 0. 1)%, respectively, being markedly higher than that of control (1. 2 ± 0. 1)% (P < 0.05). IC(50) was 24. 6 nmol/L of bortezomib treated for 24 h. When K562 cells were treated with 30 nmol/L of bortezomib for 12 - 48 h, the inhibition rates were (29. 1 ± 0. 9)% to (59. 8 ± 1. 2)%, respectively, the differences being statistically significant (P < 0.05) between 12 h group and 24 h group, while there was no statistical difference between 24 h, 36 h and 48 h groups. K562 cells treated with 30 nmol/L bortezomib for 24 h showed nuclear condensation, nuclear margination, nuclear fragmentation, cytoplasmic vacuoles and a large number of apoptotic body formation. The apoptotic cells rate was 83. 67% in bortezomib treated group, and 2. 33% in untreated group (P < 0.05). The expression of XIAP mRNA was decreased in a dose-dependent manner, and the expression of its protein was down-regulated.</p><p><b>CONCLUSION</b>Bortezomib can inhibit the proliferation of K562 cells, and induce apoptosis by down-regulating the expression of XIAP, providing the laboratory evidence for the targeted therapy in acute leukemia.</p>

Clinical features and survival analysis of patients with CD20 positive adult B-lineage acute lymphoblastic leukemia / 中国实验血液学杂志

The aim of this study was to explore the clinical features and survival of adult patients with CD20 positive B-lineage acute lymphoblastic leukemia (B-ALL). The clinical manifestations, examination results, therapeutic effect and survival rate of 119 adult B-ALL patients diagnosed and treated in our hospital from May 2004 to January 2008 were analyzed retrospectively. The results showed that among 119 cases, CD20 positive B-ALL accounted for 40 cases (33.61%), CD20 negative B-ALL patents accounted for 79 cases (66.39), the percentage of male patients in CD20 positive and negative groups were 72.50% and 50.63%, the leukocyte counts at diagnosis in these two groups were (27.35+/-30.29)x10(9)/L and (0.11+/-81.72)x10(9)/L, respectively, there were significant differences (p<0.05), whereas the distribution of age, infiltration of liver, spleen, lymph nodes and central nervous system, the hemoglobin and platelet levels, the expression of myeloid lineage marker, the incidence of Ph chromosome, the ratio of hyperdiploid and normal karyotype, the complete remission rate within 4 weeks, induction death rate and relapse rate and so on in CD20 positive and negative groups showed no significant differences (p>0.05). The analysis of Kaplan-Meier curve on survival rate demonstrated that the median overall survival time and 3-year overall survival rate of adult B-ALL patients in CD20 positive and negative groups were 11.0 months and 12.0 months, 28% and 20% respectively, there were no statistical differences (p=0.832). It is concluded that the expression of CD20 in adult B-ALL appears to be associated with sex and leukocyte count, but not associated with other clinical features, which indicates no significant influence on the prognosis of patients.

Immunophenotyping characteristics of adult patients with acute lymphoblastic leukemia in different ages / 中国实验血液学杂志

The purpose of this study was to investigate the immunophenotyping characteristics of adult acute lymphoblastic leukemia (ALL) patients in groups of different ages. Immunophenotyping was performed in 260 ALL patients by flow cytometry using a panel of monoclonal antibodies and CD45/SSC gating. The results indicated that (1) all the 82 cases of T-cell acute lymphoblastic leukemia (T-ALL) expressed CD7 (100%) while the positive rate of CD2 remarkably decreased with aging. The positive rate of CD2 in patients aged 14 to 18 years (adolescents) was 91.67%, which is significantly higher than that in cases aged 19 to 35 years (young adults) and > 35 years (older adults) (65.71% and 43.48% respectively, p < 0.05); the positive rate of CD34 and HLA-DR increased with aging, there was significant difference of the HLA-DR expression between the older adults group (39.13%) and the other two groups (4.17% in adolescents and 11.43% in young adults respectively (p < 0.05). Moreover, there were significant differences of the myeloid antigen (MyAg) and CD13 expression between the older adults and younger adults (p < 0.05). (2) As to adult B-cell acute lymphoblastic leukemia (B-ALL), the positive rates of CD19 and HLA-DR in 178 cases were 100%; the positive rate of CD33 in young adults was significant higher than that in adolescents (p < 0.05), the differences of the other marker expressions failed to reach statistical significance in adult B-ALL patients. It is concluded that the immunophenotypes of adult T-ALL are evidently heterogeneous in different ages, and expression with more aberrant phenotypes indicates poor prognostic significance in patients older than 35 years. There is no significant association of immunophenotypes with ages among different age groups of adult B-ALL.

Expression and role of beta-catenin in acute leukemia bone marrow cells / 中国实验血液学杂志

The aim of this study was to explore the expression of beta-catenin in acute leukemia bone marrow cells and its role in the development of acute leukemia. The expression of beta-catenin of bone marrow cells was detected by immunocytochemistry and flow cytometry in 20 cases of newly diagnosed acute leukemia. Meanwhile the expression of the proliferating index Ki-67 was detected by immunocytochemistry. The results showed that the expression of beta-catenin and Ki-67 in acute leukemia bone marrow cells was significantly higher than those in control group (P < 0.05). The migration of beta-catenin from cytoplasm to nuclear was observed in acute leukemia bone marrow cells. There was a positive correlation between the expressions of beta-catenin and Ki-67 in all the cases and the Pearson correlation coefficient was 0.845. In conclusion, the expression of beta-catenin was significantly high in acute leukemia bone marrow cells and showed a positive correlation with the cell proliferation. It suggests that beta-catenin may be involved in the development of acute leukemia through promoting the excessive proliferation of cells.

Influence of chemotherapy on expression of TRAIL in primary acute leukemic cells / 中国实验血液学杂志

In order to explore the expression of TRAIL in primary acute leukemic cells and the effect of chemotherapeutic drug on TRAIL expression in acute leukemic cells, the expression of TRAIL was assessed by flow cytometry on day 0, day 1, day 3 and day 5 in 16 patients with acute leukemia received chemotherapy. Meanwhile, the bone marrow mononuclear cells of acute leukemia patients were cultured in vitro with VP-16 and INFalpha-2a. Expression of TRAIL was analyszed by flow cytometry at 24, 48 and 72 hours after treatment. The results showed that the expression of TRAIL in the peripheral blood mononuclear cells was upregulated significantly from day 1 after chemotherapy (P < 0.05). In in vitro culture test, VP-16 upregulated the expression of TRAIL on acute leukemia bone marrow mononuclear cells (P < 0.05). Compared with VP-16 alone, the combination of VP-16 with IFNalpha-2a showed no synergic effects on the expression of TRAIL. It is concluded that the expression of TRAIL increases after chemotherapy in vivo and after treatment with VP-16 and IFN in vitro, which suggests that the apoptosis induced by TRAIL may play an important role in chemotherapy of leukemia.

Effects of interferon on the expressions of TRAIL and other apoptosis- inducing genes in peripheral blood mononuclear cells from polycythemia rubra vera / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of interferon (IFN) on the natural killer (NK) cytotoxicity and the expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and other apoptosis-inducing genes in peripheral blood mononuclear cells (PBMNC) from polycythemia rubra vera (PV).</p><p><b>METHODS</b>PBMNC were collected from 12 PV patients. The NK cytotoxicity was assessed by lactate dehydrogenase release assay and lytic units (LU) were calculated based on specific lysis. The TRAIL mRNA and other apoptosis-inducing genes were determined by RNase protection assay.</p><p><b>RESULTS</b>The NK cytotoxicity of untreated PBMNC from PV was (152.0 +/- 146.6) LU. And of IFNalpha1b- and IFNalpha2b-treated PBMNC were up to (250.9 +/- 197.4) LU and (355.9 +/- 249.9) LU, respectively (P < 0.05 and P < 0.01, respectively). The expression of TRAIL mRNA was upregulated in IFNalpha1b and IFNalpha2b stimulated-PBMNC. Up-regulation of mRNA levels of FLICE, DR3, DR4 and TNFRp55 genes was observed in PBMNC after stimulated with IFNalpha, which also induced the mRNA expressions of FasL, Fas, TRADD and RIP. To explore the role of TRAIL in IFNalpha-augmented NK cytotoxicity, neutralizing assay was used and the results showed that IFNalpha-augmented NK activity could be blocked partially by peptides DR4-Fc and DR5-Fc.</p><p><b>CONCLUSION</b>IFNalpha induced/upregulated the expression of TRAIL and other apoptosis-inducing genes in PBMNC from PV patients, which partially contribute to the IFNalpha-augmented NK cytotoxicity augmentation. This may be one of the mechanisms of IFNalpha therapy for PV.</p>