Analysis of the international staging system of multiple myeloma and its comparison with the DS and IFM staging system in 122 Chinese patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To verify applicability of the International Staging System (ISS) for multiple myeloma (MM) to 112 Chinese MM patients and compare ISS with Durie-Salmon (DS) and Intergroup Francophone du Myeloma (IFM) staging system in predicting prognosis.</p><p><b>METHODS</b>112 previously untreated MM patients in Blood Diseases Hospital of CAMS were analyzed according to ISS retrospectively.</p><p><b>RESULTS</b>1) Serum beta2-microglobulin (beta2-MG) > or = 3.5 mg/L was an independent adverse prognostic factor for overall survival (OS), and serum albumin <35 g/L predicted for time to progression (TTP), 2) In the 58 cases having cytogenetic data, chromosome 13 aberration (Delta 13) was the only independent adverse prognostic factor for OS; 3) Factors significantly related to serum beta2-MG were serum creatinine, 24h urinary protein,body mass index (BMI) and performance status (PS); and those related to serum albumin were hemoglobin level, percentage of bone marrow plasma cells, lactate dehydrogenase(LDN), fever, PS, class of M-protein, serum phosphorus and BMI; 4) All traditional prognostic factors had no statistical difference between ISS stage II and III excepting for serum beta2-MG and creatinine, and 5/6 Delta 13 patients were classified to ISS stage II; 5) The median OS of ISS stage I, II, III were 69, 23 and 26 months (m) respectively, being no statistical difference between stage II and III; for DS system, 89.5% of patients were classified in stage III, being no statistical difference for OS between the stage I/II and III; while for IFM system, the median OS of low-, intermediate- and high-risk group were 69, 40 and 8 months respectively, being statistically different between high-risk and intermediate/ low-risk groups.</p><p><b>CONCLUSIONS</b>From the result of our limited analysis, the staging of ISS II and III seems unsuitable for Chinese MM patients. The IFM staging system ,which incorporates delta 13, is more effective than ISS, and DS staging system in predicting prognosis.</p>

Effects of beta-catenin-specific siRNA interference on Jurkat and K562 cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To inhibit the expression of beta-catenin and investigate the effect of the beta-catenin gene on Jurkat and K562 cells.</p><p><b>METHODS</b>siRNA specifically knocking down the expression of beta-catenin was used to testify the function of beta-catenin in Jurkat and K562 cells. Real time polymerase chain reaction and Western blot were performed respectively to testify the mRNA level and protein level of beta-catenin. Growth curve was determined by counting viable cells using trypan blue refusal-dyed method. The proliferation of cells was assayed by clonogenic counting and MTT method. The apoptotic cells were measured by Annexin V/PI staining. The cell cycle analysis was performed based on propidium iodide staining.</p><p><b>RESULTS</b>Compared with the control group (transfected with siRNA directed against scramble gene), the survival, colonogenicity, and proliferation of the Jurkat and K562 cells were significantly decreased in experimental group transfected with beta-catenin siRNA. The colonogenicity was decreased from 31.9 +/- 5.55 (siRNA) to 25.0 +/- 5.13 (control) in Jurkat cells, and from 47.33 +/- 8.52 (siRNA) to 39.33 +/- 6.26 (control) in K562 cells (both P <0.05). The inhibition rate was (49.3 +/- 9.86)% (siRNA) and (15.1 +/- 6.55)% (control) respectively in Jurkat cells, and (39.4 +/- 7.56)% (siRNA) and (10.1 +/- 6.89)% (control) in K562 cells (both P <0.05). In addition, the apoptotic rate increased from (23.5 +/- 2.82)% (control group) to (55.9 +/- 2.22)% (experiment group) in Jurkat cells and from (14.9 +/- 8.54)% (control group) to (27.9 +/- 15.3)% (experiment group) in K562 cells. However, cell cycle analysis revealed no obvious phases change both in Jurkat and in K562 cells.</p><p><b>CONCLUSION</b>Knock-down of beta-catenin gene may decrease the proliferation, survival, and clonogenicity in Jurkat cells and K562 cells.</p>

Expression of beta-Catenin in Leukemic Cell Lines / 中国实验血液学杂志

This study was aimed to investigate the expression of beta-catenin in leukemic cell lines and its relationship with pathogenesis of leukemia, semi-quantitative RT-PCR and Western blot were performed to detect the expression of beta-catenin in a panel of 15 human hematopoietic cell lines (U937, KG1a, Jurkat, K562, Namalwa, HEL, HUT78, Raji, Daudi, CEM, LCL-H, HL-60, NB4, J6-1, Ramos). Immunocytochemistry was performed in some of these cell lines to detect the location of beta-catenin. The results showed that the beta-catenin gene was widely expressed in most leukemic cell lines in various degree, the high expression of beta-catenin was found is U937, KG1a, Jurkat, K562 and Namalwa cells, middle expression of beta-catenin was observed in HEL, HUT78, Raji, Daudi and CEM cells, lower expression of beta-catenin was observed in LCL-H, HL-60, NB4, J6-1 and Ramos cells. The expression level of beta-catenin protein was identical to the expression level of beta-catenin mRNA. The expression of beta-catenin could be found in nuclei of all cells mentioned above, but their levels were different between them. Abundant beta-catenin also could be observed in nuclei of some leukemic cells by immunocytochemistry. It is concluded that overexpression of beta-catenin in leukemia cells, as a key mediator of Wnt signaling transduction pathway, indicates that the Wnt signaling transduction pathway may be aberrantly activated in leukemia.

Expression of beta-Catenin Gene in CML and its relationship with bcr/abl / 中国实验血液学杂志

This study was aimed to quantitatively detect the expression level of beta-catenin and bcr/abl in different phases of chronic myeloid leukemia (CML) and to analyze their potential relationship and significance in the progression of CML. First, the total RNA isolated from BMMNC of patients with CML and donors was reversely transcribed into cDNA. The real-time quantitative PCR method was used to analyze the expression level of beta-catenin and bcr/abl. The expression level of beta-catenin and bcr/abl in different phases of CML was compared and the correlation was analyzed between the two genes. The results showed that the beta-catenin gene in BMMNC of blast crisis of CML patients was expressed significantly higher than that in chronic phase (p < 0.001) and accelerated phase (p = 0.016) of CML patients and in normal donors (p = 0.004). The expression of bcr/abl in blast crisis of CML was statistically higher than that in chronic phase of CML (p = 0.001). The expression levels of beta-catenin and bcr/abl were correlated with each other in CML patients (r = 0.620, p < 0.001). It is concluded that the beta-catenin gene in blast crisis of CML patients express higher than that in chronic phase and accelerated phase of CML, and its expression level is correlated with the level of bcr/abl expression. The increased expression of beta-catenin may be account partly for the blast crisis of CML.

The analysis of prognostic variables in 123 patients with multiple myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To assess the prognostic value of biological features and therapy-related factors in multiple myeloma (MM).</p><p><b>METHODS</b>123 patients with newly diagnosed MM between January 1998 and May 2005 were enrolled in this retrospective study. Biological features at presentation and therapy-related factors were analysed. The overall survival (OS) and time to progression (TTP) were estimated by Kaplan-Meier analysis and the distribution of OS and TTP were compared using log-rank test. Cox regression was used to identify the independent prognostic factors.</p><p><b>RESULTS</b>(1) The univariate analysis indicated that more immature plasma cells in bone marrow biopsy, C-reactive protein >8. Omg/L, CD117 expression, serum beta2-microglobulin (beta2-MG) (3.5 approximately 5.5 mg/L), abnormal cytogenetics aberration of chromosome 13 (Delta13), hypodiploid, poor response to chemotherapy, interferon(IFN) therapy less than 6 months were associated with shorter OS(P <0.05). Lytic bone lesions at presentation, more immature plasma cells in bone marrow biopsy, serum beta2-MG (3.5 approximately 5.5 mg/L), poor response to chemotherapy, and IFN therapy less than 6 months as well as abnormal cytogenetics, hypodiploid and Delta13 were associated with shorter TTP (P <0.05). (2) Multivariable COX analysis indicated IFN therapy more than 6 months was a protective factor for OS and TTP, and more immature plasma cells in bone marrow biopsy was an independent poor prognostic factor for TTP.</p><p><b>CONCLUSION</b>The morphology of myeloma cells is useful for assessing the prognosis. And IFN therapy more than 6 months could lengthen OS and TTP.</p>

The expression of beta-catenin and its significance in leukemia cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of beta-catenin in patients with leukemia and explore its significance in leukemias.</p><p><b>METHODS</b>RT-PCR was used to detect the expression of beta-catenin in bone marrow mononuclear cells (BMMNCs) from patients with leukemia. Immunocytochemistry was in some of patients to detect the distribution of beta-catenin at the same time. The clinical significance of beta-catenin was analyzed in combination with patients' clinical information.</p><p><b>RESULTS</b>Expression of beta-catenin was statistically higher in acute myeloid leukemia (AML) and acute lymphocytic leukemia (ALL) samples than in normal donors (P = 0.001 and 0.016 respectively) and chronic phase chronic myeloid leukemia (CML) patients (P = 0.001 and P = 0.008 respectively), while there was no statistic difference between AML and ALL patients (P = 0.58). In addition, beta-catenin expression in chronic phase CML patients was like that in normal donors (P = 0.49), but increased significantly in blast crisis and accelerated phase. Immunocytochemical analysis revealed that BMMNCs from normal donors expressed beta-catenin on the plasma membrane and cytoplasma, while those from acute leukemia expressed beta-catenin to varying degrees in the nucleus as well. The expression of beta-catenin gene statistically showed the highest level in M5 (n = 15) and the lowest level in M3 (n = 18). No clinical features, such as, age, initial WBC count, therapy response rate, blast cell numbers or cytogenetic risk was found to be correlated with the expression of beta-catenin excepting for CD34+ positive rate (P = 0.004) in AML.</p><p><b>CONCLUSION</b>As a key mediator of Wnt signal transduction way, overexpression of beta-catenin in leukemia cells indicates that it might be aberrantly activated in acute leukemia, accelerated or blastic phase of CML.</p>

The outcome analysis of different treatment regimens in 206 patients with multiple myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyse the outcome of different regimens for the treatment of patients with multiple myeloma (MM).</p><p><b>METHODS</b>Response rate, median survival time and overall survival rate of 206 MM patients treated with different protocols were retrospectively analysed.</p><p><b>RESULT</b>The median survival time, 3- and 5-year overall survival (OS) of 200 MM patients treated with conventional therapy were 30.5 months, 32.0% and 15.8%, respectively. The total response rate and complete response (CR) rate of 195 patients treated with MP regimen and combination chemotherapy (CCT) were 45.6% and 14.9%, respectively. The response rates were higher for the patients treated with CCT than for those treated with MP (50.3% versus 30.4%, P < 0.05). The median survival time, 3- and 5- year OS in MP versus CCT group were 30.0 versus 30.5 months, 22.0% versus 35.0%, 13.2% versus 16.7%, respectively, but all of them have no statistical difference. Compared with those without IFN alpha maintenance therapy, patients received IFN alpha therapy showed a higher response rate (34.4% versus 53.6%, P < 0.05) and a longer median survival time (27 versus 52 months, P < 0.01). The total response in patients received thalidomide was 65.5%. Of the 6 patients received hematopoietic stem cell transplantation (HSCT), 5 remained alive in CR or PR with a mean survival time of (73.0 +/- 12.5) months.</p><p><b>CONCLUSIONS</b>CCT yields higher response rates, but not longer survival time than MP does for the treatment of MM. The response rate as well as the overall survival rate increased when IFN alpha was used as maintenance therapy. Thalidomide can improve response rate as well. HSCT could prolong survival time in patients aged < 60 years with good status.</p>

Preliminary study on extensive amplification of human dendritic cells differentiated from cord blood CD34+ progenitor cells by two-step culture / 中华血液学杂志

<p><b>OBJECTIVE</b>To Explore a two-step culture system to generate a large number of dendritic cells (DC) differentiated from cord blood (CB) CD(34)(+) cells.</p><p><b>METHODS</b>Enriched CB CD(34)(+) cells with immunoadsorption were primarily cultured in the presence of stem cell factor (SCF), Flt-3 ligand (FL), thrombopoietin (Tpo) and interleukin-3 (IL-3) for 7 (group I), 10 (group II) or 14 days (group III) respectively, and then further cultured with GM-CSF, IL-4 and TNF-alpha for 5 - 8 days to induce DC. The expansion and cell function were evaluated by flow cytometry (FCM) and mix-lymphocyte reaction (MLR), and detection of IL-12 in the supernatant by using ELISA.</p><p><b>RESULTS</b>The total nucleated cells with 53.39 +/- 20.59-, 307.17 +/- 119.59- and 1117.25 +/- 335.49-folds expansion could be respectively obtained after 7 - 14 days of expansion culture. After DC induction, CD(1a)(+) cells were 21.40 +/- 16.70-, 143.2 +/- 60.35- and 150.8 +/- 42.16-fold increase as compared to the initial nucleated cells. Comparing with that in group I, the CD(1a)(+) cells were much more in groups II and III; but there was no difference between the latter two groups (P > 0.05). The cultured cells in the three groups showed almost the same allo-stimulatory capability and IL-12 excretion when the second culture duration maintained 8 days, while the capability and excretion were greatly decreased when the duration shortened to 5 days (P < 0.05).</p><p><b>CONCLUSION</b>A plenty of functionally mature DC could be obtained from the CD(34)(+) cells in the two-step culture system of 7 - 10 days HSC expansion followed by 8 days DC induction.</p>

The expression of CD133 in acute leukemia and its clinical significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the expression of CD133 and its clinical significance in acute leukemia (AL) patients.</p><p><b>METHODS</b>The expression of CD133 and CD133 mRNA in leukemic blasts from 76 AL patients were detected by three-color flow-cytometry and hemi-quantitative RT-PCR respectively.</p><p><b>RESULTS</b>(1) CD133 mRNA expression was highly correlated with CD133 expression in both of normal donors and AL patients groups. The expression of CD133 in AL patients was significantly higher than that in control group (P < 0.01). (2) The positive rates of CD133 and CD133 mRNA in AL group were 42.1% (32/76) and 46.1% (35/76) respectively. There was no significant difference in CD133 expression between AML-M(3) and normal control, AML and ALL, as well as T-ALL and B-ALL. The expression of CD133 in AML-M(4) were significantly higher than those in other AML subtypes (81.8% vs 43.7% and 81.8% vs 46.9% at CD133 and CD133 mRNA level, respectively, P < 0.01). (3) The expression of CD133 in AML was significantly correlated with the expression of CD34 and HLA-DR (P < 0.001). (4) The expression of CD133 had no relationship with the clinical prognostic factors such as cytogenetic or molecular aberrations, WBC counts, LDH, mdr1 expression and age. (5) There was a trend toward lower CR rate in CD133(+) cases, but only CD34/CD133(+) double positive cases had significant lower CR rate than that of negative ones (44.4% vs 71.4%, P < 0.05).</p><p><b>CONCLUSIONS</b>AL had significantly higher CD133 expression compared to normal control. The detection of CD133 expression might help to identify AL type and predict therapeutic outcomes. Co-expression of CD133/CD34 might convey adverse prognosis of AL.</p>