BAFF level in bone marrow and expression of BAFF receptor on B cells in multiple myeloma patients / 中国实验血液学杂志

This study was purposed to investigate the B cell-activating factor belonging to the TNF family (BAFF) and a proliferation-inducing ligand (APRIL) levels in bone marrow, and the BAFF receptor expression level on B cells in multiple myeloma (MM) patients, in order to explore the characteristics of B cells in bone marrow of MM patients. MM patients were studied before treatment (newly diagnosed group, 19 patients) and after treatment with improvement (stable group, 17 patients), 10 non-hematologic patients were selected as control (control group). The BAFF receptors (BAFF-R) and transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) on B cell (CD19(+)), naive B cell (CD19(+)IgD(+)) and memory B cell (CD19(+)CD27(+)) of bone marrow in all groups were detected by flow cytometry. The BAFF, APRIL level in bone marrow supernatant were tested with ELISA. The results showed that the BAFF-R expression level on CD19(+) cells in newly diagnosed group were higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group and stable group, but BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group was higher than that in control group; the BAFF-R expression level on CD19(+)CD27(+) cells in newly group was higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in stable group and control group. There was no significant difference among the TACI expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in newly diagnosed group, stable group and control group. The bone marrow supernatant BAFF level in newly diagnosed group was higher than that in stable group and control group, but there was no significant difference between stable group and control group. There was no significant difference among the bone marrow TACI levels in newly diagnosed group, stable group and control group. It is concluded that both the bone marrow BAFF level and the BAFF-R expression level on CD19(+) cell, CD19(+)IgD(+) cells and CD19(+)CD27(+) cells in MM patients increase, which may help to stimulate B cells, thereby may relate with to MM pathogenesis.

Correlation of CD19 positive cell counts in bone marrow with therapeutic efficacy in patients with multiple myeloma / 中国实验血液学杂志

This study was aimed to analyze the correlation of CD19 positive cell counts in bone marrow of multiple myeloma(MM) patients with therapeutic efficacy and investigate the characteristics of CD19 cell change in MM bone marrow. The CD19(+) and CD38(++)CD45(-), CD38(++)CD45(-)CD56(+) cells in bone marrow of 63 MM patients were detected by flow cytometry. The difference of CD19(+), CD38(++)CD45(-), CD38(++)CD45(-)CD56(+) cell counts at different stages and types, as well as their relation with results of 4 course of VADM or VD chemotherapy were analyzed. The results showed that in 63 MM patients, CD19(+) cell ratio at stage II were higher than those at stage III; CD38(++)CD45(-)CD56(+) cell ratio at stage II were lower than those at stage III; CD19(+) cell ratio in type IgA were higher than those in type IgD; the CD38(++)CD45(-), CD38(++)CD45(-)CD56(+) cell counts in type IgA were obviously lower than those in type IgG, IgD and light chain which showed a negative correlation between cell counts of CD19(+) against CD38(++)CD45(-), CD38(++)CD45(-)CD56(+). CD19(+) cell counts in effective treatment group of all 43 patients and the effective treatment group with VD were both higher than those in the ineffective treatment group; CD38(++)CD45(-) cell counts in effective treatment group with VD was obviously lower than those in ineffective treatment group, and CD38(++)CD45(-), CD38(++)CD45(-)CD56(+) in effective treatment group of all 43 patients were lower than those in ineffective treatment group. It is concluded that CD19(+) cell counts in bone marrow may be related to disease status and development stage of MM, which may be useful to predict treatment efficacy and prognosis.

The role of B cell-activating factor secreted by peripheral blood monocyte-derived dendritic cell in chronic idiopathic thrombocytopenic purpura / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the characteristics of B cell-activating factor (BAFF) secreted by peripheral blood monocyte-derived dendritic cell (MoDC) in chronic idiopathic thrombocytopenic purpura (cITP) and the function of MoDC on B cell proliferation.</p><p><b>METHODS</b>Ten cITP patients were studied dynamically before and after treatment. The BAFF levels in serum and the supernatant of LPS stimulated MoDC were tested with ELISA. The BAFF gene expression in LPS stimulated MoDC was tested with RQ-PCR, the B cell proliferation co-cultured with the supernatant of LPS stimulated MoDC for 5 days was tested with flow cytometry for CFSE and (3)H thymidine incorporation.</p><p><b>RESULTS</b>The BAFF level in serum (serum BAFF) \[(2461 ± 483) ng/L\], and supernatant of LPS stimulated MoDC (supernatant BAFF) \[(1113 ± 113) ng/L\] and BAFF mRNA in LPS stimulated MoDC (BAFF mRNA) (1.70 ± 0.23) before treatment were higher than that after treatment \[(621 ± 53) ng/L, (490 ± 49) ng/L and 0.37 ± 0.12\] and normal group \[(742 ± 77) ng/L, (582 ± 63) ng/L and 0.52 ± 0.08\]. There was a positive correlation among serum BAFF, supernatant BAFF and BAFF mRNA, and a negative correlation among serum BAFF, supernatant BAFF and BAFF mRNA and blood platelet count (BPC) in all ITP patients. The supernatant of LPS-stimulated MoDC from untreated patients enhanced B cell proliferation as compared with the supernatant of LPS-stimulated MoDC from treated patients and normal group.</p><p><b>CONCLUSION</b>BAFF might contribute to disease development in cITP. MoDC may directly increase B cell proliferation by secreting BAFF without T cell help, playing an important role in the antibody production in cITP.</p>

Dynamic observations of beta-catenin in chronic myeloid leukemia and its relationship with cytogenetic response / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the changes in the expression of beta-catenin in patients with chronic myeloid leukemia (CML) in different phases, and explore the relationship between beta-catenin and the cytogenetic response to imatinib mesylate.</p><p><b>METHODS</b>Beta-catenin mRNA and protein expressions were detected by RT-PCR and Western blotting in the bone marrow mononuclear cells (BMMNCs) from 99 CML patients. The expressions of BCR-ABL fusion gene at both the mRNA and protein levels were detected by fluorescence in situ hybridization (FISH) in 94 patients before and during the one-year treatment with imatinib mesylate at the interval of 3 months, and the relationship between beta-catenin and cytogenetic response to imatinib mesylate was analyzed.</p><p><b>RESULTS</b>The expression of beta-catenin increased significantly in patients with blast crisis and accelerated phase (P<0.001), but showed no significant difference between normal subjects and CML patients in the chronic phase (P>0.05). The main cytogenetic remission rate was significantly higher in patients who were consistently negative for beta-catenin than in those consistently positive for beta-catenin or those with a positive transformation (P<0.001).</p><p><b>CONCLUSION</b>Beta-catenin overexpression in the progression of CML, consistent high level of beta-catenin or a positive transformation may indicate a poor response to imatinib, and early measures should be taken to increase the remission rate.</p>

Significance of sRANKL/OPG ratio in diagnosis of multiple myeloma bone disease / 中国实验血液学杂志

This study was purposed to investigate the relationship between the levels of soluble receptor activator of nuclear factor kappa B ligand (sRANKL) and osteoprotegerin (OPG) in serum of the patients with multiple myeloma (MM) and multiple myeloma bone disease (MBD). The serum levels of sRANKL, OPG, tartrate-resistant acid phosphatase-5b (TRAP-5b) and C-terminal telopeptide of collagen I (CTP-I) which both are indexes for metabolism of osteoclast (OC) in newly diagnosed MM patients (n=42, experimental group) and healthy persons (n=25, control group) were detected by enzyme-linked immunosorbent assay. The roentgenography was used to determine bone damage in MM patients at the same time. According to these results acquired, the correlation of sRANKL/OPG ratio with levels of TRAP-5b/CTP-I, the incidence and degree of bone destruction were analyzed. The results indicated that the level of sRANKL (median value 9.33 microg/L) increased and level of OPG (median value 4.93 microg/L) decreased and the sRANKL/OPG ratio (2.65) increased significantly in experimental group. Compared with control group, the differences in all the corresponding indicators were statistically significant (p<0.05). The sRANKL/OPG ratio was closely related to levels of TRAP-5b (r=0.512, p<0.05) and CTP-I (r=0.481, p<0.05) in MM patients. After all patients in experimental groups were divided into group with bone destruction (n=29) and without bone destruction (n=13), the sRANKL/OPG ratio in the group with bone destruction was 5.13 and much higher than that in group without bone destruction (1.12) (p<0.05). A close correlation between the sRANKL/OPG ratio and degree of bone destruction (r=0.445, p<0.05) was acquired when all MM patients were divided into three groups according to degree of bone destruction, but no difference between the ratio and clinical classification and International Staging System (ISS) in MM patients was found. It is concluded that the sRANKL/OPG ratio in serum of MM patients is significantly elevated, which may be closely related to increase metabolism of OC along with the incidence and degree of bone destruction. In short, the sRANKL/OPG ratio can be used as a reference index for the diagnosis of MBD.

Osteoclast differentiation regulated by receptor activator of nuclear factor kappaB probably through a novel signaling pathway / 中国实验血液学杂志

This study was aimed to investigate the signaling pathways regulating osteoclast (OC) differentiation by receptor activator of nuclear factor kappa (RANK) under physiological condition so as to provide some theoretical basis for clarifying mechanism of bone destruction in multiple myeloma. A mutant TNFR(1)/RANK(2) (named RANK-Mu) chimera consisting of tumor necrosis factor receptor 1 (TNFR(1)) and RANK intramembrane domain was constructed by using deletion mutation for deleting IVVY amino acids in RANK intramembrane domain in accordance with (535-)IVVY(-538) as specific domain regulating OC differentiation by RANK. The RANK-Mu and TNFR(1)/RANK chimera without mutation (RANK-WT) were packaged by using plat E cell line to produce the retrovirus, which were transfected into bone marrow macrophages (BMMs) of TNFR(1)/TNFR(2) double knockout mice. After stimulation of these transfected BMMs with TNF-alpha, the differentiation of BMMs into OCs were observed, meanwhile the phosphorylation of NF-kappab, JNK, p38 and ERK was detected by Western blot after stimulation of these BMMs with TNF-alpha. The results showed that BMMs transfected with RANK-WT could be differentiated into OCs and phosphorylation of NF-kappaB, JNK, P38 and ERK were activated at 5 - 10 minutes after being stimulated by TNFalpha. BMMs transfected with RANK-Mu could not be differentiated into OCs, but phosphorylation of NF-kappaB, JNK, P38 and ERK were activated also. It is concluded that RANK regulates osteoclast differentiation probably not through 4 typical signaling pathways, named as NF-kappaB, JNK, P38 and ERK, in this process other new signaling pathways maybe participate.

Therapeutic effects of imatinib on chronic myeloid leukemia in different phases and the factors affecting the effects / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of imatinib on chronic myeloid leukemia (CML) in different phases and analyze the factors that may affect the effects.</p><p><b>METHODS</b>Eighty-five patients with CML in chronic phase, 24 in accelerated phase and 19 in blastic phase patients were treated with imitinib. The hematologic response, cytogenetic response, molecular response, overall survival (OS), progression-free survival (PFS) and adverse events were analyzed in these groups.</p><p><b>RESULTS</b>The rates of complete hematologic response (CHR), complete cytogenetic response (CCyR) and complete molecular response (CMoR) of the patients in chronic phase were 100%, 82.4% and 21.2%, respectively, and the 5-year OS and PFS of these patients were 92.1% and 84.7%. All these rates were significantly higher than those in patients in accelerated and blastic phases (P<0.0001). The CCyR, CMoR, 5-year OS and PFS in the 42 newly diagnosed patients in chronic phase were 92.9%, 26.3%, 100% and 95.2%, respectively, all significantly higher than those in patients with interferon therapy failure (P<0.001). Severe leukocytopenia and thrombocytopenia occurred at greater frequencey in AP and BP patients than in chronic phase patients (P<0.0001). Non-hematologic toxicity was rarer and milder in patients in chronic phase. Multivariate analysis showed that interferon therapy prior to imitinib treatment and prolonged drug cessation were the independent factors that affected the achievement of cytogenetic response and PFS.</p><p><b>CONCLUSION</b>Early imitinib therapy can be effective and safe, and should be used as the first line drug for CML.</p>

Imatinib induces c-kit positive myeloma cells apoptosis / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the influence of Imatinib on multiple myeloma cells expressing c-kit in vitro and its mechanism.</p><p><b>METHODS</b>KM3 cells were treated with Imatinib at different concentrations, and cell growth index were evaluated by XTT assay, cell cycle by flow cytometry, apoptosis by Annexin V/ PI and DNA ladder, and change in protein level by Western blot.</p><p><b>RESULTS</b>Imatinib inhibited proliferation of KM3 cells at concentrations more than 0.25 micromol/L in a dose-dependent manner, and the 48 h IC50 was 0.33 micromol/L (P < 0.01). Imatinib arrested cell in C0/G1 phase. Annexin V/PI staining and DNA ladder indicated that Imatinib had a substantial effect on inducing apoptosis of KM3 cells in a dose-dependent manner and induced pro-caspase-3 and poly ADP-ribose polymerase (PARP) cleaved. Imatinib inhibited expression of c-kit and provoked a decrease of IL-6 induced c-kit phosphorylation in vitro.</p><p><b>CONCLUSION</b>Imatinib inhibits KM3 cells proliferation and induces the cells apoptosis by inhibiting c-kit signalling transduction.</p>

Effects and prognostic factors of HLA-matched sibling donor allogeneic hematopoietic stem cell transplantation for chronic myelogenous leukemia / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To retrospectively analyze the curative effects and prognostic factors of HLA-matched sibling donor allogeneic hematopoietic stem cell transplantation (allo-HSCT) for chronic myelogenous leukemia patients (CML).</p><p><b>METHODS</b>Of the 35 CML patients, 26 were males and 9 were females, with a median age of 32 (12 - 50) years. 30 patients were in chronic phase of CML, 5 patients were in accelerated phase. Allo-HSCT from HLA identical siblings was performed for 35 patients, of whom 11 received bone marrow transplantation (BMT) and 24 peripheral blood stem cell transplantation (PBSCT). Conditioning regimens was TBI (total-body irradiation) + CY (CTX) protocol in 8 patients and BU/CY protocol in 27 patients. The average follow-up was 48 months (range 7 - 108 months).</p><p><b>RESULTS</b>34 (97.1%) patients were successfully engrafted. Among them, 21 patients (60.0%) had three years disease-free (DFS) survival. The overall 5-year survival (OS) was 57.1%. Two patients (5.7%) relapsed. Transplant-related mortality occurred in 12 patients. Hemorrhagic cystitis (HC) occurred in 5 patients and HVOD was observed in 1 patient. Acute graft-versus-host disease (aGVHD) occurred in 18 patients (51.4%), among them 7 patients (20.0%) were of grade III-IV. Chronic GVHD was in 17 patients (48.5%). There was no significant difference in 3-years DFS between BMT group and PBSCT group (54.5% vs. 62.5%, P > 0.05). The 3-year disease-free survival (DFS) was 42.9% in TBI/CY group and 55.6% in BU/CY group (P > 0.05). In univariate prognostic analysis model, the DFS at 3 years is 75% and 47.4% for < or =30 years patients and >30 years patients, respectively, P < 0.05. The 3-year DFS of patients with first chronic phase is higher than patients with advanced diseases (61.3% vs. 40%, P < 0. 05). The 3-year DFS in patients of grade I - II GVHD was higher than that in patients of grade III-IV GVHD (81.8% vs. 14.3%, P < 0.05).</p><p><b>CONCLUSION</b>The patients who had transplantation done within 1 year after diagnosis during their first chronic phase of disease and who had low-grade GVHD have better prognosis. Those patients who had III-IV acute GVHD are prone to incorporate severe infection, which was a worse prognostic factor of allo-HSCT for chronic myelogenous leukemia.</p>

Study of TRAIL receptors expression on the mononuclear cells from multiple myeloma patients and KM3 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the differential expression of four TRAIL receptors on bone marrow mononuclear cells (BMMNC) from multiple myeloma (MM) patients and myeloma cell line KM3 cells, to compare their altered expressions after chemotherapy and to explore the mechanisms by which TRAIL selectively kills tumor cells.</p><p><b>METHODS</b>Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and flow cytometry were used to investigate the expression of four TRAIL receptors on BMMNCs in 23 MM patients, KM3 cells and 15 controls, and the changes of their expression pattern after chemotherapy and after incubation of KM3 cells with sub-clinical concentration of doxorubicin.</p><p><b>RESULTS</b>DR4 and DR5 were highly expressed on KM3 cells with no expression of DcR1 and DcR2. Expressions of DR4 and DR5 on BMMNCs from MM patients were higher and expression of DcR1 and DcR2 were lower than that of controls (P < 0.05). The expression of DR5 on MM and KM3 cells was up-regulated after chemotherapy and exposure to doxorubicin (P < 0. 05).</p><p><b>CONCLUSIONS</b>The expressions of four TRAIL receptors on myeloma cells and normal controls were different, which might account for the selective killing effect of TRAIL on MM cells. Up-regulated DR5 on KM3 cells after incubating with doxorubicin and after chemotherapy suggests the cytotoxic agents might enhance the apoptosis of MM cells.</p>

Values of soluble thrombomodulin and von Willebrand factor judging reject reaction in liver transplantation / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To find sensitive and specific laboratory examination items for early diagnosing and monitoring liver transplantation reject reaction.</p><p><b>METHODS</b>Randomly investigate 41 liver transplantation patients, among them there were 16 patients with reject reaction (including 12 with acute rejection, 4 with chronic rejection). Plasma soluble thrombomodulin (STM) and von Willebrand factor (vWF) levels were measured before operation and every other day after operation.</p><p><b>RESULTS</b>Plasma STM level increased significantly after operation, two days before rejection and after acute rejection (5.58 ng/ml +/- 0.42 ng/ml, 5.93 ng/ml +/- 0.45 ng/ml, and 7.88 ng/ml +/- 0.29 ng/ml, respectively), so did vWF level (101.2% +/- 4.68%, 104.3% +/- 5.78%, and 127.7% +/- 5.74%, respectively). STM level was much higher in acute rejection than that in chronic rejection (7.88 ng/ml +/- 0.29 ng/ml vs. 6.35 ng/ml +/- 0.54 ng/ml, t = 2.46, P < 0.05), in no reaction group after impacting therapy than in effective group (8.30 ng/ml +/- 0.19 ng/ml vs. 3.82 ng/ml +/- 0.22 ng/ml, t = 12.98, P < 0.01), and in dead group after treatment than in living group (7.98 ng/ml +/- 0.18 ng/ml vs. 6.51 ng/ml +/- 0.41 ng/ml, t = 3.39, P < 0.01).</p><p><b>CONCLUSIONS</b>Plasma STM and vWF can be taken as laboratory items for monitoring liver transplantation rejection. Plasma STM can act as not only an early prognosticating marker, but also suitable to distinguish acute from chronic reject reaction, and as a marker for monitoring impacting therapy effect and judging prognosis.</p>

In vitro inhibition and mechanism of multiple myeloma cells growth by thalidomide / 中国实验血液学杂志

To investigate the influence of the thalidomide on the growth of multiple myeloma cells from untreated, relapsed or refractory patients and summarize its mechanisms, thalidomide influence on colony growth of untreated, relapsed or refractory multiple myeloma cells cultured by semisolid methylcellulose was observed. The level of interleukin-6 (IL-6) autosecreted by myeloma cells was tested by IL-6-dependent cell line when myeloma cells were treated with thalidomide at 200 microgram/ml, and in the same concentration of thalidomide the expression of IL-6 receptor were tested by flow cytometry. Results showed that colony growths of myeloma cell from untreated and relapsed or refractory patients were all colonies were inhibited when treated by thalidomide up to 75 microgram/ml or 100 microgram/ml concentration. The inhibition was concentration-dependent, higher concentration cause more inhibition. After treatment with thalidomide at 200 microgram/ml, the concentrations of IL-6 secreted by myeloma cells were (148.5 +/- 96.7) microgram/ml, and the levels of IL-6 receptor expressed on the cell surface were 16.7% and 20.2% in untreated and relapsed or refractory patients, respectively, and those were significantly lower than those levels in the cells before exposure to thalidomide. It was concluded that thalidomide can inhibit growth of both relapsed or refractory cells and untreated myeloma cells in vitro. Therefore, it can be used to treat untreated multiple myeloma patients. Inhibiting tumor cells secreting level of IL-6 and reducing the expression of IL-6 receptor on myeloma cell surface is one of the mechanisms for thalidomide to remedy multiple myeloma patients