[The clinical features, chemotherapy responses and survival of 223 patients with newly diagnosed multiple myeloma].

OBJECTIVE: To explore the proportion, clinical and laboratory features, chemotherapy responses and long term survival of different kinds of newly diagnosed multiple myeloma (MM) in China. METHODS: The clinical data of 223 cases of newly diagnosed MM patients were gathered in the First Affiliated Hospital of Sun Yat-sen University from Jan, 2000 to Seb, 2007 were retrospectively analyzed. RESULTS: The proportions of each kind of MM including IgG, IgA, light chain, IgD, IgM and biclonal MM were 48.0%, 20.6%, 25.6%, 4.0%, 0.9% and 0.9% respectively. No IgE and nonsecretory myeloma was found. The median age of onset was 58 years, of which that of the IgA type was the oldest one and the light chain type was the youngest (P = 0.004). Bone pain, renal insufficiency, and anemia were the most common symptoms which accounted for 67.7%, 61.0% and 45.3% respectively. The incidences of renal inadequacy, hypercalcemia and pathological fracture in light chain type were higher than those in IgG and IgA types. Besides, no M protein were found in serum protein electrophoresis and no elevation of total globulin. The clinical features of IgD type were similar to that of the light-chain type. The total chemotherapy efficacy rate of 89 patients who were treated with more than 3 cycles in our hospital is 61.8%, which has no difference in all types. Median overall survival of the 89 patients was 33.0 months. CONCLUSION: IgG is the most common type in MM. Bone pain, anemia, hypercalcemia and renal insufficiency are common symptoms. Immunofixation electrophoresis should be performed routinely to avoid missed diagnosis of light-chain and IgD types of MM.

[Effects and prognostic factors of HLA-matched sibling donor allogeneic hematopoietic stem cell transplantation for chronic myelogenous leukemia].

OBJECTIVE: 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). METHODS: 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). RESULTS: 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). CONCLUSION: 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.

[The relation between Bence Jones protein gene polymorphism and function of renal tubular-epithelial cell in multiple myeloma].

OBJECTIVE: Bence Jones protein (BJP) plays an important role in multiple myeloma (MM) renal lesion, we try to study the relation between the characteristics of BJP variable gene and the function of renal tubular-epithelial cell (TEC). METHODS: MM patients whose function of TEC was abnormal at diagnosis constituted a group damage and patients whose function of TEC was normal for a long period a group normal. We also collected MM patients and divided them into a group BJPkappa and a group BJPlambda. Total RNAs were isolated from the mononuclear cells of bone marrow and reverse transcription was carried out with an oligo dT18 primer; these cDNAs were then amplified with PCR, cloned and sequenced, the comparison and analysis of the sequences were made according to current ESBC/Gen Bank sequence directories. RESULTS: 3/5 cases of BJPlambda use V3-4 gene in the group damage, the replacement R mutation ratio of CDRS in the group damage (7.57 +/- 3.40) was higher than that in the group normal (4.25 +/- 1.90) and higher than that of FWRS in the group damage (3.29 +/- 1.25); R mutation ratio of CDRS region in the group BJPlambda (6.64 +/- 2.38) was higher than that in the group BJPkappa (4.10 +/- 2.13) and higher than that of FWRS in the group BJPlambda (2.91 +/- 0.94), R mutation ratio of CDRS both in the group BJPkappa or BJPlambda was higher than that of FWRS respectively. CONCLUSIONS: The higher ratio of renal lesion in some MM patients and BJPlambda patients is correlated with the high ratio of their variable gene R mutation which result in changing organization and physicochemical activity of BJP. BJP translated by some subtype genes may incline to injure the function of TEC.

[Allogeneic umbilical cord blood stem cell transplantation in Duchenne muscular dystrophy].

OBJECTIVE: To study the feasibility of treatment of Duchenne muscular dystrophy (DMD) with umbilical cord stem cell transplantation. METHODS: HLA matching was conducted for a 11-year-old DMD boy with family history was underwent umbilical cord blood stem cell transplantation and a sample of umbilical cord stem cells with 5 matched HLA sites was found in the cord blood bank with 27.32 x 10(8) nucleated cells, about 2.6 times that of the treatment dosage for him. After pretreatment with busulfan 14 mg/kg.d, cyclophosphamide 50 mg/kg.d, and rabbit anti-human thymocyte globulin 10 mg/kg.d, the allogeneic cord blood stem cells were transplanted intravenously. Cyclosporin A, methylprednisolone and MMF were used after the transplantation so as to prevent graft versus host reaction. Prostaglandin E1 was used to prevent Budd-Chiari syndrome, and ganciclovir was used to prevent cytomegalovirus infection. At the same time, Gran, granulocytic cell stimulating factor, and gammaglobulin were also used. Biochemistry test, including serum creatine kinase (CK), was conducted. Evidence of reconstruction of blood making, including conversion of blood type, was observed. PCR-STR analysis was used to observe the status of implantation of the donor umbilical cord blood stem cells. RESULTS: (1) 12 days after transplantation, the white blood cells (WBC) of peripheral blood were 0.5 x 10(9)/L, 14 days after, the numbers of WBC and neutrophils were 1.0 x 10(9)/L and 0.6 x 10(9)/L respectively. In 37 days, granulocytic cell stimulating factor was no more used, the peripheral blood WBC fluctuated around 3.34 approximately 12.2 x 10(9)/L. In the 27th day, the number of blood platelets was more than 20 x 10(9)/L and hemoglobin rose to 88 g/L. On the 24th day red blood cells transfusion was stopped. (2) In the 42nd day, the blood type of the patient transformed from type A before transplantation to type AB (the blood type of transplanted stem cells is type B). (3) PCR-STR test of the peripheral blood made 17, 26, and 42 days after transplantation showed that the gene type of the patient was mixed mosaic: The ratio of donor gradually increased from 40% approximately 45% to 55% approximately 65%. (4) In the 38th day I degrees GVHD appeared. (5) serum CK level declined from 6000 U/L to 600 approximately 2200 U/L. (6) In the 42nd day, physical examination revealed obviously improvement in walking, turning the body over, and standing up. CONCLUSION: This is first case of prospective clinical transplantation on DMD by allogeneic cord blood stem cell. Umbilical cord stem cell transplantation helps re-build blood-making function, and improve locomotive function with a mild GVHD reaction. The genotype of rebuilt blood is mosaic but the ratio of gene mosaic gradually turn from recipient gene type > donor gene type to recipient gene type < donor gene type. The serum CK level decreases significantly after transplantation, which may slow down the necrosis of muscle cell. DMD patient will be benefited by stem cell transplantation.

[Values of soluble thrombomodulin and von Willebrand factor judging reject reaction in liver transplantation].

OBJECTIVE: To find sensitive and specific laboratory examination items for early diagnosing and monitoring liver transplantation reject reaction. METHODS: 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. RESULTS: 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). CONCLUSIONS: 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.

[BAFF level in bone marrow and expression of BAFF receptor on B cells in multiple myeloma patients].

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].

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].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Dynamic observations of beta-catenin in chronic myeloid leukemia and its relationship with cytogenetic response].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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). CONCLUSION: 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.

[Significance of sRANKL/OPG ratio in diagnosis of multiple myeloma bone disease].

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].

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.

[Imatinib induces c-kit positive myeloma cells apoptosis].

OBJECTIVE: To explore the influence of Imatinib on multiple myeloma cells expressing c-kit in vitro and its mechanism. METHODS: 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. RESULTS: 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. CONCLUSION: Imatinib inhibits KM3 cells proliferation and induces the cells apoptosis by inhibiting c-kit signalling transduction.

[Therapeutic effects of imatinib on chronic myeloid leukemia in different phases and the factors affecting the effects].

OBJECTIVE: To evaluate the therapeutic effect of imatinib on chronic myeloid leukemia (CML) in different phases and analyze the factors that may affect the effects. METHODS: 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. RESULTS: 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. CONCLUSION: Early imitinib therapy can be effective and safe, and should be used as the first line drug for CML.

Aurora kinase inhibitory VX-680 increases Bax/Bcl-2 ratio and induces apoptosis in Aurora-A-high acute myeloid leukemia.

Previously, we and others showed that mitotic Aurora-A kinase (Aur-A) was required for accurate mitotic entry and proper spindle assembly. In this study, we found that expression of Aur-A was markedly elevated in bone marrow mononuclear cells (BMMCs) obtained from a significant portion of de novo acute myeloid leukemia (AML) patients. Targeting human primary AML cells with Aur-A kinase inhibitory VX-680 led to apoptotic cell death in a dose-dependent manner. Importantly, VX-680-induced cell death was preferentially higher in Aur-A-high primary leukemic blasts compared with Aur-A-low AML (P < .001) or normal BMMCs (P < .001), suggesting the possible pharmacologic window in targeting Aurora kinase among Aur-A-high VX-680-sensitive leukemia patients. VX-680-induced cell death in AML cell lines was accompanied by formation of monopolar mitotic spindles, G(2)/M phase arrest, decreased phosphorylated(p)-Akt-1, and increased proteolytic cleavage of procaspase-3 and poly(ADP)ribose polymerase. Notably, VX-680 increased Bax/Bcl-2 expression ratio, a favorable proapoptotic predictor for drug response and survival in AML. Lastly, VX-680 enhanced the cytotoxic effect of the chemotherapeutic agent etoposide (VP16) on AML cells. Together, we concluded that Aurora kinases were potentially therapeutic targets for AML and that Aur-A-high expression may serve as a differential marker for selective treatment.

[Efficacy of bortezomib combined dexamethasone in 24 patients with multiple myeloma].

BACKGROUND & OBJECTIVE: Bortezomib, a potent and reversible proteasome inhibitor, induces apoptosis of myeloma cells, resulting in durable responses in patients with multiple myeloma (MM). This study was to explore the medical effects and side effects of bortezomib combined dexamethasone in treating newly diagnosed and relapsed or refractory MM, and evaluate the safety of this regimen in the patients with renal impairment. METHODS: Twenty-four MM patients were treated with bortezomib and dexamethasone in a 21-day cycle. The patients received a median of 3 cycles (range, 1-8 cycles) of the treatment. Response to bortezomib was evaluated according to the criteria of the European Group for Blood and Marrow Transplantation (EBMT); adverse events were graded according to the National Cancer Institute Common Toxicity Criteria. RESULTS: During the follow-up with a median of 4 months, 19 (79.2%) patients responded to the treatment. The complete remission (CR) rate was significantly higher in the patients of light-chain type than in those of non-light-chain type (57.1% vs. 5.9%, P=0.014). The response rates of the patients with and without renal impairment were similar (100% vs. 70.6%, P=0.272), and the renal functions were ameliorated in the patients with renal impairment during chemotherapy. Grade III-IV adverse events, including leucocytopenia (8.3%), thrombocytopenia (33.3%), diarrhea (8.3%) and debility (4.2%), could be relieved by symptomatic treatment or delayed chemotherapy. CONCLUSIONS: The combination of bortezomib and dexamethasone shows obvious effects on MM, especially in the patients with light-chain type. The adverse events can be tolerant in most patients, and this regimen is also safe in the patients with renal impairment.

[Expression and clinical significance of vascular endothelial growth factor and its receptors in multiple myeloma].

BACKGROUND & OBJECTIVE: Bone marrow angiogenesis plays an important role in the development and prognosis of multiple myeloma (MM). Vascular endothelial growth factor (VEGF) is the most potent stimulatory factor in angiogenesis. This study was to examine the expression of VEGF and its receptors in MM, and analyze their correlations to the tumorigenesis and development of MM. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect mRNA expression of VEGF, Flt-1, and KDR in bone marrow samples from 12 newly diagnosed MM patients, 23 relapsed/refractory MM patients, 16 control people, and in KM3 cells. RESULTS: The positive rates of VEGF and Flt-1 were significantly higher in MM patients than in control people (62.9% vs. 18.8%, P<0.01; 80% vs. 31.3%, P<0.01). The mRNA levels of VEGF and Flt-1 were significantly higher in MM patients than in control people (0.41+/-0.19 vs. 0.06+/-0.01, P<0.05; 0.60+/-0.33 vs. 0.08+/-0.03, P<0.01). The positive rates of VEGF and Flt-1 mRNA were not significantly different between newly diagnosed and relapsed/refractory MM patients (66.7% vs. 60.9%, P>0.05; 83.3% vs. 78.3%, P>0.05), and between stage II MM and stage III MM (50% vs. 73.7%, P>0.05; 81.3% vs. 78.9%, P>0.05). The mRNA levels of VEGF and Flt-1 were significantly higher in relapsed/refractory MM patients than in newly diagnosed MM patients (0.49+/-0.20 vs. 0.28+/-0.04, P<0.05; 0.70+/-0.38 vs. 0.41+/-0.06, P<0.05), and were significantly higher in stage III MM than that in stage II MM (0.48+/-0.19 vs. 0.28+/-0.09, P<0.05; 0.75+/-0.35 vs. 0.41+/-0.21, P<0.05). KDR was detected only in 3 MM patients and was not detected in control group. CONCLUSION: High expression of VEGF and Flt-1 mRNA is associated with the development of MM.

[Expression and clinical significance of beta-catenin in multiple myeloma].

BACKGROUND & OBJECTIVE: beta-catenin is the pivotal regulator in Wnt pathway and in charge of cellular adhesion and signal conduction. Overexpression of beta-catenin has been observed in various human tumors. This study was to investigate the expression and clinical significance of beta-catenin in multiple myeloma (MM). METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect mRNA and protein expression of beta-catenin in bone marrow samples from 12 newly diagnosed MM patients, 14 relapsed/refractory MM patients, and 11 healthy donors. The clinical data and treatment outcomes of the MM patient were also analyzed. RESULTS: The positive rate and the expression level of beta-catenin mRNA were significantly lower in healthy donors than in newly diagnosed patients and relapsed/refractory patients (27.3% vs. 75.0% and 100%, P=5.01e(-4); 0.35+/-0.17 vs. 0.72+/-0.11 and 0.85+/-0.16, P=5.88e(-5)); the mRNA level of beta-catenin was significantly lower in newly diagnosed patients than in relapsed/refractory patients (P=0.045). beta-catenin protein was not detected in healthy donors; while the positive rate and the expression level of beta-catenin protein were significantly lower in newly diagnosed patients than in relapsed/refractory patients (50.0% vs. 85.7%, 0.32+/-0.11 vs. 0.21+/-0.08, P=0.039). In the 10 newly diagnosed MM patients with evaluable treatment outcomes, the positive rate of beta-catenin protein was significant lower in the 7 patients without response than in the 3 patients showed response (14.3% vs. 100%, P=0.033). The positive rate of beta-catenin protein was significant higher in the 16 Durie/Salmon stage III patients than in the 10 stage II patients (87.5% vs. 40.0%, P=0.026), and significant higher in ISS stage III patients than in stage I and II patients (100% vs. 45.5% and 33.3%, P=0.006, P=0.032). The protein level of beta-catenin was positively correlated to serum levels of beta2-MG (r=0.688, P=0.002) and lactate dehydrogenase (LDH) (r=0.502, P=0.034). CONCLUSION: The expression of beta-catenin is related with treatment outcome, Durin-Salmon stage and ISS stage of MM, and is positively correlated to serum levels of LDH and beta2-MG.

[Efficacy and toxicity of intravenous busulfan-based conditioning before allogeneic peripheral blood stem cell transplantation for leukemia].

BACKGROUND & OBJECTIVE: Busulfan (Bu) is commonly used as a component of conditioning regimens for hematopoietic stem cell transplantation. Erratic gastrointestinal absorption as a result of oral administration of Bu not only affects the efficacy, but also increases the risk of toxicity. This study was to analyze the efficacy and toxicity of intravenous Bu and cyclophosphamide (Cy) conditioning before allogeneic peripheral blood stem cell transplantation (allo-PBSCT) for leukemia. METHODS: Fifteen leukemia patients received intravenous Bu/Cy conditioning before allo-PBSCT, while 20 patients received oral Bu/Cy conditioning. The responses and adverse events of the 2 groups were assessed. RESULTS: All 15 patients in intravenous Bu/Cy group had hematopoietic engraftment. The median time of engraftment was 12 (9-15) days for neutrophils and 15 (11-24) days for platelets. Of the 15 patients, 6 (40.0%) developed acute graft-versus-host disease (aGVHD), including 4 cases of grade I-II aGVHD and 2 cases of grade III-IV aGVHD; during conditioning, 7 (46.6%) had vomiting, 1 (6.7%) had oral mucositis, 1 (6.7%) had hemorrhagic cystitis, 2 (13.3%) had hepatic damage, none developed seizure. With a median follow-up of 180 days (range, 35-420 days), 14 (93.3%) patients were alive, 1 died of severe aGVHD accompanied fungal infection of the lung and central nerve system. The occurrence rates of hepatic damage and oral mucositis were significantly lower in intravenous Bu/Cy group than in oral Bu/Cy group (13.3% vs. 60.0%, 6.7% vs. 80.0%, P<0.01). There were no significant differences in hematopoietic reconstruction, aGVHD, stomatitis, gastrointestinal reaction, and hemorrhagic cystitis between the 2 groups (P>0.05). CONCLUSION: The intravenous Bu/Cy conditioning before allo-PBSCT for leukemia has clear efficacy with low extramedullary toxicity.

[Effect of silencing survivin gene on sensitivity of myeloma cells to adriamycin].

BACKGROUND & OBJECTIVE: Survivin, a member of the inhibitors of apoptosis protein family (IAPs), is overexpressed in many cancers, but not in normally differentiated adult tissues. It is known to be involved in poor prognosis and resistance to chemotherapy and radiotherapy in many cancers. This study was designed to use RNA interference technique to down-regulate the expression of survivin gene in human myeloma cell line KM3, observe its effects on apoptosis of KM3 cells and sensitivity of KM3 cells to adriamycin (ADM). METHODS: Small interfering RNA (siRNA) targeting survivin mRNA was designed and in vitro chemosynthesized, and transfected into KM3 cells. Survivin mRNA and protein levels in KM3 cells were detected using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot 24, 48, and 72 h after transfection. The apoptosis of KM3 cells was observed under fluorescent microscope before and after transfection. The sensitivity of KM3 cells to ADM before and after transfection was assessed. RESULTS: The mRNA level of survivin was down-regulated by (47.0+/-4.3)%, (81.4+/-6.2)%, and (49.1+/-7.9)% of control at 24, 48, and 72 h after siRNA transfection, respectively; while the protein level of survivin was down-regulated by (39.6+/-3.8)%, (56.4+/-6.7)%, and (68.2+/-5.1)% of control, respectively. Under fluorescent microscope, the apoptosis rate of KM3 cells was (28+/-7)% at 48 h after transfection of survivin siRNA, which was significantly higher than that of control cells after transfection of negative siRNA (P<0.05). The 50% inhibition concentration (IC(50)) of ADM to KM3 cells was decreased from (1.12+/-0.14) micromol/L to (0.31+/-0.03) micromol/L. CONCLUSION: Down-regulation of survivin expression in KM3 cells by siRNA could effectively induce apoptosis of KM3 cells and increase their sensitivity to ADM.

[Preliminary report of fludarabine, mitoxantrone and dexamethasone in treating refractory or relapsed multiple myeloma].

BACKGROUND & OBJECTIVE: Multiple myeloma has low complete remission rate and high recurrence rate. Recurrence or relapse of the disease is almost inevitable for most of the patients after several cycles of combined chemotherapy. This study was designed to compare efficacy of fludarabine-based regimen (fludarabine, mitoxantrone and dexamethasone) with that of pirarubicin, vincristine and dexamethasone (VAD) on refractory or relapsed multiple myeloma, and analyze their toxicities. METHODS: Twenty-two patients who had received VAD regimen were taken as a historical control group. A total of 11 patients received FND regimen. The differences between FND group and VAD group were observed and compared. The following indexes were assessed before, during, and after treatment: the partial remission (PR) rate, total response rate, time to achieve PR, the number of patients and time of serum M-component drop to more than 50% of the pre-treatment value, the ratio of myeloma cells in bone marrow drop to less than 5% or more than 80% of pre-treatment level, and the hemoglobin level increase to more than 20 g/Lû the white blood cell and platelet count of the peripheral blood, serum calcium, creatinine, beta2-microglobin and glutamic-oxaloacetic transaminase (GPT) level, adverse events were also analyzed. RESULTS: The PR rate was significantly higher in FND group than in VAD group (45.5% vs. 22.7%, P<0.05). The median time to achieve PR was significantly longer in FND group than in VAD group (76 days vs. 68 days, P<0.05). The occurrence rates of M-component decrease and hemoglobin elevation were significantly higher in FND group than in VAD group [45.5% vs. 22.7%, and 54.5% vs. 18.2%, P<0.05]. There were no significant differences in serum calcium, creatinine and GPT level pre- and post-treatment between the 2 groups. The level of serum beta2-microglobin after treatment was significantly lower than that before treatment in FND group [(1 042.8+/-72.3) microg/L vs. (2 350.2+/-184.0) microg/L, P<0.05]. The nadir white blood cell count was significantly lower in FND group than in VAD group [(0.9+/-0.46)x10(9)/L vs. (2.09+/-0.6)x10(9)/L, P<0.05], and the occurrence rates of fever and cough was significantly higher in FND group than in VAD group (36.4% vs. 4.5%, 45.5% vs. 9.0%, P<0.05). CONCLUSIONS: Compare with VAD regimen, FND regimen may enhance the PR rate of refractory or relapsed multiple myeloma patients, but it takes longer time to achieve PR, and shows obvious bone marrow inhibition, with no significant renal or hepatic toxicity. FND regimen is effective and safe in treating refractory or relapsed multiple myeloma.