Inhibit the proliferation of leukemia cell line NB_4-R2 with all-transretinoic acid-resistant in vitro by bortezomib / 白血病·淋巴瘤

Objective To investigate the effect of bortezomib on the proliferation and apoptosis in leukemia cell line NB_4-R2 in vitro and provide some new evidences for the treatment of acute promyelocytic leukemia APL with ATRA-resistant using bortezomib. Methods NB_4-R2 cells were incubated with bortezomib at different does for 48 h. The proliferation capacity was measured by MTT assay, the morphology of cell apoptosis observed with Hoechst33342 staining by fluorescence microscopy and the percentage of apoptosis calculated by flow cytometry. The expression of apoptosis protein of cleaved (poly ADP-ribose polymerase, PARP) and Caspase-3 were determined by Western blotting. Results The proliferation of NB_4-R2 cells were obviously inhibited by bortezomib in vivo and the role of inhibition was a does-dependant manner within the scope of the bortezomib concentration from 1-5 μg /L.The incidence of inhibition was up to 74.9 % at the bortezomib concentration of 5 μg/L. Within this scope of the bortezomib concentration mentioned above, the role of inhibition of proliferation of NB_4-R2 cells mainly showed an increase of the late apoptosis, and the percentage of apoptosis was up to 78.9 %. In the meaning time, the expressions of the apoptotic protein of cleaved PARP and Caspase-3 were up-regulated in NB_4-R2 cells after treated with bortezomib by Western blotting assay. Conclusion Bortezomib can inhibit the proliferation of NB_4-R2 cells in vivo by inducing cell apoptosis.

Expression of β-catenin in patients with chronic myeloid leukemia / 中国病理生理杂志

AIM: To observe the expression of β-catenin in patients with chronic myeloid leukemia (CML) at different disease phases, and to analyze the relationship between BCR-ABL and cytogenetic response to imatinib mesylate. METHODS: RT-PCR and Western blotting were used to detect β-catenin mRNA and protein expression in bone marrow mononuclear cells (BMMNCs) from 99 patients with CML. The association with BCR-ABL and BCR-ABL fusion was determined by FISH in 94 patients after one year treatment with imatinib mesylate, and the relationship between β-catenin and cytogenetic response to imatinib mesylate was analyzed. RESULTS: The expression of β-catenin was increased significantly in patients with blast crisis and accelerated phase (P<0.01), while the expression of β-catenin between normal person and chronic phase of CML patients was not statistically different (P>0.05). No significant relation between β-catenin and BCR-ABL expression (r=0.314, P>0.05) was observed. The expression of β-catenin was increased significantly in the patients who did not reach main cytogenetic remission (P<0.01). CONCLUSION: The patients in progression phases of CML over-express β-catenin. The expression of β-catenin is not significantly related to BCR-ABL expression, but related to the therapeutic response of imatinib. Beta-catenin may be involved in the mechanism of CML progression and could be used as a new therapeutic target.

Reversal of multidrug-resistance in human leukemia cell line K562/A02 by a cyclosporin D analogue PSC 833 / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the efficacy of PSC 833 on multidrug resistance (MDR) reversal and its mechanism.</p><p><b>METHODS</b>Human erythroleukemic cell line K562 and its doxorubicin-resistant counterpart K562/A02 were used in the study. Cytotoxicity was assessed by MTT assay, P-gp expression by direct immunofluorescence and mdr1 mRNA expression by reverse transcriptase polymerase chain reaction (RT-PCR) with beta-actin as internal control. Intracellular DNR retention was measured with flow cytometry.</p><p><b>RESULTS</b>K562/A02 cells displayed high levels of mdr1 mRNA and P-glycoprotein and reduced DNR retention compared to their parental K562 cells. 1 micromol/L of PSC 833 had no effect on the levels of mdr1 mRNA and P-gp expression in K562/A02 cells (P > 0.05). PSC 833 conferred a dose-dependent increase on chemosensitivity of K562/A02 to DNR, and its effect was at least 3-fold more potent than that of CsA or Ver. PSC 833 could increase DNR retention in K562/A02 cells. A 100.9% restoration of intracellular DNR retention of the level of K562 cells was gained by PSC 833 at 1.0 micromol/L in K562/A02 cells, whereas only a 86.9% restoration of DNR retention was obtained by CsA at 10 micromol/L in the K562/A02 cells. No effect on DNR sensitivity and retention was found in K562 cells (P > 0.05).</p><p><b>CONCLUSION</b>PSC 833 is at least 3 approximately 10 fold more potent than CsA or Ver with respect to MDR reversing activity, and it may function by inhibiting the function of P-gp and not reducing the levels of mdr1 mRNA and P-gp directly.</p>

Influence of thalidomide on interleukin-6 and its transmission in multiple myeloma patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the mechanism and influence of thalidomide on interleukin-6 (IL-6), IL-6 receptor (IL-6R) and its transmitting chain in multiple myeloma patients.</p><p><b>METHODS</b>Serum level of IL-6, expression of IL-6R on myeloma cells and IL-6R beta mRNA in multiple myeloma patients were measured by enzyme linked immunosorbent assay (Elisa), flow cytometry and reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Serum level of IL-6 in multiple myeloma patients was 564.8 +/- 319.4 ng/L, with a positive rate on the myeloma cells of 33.6% before oral 200 mg/d thalidomide. They were 560.3 +/- 414.8 ng/L and 31.8% on D14 after oral 200 mg/d thalidomide, which were not significantly different as compared with those before (P > 0.05). On D14, 28, 42, 56 and 84 after oral 400 mg/d thalidomide, the serum level of IL-6 in multiple myeloma patients were 516.7 +/- 131.9 ng/L, 426.7 +/- 180.4 ng/L, 387.9 +/- 187.4 ng/L, 350.1 +/- 85.5 ng/L and 212.3 +/- 92.5 mg/L, with positive rates on the myeloma cells of 28.5%, 24.3%, 21.3%, 12.6% and 10.1%, which were all lower than those before oral 200 mg/d thalidomide (P < 0.05 or P < 0.01). Ratios before and on D14 after oral 200 mg/d thalidomide were 7.8 and 6.9, with no statistical significance (P > 0.05). Ratios on D14, 28 after oral 400 mg/d thalidomide were 5.3 and 2.7, which were lower than those before oral 200 mg/d thalidomide (P < 0.01).</p><p><b>CONCLUSION</b>Reduction of serum level of IL-6 in multiple myeloma patients and decrease in IL-6R expression on the myeloma cells and IL-6R beta mRNA occur on D14 after oral 400 mg/d thalidomide. These changes become more obvious with time. The antitumor mechanism of thalidomide may be related to reduction of IL-6 serum level in multiple myeloma patients and decrease in IL-6R expression on the myeloma cells and IL-6R beta mRNA.</p>

The expression of ?_2 integrins and L-selectin on acute lymophocytic leukemic cells and its clinical implications / 中国病理生理杂志

AIM and METHODS: To investigate the expression of adhesion molecule ? 2 integrins (CD11a、CD11b) and L-selectin(CD62L )on Acute Lymophocyte Leukemia(ALL) cells and its Clinical Implications. Adhesion molecules CD11a、CD11b、 CD62L of 45 ALL patients and 25 health people were measured by flow-cytometric analysis. RESULTS:①CD11a and CD11b expression were lower on ALL cells than the normal hematopoietic cells. The rate of low expression was 100% for CD11b, 50% for CD11a,respectively. CD62L expression were higher on ALL cells than the normal hematopoietic cells.②The CD11a was lower expressed on B-ALL than T-ALL. CD62L was higher on T-ALL than B-ALL. ③ The expression of CD11a in the invasion group was much higher than that in the non-invasive group(P

The expression of β2 integrins and L - selectin on acutelymophocytic leukemic cells and its clinical implications / 中国病理生理杂志

AIM and METHODS: To investigate the expression of adhesion molecule β2 integrins (CD11a、 CD11b) and L-selectin(CD62L )on Acute Lymophocyte Leukemia(ALL) cells and its Clinical Implications. Adhesion molecules CD11a、CD11b、 CD62L of 45 ALL patients and 25 health people were measured by flow - cytometric analysis. RESULTS :①CD11a and CD11b expression were lower on ALL cells than the normal hematopoietic cells. The rate of low expression was 100% for CD11b, 50% for CD11a,respectively. CD62L expression were higher on ALL cells than the normal hematopoietic cells.②The CD11a was lower expressed on B - ALL than T- ALL. CD62L was higher on T- ALL than B- ALL. ③The expression of CD11a in the invasion group was much higher than that in the non - invasive group( P < 0.05).④The levels of CD11a,CD11b were returned to normal levels at remission. CONCLUSION: These results suggest that there are abnormalities in the expression of cell adhesion molecules in ALL which may help identify ALL subtypes and the treatment effect.

In Vitro Expansion of Bone Marrow Stem/Progenitor Cells by Human Placental Cell-Free Suspension / 中国实验血液学杂志

In order to explore the expansive effect of human placental cell-free suspension (HPCFS) on bone marrow hematopoietic stem/progenitor cells, and to compare the effect of HPFCS with some cytokines and their combination, human marrow CFU-GM, CFU-GEMM and BFU-E were assayed in a semisolid methyl cellulose culture system using HPCFS, IL-3, GM-CSF, and IL-3 + IL-6 + GM-CSF + EPO as colony stimulating factors, respectively. The results showed that HPCFS stimulated the growth of CFU-GM, CFU-GEMM and BFU-E and the optimal concentrations for stimulating effect were 200 - 300 micro g protein/L, and the yield of 3 kinds of colony in HPCFS group was higher than that in IL-3, GM-CSF and IL-3 + IL-6 + GM-CSF + EPO groups. The expansive effect of HPCFS on marrow progenitors was superior to IL-3, GM-CSF and IL-3 + IL-6 + GM-CSF + EPO. Human placental cell-free suspension contained a variety of cytokines to stimulate proliferation of hematopoietic stem/progenitor cells, and it could be used as an efficacious and inexpensive agent to expand hematopoietic stem/progenitor cells in vitro.

Study on bone marrow angiogenic mediators and inhibitors in aplastic anemia / 中国病理生理杂志

AIM: Through detecting bone marrow angiogenic mediators and inhibitors in aplastic anemia(AA) patients,the value of angionesis in AA pathogenesis was elucidated.METHODS: The patients were divided into severe AA group(SAA,8 patients),non severe AA group(NSAA,10 patients),and normal control group(7 persons),5 patients were observed before treating(group beginning) and getting improvement(group improving).The angiogenic mediators vascular endothelial growth factor(VEGF) and bFGF were detected by ELISA,angiogenic inhibitors IFN? and TSP were detected by ELISA and flow cytometry,respectively.RESULTS: The levels of VEGF were lower in SAA group and NSAA group than those in control group significantly(P

Bone marrow hematopoietic microenvironment in patients with systemic lupus erythmatosus / 中国病理生理杂志

0.05).We did not find any difference of the expression of fibronectin,laminin and type IV collagen in them.Expression of ICAM and VCAM were(56.4?14.8)% and(55.6?12.2)%,respectively,obviously higher than those in control group(P

Study of cytotoxicity of anti-CD20 monoclonal antibody against myeloma cells in vitro / 中国病理生理杂志

AIM: To observe the cytotoxicity on myeloma cells mediated by anti-CD20 monoclonal antibody-mabthera, after heightening level of CD20 expression on myeloma cells membrane by ?-interferon. METHODS: 10 untreated(UT) and 10 relapsed or refractory(RR) MM patients'myeloma cells were cultured with human recombinant ?-interferon (hr?-IFN) at concentrations of (0-800)?10~3 U/L to heighten level of CD20 expression, then complement-dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) on myeloma cells mediated by mabthera were studied through MTT methods. RESULTS: When CD20 expression of UT MM and RR MM patients' myeloma cells increased after treated by hr?-IFN, 12 mg/L and 16 mg/L mabthera mediated ADCC and CDC (against) myeloma cells in group UT patients and group RR patients, respectively. CONCLUSION: After heightened level of CD20 expression on myeloma cells membrane by hr?-IFN, mabthera mediated ADCC and CDC against myeloma cells in vitro.

Clinical and molecular biological characteristics of subcutaneou panniculitic T-cell lymphoma / 中国免疫学杂志

Objective:To study the clinical pathologic,immunophenotypic and clinical and molecular biological characteristics of subcutaneous panniculitic T-cell lymphoma.Methods:The clinical and pathologic features were studied by clinical observation,laboratory test and clinical pathology.Immunophenotypic features were measured by paraffinnimmunoperoxidase with the antibodies of LCA?CD3?UCHL?L26,by freeze section immunoperoxidase with the antibodies of ??TCR???TCR and analysis of the subtype with the antibodies of V 1 ??V 2 ?.T cell receptor-? gene rearrangements were studied by polymerase chain reaction(PCR).Results:Cutaneous node,high fever,loss of body weight appeared in seven patients and hemophagocytic syndrome in five patients.Imbalance of dielectric,acid and basic,abnormality of the enzyme involvement of the subcutaneous fat in a lacelike pattern with neoplastic cells were seen in all patients.The expression of LCA?CD 3?UCHL occurred in all patients,but no L26.The expression of ??TCR was found in three patients whose ??TCR was V 2+ ? and the expression of ??TCR was found in one of four patients.T cell receptor-? gene rearrangements were seen in three patients.Conclusion:SPTCL is a critical disease.The tumor cells origin from V 2+ ? of T-lymphocyte relating to cutaneous lymphotissue.T cell receptor-? gene rearrangements may happened in SPTCL patient.

Decreased effect of Rituximab on inhibition of granulocyte-macrophage colony forming unit of normal bone marrow and SLE patients by SLE's B-lymphocyte / 中国免疫学杂志

0.05 and P values in later four groups were

The expression of adhesion molecule CD11a,CD11b,CD62L on malignant lymphoproliferative disorders and its clinical implications / 中国免疫学杂志

Objective:To investigate the expression of adhesion molecule including CD11a、CD11b、CD62L on malignant lymophoproliferative disorders and its clinical implications.Methods:Adhesion molecule CD11a、CD11b、CD62L of 35 Acute Lymophocytic Leukemia(ALL)、30 multiple myeloma(MM)、4 Chronic Lymophocytic Leukemia(CLL)、14 lymphosarocoma cell leukemia patients and 25 health people were measured by flow cytometric analysis.Results:①CD11a and CD11b expression were lower on ALL、MM、CLL cells than the normal hematopoietic cells.CD62L expression were lower on CLL、MM、lymphosarocoma cell leukemic cells than the normal hematopoietic cells.②The CD11a was lower expressed on ALL than lymphosarocoma cell leukemic cells,CD62L was higher on ALL than lymphosarocoma cells leukemia.③The expression of CD11a in the ALL invasion group was much higher than that in the noninvasive group(P

Effects of Bene Jones protein and TGF-?_1 on the proliferation of rat renal proximal tubular cells in vitro / 中国病理生理杂志

AIM: To study the effect of Bene Jones protein (BJP) from multiple myeloma(MM) patient and TGF-? 1 on cultured renal proximal tubular cell(PTC) proliferation. METHODS:[ 3H] TdR incorporation was used to study the effect of ?BJP and TGF-? 1 on cultured rat NRK.52E PTC proliferation,the expression of TGF-? 1 in the supernatant of PTC cultured with BJP was assessed with ELISA. RESULTS:① [ 3H] TdR incorporation of PTC was inhibited by BJP in a dose-dependent manner,when co-cultured with 100-800 ?mol/L BJP and 2.0 ?g/L TGF-? 1, the [ 3H] TdR incorporation was lower than that of BJP alone, especially when BJP≥400 ?mol/L; ②The expression of TGF-? 1 in the supernatant of PTC cultured with BJP was increased ,especially when BJP≥400 ?mol/L( P

Idarubicin combined chemotherapy regimen for 22 cases with refractory or relapsed multiple myeloma / 中国癌症杂志

Purpose:To study the efficacy and toxicity of VIDM regimen (vincristine,idarubicin,dexamethasone and melphalan) for refractory or relapsed multiple myeloma(MM).Methods:45 refractory or relapsed MM patients were randomized into two groups: VIDM group(treatment group) had 22 patients treated with VIDM regimen, VAD group(control group) had 23 patients with VAD regimen.Results:The effective rate with VIDM regimen (59.1%) was significantly higher than that of VAD regimen (26.1%) (P

Suppression of Hematopoietic Stem /Pro genitor Cells by Sera from Patients with Systemic Lupus Erythe-matosus / 中华皮肤科杂志

Objective To explore the mechanism of hematolo gic abnormality in patients with systemic lupus erythematosus(SLE).Methods Suppression of granulocytic and ery throid colony formation of bone marrow cells fromhealthy individuals was examined in vitro by using methylcell ulose culture with sera or IgGdelete d sera from patients with SLE.Results①Both colony-forming units of erythr ocytes(CFU-E)(in 66.7%of samples tested)and granulocytes(CFU-GM)(in 70%of samples tested)of normal bone marrowcells were sign ificantly inhibited by sera frompatients with SLE(P

Re-identification of special motif regulating osteoclast differentiation in RANK / 中国病理生理杂志

AIM: To re-identify the special motif regulating osteoclast(OC)differentiation in receptor activator of nuclear factor kappa B(RANK)to provide evidences for studying the mechanism of OC differentiation.METHODS: Eight amino acids were mutated(from DIIVVYVS into ELLAAFAA)in the fragment between the 533th and the 540th amino acids in RANK cytoplasmic domain.Eight mutant TNFR1/RANK chimeras,each consists of TNFR1(tumor necrosis factor receptor 1)extracellular domain linked to transmembrane domain and cytoplasmic domain of RANK with one amino acid mutated in cytoplasmic domain was constructed by point mutation method.After the eight mutant chimeras were finished,they were packed with plat E cell line to produce the retrovirus expressing mutant TNFR1/RANK.The bone marrow macrophages(BMMs),isolated from TNFR1/R2 double knockout mice,were infected with retrovirus derived from different mutants and infected BMMs which did not differentiated into OCs were inspected after stimulated by TNF-? and M-CSF.The fragment consisted of different amino acids in TNFR1/RANK chimeras,which couldn't induce OC formation after mutated,may be the special motif regulating OC differentiation.RESULTS: We found that all BMMs transfected by TNFR1/RANK-533,TNFR1/RANK-539 or TNFR1/RANK-540 differentiated into OCs,indicating that none of amino acids D533,V539 or S540 had an effect on OC differentiation.A fewer of BMMs transfected by TNFR1/RANK-534 differentiated into OCs,indicating that I534 had a partial effect on OC formation.Most importantly,BMMs transfected TNFR1/RANK-535,TNFR1/RANK-536,TNFR1/RANK-537 or TNFR1/RANK-538 did not differentiated into OCs,indicating each of amino acids I535,V536,V537 and Y538 played a pivotal role in OC differentiation.CONCLUSION: The amino acid fragment consists of I534,I535,V536,V537 and Y538 may be the special motif regulating OC differentiation in RANK.

Influence of Fludarabine on lupus activity of BXSB mice / 中国免疫学杂志

Objective:To investigate influence of Fludarabine to lupus activity of lupus mice and the effectiveness and safety of Fludarabine on the treatment of BXSB lupus mice and their possible methanism Methods:Fludarabine of 30 mg/(m 2?d) was injected into BXSB by tail vein,3 days continuously Results of treatment was observed through counting number of peripheral blood WBC Anti ds DNA and anti nuclear antibody in BXSB serum was measured by ELISA Pathologic transformation of kidney or urine protein was measured by immunofluorescence or urine protein test paper after treated by Fludarabine Expression of CD4 +Fas +?CD8 +Fas +?CD45RO + Fas + on T lymphocyte were measured by flowcytometry Results:The number of peripheral blood WBC of BXSB mice began to decline from the third day after treated by Fludarabine The lowest value 〔0 5?0 2)?10 9 L -1 〕of the number of peripheral blood WBC appeared on the seventh day and on the nineteenth day the number of peripheral blood WBC was up to 1 0?10 9 L -1 after treated by Fludarabine The falling of anti ds DNA and anti nuclear antibody in BXSB serum appeared on the fourteenth and twentieth one day after treated by Fludarabine, respectively Reduction of from +～++ turning?kindey immunofluorescence were 72 7% in group of Fludarabine Reduction of protein urine from ++～+++ turning ?～ - appeared on the twentieth one and twentieth eight days Reduction of expression of CD4 +Fas +?CD8 +Fas +?CD45RO + Fas + on T lymphocyte was obvious after treated by Fludarabine Conclusion:Fludarabine can evidently reduce the serum level of anti ds DNA and anti nuclear antibody of BXSB and reduce injury of kidney and formation of protein urine So Fludarabine may have good curative effect to severe SLE Fludarabine can reduce expression of CD4 +Fas +?CD8 +Fas +?CD45RO + Fas + on T lymphocyte,which may be one of methanism of Fludarabine to SLE

Influence and mechanism of thalidomide on bone marrow hematopoietic progenitor cells in vitro / 中国免疫学杂志

Objective:To study the influence and mechanism of thalidomide on bone marrow hematopoietic progenitor cells in multiple myeloma patients and in normal controls.Methods:Bone marrow GM CFU, E CFU and MK CFU from multiple myeloma patients and from normal controls were cultured in methylcellulose semisolid medium in vitro after being treated by different concentration of thalidomide. IFN ? and TNF ? concentration in cell suspension of bone marrow hematopoietic progenitor cells from multiple myeloma patients and from normal controls were measured by ELISA.Results:Thalidomide inhibited GM CFU in multiple myeloma patients and in normal controls at concentration higher than 200 and 100 ?g/ml, respectively, and inhibited MK CFU in multiple myeloma patients and in normal controls at concentration higher than 300 ?g/ml and 150 ?g/ml, respectively. However, there was no significantly influence of thalidomide on E CFU in multiple myeloma patients and in normal controls at concentration between 10 and 400 ?g/ml. Thalidomide increased IFN ? concentration in cell suspension of bone marrow hematopoietic progenitor cells from multiple myeloma patients and from normal controls at concentration greater than 100 and 200 ?g/ml, respectively. The level of thalidomide of higher than 300 ?g/ml reduced TNF ? concentration in cell suspension of bone marrow hematopoietic progenitor cells from multiple myeloma patients.Conclusion:Different concentrations of thalidomide have different influences on the hematopoietic function of bone marrow hematopoietic progenitor cells in multiple myeloma patients and in normal controls, probably mediated through increased IFN ? by thalidomide.

The modulation of expression of adhesion molecules ? integrins and L-seletin on CD34~+ cells during peripheral blood mobilization with cytotoxic chemotherapy and G-CSF / 中国免疫学杂志

Objective:To study whether the mobilization process is associated with change in expression of adhesion molecules on CD34 +cells Methods:Two colour fluorescence analysis was used to study the expression of adhesion molecule CD62L,CD49d,CD11a,CD11b on CD34 + cells of peripheral blood stem cells(PBSC) before and after mobilizing with G CSF and cytotoxic chemotherapy in 15 cancer patients undergoing PBCST Results:① The expression of adhesion molecules on CD34 +cells revealed a significant reduction of CD11a, CD49d and CD62L at days 7 compared to the baseline level (P