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Primary central nervous system lymphoma (PCNSL) is a rare and special kind of non-Hodgkin lymphoma. Chemotherapy based on high-dose methotrexate (HD-MTX) is considered a standard therapy for newly diagnosed PCNSL, but the prognosis of PCNSL is poor due to its high invasiveness and recurrence rate. Risk stratification and prognosis assessment of PCNSL in diagnosis is particularly important in clinical treatment. At present, MSKCC (the Memorial Sloan Kettering Cancer Center), the International Extranodal Lymphoma Study Group (IELSG) and Nottingham/Barcelona scoring models are mostly used to classify the risk and evaluate the prognosis of PCNSL. With the further understanding of immunohistochemistry and molecular genetics of PCNSL, potential prognostic factors continue to emerge.
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Objective@#To investigate the effects of artesunate combined with bortezomib on the proliferation, apoptosis and autophagy of human acute myeloid leukemia cell lines MV4-11, and its mechanisms.@*Methods@#MTT method was used to determine the anti-proliferation effect of different concentrations of artesunate, bortezomib and their combination on MV4-11 cells. The cell apoptosis were analyzed by flow cytometry. The expression of cleaved-Caspase-3, Bcl-2 family protein (Bcl-2, Mcl-1, Bim, Bax) and autophagy-related protein LC3B were assayed by Western blot.@*Results@#Artesunate displayed a proliferation inhibition effect on MV4-11 with dose- and time-dependent manner, the IC50 of artesunate on MV4-11 after 48 hours was 1.44 μg/ml. Bortezomib displayed a proliferation inhibition effect on MV4-11 with dose-dependent manner, the IC50 of bortezomib on MV4-11 after 48 hours was 8.97 nmol/L. The combination of artesunate (0.75, 1.0 μg/ml) and Bortezomib (6, 8 nmol/L) showed higher inhibition on MV4-11 than artesunate or bortezomib alone in the same concentration gradient after 48 hours (P<0.05) . The cooperation index of the two drugs were all less than 1. The 48 h apoptotic rate of artesunate (1.5 μg/ml) on MV4-11 was (15.27±2.18) %, (19.85±3.23) % of bortezomib (8 nmol/L) , (81.67±5.96) % of combination of the two drugs, significantly higher than the single group (P<0.05) . When combination of the two drugs on MV4-11 after 24 hours, the levels of pro-apoptotic protein Bim and the cleaved activation of Caspase-3 and autophagy-related protein LC3B were up-regulated and the anti-apoptotic protein Bcl-2 expressions was down-regulated.@*Conclusion@#Combination of artesunate with bortezomib shows a significant synergistic effects on proliferation, apoptosis and autophagy of MV4-11 cell lines, which may be associated with Bcl-2 family proteins expression.
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The aim of this study was to construct the lentivirus expression vector of human FLT3-ITD mutation and to screen the AML cells for stable expression of FLT3-ITD.We screened the AML patient with FLT3-ITD mutation by peripheral blood DNA extraction and PCR,and then the ORF of FLT3-ITD was constructed using the method of homologous recombination,which used the BamH Ⅰ and Not Ⅰ double enzyme digestion of the ORF and the FIV vector.Results were confirmed by restriction enzyme identification and PCR.The constructed recombinant vector was co-transfected with VSVG and g/p into HEK293T cells to produce the lentiviral parti cles by transfection reagent.The lentiviral particles were transduced into K562 cells and the infection efficiency was measured by fluorescence microscope.The cells were sorted by the Flow Cytometer.The expression of FLT3 in the stable cell lines was detected by Western blot.There were 42 patients with FLT3-ITD mutation among the 207 patients we collected.We successfully amplified the ORF of FLT3 of the patient with highest mutant/wt ration.Then constructed the lentivirus expression vector FLT3-ITD/FIV and overexpressed it in K562 cell lines.We screened the cells expressed FLT3-ITD/FIV by fluorescence activated cell sorting,and Western Blot results proved the expression of FLT3-ITD.The over-expression 1 entiviral vector of human FLT3-ITD/FIV has been successfully constructed and cell lines for stable expression of FLT3-ITD have been successfully established.This will shed a light on the future study of FLT3 function in AML and provide a new in vitro model for AML.
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Objective To investigate the clinical characteristics and prognosis of primary diffuse large B-cell lymphoma of central nervous system ( PCNS DLBCL). Methods The data of 70 patients with PCNS DLBCL confirmed by pathology were retrospectively analyzed. Survival and prognostic analyses were further conducted in the 66 follow-up patients. Results Median age at diagnosis was 57 years old. The ratio of male and female was 1.3∶1. The time from having symptoms to seeking medical advice was less than 2 months in 54 (77.1%) patients. 44 (62.9 %) patients had increased intracranial pressure, and 26 (37.1 %) patients had limb weakness or hemiplegia symptoms. 37 (52.9%) patients were multiple lesions, 59 (84.3%) cases were supratentorial, and 46 (65.7%) cases showed involvement of deep-brain tissues. Among 66 follow-up patients 7 cases received supportive and palliative care, 27 cases received surgery, 6 cases received radiotherapy, 9 cases received chemotherapy alone, and 21 cases received radiotherapy in addition to chemotherapy. The median overall survival (OS) was 9 months (95 % CI 1-16 months), and the 2-year survival rate was 36.1 %. The median OSs of the supportive and palliative therapy group and the surgery group were 2 months and 3 months, respectively. The median OS of the chemotherapy, radiotherapy or combination group was 33 months (95%CI 22-43 months) and the 2-year OS rate was 56.9 %. The Cox multivariate regression analysis showed that the involvement of deep-brain tissues (P=0.04) and not receiving radiotherapy or chemotherapy (P=0.00) were related to poor prognosis. Conclusions PCNS DLBCL is a highly aggressive and malignant tumor. Patients undergoing only surgery have poor effect and short survival. The patients with involvement of deep brain tissues have a poor prognosis. The chemotherapy, radiotherapy or combination of them may improve the prognosis.
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Objective:To study the curative efficacy of decitabine combine with CAG regimen in the treatment of acute myeloid leukemia and its effects on the serum Interferon-γ (IFN-γ),alpha hydroxybutyrate dehydrogenase (HBDH) and lactate dehydrogenase (LDH) levels.Methods:70 cases of patients with acute myeloid leukemia who were treated from February 2013 to October 2016 in our hospital were selected as research objects.According to the random number table,the patients were divided into the observation group (n=35) and the control group (n=35).Both groups of patients were treated conventional treatment.The control group was treated with CAG scheme,intravenous injection of aclarubicin,20 mg each time,granulocyte colony stimulating factor,300 μg each time,subcutaneous intravenous cytarabine,10～15 mg/m2 each time,while the observation group was treated with intravenous drip of decitabine on the basis of control group,15 mg/m2 each time.Then the therapeutic effect,serum interferon-γ (IFN-γ),alpha hydroxybutyrate dehydrogenase (HBDH),lactate dehydrogenase (LDH) levels before and after treatment,incidence of adverse reactions were compared between two groups.Results:After treatment,the total effective of observation group was significantly higher than that of the control group [91.42％(32/35)vs68.57％(24/35)] (P ＜0.05);the serum IFN-γ,HBDH,LDH levels were significantly higher than those of the control group [(3.21± 1.01)pg/ml vs.(5.13 ± 1.90)pg/mL,(103.62± 26.39)U/L vs.(118.80± 28.60)U/L,(101.36± 27.32)U/L vs.(123.08 ± 30.59)U/L] (P ＜0.05);the incidence rate of adverse reactions was significantly lower than that of the control group [20.00％(7/35) vs.42.85％(15/35)(P ＜0.05)].Conclusion:Decitabine combined with CAG regimen was more effective for acute myeloid leukemia than CAG regimen alone,which might be related to reduce the serum levels of IFN-γ, HBDH and LDH.
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Objective: In this study, we aimed to investigate changes of peripheral Th17 and Treg cells frequencies in the newly-diagnosed Chronic Lymphocytic Leukemia [CLL] patients for 12 months
Methods: In this research,50 CLL patients were enrolled
Circulating Th1, Th17 cells and CD4+CD25+Foxp3+Treg cells were analyzed by flow cytometry. Plasma levels of related cytokines were detected by enzyme-linked immuno sorbent assay [ELISA]
The study was carried out from January 2012 to October 2013 at Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, P.R. China
Results: Compared with healthy controls, Th17 cells related cytokines were significantly increased in CLL patients, while Treg cells related cytokines were significantly lowered. In the follow-up, we found that the frequency of Treg cells was irregular, while the frequency of Th17 cells was gradually decreased
Conclusion: Our study suggested that Th17 cells may play important role in the immune regulation of CLL, and may become a new target in CLL therapy
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The 55th ASH annual meeting hold special education sessions and oral and poster presentations focused on Hodgkin lymphoma (HL).Experts from world around made highly of the monoclonal antibody to CD30 (BV),for it' s effectiveness on refractory or relapsed HL.The targeted chemotherapy which integrated BV with standard chemotherapy appeared promising as first line therapy in a few centers.The applications of other molecular inhibitors,anti-CD20 antibody and involved field radiation therapy had been discussed.A broad presentations and discussions had been made on HL of the gene expression profiling,molecular susceptibilities,the long non-coding RNA,the miRNA,the molecular diagnosis with peripheral blood,as well as prognosis related factors.
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ObjectiveTo investigate the efficacy of co-infusing cord blood (CB) as the third party cells on graft versus host disease (allo-GVHD) prophylaxis after unrelated or haploidentical donor allogeneic hematopoietic stem cell transplantation (allo-HSCT).MethodsFrom 2007 to 2011,41 patients receiving unrelated or haploidentical donor allo-HSCT were analyzed retrospectively.Twenty-five patients received one unit of HLA 4/6-6/6 matched CB one day before SCT as CB group,and median MNC dose was (1.64 ± 0.49) × 107/kg.Sixteen cases not receiving CB served as control group.All patients received antithymocyte globulin,cyclosporine,methotrexate,and mycophenolate mofetil as GVHD prophylaxis.The incidence and severity of aGVHD,and treatment-related mortality were compared between two groups.ResultsThe main clinical characteristics in both groups were comparable.The cumulative incidence of aGVHD in CB group and control group was 44.0％ versus 68.8％ respectively (x2 =2.403,P＞0.05).The cumulative incidence of grades Ⅲ to ⅣV aGVHD in CB group and control group was 16.0％ and 37.5％ respectively (x2 =2.445,P＞0.05).The 100-day treatment-related mortality in CB group and control group was 12.0％ and 12.5％ respectively (x2 =0.002,P＞0.05).ConclusionCord blood as the third party cells might reduce the incidence and severity of aGVHD in unrelated or haploidentical donor HSCT.The efficacy and the mechanism of this strategy need to be further explored by prospective randomized controlled trials.
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Objective To evaluate the relationship between plasma imatinib and its effect in the treatment of chronic myeloid leukemia(CML). Methods Fifty-one CML patients were included in this study,who began taking imatinib from July 2005 to February 2008, with 34 cases of male, and 17 cases of female.Nine patients took imatinib at dose of 300 mg/d, 37 patients took imatinib at dose of 400 mg/d, and 5 patients took imatinib at dose of 600 mg/d. High-performance liquid chromatography was used to test imatinib plasma levels. Results The imatinib plasma levels was imatinib dose-related, and the imatinib plasma trough levels significantly varied between individuals[(342-4688)ng/ml]. The imatinib plasma levels was significant lower in 300 mg/d dose group [(1037±514) ng/ml] than 400 mg/d dose group [(2123±1016) ng/ml] (t =2.34, P =0.032),and the effective rate was 66.7 % (6/9) in 300 mg/d dose group, which was lower than 400 mg/d dose group of 89.19 % (33/37) (χ2=7.14, P =0.008). In 300 mg/d and 400 mg/d dose groups, 39 patients achieved effective treatment, and their imatinib plasma levels was significant higher than that of 7 patients who did not achieved effective treatment (t =2.25, P =0.037). The ROC curve results suggested that clinical treatment may be poor when the imatinib plasma level was lower than 1050 ng/ml (sensitivity was 84.6 %, specificity was 71.1 %).Conclusion The imatinib plasma levels was dose-related, and significantly varied between individuals.Clinical treatment effect may be poor when the imatinib plasma level was lower than 1050 ng/ml.
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Objective Comparative evaluation of flow cytometric immunophenotyping in the diagnosis and differentiation of lymphadenopathy,lymphoma and reactive lymphoid hyperplasia. Methods Ninty-nine fine-needle aspiration specimens from patients with tentative clinical lymphoprofierative disorders were consecutively analyzed by both cytology and flow cytometry with histology results as the gold standard. The three color antibodies including CD3,CD3,CD4,CD5,CD10,CD19,CD20,CD23,CD45,K,λ,FMC7 and CD34 were used for cell composition evaluation and cells with abnormal phenotype. Lymphoma cases were classified according to new WHO classification and subtypes were categorized by immunophenotypic analysis. The results from flow cytometry and cytology were compared. Results By cytological study, 40 of 99 cases were diagnosed with lymphoma, 29 cases were diagnosed with metastatic carcinoma, and 30 cases were diagnosed with reactive lymphoid hyperplasia, necrosis or tuberculosis. Among them, 2 non-Hodgkin lymphoma(NHL) cases were misdiagnosed as reactive lymphoid hyperplasia by cytology. Biopsy was performed in 18 cases of NHL including 16 B-NHL and 2 T-NHL By flow cytometry study, 35 of 99 eases were diagnosed with lymphoma, including 4 cases of lymphoblast lymphoma, 1 case of T-cell lymphoma, and 30 eases of other B-NHL For those 30 cases of B-NHL, 28 cases showed monoclonal light chain expression, and k: λ orλ: k atios exceed 3: 1, and B-cell proportion was (73. 2±27. 2)%. Twenty-six cases could be sub-classified by immunophenotyped. Among 16 histologically confirmed B-NHL cases, only 2 cases diagnosed with follicular lymphoma showed discrepancy with flow cytometry results. In all cases diagnosed with reactive lymphoid hyperplasia and metastasis carcinoma , no abnormal lymphocytes can be found, and k: λ or k: λ ratios were less than 3: 1. Conclusions Fine-needle aspiration analysis with flow eytometrie immunophenotyping can be helpful in diagnosis and differential diagnosis as well as sub-classification of NHL
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Objective To evaluate the methods of measuring platelet-associated antibody PAIgG/ PAIgA/ PAIgM by flow cytometry(FCM) and enzyme linked immunosorbent assay(ELISA), and to investigate their diagnostic value for patients with idiopathic thrombocytopenic purpura(ITP).Methods With FCM and ELISA respectively, PAIg on the platelet membrane and in plasma were measured in 19 patients with ITP and 17 healthy volunteers, and were compared with each other in order to find out whether there were differences in these groups.Results FCM and ELISA measurement in patients with ITP were significantly higher than those in control group (P0.05). Compared with the results of ELISA, the positive percentage of PAIgG measured by FCM(84%) in ITP patients was slightly higher than that by ELISA(79%).Conclusion The platelet-associated antibodies of PAIgG/ PAIgA/ PAIgM, especially PAIgG,are important for diagnosing ITP. FCM, in combination with ELISA, may improve the reliability and the positive percentage of detection in ITP patients.
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Objective: To investigate the changes and significance of microvessel density(MVD) in bone marrow of patients with aplastic anemia. Methods: 30 aplastic anemia cases,20 hyperplastic anemia cases,and 20 normal individuals′ paraffin-embedded bone marrow biopsys were stained with HE and CD34 by immunohistochemistry method respectively;and the vol% of hematopoietic tissue area in bone marrows were detected accordingly. Results:The difference between the MVD of HE and CD34 was not significant in aplastic anemia cases ,and the difference between the MVD of HE and CD34 was not significant in normal individuals either. While the MVD of HE was significantly lower than that of CD34 in hyperplastic anemia cases. The difference of the MVD between light and heavy pathogenetic condition in aplastic anemia cases was significant.The MVD of CD34 were significantly different among the cases of aplastic anemia, hyperplastic anemia,and normal individuals.The MVD showed significant linear correlation with the vol% of hematopoietic tissue area (r=0.74,P
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<p><b>OBJECTIVE</b>To explore the relationship between NB4 cell apoptosis induced by tanshinone IIA (TanIIA) and the cell mitochondrial transmembrane potential (DeltaPsim).</p><p><b>METHODS</b>NB4 cells were treated with TanIIA, As(2)O(3), TanIIA plus 1.0 micro g/ml CsA and As(2)O(3) plus 1.0 micro g/ml CsA, respectively. Morphological changes were observed under light microscope and transmission electron microscope. The percentages of sub-G(1) cells and DeltaPsim of cells doublely stained with PI and Rh123 were assayed by flow cytometry.</p><p><b>RESULTS</b>The percentages of sub-G(1) cells after treatment with 1.0 micro g/ml and 2.0 micro g/ml TanIIA had no significant difference but was higher than that of 0.5 micro g/ml. After treatment with TanIIA, NB4 cells appeared the classical apoptotic morphology. The percentages of sub-G(1) cells were increased, while the DeltaPsim reduced (P < 0.01) and there was a linear correlation between them. The increment of sub-G(1) cell percentages and decrement of DeltaPsim induced by TanIIA were partly inhibited by CsA (P < 0.01).</p><p><b>CONCLUSIONS</b>TanIIA can induce NB4 cells apoptosis through opening the mitochondrial permeability transition pore and reducing DeltaPSgr;m, and this effect could be inhibited by CsA.</p>