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1.
Article in Chinese | WPRIM | ID: wpr-880135

ABSTRACT

OBJECTIVE@#To observe the clinical efficacy of allogeneic peripheral blood stem cell transplantation(allo-HSCT) on the treatment of adult acute leukemia patients, moreover, to establish and evaluate a Logistic model to predict the risk of relapse in adult acute leukemia patients after allo-HSCT.@*METHODS@#The clinical data of 145 adult acute leukemia patients treated by peripheral blood stem cell transplantation in the First Affiliated Hospital of Xi'an Jiaotong University from January 2010 to December 2019 was enrolled and analyzed retrospectively. Complications and survival of patients were observed. The relationship between patients' age, diagnosis, leukocyte count at onset, risk stratification, time of diagnosis to transplantation, HCT-CI, minimal residual disease pre-transplantation, donor-recipient sex relationship, HLA match degree, prophylaxis of graft versus host disease(GVHD), donor age, number of transfused mononuclear cells, CD34 positive cells, engraftment time, acute and chronic GVHD, CMV, EBV infection, and hemorrhagic cystitis and recurrence after transplantation were analyzed by logistic regression. Relapse prediction model was established and evaluated according to the results.@*RESULTS@#Among 145 acute leukemia patients, 81 with acute myeloid leukemia, 64 with acute lymphocytic leukemia, 18 with EBV infection, 2 with post-transplant lymphoproliferative disorder(PTLD), 85 with CMV, 26 with hemorrhagic cystitis, 65 patients developed acute GVHD, 51 patients developed chronic GVHD and 45 patients relapsed. The overall survival (OS) rates in one and three years were 86.4% and 61.8%, and the progress-free survival (PFS) rates in one and three years were 67.5% and 62.4%, respectively. There were significant differences in OS and PFS between relapsed and non-relapsed patients, as well as AML and ALL patients. Univariate analysis revealed that patient's age, risk stratification, time to transplantation, HCT-CI index, ATG based GVHD prophylaxis, minimal residual disease pre-transplantation, GVHD prophylaxis, and acute and chronic GVHD were associated with the relapse of disease, multivariate logistic regression analysis showed that pre-transplantation minimal residual disease showed positively correlation with relapse of the disease, while chronic GVHD showed negatively correlation.@*CONCLUSION@#The relapse rate of adult acute leukemia patients treated with allo-HSCT in our hospital is 31.0%, and OS of AML patients is better than ALL patients'. OS of relapsed patients is significantly lower than non-relapsed patients'. Pre-transplantation minimal residual disease is a risk factor of relapse. The risk of relapse is reduced in patients with chronic GVHD.


Subject(s)
Adult , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Humans , Leukemia, Myeloid, Acute/therapy , Peripheral Blood Stem Cell Transplantation , Recurrence , Retrospective Studies , Transplantation Conditioning
2.
Article in Chinese | WPRIM | ID: wpr-880121

ABSTRACT

OBJECTIVE@#To analyze the risk factors affecting hemorrhagic cystitis(HC) after allogeneic hematopoietic stem cell transplantation(allo-HSCT).@*METHODS@#The clinical data of 153 patients underwent allogeneic hematopoietic stem cell transplantation in the First Affiliated Hospital of Xi'an Jiaotong University from January 2010 to December 2018 were selected and retrospectively analyzed. The incidence, median time and treatment outcome of HC should be observed. Multivariate analysis was used to observe the risk factors of HC in patients, including sex, age, diagnosis, disease status before transplantation, transplantation type, ATG and CTX in the pretreatment scheme, stem cell source, neutrophil and platelet implantation time; CMV, EBV and BKV infection, and acute graft-versus-host disease(aGVHD).@*RESULTS@#Among 153 patients underwent allogeneic hematopoietic stem cell transplantation, 25 (16.34%) patients had HC, the median occurance time was 31 days, all patients achieved complete remission after treatment, no bladder irritation and bladder contracture were left. The results of univariate and multivariate Logistic regression analysis showed that the type of transplantation, ATG, CMV viremia before treatment, aGVHD (r=1.036, 3.234, 3.298 and 2.817, respectively) were the independent risk factors of HC.@*CONCLUSION@#The urinary BKV detections in the patients with HC are positive, mainly occured during the period from day +13 to days +56. HLA haplotype, pretreatment including ATG, and CMV viremia, and aGVHD are the independent risk factors for HC after allo-HSCT.


Subject(s)
Cystitis/etiology , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation/adverse effects , Humans , Retrospective Studies , Risk Factors
3.
Journal of Experimental Hematology ; (6): 1272-1277, 2020.
Article in Chinese | WPRIM | ID: wpr-827127

ABSTRACT

OBJECTIVE@#To explore the renal pathology and cytogenetic features in the multiple myeloma (MM) patients with renal impairment.@*METHODS@#The clinical data of newly diagnosed MM patients with renal impairment in our hospital from January 2009 to January 2019 were analyzed retrospectively, and the relationship between FISH results and results of renal pathological exanimation was analyzed statistically by using SPSS 20.0.@*RESULTS@#A total of 20 patients underwent renal biopsy, included 12 males and 8 females. FISH result showed that out of 20 patients, 7 cases presented interstitial nephritis, among which 3 cases were negative for FISH, and in the remaining cases the rate of IgH rearrangement, 1q21 amplification, RB1 deletion, D13S319 deletion, and P53 deletion detection was 42.86%, 28.57%, 28.57%, 28.57% and 14.29% respectively, the detection positive rate was statistically significantly lower as compared with total probe positive rate (P<0.01). There were 6 cases of cast nephropathy, among which IgH rearrangement, the rate of 1q21 amplification, RB1 deletion, D13S319 deletion, and P53 deletion detection was 66.67%, 50%, 66.67%, 50% and 0% respectively. Compared with the total probe positive rate, there was no statistical significance (P>0.05). There were 4 cases of acute tubular necrosis, among which the detection rates of IgH rearrangement, 1q21 amplification, RB1 deletion, D13S319 deletion, and P53 deletion was 100%, 50%, 50%, 25% and 25%, respectively. Compared with the total probe positive rate, there was no statistical significance (P>0.05). There were one case of amyloidosis, and one case of tubular nephropathy with amyloidosis, the detection with 5 probes were all positive. One case of light chain deposition disease was positive for RB1 gene deletion + D13S319 gene deletion.@*CONCLUSION@#FISH in the MM patients with different renal pathological changes is characterized by heterogeneity, which can be used to predict the risk of renal damage and speculate possible renal pathological types to guide prognosis.


Subject(s)
Chromosome Aberrations , Cytogenetic Analysis , Cytogenetics , Female , Humans , In Situ Hybridization, Fluorescence , Male , Multiple Myeloma , Retrospective Studies
4.
Journal of Experimental Hematology ; (6): 1149-1153, 2019.
Article in Chinese | WPRIM | ID: wpr-775750

ABSTRACT

OBJECTIVE@#To investigate the effect of decitabine on proliferation and apoptosis of multiple myeloma KMS-18 cells and its possible mechanism.@*METHODS@#CCK-8 was used to detect cell proliferation, flow cytometry was used to detect the changes of apoptosis, real-time quantitative PCR was used to detect the expression of P53 gene mRNA in myeloma KMS-18 cells, and MSP assay was used to detect the methylation status of P53 gene promoter.@*RESULTS@#The proliferation inhibition and apoptosis of KMS-18 cells significantly increased after treatment by decitabine (P<0.05). The expression of P53 mRNA increased in KMS-18 cells after treatment of decitabine (P<0.05). The methylation status of the P53 gene promoter in KMS-18 cells could be partially reversed by decitabine.@*CONCLUSION@#Decitabine can inhibit the proliferation of KMS-18 cells and induce their apoptosis, its mechanism ralates with partially reversing the methylation of P53 gene promoter in KMS-18 cells.


Subject(s)
Apoptosis , Cell Line, Tumor , Cell Proliferation , DNA Methylation , Decitabine , Humans
5.
Journal of Experimental Hematology ; (6): 1374-1379, 2019.
Article in Chinese | WPRIM | ID: wpr-775711

ABSTRACT

OBJECTIVE@#To investigate the influence of oridonin on the killing activity of NK-92 MI cells targeting THP1 and the related mechanism.@*METHODS@#The killing activity of NK-92 MI to THP1 before and after oridonin treatment was detected by LDH release assay; the expression of natural killer cell ligands activating receptor D (NKG2D, including MICA, MICB, ULBP1, ULBP2 and ULBP3) was detected by real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot respectively; the expression of cytokine TNF-α, TNF-β and IFN-γ in the co-culture supernatant of NK-92 MI cells and THP1 cells were measured by ELISA.@*RESULTS@#The killing efficiency after oridonin treatment at different effector-target ratio (1:1, 5:1, 10:1) was all significantly up-regulated in comparison with that before oridonin treatment (P<0.05). QRT-PCR and Western blot showed that the expressions of mRNA and protein levels of MICB, ULBP1, ULBP2 increased to varying degree (P<0.05), but the expression levels of MICA and ULBP3 were not statistically significant between experimental group and control group (P>0.05). ELISA results indicated that IFN-γ and TNF-β release were significantly increased after oridonin treatment (P<0.05), however, the TNF-α release was not statistically different in comparison with control group (P>0.05).@*CONCLUSION@#Oridonin can significantly improve killing efficiency of NK-92 MI on THP1, that might be related with up-regulation of MICB, ULBP1 and ULBP2 expression and promotion of IFN-γ and TNF-β release.


Subject(s)
Cell Line, Tumor , Diterpenes, Kaurane , Pharmacology , GPI-Linked Proteins , Histocompatibility Antigens Class I , Humans
6.
Journal of Experimental Hematology ; (6): 1402-1408, 2019.
Article in Chinese | WPRIM | ID: wpr-775707

ABSTRACT

OBJECTIVE@#To investigate the mechanism of rapamycin-induced apoptosis of chronic myelogenous leukemia cells.@*METHODS@#The chronic granulocytic leukemia K562 cells were divided into 3 groups: A, B and C group were treated with rapamycin of 10, 15 and 20 nmol/L, repectively for 24 h, while the K562 cells in control group were not treated with rapamycin. The effect of rapamycin on the proliferation of K562 cells was detected by MTT, and the effect of rapamycin on the apoptosis of K562 cells was detected by AnnexinV-FITC/PI double staining. The expression level of EZH2/Hedgehog signaling pathway genes in K562 cells was detected by RT-PCR, and Western blot was used to detect the levels of apoptotic protein and the related signaling pathway proteins in K562 cells.@*RESULTS@#The MTT assay showed that the different concentration of rapamycin had obvious inhibitory effects on the cells, and the survival rate of cells in group C was 37.6%±3.4%, which was significantly lower than that of the other groups (P<0.05). The apoptosis rate of cells in group C was 93.1%±8.1%, which was significantly higher than that of the other groups (P<0.05). By Western blot, it was found that the relative expression levels of Caspase-3 and BAX protein in group C were 0.36 ± 0.04 and 0.39±0.06, respectively, which were significantly higher than those in other groups (P<0.05), and the level of BCL-2 protein was 0.17±0.03, which was significantly lower than that of other groups (P<0.05). By RT-PCR, it was found that the mRNA levels of EZH2 and Hedgehog genes in A, B and C groups were significantly lower than those in the control group (P<0.05), but mRNA level of Ptch1 gene was significantly higher than that of the control (P<0.05). By Western blot, it was found that the expression levels of EZH2 and Hedgehog protein in A, B and C groups were significantly lower than that in the control group (P<0.05), but the level of Ptch1 protein was higher than that of the control (P<0.05). The relative levels of EZH2 and Hedgehog protein in group C were 0.21 ±0.03 and 0.16±0.05 respectively, which were significantly lower than those in other groups (P<0.05), and Ptch1 protein level were 0.46 ±0.06, significantly higher than that of other groups (P<0.05).@*CONCLUSION@#Rapamycin can inhibit the protein expression of EZH2 in leukemic cells, thus interfere with the activation of Hedgehog signaling pathway, promote the expression of apoptotic protein, reduce the level of anti apoptotic protein, and eventually induce apoptosis of leukemia cells.


Subject(s)
Apoptosis , Cell Proliferation , Enhancer of Zeste Homolog 2 Protein , Hedgehog Proteins , Humans , K562 Cells , Signal Transduction , Sirolimus
7.
Article in Chinese | WPRIM | ID: wpr-698246

ABSTRACT

Objective To investigate the influences of donor HBV infection on allogeneic hematopoietic stem cell transplantation recipients.Methods We made a retrospective analysis of data of four patients without HBV infection who underwent allogeneic hematopoietic stem cell transplantation from January 2015 to December 2016. Among them donors of these patients all had HBV infection.We then observed the influences of HBV infection on hematopoietic reconstruction,hepatic vein occlusive disease and HBV infection.Results HBV serological conditions of two donors were HbsAb,HbeAb and HbcAb positive,and quantitative of HBV-DNA was negative;the donor and the recipient did not use anti-HBV drugs.One donor was HbsAg,HbeAb and HbcAb positive,and the quantitative of HBV-DNA was also positive.Another donor was HbsAg and HbcAb positive,and the quantitative of HBV-DNA was also positive.These two donors received oral nucleoside therapy one month before stem cell collection and the recipients of these two donors also took nucleoside drugs one week before the conditioning.Hepatitis B immune globulin was given after transfusion of stem cells and the third day and seventh day after transplantation.Quantitative of HbsAb was detected each month and if it was less than 150 IU,hepatitis B immune globulin would be given.All the recipients had hematopoietic reconstruction and no VOD or hepatitis B virus infection occurred.Conclusion Oral administration of nucleoside drugs combined with hepatitis B immunoglobulin can effectively prevent HBV infection in recipients with HBV infection donors.

8.
Article in Chinese | WPRIM | ID: wpr-690966

ABSTRACT

<p><b>OBJECTIVE</b>To explore the characteristics and diagnostic values of bone marrow cell morphology and immunophenotyping in lymphoma cell leukemia patients.</p><p><b>METHODS</b>Data of the bone marrow cell morphology and immunophenotyping of 35 patients with lymphoma cell leukemia admitted from January 2012 to January 2017 were analyzed retrospectively, and the value of bone marrow cell morphology and immunophenotype in the diagnosis of lymphoma cell leukemia was evaluated.</p><p><b>RESULTS</b>Bone marrow cell morphological examination showed the typical lymphoma cells in all the patients. The expression of differentiation antigens in lymphoma cell leukemia was consistent with that of original pathological diagnosis. In T-cell lymphoma cell leukemia, the expression of CD7, CD3, CD2, CD5, CD11b, CD34, and HLA-DR were present predominantly, among them the CD7 was the most sensitive antigen and its positive expression rate was 69.2%. In B-cell lymphoma cell leukemia, the expression of CD19, CD20, CD22, CD79a, Skappa, and early antigen HLA-DR were observed predominantly, among them the positive expression rate of CD19 was the highest (89.5%). Out of 35 cases, 28 cases showed that the percentage of lymphoma cells on bone marrow smears was consistent with that of bone marrow immunophenotyping, and 7 cases showed that the percentage of lymphoma cells between bone marrow smears and immunophenotyping differed by more than 1.5-fold.</p><p><b>CONCLUSION</b>Bone marrow slides combined with immunophenotyping may be helpful for judging lymphoma cell marrow invasion and making early diagnosis of lymphoma cell leukemia.</p>


Subject(s)
Bone Marrow Cells , Flow Cytometry , Humans , Immunophenotyping , Leukemia , Lymphoma , Retrospective Studies
9.
Journal of Experimental Hematology ; (6): 1317-1322, 2018.
Article in Chinese | WPRIM | ID: wpr-689937

ABSTRACT

<p><b>OBJECTIVE</b>To expolore the effect of programmed death receptor ligand 1 (PD-L1) expression level on killing effect of different cell lines of acute myeloid leukemia (AML) and its possible mechanism.</p><p><b>METHODS</b>Peripheral blood from healthy individuals was collected routinely; NK cells were isolated using immunomagnetic beads; PD-L1 expression level was detected by flow cytometry; the killing effect of NK cells on acute myelogenous leukemia cell lines was evaluated with LDH release method and monoclonal antibody blocking experiment; the expression levels of IFN-γ and IL-2 in the supernatants from the co-cultured effector/targer cells were measured by ELISA.</p><p><b>RESULTS</b>The ratio of CD3CD56NK cells increased from (12.44±3.48)% to (71.29±5.65)%. The flow cytometry showed that KG-1a cells lowly expressed PD-L1 (8.35±4.12)%, but the THP cells a highly expressed PD-L1 (76.42±26.54)%. Meanwhile, the NK cells displayed a more efficient killing effect on KG-1a cells than that of THP1 cells (P<0.05). Moreover, PD-L1 monoclonal antibody could reinforce NK cell killing effect and, promote the secretion of IFN-γ and IL-2 in 5 acute myelogenous leukemia cell lines to varying degree.</p><p><b>CONCLUSION</b>The killing effect of NK cells on acute myelogenous leukemia cell line is inversely proportional to PD-L1 expression; blocking PD1/PD-L1 binding can significantly enhance the killing efficiency of effector-target cells, which way be related with promoting the release of IFN-γ and IL-2.</p>

10.
Journal of Experimental Hematology ; (6): 1431-1435, 2017.
Article in Chinese | WPRIM | ID: wpr-301711

ABSTRACT

<p><b>OBJECTIVE</b>To study the expression of DNMT3b gene in myeloma RPMI8226 cells and its biological significance.</p><p><b>METHODS</b>The activity of DNA methyltransferase was detected by ELISA, and the expression of DNMT3b in RPMI8226 cells was analyzed by semi-quantitative RT-PCR and real-time fluorescent quantitative PCR. The proliferation and expression of DNMT3b gene in RPMI8226 cells intervened with capecitabine for 24 hours were detected.</p><p><b>RESULTS</b>The activity of DNMT and expression of DNMT3b in RPMI 8226 cells increased. The proliferation of RPMI8226 cells was inhibited, and the apoptosis occurred in RPMI 8226 cells intervened with capecitabine for 24 hours. The expression level of DNMT3b gene was decreased after being intervened with capecitabine for 24 hours.</p><p><b>CONCLUSION</b>The expression level of DNMT3b in myeloma RPMI 8226 cells increase, and capecitabine can inhibit the proliferation of RPMI 8226 and induce apoptosis by inhibiting the expression of DNMT3b gene. Therefore, DNMT3b is expected to be a new target for myeloma therapy.</p>

11.
Article in Chinese | WPRIM | ID: wpr-271901

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the risk factors and therapeutic outcome of acute graft versus host disease (aGVHD) in patients with acute leukemia after haploidentical peripheral hematopoietic stem cell transplantation.</p><p><b>METHODS</b>The clinical data of 19 cases of acute leukemia underwent haploidentical hematopoietic stem cell transplanttion during January 2010 and December 2010 were retrospectively analyzed. The effects of patients sex, donor-recipient sex difference, donor age, conditioning regimen, dosage of anti-thymocyte globulin(ATG), mononuclear cell and CD34cell counts on the intestinal aGVHD were analyzed by Logistic regression.</p><p><b>RESULTS</b>Intestinal aGVHD occurred in 5 cases with 1 case at stage II 3 cases at stage III and 1 case at stage IV on the 7th, 22th, 27th, 70th and 154th day after transplantation, respectively. Single factor analysis showed that the patient's sex, donor-recipient sex difference, donor age, dosage of ATG, mononuclear cell and CD34cell counts were not related with the occurrence of the intestinal aGVHD, and the conditoning regimen was the risk factor for the intestinal aGVHD. 2 cases among 5 cases with intestinal aGVHD were treated with methylprednisolone at dosage of 1 mg/kg per day, 1 case was treated with methylprednisolone therapy combined with tacrolimus. 2 cases of methylprednisolone-resistance were treated with CD25 monoclonal antibody. Intestinal aGVHD of all patients was improved after the above-mentioned treatment.</p><p><b>CONCLUSION</b>Conditioning regimen of haploidentical peipheral hematopoieitc stem cell transplantaion has effects on the intestinal aGVHD, which needs to be confirmed by further research.</p>

12.
Article in Chinese | WPRIM | ID: wpr-272508

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of silencing SET gene on the biological characteristics of acute promyelocytic leukemia NB4-R1 cells.</p><p><b>METHODS</b>The expression vector of pGCSIL containing SET-shRNA were transfected into 293T cells by using other packaging plasmids. The supernatant of the 293T cells was harvested for lentivirus. The SET-shRNA lentiviral vector was transfected into acute promyelocytic leukemia NB4-R1 cells and a stably transfected cell line was established. Real-time quantitative PCR and Western blot were used to assay the silencing efficiency on SET gene and the expression of PP2A. The cell cycle distribution was tested by flow cytometry.</p><p><b>RESULTS</b>The expression of SET in experimental group statistically decreased as compared with that of the control group. The expression of PP2A was obviously raised at the level of mRNA and protein. The percentage of NB4-R1 cells in G0/G1 phase significantly increased, while the percentage of cells in S phase significantly decreased.</p><p><b>CONCLUSION</b>The silencing gene in acute promyelocytic leukemia NB4-R1 cells using SET-shRNA lentiviral vector can increase the expression of PP2A and interfere of the cell cycle in NB4-R1 cells. This study has laid a experimental base for targed therapy of patients with acute promyelocytic leukemia.</p>


Subject(s)
Cell Cycle , Cell Line, Tumor , Gene Silencing , Genetic Vectors , HEK293 Cells , Histone Chaperones , Genetics , Humans , Lentivirus , Leukemia, Promyelocytic, Acute , Genetics , Pathology , Protein Phosphatase 2 , Metabolism , RNA, Messenger , RNA, Small Interfering , Transcription Factors , Genetics , Transfection
13.
Journal of Experimental Hematology ; (6): 1397-1403, 2016.
Article in Chinese | WPRIM | ID: wpr-332680

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of hepatovirus B(HBV) infection on the hematopoietic stem cell collection and implantment in lymphoma patients received autologous peripheral hematopoietic blood stem cells transplantation.</p><p><b>METHODS</b>Clinical data of 40 lymphoma patients who received autologous peripheral hematopoietic blood stem cell transplantation between January 2006 and October 2014 was analyzed retrospectively. Among 40 patients with lymphoma 8 patients combined with HBV infection were prophylacticly given nucleoside analogues and 32 patients without HBV infection. The counts of mononuclear cells(MNC) and CD34 positive cells were collected and the hematopoietic reconstitution as well as overall survival rates and progress-free survival rates were detected and counted between patients with or without HBV infection.</p><p><b>RESULTS</b>The counts of MNC and CD34 positive cells in all patients were standard, and there was no significant difference between patients with or without HBV infection. HBV wasn't reactivated among the 8 patients with HBV infection. The 1, 3 and 5 years' overall survival rates and progress-free survival rates of patients with HBV infection were 100%, 85.7%, 57.1% and 100%, 80%, 53%, respectively and the 1,3 and 5 years' overall survival rates and progress-free survival rates of patients without HBV infection were 100%, 88.9%, 82.1% and 90%, 90%, 90%, respectively.</p><p><b>CONCLUSION</b>HBV infection may have no effect on the collection of stem cells and hematopoietic reconstitution. Prophylactic use of nucleoside analogues can effectively prevent the hepatitis B virus reactivation, moreover had no effect on the collection and hematopoietic reconstitution.</p>

14.
Journal of Experimental Hematology ; (6): 1529-1532, 2016.
Article in Chinese | WPRIM | ID: wpr-332657

ABSTRACT

<p><b>OBJECTIVE</b>To observe the efficacy and adverse reactions of autologous PBSC collection when the autoPBSC procedure and MNC procedure of COBE Spectra cell separator and the MNC procedure of Spectra Optia cell separator were used.</p><p><b>METHODS</b>The autologous perepheral blood hematopoietic stem cells from 41 patients were collected by using autoPBSC procedure and MNC procedure of COBE Spectra blood cell separator and MNC procedure of Spectra Optia blood cell separator. The numbers of MNC and CD34cells collected by 3 collected procedure, the difference of hemoglobin (Hb) drop and platelet decrease, and the adverse reaction of patients were observed.</p><p><b>RESULTS</b>When the whole blood processing and the collection time were basically same among these 3 groups, the MNC counts collected by MNC procedure of COBE Spectra and Spectra Optia were higher than that of AutoPBSC procedure of COBE Spctra, but the CD34cell count was lower than that collected by AutoPBSC procedure (P< 0.05). The final product volume collected by MNC procedure of COBE Spectra and Spectra Optia was bigger than that collected by AutoPBSC procedure. In comprission with MNC procedure of COBE Spectra cell seperator, the CD34count collected by MNC procedure of Spectra Optia Seperator did not show significant difference, but the CD34cell count collected by MNC procedure of Spectra Optia was higher than that collected by MNC procedure of COBE Spectra cell separator (P<0.05). The platelet count and hemoglobin level collected by MNC procedure of Spectra Optia were lower than those before collection. The adverse reactions in the 3 procedures were similar, and the patients could tolerate them.</p><p><b>CONCLUSION</b>The AutoPBSC procedure of COBE Spectra and MNC procedure of Spectra Optia are better than MNC procedure of COBE Spectra for autologous peripheral blood hematopoietic stem cells collection. The loss of blood platelet and hemoglobin after collection is lowest in MNC procedure of Spectra Optia.</p>

15.
Article in Chinese | WPRIM | ID: wpr-259642

ABSTRACT

<p><b>OBJECTIVE</b>This study was to establish a stable, effective and reproducible human acute promyelocytic leukemia model in severe combined immunodeficient (SCID) mice by using NB4 cell line, and to investigate the disease course character and biological behaviors.</p><p><b>METHODS</b>Three-five-week-old SCID beige mice were divided randomly into two groups: experimental and control group. SCID mice of experimental group were transplanted by tail vein (iv) injection of 5×10(6) NB4 cells. The WBC cell count and the positive rate of promyelocytes in peripheral blood were dynamically monitored by using smears. Morphological examination and histopathological assay were employed to confirm NB4 cell infiltration in organs (liver, spleen, lung, kidney and brain). The expression level of PML-RARα fusion protein was detected by Western blot.</p><p><b>RESULTS</b>Within two weeks there was no significant difference in peripheral blood WBC count between two groups (P > 0.05), meanwhile, NB4 cells were not found. At the day 21 and 28 after inoculation, the peripheral blood white blood cell count of experimental group reached to (4.79 ± 1.13)×10(9)/L and (7.62 ± 2.24)×10(9)/L respectively, which were significantly higher than that in control group (P < 0.05); simultaneously, the positive rates of promyelocytes on smears were (2.14 ± 0.63)% and (6.6 ± 2.76)%, respectively. Morphological observation showed single or multiple tumor lumps at day 21 after inoculation; HE staining of tissue biopsies demonstrated a large number of promyelocyte in the liver, spleen, lung, kidney and brain tissue. Cell immunofluorescence results showed that the CD33 expression of bone marrow cells in mice of experimental group was strongly positive (P < 0.05). Western blot confirmed that the PML-RARα fusion protein was expressed variously in liver, kidney and brain tissue.</p><p><b>CONCLUSIONS</b>The human acute promyelocytic leukemia SCID mouse model is succesfully established by tail vein injection of NB4 cells. This model can mimic the characters of involved bone marrow and diffuse growth of cells. This model is a useful tool to explore the pathogenic mechanism and experimental treatment of human leukemia.</p>


Subject(s)
Animals , Cell Line , Granulocyte Precursor Cells , Humans , Leukemia, Promyelocytic, Acute , Mice , Mice, SCID , Oncogene Proteins, Fusion
16.
Article in Chinese | WPRIM | ID: wpr-259636

ABSTRACT

<p><b>OBJECTIVE</b>This study was to investigate the apoptosis-inducing effect of As(4)S(4) on the retinoic acid-resistant acute promyelocytic leukemia (APL) NB4-R1 cells and its potential mechanisms.</p><p><b>METHODS</b>The leukemia cell line NB4-R1 was cultured in vitro and divided into control group and treatment group. The apoptosis rate and cell cycle were detected by flow cytometry. The apoptotic DNA fragments were analyzed by agarose gel electrophoresis. The changes of BCL-2, BAX and Caspase-3 were determined by Western blot.</p><p><b>RESULTS</b>After NB4-R1 cells were treated with As(4)S(4)(25 µmol/L) for 0 h, 24 h, 48 h, the percentage of early apoptotic cells was obviously raised from 0% to 24.49% and 47.41%, the percentage of late apoptotic cells were elevated from 0.08% to 14.72% and 20.70%. Compared with control group, the DNA degradation revealed a characteristic DNA ladder during agarose gel electrophoresis after treatment for 24 h. The drug significantly induced an accumulation of the S phase cell population from 31.85% of the untreated cells to 42.53% and 55.12% treated with the different time whereas the NB4-R1 cells in G0/G1 phase decreased from 57.30% to 37.56% and 28.51%. As(4)S(4) could decrease the expression of BCL-2 and increase the level of BAX. Pro-caspase-3 could be cleaved into small active fragments under the apoptotic stimulation.</p><p><b>CONCLUSION</b>As(4)S(4) can efficiently induce NB4-R1 cell apoptosis, which may be related with the down-regulation of BCL-2 and the up-regulation of BAX, as well as the activation of Caspase-3.</p>


Subject(s)
Apoptosis , Caspase 3 , Cell Cycle , Cell Line, Tumor , Down-Regulation , Drug Resistance, Neoplasm , Humans , Leukemia, Promyelocytic, Acute , Tretinoin , Up-Regulation
17.
Article in Chinese | WPRIM | ID: wpr-302409

ABSTRACT

This study was aimed to explore the effect of realgar (As4S4) on growth inhibition and apoptosis induction of DLBCL cell line SU-DHL-4 and its mechanisms. The inhibitory effect of realgar on the cell growth were detected by MTT method. The morphological changes of SU-DHL-4 were observed by transmission electron microscopy (TEM). The apoptosis of SU-DHL-4 cells treated with realgar were detected by flow cytometry with Annexin V-FITC/PI double staining and DNA agarose gel electrophoresis. The cell cycle was examined by flow cytometry with PI staining. The expressions of apoptosis-related proteins (BCL-2 , Caspase-3,BAX) were detected by Western blot. The results showed that the realgar at the concentration of 20, 40, 80 µmol/L all could inhibit the proliferation of SU-DHL-4 (P < 0.05), and in a certain time and concentration range, the inhibition rate was enhanced in a time and dose dependent manner(r = 0.982). Flow cytometric test results showed that realgar could induce SU-DHL-4 cell apoptosis after treating for 48 hours, and the apoptosis rate increased with the increasing of drug concentration (P < 0.05). After treating SU-DHL-4 cells with Realgar for 48 h, the cell cycle was blocked in the S phase (P < 0.05). TEM results revealed that when treated with realgar for 48 h, the typically apoptosis morphology-apoptotic bodies were observed in all drug-treated group, furthermore, some necrotic cells in the 80 µmol/L group were observed. After intervened by realgar for 48 h, the DNA Ladder pattern was seen according to agarose gel electrophoresis. Western blot showed that the expression of Bcl-2 protein was down-regulated while the expressions of BAX and Caspase-3 protein were up-regulated when treating SU-DHL-4 cells with realgar for 48 h. It is concluded that realgar can inhibit cell growth and induce cell apoptosis, which may be related with up-regulation of Caspase-3 and BAX expression and down-regulation the of BCL-2 expression.


Subject(s)
Apoptosis , Arsenicals , Pharmacology , Caspase 3 , Metabolism , Cell Line, Tumor , Cell Proliferation , Flow Cytometry , Humans , Leukemia, Lymphocytic, Chronic, B-Cell , Metabolism , Pathology , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Sulfides , Pharmacology , bcl-2-Associated X Protein , Metabolism
18.
Chinese Medical Journal ; (24): 4078-4082, 2013.
Article in English | WPRIM | ID: wpr-236102

ABSTRACT

<p><b>BACKGROUND</b>Decreasing the intracranial pressure has been advocated as one of the major protective strategies to prevent spinal cord ischemia after endovascular aortic repair. However, the actual changes of cerebrospinal fluid (CSF) pressure and its relation with spinal cord ischemia have been poorly understood. We performed CSF pressure measurements and provisional CSF withdrawal after thoracic endovascular aortic repair, and compared the changes of CSF pressure in high risk patients and in patients with new onset paraplegia and paraparesis.</p><p><b>METHODS</b>Four hundred and nineteen patients were evaluated for the risk of spinal cord ischemia after thoracic endovascular aortic repair. Patients with identified risk factors before the procedure constituted group H and received prophylactic sequential CSF pressure measurement and CSF withdrawal. Patients who actually developed spinal cord ischemia constituted group P and received rescue CSF pressure measurements and CSF withdrawal.</p><p><b>RESULTS</b>Among the 419 patients evaluated, 17 were graded as high risk. Four patients actually developed spinal cord ischemia after endovascular repair. The incidence of spinal cord ischemia in this investigation was 0.9%. The patients who actually developed spinal cord ischemia had no identified risk factors and had elevated CSF pressure, ranging from 15.4 to 30.0 mmHg. Six of the 17 patients graded as high risk had elevated CSF pressure: >20 mmHg in two patients and >15 mmHg in four patients. Sequential CSF pressure measurements and provisional withdrawal successfully decrease CSF pressure and prevented symptomatic spinal cord ischemia in high-risk patients. However, these measurements could only successfully reverse the neurologic deficit in two of the patients who actually developed spinal cord ischemia.</p><p><b>CONCLUSIONS</b>Cerebrospinal fluid pressure was elevated in patients with spinal cord ischemia after thoracic endovascular aortic repair. Sequential measurements of CSF pressure and provisional withdrawal of CSF decreased CSF pressure effectively in high risk patients and provided effective prevention of spinal cord ischemia. Risk factor identification and prophylactic measurements play the key role in prevention of spinal cord ischemia after thoracic endovascular aortic repair.</p>


Subject(s)
Aged , Aorta, Thoracic , General Surgery , Cerebrospinal Fluid Pressure , Physiology , Female , Humans , Male , Middle Aged , Spinal Cord Ischemia
19.
Journal of Experimental Hematology ; (6): 1530-1534, 2013.
Article in Chinese | WPRIM | ID: wpr-264981

ABSTRACT

This study was aimed to investigate the relation of reinfused hematopoietic stem cell volume and recipient's leukocyte count at reinfusion with prognosis of disease in allo-hematopoietic stem cell transplantation (allo-HSCT). The clinical data of 37 patients received allo-HSCT in our hospital were analyzed retrospectively. The 37 patients were divided into agranulocytosis and non-agranulocytosis groups according to the recipient's leukocyte count at reinfusion, and were divided into the high dose and low dose groups according to the median number of reinfused mononuclear cells (MNC) and CD34(+) cells. Then, hematopoietic reconstructions,GVHD, relapse and death rates of patients were compared. The results showed that the hematopoietic reconstruction of patients in non-agranulocytosis group and high dose MNC group were earlier than that in agranulocytosis group and low dose MNC group. There was no significant difference of hematopoietic reconstruction between the groups of high dose CD34(+) cells and low dose CD34(+) cells. The GVHD incidence was higher in high dose MNC group and non-agranulocytosis group than that in low dose MNC group and agranulocytosis group (P < 0.05). There were no statistical differences of relapsed and death rates between different reinfused number of HSC and recipient's leukocyte count at reinfusion.It is concluded that the infused MNC number and the recipient's leukocyte count at reinfusion in allo-HSCT correlated with hematopoietic reconstruction, the GVHD incidence is high in high dose MNC and non-agranulocytosis groups, the reinfused HSC volume and the recipient's leukocyte count at reinfusion not significantly relate with relapse and death rates.


Subject(s)
Adolescent , Adult , Child , Female , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Methods , Hematopoietic Stem Cells , Cell Biology , Humans , Leukocyte Count , Male , Middle Aged , Prognosis , Recurrence , Retrospective Studies , Young Adult
20.
Article in Chinese | WPRIM | ID: wpr-332782

ABSTRACT

To explore the mechanism of autophagic death of acute myelocytic leukemia cell U937 induced by clofarabine, the MTT bioassay was used to analyze the growth inhibitory effect and half inhibition concentration on U937 incubated in vitro with different concentrations of clofarabine for 24 and 48 hours, and the flow cytometry was used to detect the autophagy rate of U937. The expression of Beclin 1 in U937 treated by clofarabine for 48h was measured by Western blot. The results indicated that when U937 cells were treated with 0.01 µmol/L and 0.15 µmol/L clofarabine for 48 hours, the proliferation inhibition rate was 46.92% ± 4.24% and 86.10% ± 1.16%, and the half inhibition concentration of clofarabine was 0.022 µmol/L. With 0.01 µmol/L and 0.1 µmol/L clofarabine on U937 for 48 hours, the autophagy rate was 11.0033% ± 1.4387% and 59.4133% ± 3.5409%, and increased in dose-dependent manner (r = 0.99). Meanwhile the Beclin 1 was upregulated along with increase of clofarabine concentration, as compared with control group, the difference was statistically significant (P < 0.05). It is concluded that the different concentrations of clofarabine can significantly inhibit the proliferation of U937 in dose-dependent manner, and the mechanism of autophagic cell death in U937 may be associated with the upregulation of Beclin 1 expression.


Subject(s)
Adenine Nucleotides , Pharmacology , Apoptosis , Apoptosis Regulatory Proteins , Metabolism , Arabinonucleosides , Pharmacology , Autophagy , Beclin-1 , Cell Proliferation , Humans , Membrane Proteins , Metabolism , U937 Cells
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