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1.
Hematology ; 28(1): 2171723, 2023 Dec.
Article in English | MEDLINE | ID: mdl-36752506

ABSTRACT

Asparaginase-associated pancreatitis (AAP) is a common and fatal complication after ASNase treatment in acute lymphoblastic leukemia(ALL). Here, a total of 1063 pediatric ALL patients treated with SCCLG-ALL-2016 regimen were collected since October 2016 to June 2020, including 35 patients with AAP. The clinical characteristics of AAP and non-AAP patients were compared. In AAP patients, the possible factors that affected the recurrence of AAP were analyzed, and the possible risk factors related to ALL-relapse were discussed. The results showed that age was a risk factor (P = .017) that affect the occurrence of AAP. In AAP patients, AAP tended to develop after the second use of PEG-ASNase (25.71%). In the follow-up chemotherapy, 17 patients re-exposed to ASNase and 7 cases developed AAP again with a percentage was 41.2%. There were no special factors that related with the recurrence of AAP. This study also found no association between the occurrence of AAP and prognosis of ALL, with the 4-year incidence of ALL relapse in AAP and non-AAP patients were 15.9% v.s.11.7% (HR: 1.009, 95% CI:0.370-2.752, P = .986), and there were no special factors that related with the ALL relapse among AAP patients. Based on the above results, the occurrence of AAP is related to age and should be vigilant after the second use of PEG-ASNase after use in pediatric ALL patients. Moreover, AAP is not associated with ALL relapse, but there is a high AAP recurrence rate when re-exposure to ASNase.


Subject(s)
Antineoplastic Agents , Pancreatitis , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Humans , Child , Asparaginase/adverse effects , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Pancreatitis/chemically induced , Pancreatitis/drug therapy , Polyethylene Glycols/therapeutic use , Prognosis , Recurrence , Antineoplastic Agents/therapeutic use
2.
Front Oncol ; 11: 797194, 2021.
Article in English | MEDLINE | ID: mdl-34988026

ABSTRACT

PURPOSE: To analyzed the outcome of ETV6/RUNX1-positive pediatric acute B lymphoblastic leukemia (B-ALL) with the aim of identifying prognostic value. METHOD: A total of 2,530 pediatric patients who were diagnosed with B-ALL were classified into two groups based on the ETV6/RUNX1 status by using a retrospective cohort study method from February 28, 2008, to June 30, 2020, at 22 participating ALL centers. RESULTS: In total, 461 (18.2%) cases were ETV6/RUNX1-positive. The proportion of patients with risk factors (age <1 year or ≥10 years, WB≥50×109/L) in ETV6/RUNX1-positive group was significantly lower than that in negative group (P<0.001), while the proportion of patients with good early response (good response to prednisone, D15 MRD < 0.1%, and D33 MRD < 0.01%) in ETV6/RUNX1-positive group was higher than that in the negative group (P<0.001, 0.788 and 0.004, respectively). Multivariate analysis of 2,530 patients found that age <1 or ≥10 years, SCCLG-ALL-2016 protocol, and MLL were independent predictor of outcome but not ETV6/RUNX1. The EFS and OS of the ETV6/RUNX1-positive group were significantly higher than those of the negative group (3-year EFS: 90.11 ± 4.21% vs 82 ± 2.36%, P<0.0001, 3-year OS: 91.99 ± 3.92% vs 88.79 ± 1.87%, P=0.017). Subgroup analysis showed that chemotherapy protocol, age, prednisone response, and D15 MRD were important factors affecting the prognosis of ETV6/RUNX1-positive children. CONCLUSIONS: ETV6/RUNX1-positive pediatric ALL showed an excellent outcome but lack of independent prognostic significance in South China. However, for older patients who have the ETV6/RUNX1 fusion and slow response to therapy, to opt for more intensive treatment.

3.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 27(4): 1058-1063, 2019 Aug.
Article in Chinese | MEDLINE | ID: mdl-31418357

ABSTRACT

OBJECTIVE: To detect the expression of CRLF2 in bone marrow mononuclear cells from children with newly diagnosed acute lymphoblastic leukemia(ALL) and to explore its clinical significance in pediatric ALL. METHODS: A total of 218 children with newly diagnosed ALL who achieveal the complete remission and had the complete follow-up information were selected, and the expression level of CRLF2 in bone marrow mononuclear cells of these children was detected by real-time fluorescent quantitative PCR, and the significance of CRLF2 expression level in clinical prognosis of ALL children was analyzed by using statistical method. RESULTS: 28 cases in 218 children with complete data showed high expression of CRLF2. The cumulative recurrence rate in the CRLF2 high expression group was significantly higher than that in the low expression group (53.6% vs 12.6%) (P<0.01). The predicted 5-year recurrence-free survival rate (RFS) of ALL children with CRLF2 high expression was significantly higher than that of low expression group (P<0.01). There was no significant difference in the predicted 5-year RFS between ALL children with CRLF2 low and high expression in the standard-risk(SR) group (P>0.05). The predicted 5-year RFS of ALL children with CRLF2 low expression was higher than that of ALL children with CRLF2 high expression in the intermediate-risk (IR) and high-risk (HR) groups. (P<0.05). Cox analysis showed that CRLF2 high expression is an independent risk factor for the relapse of children with ALL. CONCLUSION: The recurrence rate of pediatric ALL with CRLF2 high expression is high, and CRLF2 high expression is an important prognostic factor for high risk of relapse in ALL children with IR and HR. It is necessary to use CRLF2 expression as an indicator of risk stratification in pediatric ALL.


Subject(s)
Bone Marrow , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Receptors, Cytokine/metabolism , Child , Humans , Prognosis , Recurrence , Risk Factors
4.
Zhongguo Dang Dai Er Ke Za Zhi ; 20(7): 538-542, 2018 Jul.
Article in Chinese | MEDLINE | ID: mdl-30022754

ABSTRACT

OBJECTIVE: To study the effects of minimal residual disease (MRD) level on day 33 of remission induction and IKZF1 genotype on the survival of children with B-lineage acute lymphoblastic leukemia (B-ALL). METHODS: A total of 152 children with newly-diagnosed B-ALL who had complete remission after the first cycle of the chemotherapy and had complete follow-up information were enrolled in this study. According to the MRD detection by flow cytometry on day 33 of remission induction, they were divided into three groups: standard-risk (SR) group (MRD <10-4; n=60), intermediate-risk (IR) group (10-4≤ MRD <10-2; n=55), and high-risk (HR) group (MRD ≥10-2; n=37). Nested RT-PCR was used to determine the IKZF1 genotype of all children before chemotherapy. The effects of MRD level on day 33 of remission induction and IKZF1 genotype on the recurrence-free survival (RFS) of children with B-ALL were analyzed. RESULTS: There were 7 common IKZF1 subtypes in all the 152 children with B-ALL: IK1, IK2/3, IK4, IK6, IK8, IK9, and IK10. Of the 152 children, 130 had functional subtypes of IKZF1 and 22 had non-functional subtypes of IKZF1. During the follow-up period, relapse occurred in 26 (17%) children, and the recurrence rate was highest in the HR group (P<0.05). However, there was no significant difference in the recurrence rate between the SR group and the IR group (P>0.05). The cumulative recurrence rate of the children with non-functional subtypes of IKZF1 was significantly higher than that of those with functional types of IKZF1 (P<0.01). The predicted 5-year RFS rates in the SR, IR, and HR groups were (94.2±2.9)%, (86.7±3.8)%, and (56.2±4.5)% respectively (P<0.05). The 5-year RFS rate of the children with functional subtypes of IKZF1 was significantly higher than that of those with non-functional subtypes of IKZF1 (P<0.01). There was no significant difference in the predicted 5-year RFS rate between the children with functional subtypes of IKZF1 and those with non-functional subtypes of IKZF1 in the SR group (P>0.05). However, the predicted 5-year RFS rate of the children with functional subtypes of IKZF1 was significantly higher than that of those with non-functional subtypes of IKZF1 in the IR group and the HR group (P<0.05). CONCLUSIONS: B-ALL children with non-functional subtypes of IKZF1 have a high recurrence rate, and the recurrence rate will be even higher in B-ALL children with non-functional subtypes of IKZF1 and MRD ≥10-4 on day 33 of chemotherapy.


Subject(s)
Ikaros Transcription Factor/genetics , Neoplasm, Residual/genetics , Neoplasm, Residual/mortality , Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics , Precursor Cell Lymphoblastic Leukemia-Lymphoma/mortality , Antineoplastic Combined Chemotherapy Protocols , Child , Child, Preschool , Female , Genotype , Humans , Male , Neoplasm, Residual/therapy , Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy , Prognosis , Recurrence , Remission Induction , Survival
5.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 25(1): 94-98, 2017 Feb.
Article in Chinese | MEDLINE | ID: mdl-28245382

ABSTRACT

OBJECTIVE: To investigate the effect of icaritin on the proliferation and apoptosis of THP-1 cells and its mechanism. METHODS: After treated with various concentrations of icaritin, cell proliferation was detected by MTS method, and apoptosis was measured with flow cytometry and Hoechst 33258 staining. Expression of BCL-2, BAX and Caspase-3 protein in THP-1 cell was detected by Western blot. RESULTS: After treatment with various concentrations (4-32 µmol/L) of icaritin for 24, 48, 72 h, the inhibition rate of cell growth significantly increased (P<0.05) in time-dose dependent manner(r=0.946); and the apoptotic rate of cells significantly increased (P<0.05) in time-dose dependent manner(r= 0.924). The expression of BCL-2 protein at 48 h decreased significantly in icaritin-treated group, compared with that in control group (P<0.05), while the expression of BAX and Caspase3 protein at 48 h increased significantly in icaritin-treated group, compared with that in control group (P<0.05). CONCLUSION: Icaritin can inhibit proliferation and induce apoptosis of THP-1 in vitro, Icaritin may induce apoptosis in THP-1 cells through the mitochondrial pathway.


Subject(s)
Apoptosis/drug effects , Flavonoids/pharmacology , Caspase 3 , Cell Line, Tumor , Cell Proliferation/drug effects , Humans , Proto-Oncogene Proteins c-bcl-2/drug effects , Proto-Oncogene Proteins c-bcl-2/metabolism , THP-1 Cells
6.
Zhongguo Dang Dai Er Ke Za Zhi ; 15(2): 91-4, 2013 Feb.
Article in Chinese | MEDLINE | ID: mdl-23428119

ABSTRACT

OBJECTIVE: To investigate the change in dendritic cells (DCs) in children with chronic immune thrombocytopenia (cITP) and the effect of glucocorticoid on DCs in children with cITP. METHODS: Fifteen children with cITP and 20 healthy controls were included in the study. Flow cytometry was used to measure the DC subsets count in the 15 children with cITP before and after glucocorticoid treatment as well as the corresponding values in the 20 healthy controls. The DCs derived from peripheral blood monocytes in children with cITP were cultured in vitro and collected, and their immunophenotypes were determined by flow cytometry. RESULTS: Before glucocorticoid treatment, the children with cITP showed no notable change in the absolute count of myeloid DCs (mDCs) but showed decreased absolute count of plasmacytoid DCs (pDCs) and increased mDC/pDC ratio compared with the healthy controls (P<0.05). After glucocorticoid treatment, the children with cITP demonstrated increased absolute count of pDCs and decreased absolute count of mDCs and mDC/pDC ratio compared with before treatment (P<0.05). Before glucocorticoid treatment, the children with cITP had significantly higher positive rates of HLA-DR, CD80, CD83 and CD86 on peripheral blood DCs than the healthy controls (P<0.01). All the positive rates were significantly decreased after glucocorticoid treatment (P<0.01), so that there was no significant difference from the healthy controls (P>0.05). CONCLUSIONS: Disproportion and functional disturbance of DC subsets is associated with the pathogenesis of cITP in children. Glucocorticoid can strengthen the immunosuppression of DCs in children with cITP, which may contribute to the effectiveness of glucocorticoid as a treatment.


Subject(s)
Dendritic Cells/drug effects , Glucocorticoids/pharmacology , Thrombocytopenia/immunology , Adolescent , Child , Child, Preschool , Chronic Disease , Dendritic Cells/immunology , Female , Humans , Immunophenotyping , Male , Thrombocytopenia/drug therapy
7.
PLoS One ; 6(8): e23720, 2011.
Article in English | MEDLINE | ID: mdl-21887305

ABSTRACT

PURPOSE: To explore the effects of Icaritin on chronic myeloid leukemia (CML) cells and underlying mechanisms. METHOD: CML cells were incubated with various concentration of Icaritin for 48 hours, the cell proliferation was analyzed by MTT and the apoptosis was assessed with Annexin V and Hoechst 33258 staining. Cell hemoglobinization was determined. Western blotting was used to evaluate the expressions of MAPK/ERK/JNK signal pathway and Jak-2/Phorpho-Stat3/Phorsph-Akt network-related protein. NOD-SCID nude mice were applied to demonstrate the anti-leukemia effect of Icaritin in vivo. RESULTS: Icaritin potently inhibited proliferation of K562 cells (IC50 was 8 µM) and primary CML cells (IC50 was 13.4 µM for CML-CP and 18 µM for CML-BC), induced CML cells apoptosis and promoted the erythroid differentiation of K562 cells with time-dependent manner. Furthermore, Icaritin was able to suppress the growth of primary CD34+ leukemia cells (CML) and Imatinib-resistant cells, and to induce apoptosis. In mouse leukemia model, Icaritin could prolong lifespan of NOD-SCID nude mice inoculated with K562 cells as effective as Imatinib without suppression of bone marrow. Icaritin could up-regulate phospho-JNK or phospho-C-Jun and down-regulate phospho-ERK, phospho-P-38, Jak-2, phospho-Stat3 and phospho-Akt expression with dose- or time-dependent manner. Icaritin had no influence both on c-Abl and phospho-c-Abl protein expression and mRNA levels of Bcr/Abl. CONCLUSION: Icaritin from Chinese herb medicine may be a potential anti-CML agent with low adverse effect. The mechanism of anti-leukemia for Icaritin is involved in the regulation of Bcr/Abl downstream signaling. Icaritin may be useful for an alternative therapeutic choice of Imatinib-resistant forms of CML.


Subject(s)
Antineoplastic Agents/pharmacology , Flavonoids/pharmacology , Janus Kinase 2/metabolism , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy , Mitogen-Activated Protein Kinase Kinases/metabolism , Signal Transduction/drug effects , Humans , Janus Kinases/metabolism , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/metabolism , Proto-Oncogene Proteins c-akt/metabolism , STAT3 Transcription Factor/metabolism , Tumor Cells, Cultured
8.
Zhonghua Xin Xue Guan Bing Za Zhi ; 37(8): 739-45, 2009 Aug.
Article in Chinese | MEDLINE | ID: mdl-20021931

ABSTRACT

OBJECTIVE: Coronary arterial plaque rupture and secondary thrombosis are the major pathogenesis of acute coronary syndrome (ACS). Metalloprotease (MMPs) secreted by monocyte/macrophage was the main predisposing factor of the plaque rupture and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is involved in a variety of inflammatory cytokine gene transcriptional regulations. We explored the possible role of PPAR-gamma in the regulation of MMP-9 and TIMP-1 expressed by peripheral monocyte-derived macrophages (MDMs) from patients with ACS. METHODS: Peripheral blood mononuclear cells were isolated from 48 patients with ACS and 28 healthy controls and stimulated by macrophage colony-stimulating factor (0.1 microg/ml for 24 hours) to form MDMs. MDMs were then incubated under various concentrations of rosiglitazone (0, 1, 10, 20 micromol/L) for 48 hours. The concentrations of MMP-9 and TIMP-1 in the supernatant were measured by enzyme linked immunosorbent assay, and the mRNA expression of PPAR-gamma, MMP-9 by RT-PCR and nuclear factor-kappaB P65 (NF-kappaB P65) expression by immunohistochemistry. RESULTS: PPAR-gamma mRNA expression was significantly lower while NF-kappaB P65 and MMP-9 expression as well as MMP-9 and TIMP-1 concentrations in supernatant were significantly higher in ACS group than those in control group (all P < 0.05). After rosiglitazone intervention, PPAR-gamma mRNA expression was significantly upregulated in both ACS and control groups in a dose-dependent manner. Both the MMP-9 concentration in the supernatant and MMP-9 mRNA expression were reduced post intervention with rosiglitazone in both groups. The TIMP-1 mRNA expression and concentration in supernatant were not affected by rosiglitazone in both groups. Rosiglitazone induced significant downregulation of NF-kappaB P65 expression in both groups. CONCLUSION: Rosiglitazone intervention may downregulate MMP-9 expression by upregulating PPAR-gamma expression, and by downregulating NF-kappaB expression in MDMs isolated from patients with ACS.


Subject(s)
Acute Coronary Syndrome/blood , Macrophages/metabolism , Matrix Metalloproteinase 9/metabolism , Thiazolidinediones/pharmacology , Tissue Inhibitor of Metalloproteinase-1/metabolism , Vasodilator Agents/pharmacology , Aged , Case-Control Studies , Cells, Cultured , Female , Humans , Male , Middle Aged , PPAR gamma/agonists , Rosiglitazone , Transcription Factor RelA/metabolism
9.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 16(3): 633-8, 2008 Jun.
Article in Chinese | MEDLINE | ID: mdl-18549644

ABSTRACT

This study was aimed to investigate the efficiency of 4 different culture media for in vitro culture and expanding adult human bone marrow mesenchymal stem cells (ahBM-MSCs) so as to establish a protocol of culturing and expanding hBM-MSCs and provide exprimental basis for hematopoietic blood stem cell transplantation combined with BM-MSCs. BM-MSCs were obtained from 16 fresh adult human bone marrow aspirate by gradient centrifugation with Ficoll Paque, then cultured in DMEM/F12 with 10% umbilical cord blood serum, 10% fetal calf serum (FCS), human blood serum, and MesenCult culture medium. The surface antigens of BM-MSCs were detected by flow cytometry. BM-MSCs were differentiated into osteoblasts and adipocytes under culture in the conditioned medium special for osteogenesis, and adipogenesis and the differentiated MSCs were identified by morphological observation, immunophenotype and immunohistochemical staining. The results showed that BM-MSCs could be isolated from adult human bone marrow and cultured by all culture media. The effect of umbilical cord blood serum on BM-MSC proliferation and their purity were similar to that of MesenCult culture medium, but better than that of FCS and human blood serum. The positive rate of CD29, CD73, CD105 on BM-MSCs cultured in umbilical cord serum and MesenCult medium was higher than that in FCS and adult human serum (p < 0.05), and the positive rate of CD31 was lower than that in FCS and adult human serum (p < 0.05). The positive rate of BM-MSCs differentiated into osteoblasts and adipocytes under culture in the conditioned medium for osteogenesis and adipogenesis with umbilical cord blood serum and MesenCult culture medium was also higher than that in FCS and adult human serum (p < 0.05). It is concluded that BM-MSCs can be obtained by all the four methods. DMEM/F12 with 10% umbilical cord blood serum and MesenCult culture medium are better than the others for the purification and differentiation potency of BM-MSCs in vitro. The medium with umbilical cord serum is valuable for clinical application in HSCT.


Subject(s)
Bone Marrow Cells/cytology , Cell Differentiation/physiology , Cell Proliferation , Culture Media , Mesenchymal Stem Cells/cytology , Adult , Cell Culture Techniques/methods , Cells, Cultured , Female , Fetal Blood , Humans , Male , Mesenchymal Stem Cell Transplantation , Middle Aged , Serum
10.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 16(2): 267-71, 2008 Apr.
Article in Chinese | MEDLINE | ID: mdl-18426646

ABSTRACT

This study was aimed to investigate the expression characteristics of HSP70 protein/mRNA, pim-1 mRNA in bone marrow mononuclear cells from leukemia patients, and to clarify whether these changes are related to leukemia type, tumor burden of leukemia, therapeutic reaction and prognosis. HSP70 mRNA and pim-1 mRNA in BMMNCs were detected with semi-quantitative RT-PCR in 40 leukemia patients and 10 controls. HSP70 protein in BMMNCs was assayed with Western blot in 34 leukemia patients and 10 controls. Relation of HSP70 and pim-1 expression with leukemia classification, the degree of tumor burden and therapeutic reaction were analyzed. The results showed that the BMMNCs from both leukemia patients and controls expressed HSP70 protein/mRNA. The mean ODR value of HSP70 mRNA in BMMNCs from leukemia patients was significantly higher than that of the controls; the mean ODR value of HSP70 protein/mRNA in acute myeloid leukemia and chronic myeloid leukemia patients both were significantly higher than that of acute lymphocytic leukemia patients; the mean ODR value of HSP70 protein/mRNA in acute leukemia patients with high-degree tumor burden was higher than that of the patients with low-degree tumor burden; the mean ODR value of HSP70 protein/mRNA in the patients after chemotherapy was significantly higher than that of the patients before chemotherapy; the BMMNCs from both leukemia patients and controls expressed pim-1 mRNA. The mean ODR value of pim-1 mRNA in BMMNCs from leukemia patients was significantly higher than that of the controls; the mean ODR value of pim-1 mRNA in BMMNCs for Acute lymphocytic leukemia patients was significantly higher than that of the patients suffered from acute myeloid leukemia and chronic granulocytic leukemia; there was a positive relationship between pim-1 mRNA and HSP70 mRNA expressions in leukemia patients (p < 0.05). It is concluded that there are high expressions of HSP70 and pim-1 in leukemia and their positive correlation is shown. The over-expressions of HSP70 and pim-1 protein/mRNA are related to tumor burden in leukemia patients.


Subject(s)
Bone Marrow Cells/cytology , HSP70 Heat-Shock Proteins/metabolism , Leukemia/genetics , Leukocytes, Mononuclear/metabolism , Proto-Oncogene Proteins c-pim-1/metabolism , Adolescent , Adult , Aged , Female , HSP70 Heat-Shock Proteins/genetics , Humans , Leukemia/metabolism , Male , Middle Aged , Prognosis , Proto-Oncogene Proteins c-pim-1/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Young Adult
11.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 29(6): 662-6, 2004 Dec.
Article in Chinese | MEDLINE | ID: mdl-16114552

ABSTRACT

OBJECTIVE: To investigate the characteristics of immunophenotype of chronic lymphocytic leukemia (CLL) and to evaluate the prognosis of CLL by using the indexs of the characteristics of immunophenotype combined with serum LDH and beta2-MG levels. METHODS: The immunophenotype of 15 CLL, 12 AML, 11 ALL, and 1 FCL (follicular cell lymphoma) patients were analyzed by the indirect immunofluorescent assay. The serum LDH and beta2-MG were detected in 13 CLL patients by the rate nephelometry. RESULTS: The mean age of episode for CLL was 62.9 years and the ratio of male to female was 4:1 in the CLL group. The immune markers (CD5, CD19, and CD20) were positive in the 15 CLL, and negative in the 11 ALL and the 12 AML. The expression of CD23 was positive in 12 of the 15 CLL and negative in the 11 ALL and 12 AML. The expression of CD38 was positive in the 15 CLL. The immunophenotype of 1 FCL was CD10+, CD19+, CD20+, CD5- and CD23-. The serum beta2-MG and LDH levels in 9 CLL patients at Stage Binet C were significantly higher than those in 3 CLL patients at Stage Binet A (P < 0.05). CONCLUSION: CD19+, CD20+ and CD5+ are specific assembly of immunophenotype in B-CLL and contribute to the diagnosis of CLL. The clinical stage of CLL patients of CD23- belongs to Stage Binet C, indicating the poor prognosis. CD23 contributes to differentiate CLL from FCL. CD38+ as a prognostic marker of CLL is not specific. The levels of LDH and beta2-MG are useful indexes for evaluating the prognosis of CLL.


Subject(s)
L-Lactate Dehydrogenase/blood , Leukemia, Lymphocytic, Chronic, B-Cell/immunology , Macroglobulins/metabolism , Aged , Antigens, CD19/blood , Antigens, CD20/blood , CD5 Antigens/blood , Female , Humans , Immunophenotyping , Male , Middle Aged , Prognosis
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