This study was purposed to investigate the expressions of miR-21, miR-155 and miR-210 in plasma of patients with lymphoma, and explore their role played in diagnosis, evaluation of chemotherapy effect and prognosis of lymphoma. The expressions of miR-21, miR-155 and miR-210 were assayed by RT-PCR in plasma of 54 cases of lymphoma, 10 cases of lymphonode inflammation and 27 cases of normal controls. The results indicated that the expressions of miR-21, miR-155 and miR-210 in plasma of lymphoma patients were higher than those of control group and lymphonode inflammation group (P < 0.05). The expressions of miR-21 and miR-210 in plasma of control group and lymphonode inflammation group had no significant differences (P > 0.05). The expression of miR-21 in plasma of lymphoma patient group significantly correlated with their serum LDH level. The expressions of miR-21 and miR-210 in plasma of previously untreated lymphoma patient group were higher than those of the patients treated for 6 or more courses (P < 0.05). The diagnostic accuracy of miR-21, miR-155 and miR-210 used for lymphoma patients was 56, 65, 48 respectively, and reached to 83 when combined three of them. It is concluded that the expressions of miR-21, miR-155 and miR-210 in plasma of lymphoma patients were significantly higher. Detection of these 3 miRNA in plasma of patients can contribute to the clinical diagnosis, treatment and prognosis evaluation of lymphoma.
Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Humans ; Lymphoma ; blood ; diagnosis ; Male ; MicroRNAs ; blood ; Middle Aged ; Plasma ; metabolism ; Prognosis ; Young Adult

This study was aimed to investigate the expression level and mechanism of microRNA-223 and LMO2 in acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL) cells and the mechanism. MicroRNA-223 mimics was transfected to increase the expression of MicroRNA-223 in the lymphocytes sorted by ficoll separation from the bone marrow mononuclear cells (BMMNC) of ALL and CLL patients. MicroRNA-223 inhibitor was transfected to decrease the expression of the MicroRNA-223 in the lymphocytes of normal controls. Then the expression of the MicroRNA-223 and LMO2 in transfected lymphocytes before and after cultivating for 72 hours were detected by RT-PCR, the apoptosis and cell cycle of these cells were measured by flow cytometery. The results indicated that before the transfection, the expression of MicroRNA-223 in ALL and CLL cells was (433.11 ± 144.88), which was significantly lower than that in the normal lymphocyte (949.59 ± 267.39); the expression of LMO2 was (807.10 ± 238.41), which was significantly higher than that in the normal lymphocytes (455.32 ± 176.83) (P < 0.05); after the transfection, the expression of MicroRNA-223 was (571.86 ± 142.00) in ALL and CLL cells, which was significantly higher than that before transfection (P < 0.05), but the expression of LMO2 was significantly lower than that before transfection (651.97 ± 230.12) (P < 0.05); in the normal control the expression of MicroRNA-223 obviously decreased (646.32 ± 172.93) (P < 0.05), the expression of LMO2 was significantly increased (541.27 ± 158.86.2) (P < 0.05). After transfection, the cell cycle G1/G2 phase and apoptosis changed in ALL and CLL cells. Before transfection the cell ratio in cell cycle G1/G2 phase was (94.75 ± 3.15)%, the cell ratio in S phase was (5.14 ± 3.12)%; after transfection the cell ratio in cell cycle G1/G2 phase was (97.03 ± 2.08)% and obviously increased (P < 0.05), the cell ratio in S phase was (2.97 ± 2.08)% and significantly decreased (P < 0.05). Before transfection the apoptosis rate was (54.47 ± 8.72)%, and obviously was higher than that after transfection (60.48 ± 8.81)%. And in the normal control, the cell ratio in G1/G2 phase was significantly higher than that after transfection [(96.73 ± 2.26)%, (94.55 ± 2.77)%, P < 0.05)], and the cell ratio in S phase was significantly increased [(3.25 ± 2.26)%, (5.45 ± 2.77)% (P < 0.05)]. The apoptotic rate in the ALL and CLL patients was significantly higher than that after the transfection [(54.47 ± 8.72)% vs (60.48 ± 8.81)%, respectively (P < 0.05)]. The apoptotic rate in the normal control was significantly lower than that after the transfection [(59.02 ± 10.20)%, (51.96 ± 10.20)%, respectively (P < 0.05)]. It is concluded that the expression of MicroRNA-223 decreases, and the expression of LMO2 increases in lymphocytic leukemia cells which leads to the lymphocytes over-proliferation and abnormal apoptosis, thus may be one of pathogenesis in lymphocytic leukemia.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Adolescent ; Adult ; Aged ; Apoptosis ; Case-Control Studies ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Female ; Humans ; LIM Domain Proteins ; genetics ; metabolism ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; metabolism ; Male ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; Transfection ; Young Adult

Adult ; Aged ; Aged, 80 and over ; Bone Marrow Cells ; physiology ; Female ; Humans ; Immune Evasion ; Male ; Middle Aged ; Myelodysplastic Syndromes ; etiology ; immunology

OBJECTIVETo investigate the function of dendritic cells (DC) of patients with immune related pancytopenia (IRP) and explore the role of DC in IRP.

METHODSThe expression of CD80 and CD86 on myeloid DC (mDC, Lin-HLA-DR(+) CD11c(+) cells) and plasmacytoid DC (pDC, Lin-HLA-DR(+) CD123(+) cells) of 65 IRP (37 untreated and 28 remitted) patients and 17 healthy controls were analyzed by flow cytometry.

RESULTSThe expression of CD86 on pDC was (82.47 ± 13.17)% in untreated group and (60.08 ± 14.29)% in remission group, which were significantly higher than that of controls (47.95 ± 18.59)% (P < 0.05), while the expression in untreated group was higher than that of remission group (P < 0.05). The expression of CD80 on pDC was (6.31 ± 4.49)% in untreated group, which was significantly higher than that of remitted patients (3.09 ± 2.93)% and controls (2.33 ± 2.25)% (P < 0.05). The expression of CD86 on mDC was (97.06 ± 4.82)% in untreated group and (91.35 ± 12.20)% in control group, while the expression in untreated group was higher than that of control group (P < 0.05). The expression of CD80 on mDC was (6.20 ± 5.44)% in untreated group and (3.97 ± 3.24)% in remission group, which were significantly higher than that of controls (1.86 ± 1.73)% (P < 0.05). The expression of CD86 on pDC was negatively correlated to Th1/Th2 (r = -0.733, P < 0.05), it was positively correlated to the antibody on membrane of BMMNC (r = 0.283, P < 0.05) and the quantity of CD5(+)B cells (r = 0.436, P < 0.05), while it was negatively correlated to the level of hemoglobin, platelets and white blood cells (r = -0.539, P < 0.05; r = -0.519, P < 0.05; r = -0.567, P < 0.05, respectively). The expression of CD80 on pDC was negatively correlated to the level of hemoglobin and platelets (r = -0.431, P < 0.05; r = -0.464, P < 0.05).

CONCLUSIONThe function of pDC in PB of IRP were strengthened, which was relevant to the immunopathogenesis of IRP.

Adolescent ; Adult ; Autoimmune Diseases ; complications ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Dendritic Cells ; metabolism ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Pancytopenia ; blood ; etiology ; pathology ; Young Adult

OBJECTIVETo observe the efficacy and safety of linezolid for the treatment of gram positive coccus infections in hematological disease patients.

METHODSOne hundred and two hematological disease patients with suspected or proven gram positive coccus bacteria infection were enrolled in this study. Linezolid was given at a dosage of 600 mg, iv, q12h. The mean treatment period was (10.82 ± 5.12) days (1 to 51 days) with 74.5% over 7 d and 51.0% over 10 d.

RESULTSAmong 102 patients, 57 were male, 45 female aged 11 to 81 years, with a mean of (45.26 ± 19.15) years. Ninety four cases were nosocomial infection (92.2%) and 8 community infection (7.8%); There were pneumonia in 80 (78.4%), septicemia in 11 (10.8%), and infection of other organsin 11 (10.8%); Forty five cases were proven gram positive coccus bacteria infection, and 57 were suspected infection; Fifty one bacteria strains were isolated from cultivated samples of proven patients, in which 22 were staphylococcus aureus with 19 methicillin resistant 13 hemolytic streptococcus, 9 staphylococcus epidermidis with 7 methicillin resistant 6 enterococcus faecom, and 1 enterococcus hirae. Seven cases were mixed with one kind gram negative bacillus infection, 4 mixed with two kinds of gram negative bacillus infection, and 12 mixed with fungal infection; Total clinical response rates by ITT (intention to treatment) analysis was 69.6%, in which 40 (39.2%) were curative and 31 (30.4%) obviously effective; PP (per-protocol) analysis was 70.9%, in which 39 (41.9%) were curative and 27 (29.0%) obviously effective. Bacteria clearance rate was 70.6%, and in this group the clinical effective rate was 88.9%; Adverse effect rate was 2.9%, being transient thrombocytopenia and increased transaminase.

CONCLUSIONLinezolid is a safe and effective antibiotic used in hematological disease patients complicated with infections of gram positive coccus.

Acetamides ; Anti-Bacterial Agents ; therapeutic use ; Cross Infection ; microbiology ; Gram-Positive Bacterial Infections ; drug therapy ; Gram-Positive Cocci ; Hematologic Diseases ; drug therapy ; Humans ; Linezolid ; Oxazolidinones ; Staphylococcus aureus

In order to analyze the prognosis and related factors of acute lymphoblastic leukemia (ALL), 53 newly diagnosed ALL patients were enrolled in this study. The therapeutic efficacy and prognosis of 53 cases of ALL were analyzed, the remission, relapse, overall survival and event-free survival were studied, and relation between different factors and prognosis of ALL were investigated by comparison of cases in same stage. The results showed that the complete remission was achieved in 36 out of 53 patients, the total remission rate was 67.9%, the total relapse rate was 37.7%, the median relapse duration was 6 months after remission. Median overall survival (OS) and median event-free survival (EFS) time were 4 and 1 months after remission respectively, OS and EFS rate of 18 month was 35.1% and 14.2%. The patients with different gender had significantly different EFS. Age was an independent risk factor of CR rate. White blood cell count and hemoglobin level of newly diagnosed patients were significantly correlated with OS and EFS. Absolute neutrophil count (ANC) at the end of the induction chemotherapy was an independent related factor of OS, the higher ANC, the lower risk of death. The patients with or without chemotherapy related infection had different relapse rate. The patients with bleeding after chemotherapy had lower OS when compared with those without bleeding. Serum glucose level was a significant negative prognostic factor. It is concluded that there is higher relapse rate, poor prognosis in adult ALL in comparison with children. In order to decrease the relapse rate and prolong the EFS, individual therapeutical regimens and prophylaxis of complicating diseases should be applied to ALL patients.
Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; therapy ; Prognosis ; Young Adult

OBJECTIVETo investigate the mechanisms underlying bone marrow damage by iron overload in pancytopenic patients with positive BMMNC-Coombs test (IRP).

METHODSTwenty-one iron overloading, 26 non-iron overloading IRP patients and 10 normal controls were enrolled in this study. The expressions of ROS, Bcl-2, Caspase-3 and apoptosis of BMMNC were analyzed by flow cytometry (FCM). Antioxidants were added to iron overloading IRP BMMNC, and then the changes of indices above were detected by FCM. The number and apoptosis of T lymphocytes of IRP patients were also detected.

RESULTSROS and apoptosis of BMMNC, myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones and normal controls (P < 0.05). The expressions of Bcl-2 on BMMNC, erythrocytes and stem cells of iron overloading IRP patients were significantly lower than those of non-iron overloading IRP ones (P < 0.05). The levels of Caspase-3 on myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones and normal controls (P < 0.05). After treatment with antioxidants, the expressions of ROS, Caspase-3 and apoptosis of iron overloading IRP BMMNC significantly decreased, but opposite for Bcl-2. The percentages of CD4(+) lymphocytes [ ( 40.86 ± 8.74）%] and CD4(+)/CD8(+) (1.44 ± 0.36) in PB of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones [(35.96 ± 7.03)% and 1.14 ± 0.37] and normal controls [(28.00 ± 6.73)% and 0.79 ± 0.21], respectively (P < 0.05), as opposite for CD8(+) lymphocytes (P < 0.05). The apoptosis of CD8(+) lymphocytes [(27.35 ± 10.76)%] and the ratio of CD8(+) apoptosis/CD4(+) apoptosis (2.51 ± 0.81) in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones [(15.47 ± 8.99)%] and normal controls (1.39 ± 0.47), respectively (P < 0.05). The apoptosis of erythrocytes and stem cells coated with auto-antibodies in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP and normal controls.

CONCLUSIONMechanisms underlying bone marrow damage by iron overload might be through the follows: ①The increased ROS induced by excessive iron deposition affected the expressions of Caspase-3 and Bcl-2, which caused more BMMNC apoptosis; ②The abnormal number and ratio of T lymphocytes caused by iron overload aggravated the abnormality of immunity of IRP; ③Iron overload may increase the damage to erythrocytes and stem cells coated with auto-antibodies.

Adolescent ; Adult ; Aged ; Bone Marrow ; pathology ; Case-Control Studies ; Caspase 3 ; metabolism ; Coombs Test ; Female ; Humans ; Iron Overload ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology ; physiopathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reactive Oxygen Species ; metabolism ; Young Adult

This study was purposed to investigate the immune state of T cells, the quantity and function of GPI(+) T cells and GPI(-) T cells in patients with paroxysmal nocturnal hemoglobinuria (PNH). 22 cases of PNH and 18 normal controls were enrolled in this study. Their T lymphocyte subsets, Th lymphocyte subsets were assayed by flow cytometry with the monoclonal antibodies concerned. The proportion of GPI(+) T cells or GPI(-) T cells in CD3(+) T cells, CD4(+) T cells, CD8(+) T cells and the expressions of CD69 on these T cells were also respectively assayed. The results showed that the proportion of CD4(+) T cells in CD3(+) T cells in PNH [(47.7670 +/- 13.91139)%] was lower than that in controls [(54.9592 +/- 7.11678)%] (p < 0.05). CD8(+) T cells in CD3(+) T cells of PNH cases [(52.2767 +/- 13.90395)%] were higher than that of controls [(45.2418 +/- 6.75306)%] (p < 0.05). The ratio of CD4(+) T cells to CD8(+) T cells was reverse in PNH. Those were more significantly in PNH-AA (0.77763 +/- 0.409153) (p < 0.05). The proportion of Th1 cells in PNH [(16.9136 +/- 6.78899)%], especially in PNH-AA [(22.8000 +/- 5.45244)%], was significantly higher than that in controls [(4.4600 +/- 1.81879)%] (p < 0.05). The proportion of Th2 cells in PNH [(4.7582 +/- 1.98441)%] had no difference from controls [(3.7960 +/- 1.13810)%]. The number of GPI(-) T cells in CD8(+) T cells and CD4(+) T cells were (14.6797 +/- 11.96718)% and (3.9241 +/- 2.46263)% respectively. The expression of CD69 on GPI(+) T cells or GPI(-) T cells in PNH [CD8(+) GPI(+) T cells (17.67881 +/- 8.562493)%, CD8(+) GPI(-) T cells (15.86575 +/- 7.279743)%, CD4(+) GPI(+) T cells (4.65431 +/- 1.984378)%, CD4(+) GPI(-) T cells (4.93181 +/- 1.730001)%]was significantly higher than that in normal controls [CD8(+) GPI(+) T cells (4.68038 +/- 1.216645)%, CD4(+) GPI(-) T cells (1.77339 +/- 0.645259)%] (p < 0.05), but the expression of CD69 on GPI(+) T cells was not different from that on GPI(-) T cells in PNH. It is concluded that high function of cytoimmunity in PNH may be responsible for bone marrow failure but not relates to the existence of PNH clone in T cell population.
Adolescent ; Adult ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Hemoglobinuria, Paroxysmal ; immunology ; Humans ; Immunophenotyping ; Lymphocyte Count ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology ; Young Adult

This study was aimed to explore whether the perforin gene 1 (PRF1) mutation is the basis of genetic susceptibility to pathogenesis of acquired severe aplastic anemia (SAA). DNA exon2 and exon3 of PRF1 gene in peripheral blood mononuclear cells in 31 SAA patients and 15 normal controls were amplified by PCR; the sequencing was performed by using ABI pRISM 373OXL sequencer; the mutation loci were sought through checking sequences with GenBank-reported sequences; after the mutation sequences were found, those were cloned into M13 phage vector, then the corresponding sequences of gained 2 chromosomes were sequenced respectively to determine the distribution of different mutations on chromosomes. The results showed that (1) one homozygous mutation (822 C > T, synonymous mutation) and one heterozygous mutation (907 G > A, methionine 303 valine) were found in PRF1 coding region of 2 SAA patients. These mutations were not detected in normal controls. (2) 1 SNP (rs885822) in the coding region was detected in SAA patients and controls, and the heterozygosity rate between the 2 groups was different (p < 0.05). It is concluded that perforin gene mutation may be one risk factor in the aberrant proliferation and activation of cytotoxic T cells in pathogenesis of a part of patients with aplastic anemia.
Adolescent ; Adult ; Aged ; Anemia, Aplastic ; genetics ; Base Sequence ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Genetic Predisposition to Disease ; Heterozygote ; Humans ; Male ; Middle Aged ; Mutation ; Perforin ; Pore Forming Cytotoxic Proteins ; genetics ; Young Adult

This study was purposed to investigate the BCL-2/IgH gene rearrangement in major break point region (MBR) and IgH gene rearrangements of patients with non-Hodgkin's lymphoma (NHL), and explore their significance for improving early diagnosis and accurately evaluating chemotherapy effect. DNA for BCL-2/IgH and IgH gene assays was extracted from bone marrow mononuclear cells in 70 cases of lymphoma (60 cases of B-NHL and 10 cases of T-NHL), 7 cases of lymph node inflammatory and 20 healthy controls. The BCL-2/IgH, IgH gene rearrangements were assayed by polymerase chain reaction (PCR), the assayed results were compared with results of pathological biopsy; the factors related with occurrence of these 2 kinds of gene rearrangement were analyzed and the dynamic changes of BCL-2/IgH and IgH gene rearrangements after chemotherapy were compared, the chemotherapy effect was evaluated. The results indicated that (1) BCL-2/IgH gene rearrangement in bone marrow mononuclear cells was observed in 10 cases out of 30 DLBCL cases (33.3%), and was more frequent than that in 30 other B-NHL cases (6.7%), 10 T-NHL cases (0%), 7 lymph nodes inflammatory cases (0%) and 20 healthy controls (5%) (p < 0.05). (2) the quantity of rearranged BCL-2/IgH gene of 8 DLBCL cases reduced from 0.59 to 0.16 (p < 0.05) after 2 courses of R-CHOP chemotherapy and completely disappeared after 6 courses of R-CHOP chemotherapy. (3) 81.8% patients with BCL-2/IgH gene rearrangement showed high serum LDH level, while it was observed in 28.6% patients without this gene rearrangement (p < 0.05). Lymphoma staging, systemic symptoms, β(2)-MG level, bone marrow involvement, infiltration of liver and spleen were not significantly correlated with BCL-2/IgH gene rearrangement. (4) IgH gene rearrangement was found in 9 cases out of 20 DLBCL patients (all newly diagnosed patients) (45%), IgH rearrangement was observed in 14 cases out of 30 other B-NHL (all newly diagnosed or relapsed patients, except patients with DLBCL) (46.7%) and there was no statistical difference between these 2 groups, however IgH rearrangement all were not observed in 20 healthy persons, 10 T-NHL cases and 7 lymph nodes inflammatory cases. (5) the quantity of rearranged IgH gene in 7 DLBCL cases was reduced from 0.42 to 0.13 after one course of R-CHOP chemotherapy (p < 0.05) and completely disappeared after 2 courses of R-CHOP chemotherapy. (6) 90% patients with IgH gene rearrangement had high serum LDH level, while it was found in 30% patients without this gene rearrangement (p < 0.05). Lymphoma staging, systemic symptoms, β(2)-MG levels, bone marrow involvement, infiltrations liver and spleen all were not significantly correlated with IgH gene rearrangement. It is concluded that the BCL-2/IgH and IgH gene rearrangements may be used as specific indicators in early diagnosis and accurate evaluation of therapy efficacy in B-NHL, these 2 kind of rearrangement correlate with LDH level. The BCL-2/IgH gene rearrangement is more specific for in DLBCL.
Adult ; Aged ; Aged, 80 and over ; Bone Marrow Cells ; Case-Control Studies ; Female ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; genetics ; Humans ; Lymphoma, Non-Hodgkin ; blood ; genetics ; Male ; Middle Aged ; Young Adult