Animals ; Humans ; Iron ; adverse effects ; metabolism ; Iron Overload

Hemoglobinuria, Paroxysmal ; metabolism ; Humans ; Membrane Proteins ; biosynthesis

OBJECTIVETo explore the pathogenetic mechanism of cytopenia in the patients with systemic lupus erythematosus (SLE).

METHODSTwenty-one patients with SLE and 10 healthy controls were enrolled in this study. Bone marrow mononuclear cells (BMMNC) Coombs test was used to determine the autoantibodies and in vitro CFU-E, BFU-E and CFU-GM yields were used to determine the proliferation function of hematopoietic stem/progenitor cell (HSC).

RESULTSPositive results of BMMNC-Coombs test were observed in 12 patients with SLE (57.1%), among them, 10 with (58.2%) and 2 without blood cytopenia. The yields of CFU-GM (49 +/- 40), and CFU-E (49 +/- 35) in SLE patients without cytopenia were higher than that in normal controls, but lower than that in cytopenia SLE patients. The BFU-E yield in SLE patients (3 +/- 4) was significantly lower than that in normal controls (36 +/- 12).

CONCLUSIONThere were auto-antibodies on BMMNC in patients with SLE. The blood cytopenia in SLE patients maybe resulted from the destructions of bone marrow hematopoietic cells by the autoantibodies. The capacities of bone marrow HSC in SLE patients for proliferation compensation of hematopoietic cell destruction were not damaged.

Adolescent ; Adult ; Autoantibodies ; blood ; Bone Marrow ; immunology ; pathology ; Bone Marrow Cells ; immunology ; pathology ; Case-Control Studies ; Female ; Humans ; Lupus Erythematosus, Systemic ; immunology ; pathology ; Male ; Middle Aged ; Young Adult

Bone Diseases ; pathology ; therapy ; Humans ; Multiple Myeloma ; pathology ; therapy

Objective: To explore the expression of SLAMF6 on CD8(+) T cells in patients with severe aplastic anemia (SAA) and its correlation with disease immune status. Methods: By flow cytometry (FCM), SLAMF6 expression level in peripheral blood CD8(+) T cells was detected in 21 patients with SAA and 15 normal controls respectively from February 2017 to April 2018. The correlation between SLAMF6 expression level and hematopoietic functions, including HGB, PLT, the neutrophil granulocyte and reticulocyte absolute value in peripheral blood, hyperplasia degree (percentage of granulocytes, erythrocytes, lymphocytes and megakaryocytes in bone marrow) and perforin, granzyme B, IFN-γ expression level in CD8(+) T cells were evaluated. To further confirm the effect of SLAMF6 on CD8(+) T cells, anti-SLAMF6 Ab was used to block SLAMF6 pathway (IgG as control), and FCM was used to detect the perforin, granzyme B, and IFN-γ production of CD8(+) T cells. Results: The expression of SLAMF6 on CD8(+) T cells in untreated SAA patients[(56.29±12.97)%]was significantly lower than that of normal controls[(80.96±7.36)%](t=-7.672, P<0.001). The expression of SLAMF6 on CD8(+) T cells in SAA patients were positively correlated with the HGB, PLT, the neutrophil granulocyte and reticulocyte absolute value in peripheral blood, percentage of granulocytes, erythrocytes in bone marrow (all P<0.05), but they were negatively correlated with the percentage of lymphocytes in bone marrow, and the expression of perforin, granzyme B, and IFN-γ of CD8(+) T cells (all P<0.05). After blocking SLAMF6 pathway by anti-SLAMF6 Ab, the expression levels of perforin, granzyme B and IFN-γ in SAA patients were significantly higher than those in the untreated group, and the differences were statistically significant (all P<0.05). Conclusions: SLAMF6 is significantly down-regulated on CD8(+) T cells in SAA patients, which may act as a negative immunoregulatory molecule participating in the mechanism of SAA by affecting the functional molecules secretion on CD8(+) T cells.
Anemia, Aplastic ; Bone Marrow ; CD8-Positive T-Lymphocytes ; Flow Cytometry ; Humans ; Perforin ; Signaling Lymphocytic Activation Molecule Family

OBJECTIVETo explore the mechanism of 'erythroblast island (EI)' formation in the bone marrow of patients with immun-related hemocytopenia (IRP).

METHODSThe category of BM-auto antibody (au Ab) in 48 patients with IRP was detected with FCM. The BM-au Ab in the 'EI' of these cases were explored with immuonhistofluorescence (IF). Clinical and laboratory characteristics of these cases were also analyzed retrospectively.

RESULTSIgG could be detected in the 'EI' on the BM smear of 14 cases (29.17%), BM-au Ab mainly deposited at the edge/membranes between macrophage and erythroblasts rather than cyto plasm. Positive reaction were seen in all the cases with GlycoAIgG. The red blood cell count [(1.8 ± 0.5) × 10(12)/L] and hemoglobin level [(59.6 ± 16.2)g/L] were significantly lower than that in the IF(-) group [(2.5 ± 0.9) × 10(12)/L and (83.4 ± 25.0) g/L] (P < 0.05). The percentage of reticulocyte [(2.0 ± 0.8)%], serum level of IBIL [(9.4 ± 4.7) µmol/L], percentage of erythroblats in sternum BM (0.441 ± 0.139) and response rate to therapy (85.7%) in IF(+) group were significantly higher than that in IF(-)group [(1.3 ± 1.0)%, (6.6 ± 6.7)µmol/L, 0.298 ± 0.082, 61.3%, respectively] (P < 0.05).

CONCLUSIONMacrophage was connected with erythroblasts through autologous IgG in the 'EI's of some patients with IRP. 'EI' were the places where macrophages devoured and destroyed erythroblasts rather than erythroid development and differentiation. The pathogenetic mechanism of IRP might be associated with macrophages phagocytosing and destroying BM hematopoietic cells.

Blood Cell Count ; Bone Marrow ; Bone Marrow Cells ; immunology ; Coombs Test ; Erythroblasts ; Humans

OBJECTIVETo investigate the variation of bone marrow complement level in cytopenia patients with positive BMMNC-Coombs test (CBCPC), and probe the role of complement in destroying hematopoietic cells of CBCPC patients.

METHODSOne hundred and twenty-four patients with CBCPC and twenty-three healthy donors as controls were enrolled in this study. The levels of CH50, C3, C4, C5b-9 were tested with ELISA. The auto-antibodies on bone marrow hematopoietic cells (BMHC) were examined with flow cytometry.

RESULTSThe level of C5b-9 in bone marrow (BM) of untreated CBCPC patients [(119.8+/-54.0) microg/L] was significantly higher than that of recovered patients [(100.7+/-33.4) microg/L] or normal controls [(93.9+/-28.8) microg/L] (P<0.05). The levels of CH50 in BM of untreated or recovered CBCPC patients [(33.3+/-11.5) kU/L, (30.8+/-10.3) kU/L] were significantly higher than that of normal controls [(24.1+/-6.4) kU/L] (P<0.05). The level of C3 in BM of untreated or recovered CBCPC patients [(4.9+/-2.2) mg/L], (5.0+/-3.5) mg/L] was significantly lower than that of normal controls [(7.0+/-5.6) mg/L] (P<0.05). The level of complement in peripheral blood was consistent with that in BM. CH50 in BM of CBCPC patients was negatively correlated with their C3 (r=-0.303, P=.0007) and positively correlated with their C5b-9 (r=0.241, P=0.003) levels. The level of C5b-9 in BM of CBCPC patients was higher in the BMHC-IgM positive group [(117.6+/-55.7) microg/L] than in the BMHC- IgM negative group [(99.2+/-26.2) microg/L] (P<0.05). The positive rate of CD34(+)-IgG or CD34(+)-IgM of CBCPC patients was positively correlated with their C5b-9 level (r=0.593, P=0.000, r=0.326, P=0.049). The reticulocyte percentage (r=0.421, P=0.000) and serum indirect bilirubin level (r=0.230, P=0.032) of CBCPC patients were positively correlated with their CH50 level.

CONCLUSIONSThe hematocytopenia of CBCPC patients might be related to the hematopoietic cells destruction caused by auto-antibody activated complements.

Adolescent ; Adult ; Aged ; Bone Marrow Cells ; immunology ; Case-Control Studies ; Child ; Complement System Proteins ; metabolism ; Coombs Test ; Female ; Humans ; Male ; Middle Aged ; Pancytopenia ; immunology ; metabolism ; Young Adult

OBJECTIVETo measure the quantities and apoptosis-related protein levels of B lymphocyte in the patients with immunorelated pancytopenia (IRP) and explore the action of B lymphocyte in the pathogenic mechanism of IRP.

METHODSQuantities of whole B lymphocytes and CD5+ B lymphocytes as well as the expressions of Fas and Bcl-2 in B lymphocytes in 35 patients with untreated IRP, 15 IRP patients in complete remission (CR), and 10 normal controls were assayed by flow cytometry.

RESULTSThe percentages of B lymphocyte and CD5+ B lymphocyte were significantly higher in untreated IRP patients than in CR IRP patients and normal controls (P < 0.05), and there was no significant difference between the latter two groups (P > 0.05). There was no significant difference of Fas expression in B lymphocyte among three groups (P > 0.05). The expression of Bcl-2 in B lymphocyte was significantly higher in untreated patients than in CR patients or normal controls (P < 0.01), and significantly higher in CR patients than in normal controls (P < 0.01). The apoptosis-related index was significantly lower in untreated patients than in CR patients or normal controls (P < 0.05), and significantly lower in CR patients than in normal controls (P < 0.05). The percentage of B lymphocyte was positively correlated with post-treated response time (r = 0.53, P < 0.01).

CONCLUSIONThe production of auto-antibodies in IRP patients probably has some relationship with the abnormal quantities of B lymphocyte and its subpopulations as well as with the inhibition of B lymphocyte apoptosis.

Adolescent ; Adult ; B-Lymphocytes ; cytology ; immunology ; Case-Control Studies ; Child ; Female ; Humans ; Lymphocyte Subsets ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology

Objective: To investigate whether insulin stimulates the translocation of glucose transporter-4 (GLUT4) and glucose uptak e in ischemic myocardium. Methods: Plasma concentration of gluc ose, lactate, free fatty acid and insulin were determined by autoanalyser, and G LUT4 was studied by Western blotting analysis. Results: Insulin increased GLUT4 significantly in sarcolemma of ischemic myocardium ［(25±4)% vs (40±6)%］, and GLUT4 content in intracellular membrane decreased proporti onally. The glucose uptake increased significantly in insulin-ischemic myocardi um. The uptake of insulin-ischemic myocardium was almost 2 times that of ischem ic myocardium. Conclusion: Insulin stimulation results in GLUT4 translocation and increases glucose uptake in ischemic myocardium. When myocardi al ischemia occurs, insulin is helpful in increasing myocardial glucose uptake a nd utilization.

Objective: To investigate whether there is additi ve effects of hyperinsulinemia and ischemia on expression of canine myocardial G LUT4 gene in vivo. Methods: The expression of myocardial GLU T4 was determined by semiquantitative immunoblotting.The expression of GLUT4 mRN A was determined by semiquantitative Northern blotting. Results: Dramatic changes were seen in GLUT4 mRNA and GLUT4 expression in the ischemic hearts.After infusing insulin for 8 h,regional GLUT4 mRNA and GLUT4 levels in is chemic hearts were 2.5, 2.3-fold that of expression in normal hearts(P<0.01 ). Myocardial glucose uptake in ischemic hearts was increased by 4-fold when co mpared with normal hearts(P<0.01). Conclusion: There are not only additive effects of hyperinsulinemia and low-flow ischemia on canine myoc ardial GLUT4 mRNA and GLUT4 expression in vivo, but also increase of myocar dial glucose uptake. Enhanced GLUT4 expression may be an important protective m echanism by which myocardial cells enhance glucose uptake and metabolism during low-flow ischemia.