The purpose of this study was to evaluate the safety of cryopreserved and thawed peripheral blood stem cell (PBSC) fractionated return infusions in children. 35 children patients with malignant tumors (13 acute leukaemias, 15 neuroblastomas and 7 malignant lymphomas) received fractionated return infusions of cryopreserved stem cells after undergoing high-dose chemotherapy without or with total body irradiation. The toxicities of 70 return infusions were evaluated. All patients were mobilized by chemotherapy plus recombination human granulocyte colony-stimulating factor (rhG-CSF), and then PBSCs were collected by a separator CS-3000 plus or COBE spectra-4. The grafts were cryopreserved in 10% dimethyl sulfoxide (DMSD) and stored in liquid nitrogen. There were totally 70 PBSC transfusions. The total volume of PBSCs transfused: 190 - 420 ml (265 +/- 73 ml or 13.7 +/- 4.2 ml/kg) with a mean of (4.43 +/- 1.91) x 10(8)/kg of PBSCs, and 0.94 +/- 0.18 g/kg of DMSO. The single dose: 90 - 300 ml (132 +/- 37 ml or 6.6 +/- 5.2 ml/kg) with a mean of 0.68 +/- 0.12 g/kg of DMSO. Symptoms occurring during the infusions were recorded. All patients were monitored for 24 hours after infusion. Pulse, blood pressure, body temperature, and respiratory rate were recorded every 15 minutes. At four hours before and 8 hours after infusion, urinalysis was performed. Serum potassium, sodium, creatinine, total bilirubin, aspartate amino transferase (AST), and alanine amino transferase (ALT) levels were examined within 24 hours before and after the first infusion. The results showed that the toxicities observed included hemoglobinuria in 54 return infusions (77.1%), headache in 28 (40.0%), nausea in 24 (34.3%), vomiting in 17 (24.3%), and abdominal pain in 8 (11.4%). Patients who received a graft > 200 ml tended to have a higher frequency of hemoglobinuria, headache, nausea, vomiting, or abdominal pain (P<0.01), and they disappeared quickly, too. Total bilirubin increased after the first return infusion (P<0.01), and there was a significant correlation between the volume of infusion and the degree of total bilirubin increase (r=0.8977, P<0.01). No renal failure or shock occurred. It is concluded that transient hemoglobinuria, headache, nausea, vomiting, and abdominal pain are common toxicities associated with PBSC autograft, and these toxicities are related with a single volume of PBSCs transfused. Total bilirubin increase is correlated with the volume of infusion. In a word, the toxicity is less frequent and lower severe in children with fractionated infusions of cryopreserved peripheral blood stem cell.
Acute Disease ; Adolescent ; Child ; Child, Preschool ; Cryopreservation ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Headache ; etiology ; Hematopoietic Stem Cell Mobilization ; methods ; Hemoglobinuria ; etiology ; Humans ; Leukemia ; therapy ; Lymphoma ; therapy ; Male ; Nausea ; etiology ; Neoplasms ; therapy ; Neuroblastoma ; therapy ; Peripheral Blood Stem Cell Transplantation ; adverse effects ; methods ; Recombinant Proteins

BACKGROUNDIn cardiology, it is controversial whether different therapy strategies influence prognosis after acute coronary syndrome. We examined and compared the long-term outcomes of invasive and conservative strategies in patients with non-ST-segment elevation myocardial infarction (NSTEMI) and characterized the patients selected for an invasive approach.

METHODSA total of 976 patients with acute NSTEMI were collected from December 2006 to October 2012 in the First Affiliated Hospital of Dalian Medical University Hospital. They are divided into conservative strategy (586 patients) and invasive strategy (390 patients) group. Unified follow-up questionnaire was performed by telephone contact (cut-off date was November, 2013). The long-term clinical events were analyzed and related to the different treatment strategies.

RESULTSThe median follow-up time was 29 months. Mortality was 28.7% (n = 168) in the conservative group and 2.1% (n = 8) in the invasive management at long-term clinical follow-up. The secondary endpoint (the composite endpoint) was 59.0% (n = 346) in the conservative group and 30.3% (n = 118) in the invasive management. Multivariate analysis showed that patients in the conservative group had higher all-cause mortality rates than those who had the invasive management (adjusted risk ratio [RR] = 7.795; 95% confidence interval [CI]: 3.796-16.006, P < 0.001), and the similar result was also seen in the secondary endpoint (adjusted RR = 2.102; 95% CI: 1.694-2.610, P < 0.001). In the subgroup analysis according to each Thrombolysis in Myocardial Infarction risk score (TRS), log-rank analysis showed lower mortality and secondary endpoint rates in the invasive group with the intermediate and high-risk patients (TRS 3-7).

CONCLUSIONSAn invasive strategy could improve long-term outcomes for NSTEMI patients, especially for intermediate and high-risk ones (TRS 3-7).

Acute Coronary Syndrome ; mortality ; pathology ; therapy ; Aged ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; mortality ; pathology ; therapy ; Prognosis ; Retrospective Studies

OBJECTIVETo evaluate the quantitative and functional changes of T helper (Th) cell subsets in the bone marrow of severe aplastic anemia (SAA) patients and the relationship between these changes and the patients hematopoietic function.

METHODSBy FACS, the quantity and ratio of Th1 and Th2 cells, the percentage of CD3(+)CD8(+) cells in the bone marrow were detected in 24 patients with SAA at active phase, 15 patients with SAA at recovery phase, and 16 normal controls. By radioimmunoassay, the serum levels of TNF-alpha, or IL-4 in 20 SAA patients at active phase, 12 at recovery phase and 16 normal controls were measured. The relationships between CD3(+)CD8(+) cells, TNF-alpha and Ret, ANC; and between Th1 cells and CD3(+)CD8(+) cells, TNF-alpha or Ret, ANC; between IL-4, balance of Th1/Th2 and Ret, ANC were evaluated.

RESULTSThe percentages of Th1 and Th2 cells, and ratio of Th1/Th2 in bone marrow of SAA patients at active phase were (4.87 +/- 2.64)%, (0.41 +/- 0.26)% and 21.22 +/- 5.07, respectively, being higher than those of normal controls [(0.42 +/- 0.30)% (P < 0.01), (0.24 +/- 0.17)% (P < 0.05) and (1.57 +/- 0.93) (P < 0.01), respectively] and all of them reduced to normal levels of SAA at recovery phase (P > 0.05). The percentage of CD3(+)CD8(+) cells significantly decreased from (32.32 +/- 8.69)% at active phase to (13.76 +/- 2.96)% at recovery phase (P < 0.01). The serum levels of TNF-alpha and IL-4 at active phase was (4.29 +/- 3.15) microg/L and (1.24 +/- 0.73) microg/L, respectively, being higher than those of normal controls (1.21 +/- 1.16) microg/L, (1.18 +/- 0.97) microg/L, but only the difference of TNF-alpha was statistically significant (P < 0.01). In recovery SAA patients, the serum levels of TNF-alpha significantly decreased to (1.46 +/- 1.41) microg/L (P < 0.01), and the levels of IL-4 increased markedly to (3.05 +/- 1.94) microg/L. The CD3(+)CD8(+) cells and TNF-alpha of patients negatively correlated with Ret (P < 0.05; P < 0.05) and ANC (P < 0.05; P < 0.05), Th1 cells correlated with CD3(+)CD8(+) cells and TNF-alpha positively (P < 0.01; P < 0.05), the Ret and ANC negatively (P < 0.01; P < 0.01), IL-4 and the balance of Th1/Th2 positively correlated with Ret and ANC (P < 0.05, P < 0.01; P < 0.01, P < 0.01).

CONCLUSIONThe bone marrow failure in SAA might be caused not only by the increase of Th1 cells, Th1 type effector cells and cytokines, but also by insufficient compensation of Th2 cells and Th2 type cytokines, which shifted the balance of Th1/Th2 favorable to Th1.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; pathology ; physiopathology ; Bone Marrow ; metabolism ; pathology ; CD3 Complex ; blood ; CD8 Antigens ; blood ; Child ; Female ; Hematopoietic System ; metabolism ; pathology ; physiopathology ; Humans ; Interleukin-4 ; blood ; Male ; Middle Aged ; Radioimmunoassay ; T-Lymphocytes, Helper-Inducer ; metabolism ; pathology ; Th1 Cells ; metabolism ; pathology ; Th2 Cells ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

OBJECTIVETo investigate the growth of endogenous erythroid colony (EEC) in polycythemia vera (PV) patients and its clinical significance.

METHODSBone marrow mononuclear cells of 26 PV patients, 2 secondary erythrocytosis (SE) and 19 normal controls were cultured by Marsh's method for EEC.

RESULTS1. EEC was present in 25/26 (96.2%) PV patients and was not found in 2 SE patients and 19 normal controls. 2. The number of EEC and the ratio of EEC/Epo-dependent CFU-E (EEC ratio) were positively correlated with the hemoglobin (Hb) levels (r = 0.608, P = 0.01) in PV patients, but did not correlate with white blood cell (WBC) counts, platelet counts and neutrophil alkaline phosphatase scores. 3. EEC did not correlate with PV patients' serum Epo levels (r = 0.518, P = 0.125). 4. Fifteen PV patients were treated with hydroxyurea and/or interferon-alpha. Their EEC ratio before treatment was correlated positively with the time required for complete remission (CR) (r = 0.651, P = 0.009) and negatively with the time before relapsing (r = -0.529, P < 0.02). 5. EECs of 7 PV patients treated with HU/IFN were decreased after their blood cell counts normalization. 6. There was a positive correlation between the EEC ratio and the attacks of vascular thrombosis (r = 0.524, P = 0.01). (7) The apoptosis of bone marrow mononuclear cells of PV patients was less than that of normal controls. PV patients' EEC was negatively correlated with the apoptosis of their bone marrow mononuclear cells (r = -0.192, P < 0.045).

CONCLUSIONEEC is peculiarly present in PV patients, and is a sensitive parameter in reflecting the abnormal hematopoietic clone burden and in diagnosing and monitoring the disease.

Adult ; Aged ; Apoptosis ; Bone Marrow Cells ; physiology ; Erythroid Precursor Cells ; physiology ; Erythropoietin ; blood ; Female ; Humans ; Male ; Middle Aged ; Polycythemia Vera ; blood ; therapy

OBJECTIVETo analyse the relapse rate and risk factors of autoimmune hemolytic anemia (AIHA) and Evans syndrome.

METHODSFifty two cases of AIHA and Evans syndrome in remission being followed up for 1 - 14 years (median time 3.8 years) were analysed for relapse rate. The risk factors of relapse were analysed by case-control study.

RESULTSThe total relapse rate of these AIHA and Evans syndrome patients was 57.7%, and the median remission duration to the first relapse was 9 months. The relapse rates in patients with negative Coombs test, warm autoantibodies and both of warm and cold autoantibodies were 30.8% (4/13), 54.0% (13/24) and 86.7% (13/15), respectively. The relapse rate in patients with cold antibody was the highest (P < 0.05). The relapse rate in patients with antibody titer >or= 100 was 92.9% (13/14) and was higher than that in patients with antibody titer < 100 [59.5% (13/22)] (P < 0.05). Patients treated with prednisone and cyclosporin relapsed less than those treated with prednisone alone, and the relapse was related to the therapy course of prednisone and CsA.

CONCLUSIONBecause of the high relapse rate, AIHA and Evans syndrome should be treated according to the class of autoantibodies, and with longer course of prednisone and cyclosporin and prophylaxis of infection.

Adolescent ; Adult ; Aged ; Anemia, Hemolytic, Autoimmune ; etiology ; immunology ; Autoantibodies ; blood ; Child ; Cyclosporine ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Recurrence ; Risk Factors ; Syndrome ; Thrombocytopenia ; etiology ; immunology

OBJECTIVETo study the clinical features of severe aplastic anemia (SAA) patients with complication of infection.

METHODSA retrospective analysis of prevalence of infection occurring in 229 SAA patients, their bacterial spectrum, and the effect of GM-CSF or G-CSF on the infection were done.

RESULTThe prevalence of infection in SAA patients was 86.0%, among which 54.2% was infected with gram-positive organisms, 40.0% with gram-negative bacilli and 5.8% with fungal infections. Septicemia occurred mostly with E. coli and Pseudomonas infection. Patient's neutropenia was significantly related to the infection. The patients with neutrophil count less than 0.2 x 10(9)/L had more frequent and severe infection. Age, hemoglobin level, subtype of T lymphocytes and antithymocyte globulin therapy were not related to infection. Prophylaxis usage of floxacin could not reduce patient' gastrointestinal infection. The total mortality of SAA patients with infection was 23.1%. Pulmonary infection and septicemia increased mortality, and GM-CSF/G-CSF therapy reduce mortality.

CONCLUSIONSAA patients were at high risk of infection which was significantly associated with severe neutropenia. GM-CSF or G-CSF therapy exerts an assistant role to antibiotics in controlling the infections.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; complications ; Anti-Bacterial Agents ; therapeutic use ; Bacteria ; isolation & purification ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infection ; drug therapy ; etiology ; Male ; Middle Aged ; Time Factors

OBJECTIVETo study the quantity and ratio of Th1, Th2 cells in the bone marrow of myelodysplastic syndromes (MDS) patients, and to evaluate the correlation between the ratio of the blast cells and the number of the Th1 cells in the bone marrow of MDS patients.

METHODSBy FACS, the quantity and ratio of IFN-gamma producing CD4(+) T cell (Th1) and IL-4 producing CD4(+) T cell (Th2) cells in the bone marrow were detected in 21 MDS patients, 18 normal controls and 13 severe aplastic anemia (SAA) patients respectively. The karyotypes of 18 MDS patients and 15 normal controls were assayed. The correlation between the ratio of the blast cells in the bone marrow and the number of the Th1 cells in the MDS patients were analyzed.

RESULTSThe percentages of Th1 cells, Th2 cells and ratio of Th1/Th2 in the bone marrow of normal controls were (0.48 +/- 0.10)%, (0.24 +/- 0.19)% and 2.31 +/- 0.76 respectively, while those of the MDS patients were (0.36 +/- 0.11)%, (0.76 +/- 0.35)% and 0.51 +/- 0.13. The percentage of Th1 cells of patients with MDS was reduced and the Th1/Th2 ratio was significantly lower than that of normal controls (P < 0.01). Those of the patients with SAA were (4.75 +/- 0.49)%, (0.40 +/- 0.28)% and 26.5 +/- 8.79 respectively, their Th1 cells and Th1/Th2 ratio were markedly higher than those of normal controls (P < 0.01). In all of the 15 normal controls the karyotypes were normal, but that of MDS patients was (50.00 +/- 0.10)%. The lower ratio of the Th1 cells in the bone marrow of the patients with MDS and the AML which progressed from MDS was negatively correlated with the higher percentage of the blast cells (r = -0.563, P < 0.01).

CONCLUSIONS(1) The immune function of T lymphocytes in MDS is abnormal: the balance between Th1 and Th2 cells is broken. (2) With descending of the number of Th1 cells in the bone marrow of the MDS patients, the disease is progressing to leukemia.

Adult ; Aged ; Bone Marrow ; immunology ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; immunology ; T-Lymphocytes, Helper-Inducer ; immunology

BACKGROUNDPolycythemia vera (PV) is a malignant disorder of hemaopoietic stem cells which is characterized by clonal hyperproliferation and a low rate of apoptosis. This study was to assess endogenous erythroid colony (EEC) formation in the bone marrow of PV patients and determine its clinical significance.

METHODSThe bone marrow mononuclear cells of 26 patients with PV, 2 patients with secondary erythrocytosis (SE), and 19 normal controls were cultured by Marsh's method for EEC evaluation, and the clinical significance was evaluated.

RESULTSEECs appeared in 25 patients with PV but not in 2 patients with SE and 19 normal controls. The number of EECs and the EEC ratio [EEC/erythropoietin (EPO)-dependent colony forming unit-erythroid (CFU-E)] in PV patients positively correlated with hemoglobin (Hb) levels. Their EEC number did not correlate with white blood cell (WBC) counts, platelet (PLT) counts, or leukocyte alkaline phosphatase (LAP) scores. Their EEC did not correlate with serum EPO levels. Fifteen patients with PV were treated with hydroxyurea (Hu) and/or interferon-alpha (IFN-alpha). Their EEC ratio before treatment positively correlated with the treatment time required for complete remission (CR) and negatively correlated with the time before relapse. The EEC numbers of 7 PV patients treated with Hu/IFN-alpha decreased after the blood cell counts dropped to normal levels. There was a positive correlation between the EEC ratio and the incidence of attacks of vascular thrombosis in PV patients. The numbers of apoptosised bone marrow mononuclear cells in PV patients were lower than those in normal controls. The EEC numbers of PV patients negatively correlated with the rate of apoptosis of bone marrow mononuclear cells.

CONCLUSIONSEEC formation is characteristic in PV patients. EEC number in PV patients positively correlates with Hb levels, the time required for CR, and the incidence of attacks of vascular thrombosis. EEC number negatively correlates with the time before relapse. Bone marrow suppressive treatment might decrease EEC number. Thus, EEC number is a sensitive and specific parameter reflecting the abnormal hematopoietic clone burden induced by polycythemia vera. EEC number is an important diagnostic parameter for PV patients.

Adult ; Aged ; Apoptosis ; Colony-Forming Units Assay ; Erythroid Precursor Cells ; physiology ; Erythropoiesis ; Erythropoietin ; blood ; Female ; Humans ; Male ; Middle Aged ; Polycythemia Vera ; blood ; therapy ; Thrombosis ; epidemiology

OBJECTIVETo study the apoptosis and proliferation of CD(34) positive (CD(34)(+)) bone marrow cells (BMC) in patients with polycythemia vera (PV).

METHODSThe expression of Annexin V and Ki67 of the CD(34)(+) BMC in 20 PV patients and control cases [10 essential thrombocythemia (ET), 12 normal persons] were assessed by bicolor flow cytometry (FCM), and the correlation between apoptosis and clinical situation was analysed in PV patients.

RESULTSThe Annexin V expressions of CD(34)(+) BMC were (15.96 +/- 1.45)% in PV patients and (15.53 +/- 1.76)% in ET patients which were lower than that in normal subjects [(23.61 +/- 3.89)%, (P < 0.05)]. The Ki67 expression of CD(34)(+) BMC was (48.79 +/- 11.68)% in PV patients and (49.60 +/- 9.98)% in ET patients, which were significantly higher than that in normal controls (33.87 +/- 6.82)%. The ratio of apoptosis/proliferation in PV patients was 0.33 +/- 0.10 and in ET patients 0.32 +/- 0.02 which were significantly lower than that in normal controls 0.72 +/- 0.11 (P < 0.01). The apoptosis of CD(34)(+) BMC was negatively correlated with the hemoglobin (Hb) levels (r = -0.481, P = 0.037), white blood cells (WBC) (r = -0.538, P = 0.026) and the numbers of endogenous erythroid colony (EEC) (r = -0.632, P = 0.50), and the ratio of apoptosis/proliferation was negatively correlated with the Hb (r = -0.537, P = 0.018) and WBC (r = -0.667, P = 0.003) in PV patients.

CONCLUSIONThere were lower apoptosis and higher proliferation in CD(34)(+) BMC of PV patients. Lower apoptosis was correlated with the severity of the disease.

Adult ; Annexin A5 ; analysis ; Antigens, CD34 ; analysis ; Apoptosis ; Bone Marrow Cells ; cytology ; Cell Division ; Female ; Humans ; Male ; Middle Aged ; Polycythemia Vera ; pathology

OBJECTIVETo investigate the expression of apoptosis receptor FAS (CD95) and apoptosis related protein Bcl-2 and Bax in CD34 positive bone marrow cells of the patients with polycythemia vera (PV).

METHODSThe expressions of apoptosis receptor FAS (CD95) and apoptosis related protein Bcl-2 and Bax in bone marrow CD34(+) cells from 21 PV patients, 8 essential thrombocythemia (ET) and 11 normal persons were assessed by bicolor flow cytometry (FCM), and the expressions of Bcl-2 and Bax mRNA were assessed by RT-PCR, and their correlation was analysed.

RESULTSThere was no difference between the expressions of CD95 in CD34(+) bone marrow cells of PV patients (42.65 +/- 15.56)%, and that of ET patients (45.31 +/- 17.62)% and of normal person (37.55 +/- 15.19)% (P > 0.05). There was no difference between the expression of Bax in CD34(+) bone marrow cells of PV patients (35.83 +/- 9.33)% and of normal persons (41.65 +/- 9.04)% (P > 0.05). The expression of Bcl-2 in CD34(+) bone marrow cells of PV patients (79.35 +/- 14.43)% was significantly higher than that of normal controls (55.84 +/- 13.43)% (P < 0.01). The ratio of Bax/Bcl-2 of PV patients (0.47 +/- 0.14) was significantly lower than that in normal controls (0.76 +/- 0.24) (P < 0.01). The expression of Bcl-2 mRNA in PV patients' bone marrow hematopoietic cells was higher than that of normal controls (P < 0.01). There was no difference between the expression of Bax mRNA in bone marrow hematopoietic cells of PV patients and that of normal controls. Bcl-2 expression was negatively correlated with Annexin V expression in CD34(+) bone marrow cells of PV patients.

CONCLUSIONOver-expression of Bcl-2, one of anti-apoptosis genes, in CD34(+) bone marrow cells might be involved in the lower apoptosis of PV patients.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; blood ; Bone Marrow Cells ; metabolism ; pathology ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Polycythemia Vera ; blood ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; bcl-2-Associated X Protein ; blood ; genetics ; fas Receptor ; blood