Establishment of A Mixed Chimeric Mouse Model of Allogeneic Bone Marrow Transplantation and Its Influencing Factors / 中国实验血液学杂志

OBJECTIVE@#To establish a mouse mixed chimerism (MC) model of nonmyeloablative allogeneic bone marrow transplantation(allo-BMT) and explore its affecting factors.@*METHODS@#The MC model was established by nonmyeloablative allo-BMT followed by high-dose post-transplant cyclophosphamide (PTCY). 123 mice in the experiments was retrospectively analyzed, and the factors related with the chimerism were explored with the univariate and multivariate logistic regression analysis. A multivariate linear regression was performed by R project to obtain a mathematical model for predicting the chimeric level with relevant affecting factors.@*RESULTS@#The model presented mixed chimerism on day 14 after transplantation, and was characterized by a donor lymphocyte infusion (DLI) which significantly promoted donor engraftment on day 15, but transfplantation of PBS in control group was failed. Among 123 mice, 47 (38.21%) mice were MC, while 76 (61.79%) mice were non-MC in 123 mice, respectively; univariate analysis showed that the baseline body weight of mice (P=0.001, 17.84±1.19 g vs 18.50±0.94 g), total body irradiation(TBI，P=0.048) and the using of cyclophosphamide (P=0.16) were affected the chimeric state of mice, while the number of infusing cells and the time of detection showed no significant effects. Multivariate regression analysis showed that under certain conditions, the body weight of mice on day 0 was an independent factor affecting chimeric levels (OR=0.493, 95% CI 0.307-0.791, P=0.003). Through R project multiple linear regression, the math model was achieved, which was chimerism=6.09-12×weight(g)+80.03×TBI(Gy)-4.4×cell-counts (× 10@*CONCLUSION@#The experiment presents a method for establishing a mixed chimeric mice model after non-myeloablative bone marrow transplantation and constructs a mathematical model with relevant factors affected chimerism status.

Clinical and laboratory characteristics of hematological diseases associated with eosinophilia / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the clinical and laboratory characteristics of hematological diseases associated with eosinophilia.</p><p><b>METHODS</b>Karyotype analysis was performed by direct method and/or short-time culture of bone marrow cells for R-banding. Fluorescence in situ hybridization (FISH) was performed using PDGFRα, PDGFRβ and FGFR1 break-apart probes.</p><p><b>RESULTS</b>The clinical and hematological findings of 44 patients were diagnosed as hematological diseases associated with eosinophilia. Abnormal karyotypes were detected in 6 cases (13.64%) with karyotyping. The efficiency of the detection of abnormal clone was markedly increased to 29.55% (13/44) with FISH techniques, including 7 cases with FIP1L1-PDGFRα (F/P, 15.91%), 3(6.82%) PDGFRα rearrangement, 2 (4.55%) aberrant PDGFRβ gene and 1(2.27%) FGFR1 rearrangement. Patients being PDGFRα, PDGFRβ or FGFR1 positive (13 cases) or negative (31 cases) showed predominant difference in clinical and laboratory features. The incidence of gut involvement, the absolute count of eosinophils in peripheral blood and the percentage of immature eosinophils in bone marrow were significantly increased in positive patients (P < 0.05).</p><p><b>CONCLUSIONS</b>The hematological diseases associated with eosinophilia are characterized by unique clinical and laboratory features. Karyotyping should be a routine approach to detect the abnormal clone in these diseases. Screening for PDGFRα, PDGFRβ and FGFR1 gene with FISH can provide more genetic information.</p>

Prognostic significance of EBMT score for chronic myeloid leukemia patients in allogeneic stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the risk factors of allogeneic stem cell transplantation (allo-SCT) for chronic myeloid leukemia (CML) in an attempt to avoid transplant risks.</p><p><b>METHODS</b>A total of 121 CML patients received allo-SCT were analyzed retrospectively. The risk analysis was based on the EBMT score (gratwohl score) which included donor type, age of patients, disease status before transplantation, donor/recipient sex match and time interval between diagnosis to allo-SCT. Patients were divided into 3 risk groups based on their EBMT score: low risk (score 0-2), intermediate risk (3-4) and high-risk (5).</p><p><b>RESULTS</b>The median follow-up duration was 37 (1 - 126) months. The estimated 5-year overall survival (5 y-OS), non-relapse mortality (5 y-NRM) and relapse rate (5 y-RR) were (56.8 ± 5.0)%, (35.6 ± 4.9)% and (12.9 ± 3.7)%, respectively. The 5y-OS, NRM and RR were (66.0 ± 6.1)%, (28.8 ± 6.0)% and (7.8 ± 3.3)% in the low risk group being significantly superior to both intermediate-risk \[(47.2 ± 8.7)%, (43.6 ± 8.5)% and (18.7 ± 8.1)%\] and high-risk group \[(16.8 ± 15.2)%, (66.7 ± 25.5)% and (50.0 ± 25.0)%\] (P = 0.0015, 0.045 and 0.0053 for OS, NRM and RR respectively).</p><p><b>CONCLUSION</b>The EBMT risk score can effectively predict the overall outcome, relapse and transplant-related mortality of allo-SCT for CML patients.</p>

Reduced intensity conditioning regimen for related and unrelated allogeneic peripheral blood hematopoietic stem cell transplantation in chronic myeloid leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the overall efficacy and transplant-related mortality (TRM) of related and unrelated allogeneic peripheral blood hematopoietic stem cell transplantation (allo-PBSCT) in chronic myeloid leukemia (CML) patients conditioned with fludarabine-busulfan (FB) reduced intensity regimen.</p><p><b>METHODS</b>Forty-four patients received FB (Flud 30 mgxm(-2)xd(-1) x 5 d, BU 4 mgxkg(-1)xd(-1) x 3 d) conditioning followed by allo-PBSCT. Of them, 29 patients were transplanted with related donor and 15 unrelated donor (URD). All patients received mycophenolate mofetil (MMF), CsA and MTX for acute GVHD (aGVHD) prophylaxis. 5 mg/kg rabbit-antithymocyte globulin (ATG-Fresenius) was incorporated in 15 URD recipients.</p><p><b>RESULTS</b>All patients were successfully engrafted. The median times to ANC above 0.5 x 10(9)/L in related (RG) and unrelated groups (URG) were 13.7 (9 - 18) d and 13.6 (12 - 17) d, and PLT above 20 x 10(9)/L were 15.3 (9 - 20) d and 14.7 (10 - 26) d, respectively. Two patients in RG. 1 in URG developed graft rejection 5 - 8 months after transplantation. One of the two patients in RG received second transplantation and engrafted. The cumulative incidence of aGVHD and cGVHD were 13.8% (4/29) and 46.4% (13/28) in RG, and were 33.3% (5/15) and 57.1% (8/14) in URG respectively. Two patients in RG relapsed after transplantation, and obtained CR again after donor stem cell infusion (DSI). Median time of follow-up was 34.7 (2 - 73) months. Thirty-four patients were alive and 10 died. The main causes of death were IP, GVHD, graft rejection and infection. The 5-year overall survival (OS) probability was 77.0%, and the disease-free-survival (DFS) was 73.9%, of which, 79.0% and 74.1% were in RG, and 73.3% and 73.3% in URG, respectively.</p><p><b>CONCLUSIONS</b>Fludarabine-busulfan based reduced intensity conditioning for allo-PBSCT with either related or unrelated donors is a safe, less toxic and curative approach to CML.</p>

A novel single nucleotide polymorphism-based method for quantitative assessment of chimerism after allogeneic stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To develop a novel single nucleotide polymorphism (SNP)-PCR based method for quantitative detection of chimerism after allogeneic haemopoietic stem cell transplantation (allo-HSCT), and to explore its feasibility, accuracy and superiority.</p><p><b>METHODS</b>18 SNP loci were sereened to identify informative markers for detecting chimerism in each donor/recipient pair before transplantation. Then the chimerism rate of each informative marker was analyzed by real-time quantitative PCR (RQ-PCR). The accuracy and sensitivity were verified by multiple proportion dilution and analogy chimerism compared with quantitative detection of short tandem repeat (STR)-PCR, fluorescence in situ hybridization (FISH) and fusion gene.</p><p><b>RESULTS</b>(1) The average slope of the 17 time amplications of the internal control plasmid was -3.39, the average intercept was 39.97, correlation coefficients were more than 0.995, which was close to the theoretical level. The intra- and interassay variability was 0.50% and 1.1%, respectively, which were both in the allowed ranges. A linear correlation with artificial mixed chimerism is above 0.99 and a sensitivity of 0.01% proved reproducible. (2) At least one informative marker could be found in over 95% of 40 donor/recipient pairs. The results of the chimerisms derived from SNP-PCR were consistent with that from STR-PCR (96.7%), FISH and fusion gene analasis (P > 0.05); the quantitative results of special fusion gene transcripts were negtive in complete chimerism samples, and positive in mixed chimerism samples.</p><p><b>CONCLUSIONS</b>This new assay which overcome the PCR competition and plateau biases of STR-PCR provides an accurate, reliable and rapid quantitative assessment of mixed chimerism after allo-transplantation. It is highly promising for of clinical application and may take the place of STR-PCR in the conventional chimerisim assessment.</p>

Relationship between the absolute lymphocyte count before or after autologous hematopoietic stem cell transplantation and the prognosis in patients with non-Hodgkin's lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the relationship of the absolute lymphocyte count (ALC) before or after autologous hematopoietic stem cell transplantation (AHSCT), and the infused CD34(+) count with the relapse, disease-free survival (DFS), overall survival (OS) in non-Hodgkin's lymphoma (NHL) patients.</p><p><b>METHODS</b>A retrospective analysis of the relationship of the absolute lymphocyte count before (ALC-Pro) or 15 days after (ALC-15) the AHSCT, and the infused CD34(+) cell count with the relapse, DFS, OS of NHL.</p><p><b>RESULTS</b>ALC-Pro and ALC-15 were related to NHL relapse. The patients with high levels of ALC-Pro and ALC-15 had lower relapse rate. By Kaplan-Meier curve and COX multi-variable regression analysis, it was found that both ALC-Pro and ALC-15 were factors associated with the DFS. Patients infused high levels of CD34(+) cells had longer DFS. Patients with high levels of ALC-Pro, ALC-15, CD34(+) counts had longer OS.</p><p><b>CONCLUSION</b>ALC-Pro, ALC-15 and CD34(+) counts were significant in predicting relapse, DFS and OS for patients with NHL.</p>

Clinical and laboratory study of myleodysplastic syndrome (MDS)/myeloproliferative neoplasm (MPN) with PDGFRβ abnormalities / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical and laboratory characteristics of myleodysplastic syndrome (MDS)/myeloproliferative neoplasm (MPN) with PDGFRβ abnormalities.</p><p><b>METHODS</b>Chromosome specimens were prepared directly and/or short-time culture of bone marrow cells. Karyotyping was performed with R-binding technique. Fluorescence in situ hybridization (FISH) was performed using PDGFRβ, PDGFRα, FGFR1 break-apart probes and whole chromosome 5 and 12 painting probes, respectively. The expression of JAK2 V617F was measured with quantitative PCR.</p><p><b>RESULTS</b>The clinical and hematological findings of 27 patients were compatible with diagnosis of MDS/MPN. PDGFRβ rearrangement was detected in 4 patients with D-FISH, and 2 of which were confirmed as t(5;12) by chromosome painting. PDGFRα, FGFR1 and JAK2 V617F mutation were not detected in these 4 PDGFRβ positive MDS/MPN patients with.</p><p><b>CONCLUSIONS</b>PDGFRβ gene rearrangement may be detected in some MDS/MPN patients. FISH is a convenient and reliable approach to detect PDGFRβ gene.</p>

Overview of chronic myelogenous leukemia and its current diagnosis and treatment patterns in 15 hospitals in China / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore demographic characteristics, current diagnosis and treatment patterns of chronic myelogenous leukemia (CML) patients in China.</p><p><b>METHODS</b>Data of hospitalized CML patients in 2005 whole year and outpatient information (July 1 through September 30, 2006) from 15 hospitals throughout China were analyzed.</p><p><b>RESULTS</b>A total of 1824 CML cases were analyzed, including 722 inpatients and 1102 outpatients. The male/female ratio was 1.78:1. The median age at diagnosis was 40.02 (2.45 - 83.29) years old, 90.41% of the patients were diagnosed at chronic phase. Proportion of accelerated phase or blast crisis patients increased to 21.66% during study period. 93.20% of the patients received blood routine and bone marrow morphologic examination at diagnosis and in monitoring; 70.29% were performed cytogenetic analysis and 51.54% performed molecular measurement in addition. The most common therapy for CML treatment was hydroxycarbamide. The proportion of patients treated with imatinib and interferon was 37.45% and 25.55%, respectively. Of 722 inpatients, 164 (22.72%) received hemotopoietic stem cell transplantation (HSCT). The proportions of accelerated phase and blast crisis patients treated with imatinib were 48.28% and 48.42%, respectively, being significantly higher than that of chronic phase patients (35.9%) (P < 0.05). The mean imatinib dosage administered in the three phases patients did not differ significantly. Imatinib resistance rates were 6.87% and 16.28% for outpatient and inpatient, respectively. In the outpatient group, the primary resistance to imatinib occurred comparably to the secondary resistance (68.75%), while primary resistance was predominant in inpatient group (65.71%). The intolerance rates of imatinib for outpatient and inpatient were 3.21%, 11.63%, respectively. The majority of patients treated with imatinb were not monitored in time: 63.38% patients evaluated hematologic response after 3 months of treatment, proportions of patients received cytogenetic examination after 6 months and 12 months of treatment were 41.41% and 27.35%, respectively. Mean cost for HSCT was 213 092 +/- 125 890 RMB.</p><p><b>CONCLUSIONS</b>CML in China tends to afflict younger population than in Western countries. Most patients were diagnosed in the chronic phase. Due to restriction of financial support, only one third of CML patients were treated with imatinib, and the majority of the treated were not monitored in time. Clinicians should pay attention to resistance and intolerance to imatinib treatment in accelerated phase or blast crisis patients.</p>

An analysis of prognostic factors in 80 myelodysplastic syndrome / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the WHO classification, clinical and hematological features and risk group of International Prognostic Scoring System (IPSS) in patients with myelodysplastic syndrome (MDS).</p><p><b>METHODS</b>The diagnosis and classification of MDS patients were defined according to the WHO classification. The clinical manifestations, hemogram, bone marrow biopsy and prognosis were retrospectively analyzed.</p><p><b>RESULTS</b>The median age at diagnosis of MDS was 47 yrs being younger than that in some foreign reports. The frequency of abnormal karyotype was 35.14% and +8 was the most frequent abnormal karyotype in our study. Eleven of 74 patients transformed into leukemia. Univariate analysis showed that age, chromosome abnormality, percentage of bone marrow blast cells and number of cytopenias were significantly related to prognosis. There was a statistical difference in cum survival rate between IPSS subcategories (P < 0.05) except that between low- and intermediate I-risk subcategory (P > 0.05). There were statistical differences for refractory anemia (RA) vs RA with excess blast (RAEB), refractory cytopenias with multilineage dysplasia (RCMD) vs RAEB and RAEB-I vs RAEB-II (P < 0.05).</p><p><b>CONCLUSIONS</b>There were differences in age of disease onset, distribution of WHO, sub-classification and abnormal karyotype in this cohort of MDS patients as compared with those in Europe and Japan. It is helpful in diagnosis, treatment and prognosis to divide RAEB into RAEB-I and RAEB-II. IPSS was well applicable in Chinese MDS patients.</p>

A clinical observation of fludarabine-containing regimens in the treatment of low grade non-Hodgkin's lymphoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the therapeutic efficiency and adverse effect of the fludarabine-containing regimens in the treatment of low grade non-Hodgkin's lymphoma.</p><p><b>METHODS</b>Thirty-two patients with low grade non-Hodgkin's lymphoma consisting of 19 primary one and 13 relapsed or refractory were treated with fludarabine-containing regimens, which included FMD (fludarabine, mitoxantrone and dexamethasone); FMC (fludarabine, cyclophosphamide and mitoxantrone) and FC ( fludarabine and cyclophosphamide).</p><p><b>RESULTS</b>The average course completed in these 32 patients was 4.1 with a complete response rate (CR), partial response rate (PR) and overall response rate (OR) of 65.6%, 18.8% and 84.4% , respectively. There were no significant difference in CR, PR and OR between primary and relapsed or refractory group (71.4%, 21.0%, 92.4% vs. 46.2%, 13.1%, 59.3%, respectively). Myelotoxicity and immunotoxicity was the dominating adverse effects. Ill to IV grade granulocytopenia and thrombocytopenia were observed in 31.3% (10/32) and 9.4% (3/32) of these patients respectively. Infection developed in 7 patients, and two of them died of pulmonary infection. The median follow-up period was 16 months (1-30 months) with 2-year overall-survival rate (OS) and progression-free survival rate (PFS) of 93.8% and 84.4%, respectively. No significant difference was observed between primary and relapsed or refractory group in OS (100% vs. 76.9%) and PFS (94.7% vs. 69.2%).</p><p><b>CONCLUSION</b>Fludarabine-containing regimens is well tolerated and effective in the treatment of low grade non-Hodgkin's lymphoma.</p>

Clinical and laboratory characteristics of two acute lymphoblastic leukemia patients with dicentric (9; 20) (p11 - 13; q11) / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the morphologic, immunophenotypic, cytogenetic and clinical features of acute lymphoblastic leukemia (ALL) patients with dicentric (9; 20) (p11 - 13; q11).</p><p><b>METHODS</b>Chromosome specimens of bone marrow cells were prepared by direct method and/or short-time culture. Karyo-typing was performed by R-banding technique. Dual-color fluorescence in situ hybridization (FISH) was performed using both chromosome 9 classical satellite probe and chromosome 20 alpha-satellite probe in one patient.</p><p><b>RESULTS</b>The two ALL patients were positive for CD10 and HLA-DR, showing of B cell origin. Both patients had dicentric (9; 20): case 1 was 45, XY, der (9) t (9; 20) (p11; q11), -20[20]; case 2 was 45, XX, der (9) t (9; 20) (p13; q11), t (9; 22) (q34; q11), -20[10]/46, idem, +8[16]/47, idem, +8, +21[14]. Mutual translocation between chromosomes 9 and 20 of the dicentric chromosome was confirmed by FISH in one patient.</p><p><b>CONCLUSIONS</b>Dicentric (9; 20) (p11 - 13; q11) is a rare recurring chromosome abnormality associated with ALL. Because of the subtle nature of the translocation, FISH is essential for the detection of this abnormality.</p>

Report of a case of hybrid acute leukemia with t (12; 22) and literature review / 中华血液学杂志

<p><b>OBJECTIVE</b>To report a hybrid acute leukemia (HAL) patient with t (12; 22) (p13; q12).</p><p><b>METHODS</b>Chromosome specimens were prepared by direct method and/or short-time culture of bone marrow cells. Karyotyping was performed by R-banding technique. Leukemia surface markers were detected by anti-biotin-biotin complex and monoclonal antibodies. Chromosome painting (fluorescence in situ hybridization, FISH) was performed by using whole chromosome 12 and 22 probes labeled with green and red fluorescence, respectively.</p><p><b>RESULTS</b>The clinical and hematological findings were compatible with the diagnosis of HAL. Lymphoid and myeloid markers were positive on the leukemia cells. Karyotype analysis showed that the patient had t (12; 22) (p13; q12) translocation. A reciprocal translocation between chromosomes 12p and 22q was proved by FISH.</p><p><b>CONCLUSIONS</b>t (12; 22) translocation is a rare chromosome abnormality in leukemia. Patients with t (12; 22) had unique clinical, cytogenetic features. This translocation as a cytogenetic marker for poor-prognosis in leukemia needs to be further studied.</p>

Pulmonary fungal infection in malignant hematological diseases: an analysis of 14 cases / 中国实验血液学杂志

The objective of this study was to evaluate the etiological factors, diagnosis and therapy of pulmonary fungal infection in hematological malignancies, 14 cases of malignant hematological disease with pulmonary fungal infection were collected and analyzed. The results showed that 11 out of 14 cases had the respiratory manifestations, sputum was sticky and difficult to be expectorated in 7 cases, X rays in chests showed shadows with features of stigma and sheet in 11 cases, Candida albicans and aspergillus infection were observed in 10 and 2 cases respectively, the numbers of neutrophil were below 0.5 x 10(9)/L in 8 cases and below 1.0 x 10(9)/L in 3 cases respectively, fluconazole was used for 12 cases and clinical symptoms of 11 cases were improved within two weeks. In conclusion, the occurrence of pulmonary fungal infection in malignant hematological diseases is associated with intensive chemotherapy, decrease of neutrophil counts and using of broad-spectrum antibiotics, the diagnosis at early stage is difficult and clinicians should pay more attention to its clinical and laboratory examinations, and give them therapy in time.

Study on graft-versus-host disease in the allogeneic peripheral blood stem cell transplantation for the treatment of leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the incidence and the effective prevention and treatment of graft-versus-host disease (GVHD) for the allogeneic peripheral blood stem cell transplantation (allo-PBSCT) for the treatment of leukemia.</p><p><b>METHODS</b>Fifty patients with acute leukemia (n = 29) and chronic myeloid leukemia (n = 21) were treated with allo-PBSCT. The conditioning regimens were TBI plus CTX and Vp16 or TBI plus CTX. Two regimens were used for prophylaxis of GVHD: one was the combination of low dose cyclosporine (CsA, 2 - 3 mg x kg(-1) x d(-)1 i.v. or 4 - 6 mg x kg(-1) x d(-1) p.o.) and short course methotrexate (MTX, 15 approximately 10 mg, +1, +3, +6, +11 d) (CsA/MTX group, 32 patients), the other was short course of mycophenolate mofetil (MMF, 1.0 bid, +1 - +28 d) in addition to CsA and MTX (MMF/CsA/MTX group, 18 patients).</p><p><b>RESULTS</b>All patients were successfully engrafted and the median times to ANC > 0.5 x 10(9)/L and to platelet > 20 x 10(9)/L were 14 (10 - 22) and 20 (10 - 68) days post PBSCT respectively. The incidence of acute GVHD (aGVHD) was 40% (20/50) and of grade III - IV was 12% (6/50). The chronic GVHD (cGVHD) occurred in 22 out of 33 (66.7%) evaluable patients (survived longer than 6 months post PBSCT) and extensive cGVHD in 11 out of 33 (33.3%) patients. Patients with aGVHD displayed significantly higher sIL-2R levels [(277.3 +/- 26.4) U/L] and CD(25)(+) cells [(8.1 +/- 3.4)%] than those without GVHD [(128.1 +/- 96.7) U/L and (3.6 +/- 1.7)%] (P < 0.05). The incidences of aGVHD (16.7%) and extensive cGVHD (9.1%) in MMF/CsA/MTX group were significantly lower than that in CsA/MTX group (53.1% and 45.5%, P < 0.05). The median follow-up duration was 30 (3 - 70) months and 33 patients were still alive. The relapse rate was significantly higher in GVHD negative group (47.1%) than in GVHD positive group (0, P < 0.05). The 3 year disease-free-survival (DFS) rate was 66%.</p><p><b>CONCLUSION</b>The incidence of aGVHD was low, but of cGVHD was high in allo-PBSCT. sIL-2R and CD(25)(+) cells after PBSCT may provide predictive markers for aGVHD. The MMF/CsA/MTX regimen for prevention of aGVHD in allo-PBSCT is more effective than the CsA/MTX one. There was a strong antileukemic effect of GVHD in the allo-PBSCT.</p>

Determination of CD34(+) cells in mobilized peripheral blood by flow cytometry / 中国实验血液学杂志

To explore a simple and effective method to determinate the volume of CD34(+) cells in the peripheral blood of donors received drug mobilization for stem cell transplantation by using flow cytometry, the mobilized peripheral blood from donors and 100 micro l fresh whole blood were labeled with monoclonal antibodies Anti-CD34-PE and Anti-CD45-FITC, after lying the red blood cells, and assessed with flow cytometer FL2 (log) vs SSC (log) and FL1 (log) vs SSC (log) were mainly used for analysis windows. The results showed that a level of CD34(+) cells in whole nucleated cells as low as 0.05% - 0.1% can be detected effectively using this method when 10(5) nucleated cells were counted. At day 5 or day 6, the level of CD34(+) cells in most samples of patients reached a peak volume, some of samples and the levels were more than one percent in. It was concluded that CD34(+) cells can be effectively determined by using this method. According to the relative rate of CD34(+) cells, the time to harvest the stem cells in blood can be determined.

Study on Relation of Early Immune Reconstitution with Acute Graft-Versus-Host Disease / 中国实验血液学杂志

To analyze the relation of early immune reconstitution with acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation (all-HSCT), the changes of CD3(+), CD4(+), CD8(+), CD25(+) and CD69(+) cells in peripheral blood from 26 patients with hematologic malignancies were assayed by flow cytometry within 2 months after allo-HSCT. All patients achieved hematopoietic reconstitution, and grade I and II - IV GVHD were developed in 9 and 5 patients, respectively. CD25(+) cells were increased in patients aGVHD at week 2 after transplantation and the peak value was appeared at week 3. The increase of CD25(+) cells was preceded the occurence of clinical signs of aGVHD. The maximal levels of CD25(+) cells increase correlated significantly with the severity of aGVHD. The increase of CD25(+) cells was declined along with remission of aGVHD signs. Our results suggest that analyzing immune reconstitution after allo-HSCT could predict occurence of aGVHD, and CD25(+) cell increase prior occurence of aGVHD is predictive marker for aGVHD.