Objective To investigate the mobilization effect of low-does granulocyte-colony stimulation factors(G-CSF) on allogenic peripheral stem cell transplantation (PBSCT) and the best acquisition time for haemopoietic stem cell. Methods G-CSF was administered subcutaneously at 5μg·kg~(-1)·d~(-1)to 108healthy donors. The mobilization effects were assayed by mononuclear cell count,flow cytometry and colonyforming units analysis.Results Onfourth day,fifty day and sixth day, the ratio of CD_(34)~+ were (0.71 ±0.08)%, (1.09±0.09) %, (0.57±0.08) %, respectively. The CFU-GM yield were (93.33±44.51)/10~5MNC, (124.61±57.85)/ 10~5MNC, (80.25±49.24)/10~5MNC, respectively. The number of CD_(34)~+ were (3.33±1.36)×10~6/kg, (4.14±1.67) ×10~6/kg, (2.79±1.47)×l0~6/kg, respectively. The ratio and number of CD_(34)~+ cell of PBSC collections of fifth day was higher than that of the fourth day (P <0.05). And the fourth day was higher than that of the sixth day(P < 0.05). Twice day (fourth day and fifth day) collection can obtain enough CD_(34)~+ cell for PBSCT. There were no significant adverse effects. Conclusion Low-does G-CSF takes good effects on the mobilization of PBSC. And it was better to collect stem cell on fourth day and fifth day rather than on fifth day and sixth day after mobilization.

Objective To analyze the outcomes and the prognostic factors of allogeneic peripheral blood stem cell transplantation (allo-PBSCT) for acute lymphoblastic leukemia (ALL).Method From Feb.2002 to Feb.2014,a total of 95 patients with ALL were treated with alloPBSCT in our hospital.Of these,73 cases obtained the first CR (CR1),11 cases obtained late CR,7 patients were in relapse and 3 patients suffered from primarily refractory disease (PRD) before transplant.The median age was 26 (4-57) years.Conditioning regimens including total body irradiation (TBI)/ etoposide/semustine/cyclophosphamide or busulfan/semustine/cyclophosphamide were used.Matched sibling transplantation was performed on 68 patients,and matched unrelated donor transplantation was performed on 27 patients.Combination of CsA,MTX and low-dose,short-course mycophenolate mofetil was used for graft-versus-host disease (GVHD) prophylaxis.The average fellow-up was 57 months.Result Hematopoietic reconstitution was achieved in all 95 patients.Five-year estimate of overall survival (OS) was 54.3％,disease free survival (DFS) was 51.2％,relapse rate (RR) was 30.2％ and transplant-related mortality (TRM) was 24.0％.The 5 year OS and DFS were significantly longer in patients with CR1 than in late CR and relapse/PRD patients before allo-PBSCT (P＜0.001).There was no significant difference in OS between the two different conditioning regimens.Multivariate analyses revealed that Ⅱ-Ⅳ aGVHD and cGVHD were correlated with higher TRM,CR1 before allo-PBSCT and TBI were associated with a lower RR,and non Ⅱ-Ⅳ aGVHD and CR1 before allo-PBSCT were favorable factors which were associated with OS and DFS.In the patients with DFS≥1 year after allo-PBSCT,DFS and OS were shorter in patients with cGVHD (P =0.008).Conclusion Allo-PBSCT in adult ALL patients should be performed in CR1.Severe acute and chronic GVHD are not associated with improved survival.

Objective To investigate the effect of allgeneic hematopoietic stem cell transplantation (allo-HSCT) for β-thalassemia major. Methods Twenty-four β-thalassemia major patients with median age of 4 years (range: 2～15 years), 18 boys and 6 girls, received allo-HSCT.They were classified into class Ⅱ-Ⅲ according to Pesaro thalassemia classification. Twenty-three transplantations were from sibling donor and 1 was from mother, either HLA-identical (n = 23) or HLA-mismatched (5/6) (n = 1). Fifteen patients received bone marrow transplantation (BMT) plus peripheral blood stem cell transplantation (PBSCT), and 9 were subjected to umbilical cord blood transplantation (UCBT). The conditioning regimen consisted of busalphan, cyclophosphamide,fludarabine, plus hydroxyurea before transplantation. Graft-versus-host disease (GVHD) prophylaxis included CsA, methotrexate, antilymphpcute globulin, and mycophenolate mofetil. The median follow-up period was 13 months (range: 3～69). Results Of 24 patients, there were 21 cases (87. 5 %) of disease-free survival, 1 (4. 2 %) transplantation-related death, and 2 cases (8. 3 %) of rejection. Three-year overall survival and disease-free survival rate was 91.7 % and 87. 5 %respectively. The cumulative incidence of grade Ⅱ -Ⅳ acute GVHD and chronic GVHD was 16. 7 %and 20. 3 %, particularly cumulative extensive chronic GVHD was 5. 0 %. Conclusion The sibling donor BMT plus PBSCT is an effective and safe way to treat β-thalassemia major. Cord blood is an important source of hematopoietic stem cells for HSCT. The protocol GVHD prophylaxis of CsA,MTX, ATG with a low-dose and short course of MMF can effectively reduce the incidence of severe acute GVHD, improve the outcome of thalassemia transplantation.

Objective To analyze the outcome and the prognostic factors of hepatic veno-occlusive disease (HVOD) after hematopoietic stem cell transplantation (HSCT). Methods A total of 797 patients receiving HSCT were analyzed retrospectively. The prophylaxis regimen of HVOD in the First Affiliated Hospital of Guangxi Medical University consisted of low molecular weight heparin and lipoprostaglandin E1 (PGE1). Results Fifty-nine patients (7.4%) developed HVOD at 3-49 days after HSCT (median 12 days). Age younger than 15 years at transplant( HR=6.47, P<0.001), busulphan conditioning ( HR=6.40, P<0.001), thalassemia major ( HR=6.35,P<0.001), allogeneic transplantation ( HR=7.74, P=0.005) were univariate risk factors for HVOD. Multivariate analyses suggested that thalassemia major and busulphan conditioning were independently correlated with the development of HVOD. Conclusion Thalassemia major and busulphan conditioning are independent risk factors for HVOD after HSCT.

Objective: To explore the diagnosis, treatment and prognosis of autoimmune hemolytic anemia (AIHA) after allo-HSCT in patients with thalassemia major (TM). Methods: A retrospective analysis of AIHA status after allo-HSCT in 291 TM patients from July 2007 to December 2017 was conducted. Results: Five of the 291 TM patients (1.72%) were diagnosed with post-transplant AIHA. The median time of AIHA was 7 (5-12) months after HSCT. All post-transplant AIHA patients were positive in direct and indirect Coombs test, the main clinical manifestations were dizziness, fatigue, pale complexion, skin and sclera yellow, and soy sauce urine. The incidence of AIHA was higher after unrelated donor transplantation (6.36%, 4/63) compared with that of sibling donor transplantation (0.43%, 1/228). One patient who received only prednison was dead. Four patients who received rituximab combined with prednisolone were alive, Coombs test in two of them were negative. Conclusions AIHA after allo-HSCT developed in 1.72% patients with TM. Monitoring of Coombs test was important for diagnosis of post-transplant AIHA. The incidence of post-transplant AIHA was higher in unrelated donors compared with that of sibling donors transplantation. Treatment of rituximab combined glucocorticoid was effective strategy for post-transplant AIHA.

OBJECTIVETo investigate the changes and mechanism of angiopoietin1 (Ang1) in murine bone marrow during G-CSF induced mobilization of hematopoietic stem/progenitor cell.

METHODSThe proportion of Lin-Sca1⁺cKit⁺ (LSK) cells in peripheral blood of C57BL/6 mice before and after G-CSF mobilization was detected by flow cytometry. Expression changes of Ang1 and osteocalcin (OCN) during HSC mobilization were determined by immunohistochemistry, enzyme linked immunosorbent assay (ELISA) and real-time fluorescence quantitative PCR. The number of osteoblasts in the bone marrow was counted under the microscope.

RESULTSAfter treated with G-CSF, the proportion of LSK cells in peripheral blood significantly increased from the controls (0.04 ± 0.01)% to (0.61 ± 0.05)% at day 5 (P<0.05). Before G-CSF mobilization, the endosteum cells expressed higher level of OCN and Ang1 than that of bone marrow nucleated cells. The mRNA expression level of OCN was significantly reduced from 28.64 ± 8.61 in the controls to 12.55 ± 7.06 on day 3 and 4.75 ± 1.62 on day 5, and the expression level of Ang1 also declined from 2.84 ± 0.95 in the controls to 0.93 ± 0.30 on day 3 and to 0.92 ± 0.22 on day 5 after G-CSF mobilization. The number of endosteum osteoblasts was significantly decreased after mobilization (P<0.05). The Ang1 expression was decreased in the BM after mobilization. The serum OCN was significantly reduced from (24.11 ± 3.17) ng/ml in the controls to (9.96 ± 2.16) ng/ml on day 3 and (8.43 ± 2.62) ng/ml on day 5, and the Ang1 also declined from (2.24 ± 0.52) ng/ml in the controls to (1.21±0.38) ng/ml on day 3 and (0.90±0.24) ng/ml on day 5.

CONCLUSIONIn G-CSFinduced HSPC mobilization, the bone marrow osteoblasts retraction causes reduction of Ang1, and the reduction of Ang1 may contribute to HSPC mobilization.

Animals ; Bone Marrow ; Bone Marrow Cells ; Granulocyte Colony-Stimulating Factor ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cells ; Mice ; Mice, Inbred C57BL ; Osteoblasts ; RNA, Messenger

Objective:To explore the predictive value of peripheral blood CD34-positive cell count for the stem cell mobilization effect of plerixafor in patients with multiple myeloma (MM).Methods:The clinical data of 12 MM patients who used plerixafor for stem cell mobilization in the First Affiliated Hospital of Guangxi Medical University from December 2019 to February 2021 were retrospectively analyzed. The changes of peripheral blood CD34-positive cell count and the collection status of stem cell in all patients before and after the mobilization of plerixafor were analyzed.Results:Twelve patients were included in this study. These patients were in international staging system (ISS) stage Ⅱ-Ⅲ, and the induction therapy was mainly VRD regimen. The CD34-positive cell count was increased after the use of plerixafor in all patients no matter which mobilization strategies were used before plerixafor. The CD34-positive cell count was 3.63/μl (0.72-13.53/μl) and 32.11/μl (8.52-53.68/μl) before and after the use of plerixafor, and the difference was statistically significant ( Z = -0.40, P<0.001); the median increasing time was 11.50 times (1.61-23.71 times). The mobilization failure occurred in 1 patient. The CD34-positive cell count in his blood was less than 1/μl before the use of plerixafor; though increased 11.83 times after the use of plerixafor, the CD34-positive cell count was still less than 10/μl. Pearson analysis showed that among the patients with CD34-positive cell count less than 4/μl before the use of plerixafor, there was a positive correlation in peripheral blood CD34-positive cell count before and after the use of plerixafor ( r = 0.80, P = 0.032). Conclusions:The peripheral blood CD34-positive cell count has a certain predictive value for the stem cell mobilization effect of plerixafor in MM patients.