Bioinformatics Analysis of the Influence of Coronavirus Infection on Hematopoietic System and Potential Intervention Drugs and Their Significance for COVID-19 / 中国实验血液学杂志

OBJECTIVE@#To analyze and predict the effect of coronavirus infection on hematopoietic system and potential intervention drugs, and explore their significance for coronavirus disease 2019 (COVID-19).@*METHODS@#The gene expression omnibus (GEO) database was used to screen the whole genome expression data related with coronavirus infection. The R language package was used for differential expression analysis and KEGG/GO enrichment analysis. The core genes were screened by PPI network analysis using STRING online analysis website. Then the self-developed apparent precision therapy prediction platform (EpiMed) was used to analyze diseases, drugs and related target genes.@*RESULTS@#A database in accordance with the criteria was found, which was derived from SARS coronavirus. A total of 3606 differential genes were screened, including 2148 expression up-regulated genes and 1458 expression down-regulated genes. GO enrichment mainly related with viral infection, hematopoietic regulation, cell chemotaxis, platelet granule content secretion, immune activation, acute inflammation, etc. KEGG enrichment mainly related with hematopoietic function, coagulation cascade reaction, acute inflammation, immune reaction, etc. Ten core genes such as PTPRC, ICAM1, TIMP1, CXCR5, IL-1B, MYC, CR2, FSTL1, SOX1 and COL3A1 were screened by protein interaction network analysis. Ten drugs with potential intervention effects, including glucocorticoid, TNF-α inhibitor, salvia miltiorrhiza, sirolimus, licorice, red peony, famciclovir, cyclosporine A, houttuynia cordata, fluvastatin, etc. were screened by EpiMed plotform.@*CONCLUSION@#SARS coronavirus infection can affect the hematopoietic system by changing the expression of a series of genes. The potential intervention drugs screened on these grounds are of useful reference significance for the basic and clinical research of COVID-19.

Clinical Analysis of Refractory Primary Central Nervouse System Lymphoma Treated with High-Dose MTX-Based Chemotherapy and Haploidentical Hematopoietic Stem Cell Infusion / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and safety of high-dose methotrexate-based chemotherapy combined with granulocyte-colony stimulating factor (G-CSF)-mobilized family related haploidentical donor peripheral blood hematopoietic stem cell (G-PBHSC) infusion for the treatment of patients with refractory primary central nervouse system lymphoma (PCNSL).</p><p><b>METHODS</b>Three patients with refractory PCNSL were treated in Department of Hematology of the General Hospital of the PLA's Rocket Force from March 2014 to September 2015. The sex ratio of male to female was 1:2 and the median age was 54(48-66)years old. All patients received programmed infusions of G-PBHSC after high-dose methotrexate-based chemotherapy without prophylaxis for graft-versus-host disease (GVHD).</p><p><b>RESULTS</b>Three patients had received initial chemotherapy or radiotherapy after diagnosis, one patient achieved complete remission (CR) after 3 courses of treatment and remained in CR until the end of follow-up, 2 cases achieved partial remission (PR) and the progression-free survival (PFS) time was 10 and 7 months, respectively. The patients generally well-tolerated this therapy. The main adverse effects of patients were neutropenia, thrombocytopenia and infection related with chemotherapy after each course of treatment, the median recovery times of neutrophils and platelets were 11 and 12.5 days, respectively after of programmed infusions of G-PBHSC. No GVHD was observed in any of the patients during treatment.</p><p><b>CONCLUSION</b>The combination of high-dose methotrexate-based chemotherapy with programmed haploidentical G-PBHSC infusion is a potential treatment alternative for refractory PCNSL patients.</p>

Clinical Analysis of 4 AML Patients Aged Over 80 Years Treated with Chemotherapy Combined with Haploidentical Hematopoietic Stem Cell Infusion / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and safety of MA (mitoxantrone and cytarabine) regimen chemotherapy combined with granulocyte-colony stimulating factor (G-CSF)-mobilized family related HLA-haploidentical donor peripheral blood hematopoietic stem cell (G-PBHSC) infusion for the treatment of acute myeloid leukemia (AML) patients aged over 80 years.</p><p><b>METHODS</b>Four elderly patients with AML were treated in Chinese Second Artillery General Hospital from August 2008 to September 2013. The proportion of male to female was 1 : 3 and the median age 83 (80-85) years. All patients received programmed infusions of G-PBHSC after MA regimen chemotherapy without graft-versus-host disease (GVHD) prophylaxis. After complete remission (CR), patients only received G-PBHSC infusion without chemotherapy.</p><p><b>RESULTS</b>Three cases achieved CR and their disease free survival (DFS) time was 18, 8, 6 months, respectively. 1 case did not reach remission after 2 cycles chemotherapy. The median overall survival (OS) time was 10 (3-20) months. No GVHD was observed in any of the patients during treatment. Concludsion: The combination of chemotherapy and programmed haploidentical G-PBHSC infusion is an alternative approach for AML patients aged over 80 years.</p>

Clinical analysis of lymphoblastic lymphoma/leukemia treated with Hyper-CVAD/MA regimen chemotherapy combined with haploidentical hematopoietic stem cell infusion / 中国实验血液学杂志

This study was aimed to investigate the efficacy of Hyper-CVAD/MA regimen chemotherapy combined with haploidentical hematopoietic stem cell infusion for the treatment of lymphoblastic lymphoma/leukemia (LBL/ALL). Seven patients with LBL/ALL were treated in Second Artillery General Hospital from August 2009 to September 2012. All patients received programmed infusions of granulocyte-colony stimulating factor (G-CSF)-mobilized family related HLA-haploidentical donor peripheral blood hematopoietic stem cell (G-PBHSC) after each of cycle of Hyper-CVAD/MA regimen chemotherapy without graft-versus-host disease (GVHD) prophylaxis. A total of four cycles of therapy were planned. The interval between each cycle of treatment was 8 to 12 weeks. By April 2014, the median follow-up time was 41 (20-57) months. The results showed that the 7 patients totally received 30 cycles of treatment, and all patients achieved complete remission (CR). The patients were generally well-tolerated to therapy, and the most significant toxicities of grade 3 to 4 neutropenia and thrombocytopenia developed in nearly all of the patients after each course of the Hyper-CVAD/MA regimen. No GVHD was observed in any of the patients during treatment. Up to now, 5 patients were still alive, 2 patients were died of relapse. It is concluded that the combination of chemotherapy and programmed haploidentical G-PBHSC infusion is a promising approach to the treatment of LBL/ALL.

Prophylactic G-CSF mobilized donor lymphocytes infusion after non-myeloablative stem cell transplantation prevents relapse in patients with high-risk leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the anti-leukemia effects of prophylactic G-CSF mobilized donor lymphocytes infusion (pG-DLI) and its relationship with the incidence of graft-versus-host disease (GVHD) in high-risk leukemia patients with non-myeloablative stem cell transplantation (NST).</p><p><b>METHODS</b>12 patients with high-risk leukemia were analyzed, including Ph⁺ acute lymphocytic leukemia (n=1), acute leukemia (AL) with persistent non-complete remission (n=2), acute myeloid leukemia (AML) with central nervous system (CNS) relapse (n=3), hybrid AL (n=1), secondary AML evolving from myelodysplastic syndrome (MDS/AML) (n=2), chronic myeloid leukemia in accelerated phase (CML-AP) (n=1), CML in blastic phase (CML-BP) (n=2). All patients received non-myeloablative conditioning and pG-DLIs were administered 30-40 days post transplantation if no signs of GVHD were present. The percentage of donor cell chimera was analyzed by short tandem repeat-polymerase chain reaction (STR-PCR) just before and after pG-DLI. The incidence of leukemia relapse and GVHD were observed.</p><p><b>RESULTS</b>12 high-risk leukemia patients with a median age of 38 (range: 29-52) years received G-DLI at a median interval of 35 (32-40) days. The median numbers of infused mononuclear cells (MNCs), CD34⁺, and CD3⁺ cells/kg recipient body weight was 2.3×10⁸/kg, 1.7×10⁶/kg, and 0.6×10⁸/kg, respectively. 10 of 12 patients had full donor chimera before pG-DLIs and conversion from mixed to full donor chimera occurred in the other 2 patients shortly after pG-DLI. Grade II acute GVHD (aGVHD) was observed in only 2 patients and chronic GVHD (cGVHD) developed in 6 patients, including involvement of skin (n=3), skin and intestine (n=2), liver (n=1). 1 patient died of cGVHD. With a median follow-up of 40 (24-64) months, 7 patients are alive in remission, with 3-year actuarial overall survival (OS) and disease-free survival (DFS) rates of the same 58.3%.</p><p><b>CONCLUSION</b>Our findings indicate that pG-DLI after NST does not increase the risk of aGVHD, but could enhance the capacity graft-vs-leukemia and prevent relapse in high-risk leukemia patients.</p>

Expression of B7-H1 gene in leukemia cells and its clinical significance / 中国实验血液学杂志

This study was purposed to investigate the expression of B7-H1 gene in leukemia cells and its clinical significance. The expression of B7-H1 mRNA was detected by SYBR Green I real-time quantitative PCR in a panel of 9 leukemia cell lines, 4 leukemia cell lines induced with IFN-γ, the bone marrow mononuclear cells (BMMNC) from 59 initial leukemia patients and 10 dendritic cells (DC) derived from BMMNC of initial leukemia patients, 2 solid tumour cell lines and BMMNC from 10 normal persons. The correlation between the clinical features of 59 acute leukemia patients and the expression level of B7-H1 mRNA in leukemia cells was analyzed. The results showed that the lower level of B7-H1 mRNA expression was found in leukemia cell lines and primary acute leukemia cells, but the expression level of B7-H1 mRNA was up-regulated significantly in the leukemia cell lines induced by IFN-γ and DC derived from BMMNC of leukemia patients. The expression level of B7-H1 mRNA in non complete remission (CR) patients after therapy was significantly higher than that in CR patients. It is concluded that the expression level of B7-H1 mRNA in leukemia cells is lower, but is up-regulated when affected by some factors. A correlation exists between the expression level of B7-H1 gene in leukemia cells and response of patients to therapy.

B7-H1, a prognostic factor for patient's response to therapy of acute myeloid leukemia / 中国实验血液学杂志

This study was aimed to detect the expression of B7-H1 gene in the acute myeloid leukemia (AML) cells and to explore its clinical significance. The B7-H1 gene expression was detected by real-time quantitative PCR in bone marrow mononuclear cells (BMMNC) from 40 newly diagnosed AML patients, 10 normal volunteers and 10 relapsed patients. The results showed that the expression of B7-H1 from de novo AML patients was lower than that from the normal volunteers, but it was higher in the relapsed patients. The expression level of B7-H1 gene before therapy was significantly higher in non CR patients than that in CR patients after therapy. It is concluded that there is a correlation between the expression level of B7-H1 gene and response to therapy.

Multicenter report of nonmyeloablative allogeneic stem cell transplantation for hematologic diseases / 中华血液学杂志

<p><b>OBJECTIVE</b>To observe the treatment effect and toxicity of nonmyeloablative allogeneic stem cell transplantation (NST) for hematologic diseases.</p><p><b>METHODS</b>A total of 243 hematologic diseases patients received HLA-identical NST were enrolled in this study from 9 transplant centers of NST Cooperative Group in China. Nonmyeloablative conditioning regimen was based on fludarabine (Flud), rabbit anti-human thymocyte globulin (ATG), cyclophosphamide (CTX) (FAC), and plus cytarabine or busulfan (BU) etc. Graft-versus-host disease (GVHD) prophylaxis included cyclosporin A (CsA) and mycophenolate mofetil (MMF).</p><p><b>RESULTS</b>Among the 243 patients, 219 (90.1%) achieved full donor chimerism (FDC), 2(0.8%) engraftment failure. 78 (32.1%) had mixture chimerism (MC) at 4 weeks after NST, out of which 56 switched to FDC, 16 remained MC and 6 (2.5%) developed graft rejection. The incidence of acute GVHD was 34.2%, including 6.6% of grade III-IV acute GVHD. Chronic GVHD developed in 78 (32.1%) patients. The follow-up durations were 3 - 99 months, 162 (66.7%) were still alive and the overall survival rates were 76.5%, 73.9%, 70.7%, and 27.8% for MDS/SAA, chronic myeloid leukemia, acute leukemia at first remission, and refractory or relapsed leukemia, respectively.</p><p><b>CONCLUSIONS</b>The nonmyeloablative allogeneic stem cell transplantation based on FAC conditioning results in sustained engraftment and mild aGVHD, providing a new feasible curative therapy for hematology diseases.</p>

Acute graft versus host disease in non-myeloablative allogeneic stem cell transplantation / 中国实验血液学杂志

The objective of this study was to explore the occurrence and clinical features of acute graft versus host disease (aGVHD) in non-myeloablative stem cell transplantation (NAST). 19 cases developed aGVHD out of 71 cases with NAST in recent years were analyzed retrospectively. Out of 19 cases, 9 males and 10 females at the median age of 38 (18-59), 16 cases with grade I-II aGVHD, 3 cases with grade III-IV aGVHD. The results indicated that the incidence of aGVHD in NAST was 26.7% (19/71), and severe aGVHD was 4.2%, the median onset time was 58 days (17-240 days) after transplantation. Skin and especially the intestine were the main target organs of aGVHD, while diarrhea occurred as the first symptom in 7 cases, 3 cases showed mixed acute and chronic GVHD involving more locations at the same time. aGVHD occurrence was 38.2% in those patients with full donor chimerism (FDC) and 16% in patients with the mixed chimerism (MC). It is concluded that aGVHD in NAST is less in occurrence, lighter in severity and later in time, but higher occurrence in those with early FDC, which intestine and skin are the main target organs. The clinical course is prolonged and easily complicated with severe infection in the later phase. Early combined therapy with powerful supportive treatment is necessary.

Construction and characterization of soluble HLA-A*0201-PR1 complex / 中国实验血液学杂志

This study was aimed to construct the soluble HLA-A*0201-PR1 complex for preparation of HLA-A*0201-PR1 tetramer. The recombinant HLA-A*0201-BSP (BirA substrate peptide) fusion protein as heavy chain and beta(2)-microglobulin (beta(2) m) as light chain were expressed highly as insoluble aggregates in Escherichia coli and then purified with gel filtration, and the final purity reached above 90%. The two subunits were refolded to form an HLA-A*0201-peptide complex by dilution method in the presence of an antigenic peptide PR1, a HLA-A2-restricted peptide from proteinase 3 (aa 169 - 177, VLQELNVTV). Refolded HLA-A*0201-PR1 complex was biotinylated using a BirA enzyme and purified by anion exchange chromatography on a Q-Sepharose (fast flow) column. The extent of reconstitution of the HLA-A*0201-PR1 complex was analyzed by HPLC gel filtration. The refolded and biotinylated products were detected by Western blot and ELISA with monoclonal antibody BB7.2 that recognized the natural conformations of HLA-A2 and streptavidin. The results showed that the refolded complex was composed of HLA-A*0201-BSP aggregate, HLA-A*0201-PR1 complex and beta(2) m, and reconstitution yields of 18% with PR1 was obtained. Refolded HLA-A*0201-PR1 complex could be confirmed by practical immunological method and biotinylated efficiently. It is concluded that the refolding and biotinylation of HLA-A*0201-PR1 complex is successfully obtained. This work provides the basis for the preparation of HLA-A*0201-PR1 tetramer.

Cloning and expression of HLA-A*0201-BSP / 中国实验血液学杂志

High-yield production of HLA-A*0201 heavy chain is a prerequisite to the preparation of HLA-A2 tetramer. The present study was aimed to construct the expression vector of recombinant HLA-A*0201-BSP fusion gene for preparing HLA-A2 tetramers. The extracellular region HLA*0201 was cloned by RT-PCR from HLA-A2(+) donor, and a 15-amino acid biotin-protein ligase (BirA) substrate peptide (BSP) for BirA-dependent biotinylation was added to the COOH-terminus of HLA-A*0201 heavy chain. Then the fusion gene was cloned into pBV220 vector at EcoRI and Bam HI sites and its sequence was confirmed by DNA sequence analysis. The recombinant plasmid pBV220-HLA-A*0201-BSP was transformed to the competent cells of E.coli DH5alpha. The results showed that the HLA-A*0201-BSP fusion protein was successfully expressed in the form of inclusion body and amounted to over 28% of total cell proteins via induction at 42 degrees C. After washed with triton X-100 and urea, the inclusion body was dissolved with 8 mol/L urea and then purified with Sepharcyl S-300 HR, and the final purity reached above 90%. It is concluded that the HLA-A*0201-BSP fusion gene was cloned successfully and expressed efficiently in E.coli DH5alpha. This work establishes a convenient approach for purification of large quantity of recombinant HLA-A*0201-BSP. This provides the basis for the preparation of HLA-A2 tetramers.

Cloning of Human beta2-microglobulin gene and efficient expression in Escherichia coli / 中国实验血液学杂志

Human beta(2)-microglobulin (beta(2)m) is the light chain of major histocompatibility complex (MHC) class I molecule. High-yield production of this protein is a prerequisite to the preparation of MHC class I tetramer. The present study was aimed to obtain recombinant human beta(2)m expressed in Escherichia coli (E. coli) for preparing MHC class I tetramers. For cloning of human beta(2)m gene, a pair of specific primers was designed based on the published sequence of this gene. A 300 bp specific DNA fragment corresponding to the encoding region of beta(2)m lack of the signal peptide sequence was obtained by RT-PCR from the total RNA of human leukocytes. The amplified cDNA was inserted into the IPTG-inducible expression plasmid pET-17b by Nde I and Bam H I sites and its sequence was confirmed by DNA sequence analysis. The recombinant plasmid pET-beta(2)m was transformed to the competent cells of E. coli BL21 (DE3). The results showed that beta(2)m was expressed in the form of inclusion body and amounted to over 32% of total cell proteins after IPTG induction. After washing with triton X-100 and urea, the inclusion body was dissolved with 4 mol/L urea and then purified with Sephacryl S-200 HR, and the final purity reached above 95%. The denatured protein was renatured by dilution method. Western blot assay indicated that the monoclonal antibody against human native beta(2)m could react specifically with the recombinant protein. In conclusion, the human beta(2)m gene was cloned successfully and expressed efficiently in E. coli BL21 (DE3). This work establishes a convenient approach for renaturation and purification of large quantity of recombinant beta(2)m. This provides the basis for the preparation of MHC tetramers.

ABO-incompatible nonmyeloablative allogeneic peripheral blood stem cell transplantation / 中国实验血液学杂志

To explore the effects of ABO incompatibility between recipient and donor on HLA-matched nonmyeloablative allogeneic peripheral blood stem cell transplantation (NAST), a retrospective, cohort study was performed. Among 24 HLA-matched NAST, 15 were major ABO-incompatible and 9 minor. Control group included 24 HLA-matched NAST with ABO-compatible grafts. Nonmyeloablative conditioning regimens consisted of CTX, Ara-C and ATG. The patients were given cyclosporine A and mycophenolate mofetile for prophylaxis of acute GVHD. The ABO-incompatible patients received grafts depleted erythrocytes by hydroxyethyl starch (HES) sedimentation. The results showed that successful and stable engraftment was established in 23 patients. No recipient developed clinically immediate hemolysis during graft infusion, but 2 recipients experienced delayed hemolysis attributable to the ABO incompatibility. The median time of granulocyte counts >0.5 x 10(9)/L and platelet >30 x 10(9)/L was 11 and 14.9 days, respectively. In ABO major incompatible group, the onset of erythropoiesis after NAST was delayed. One out of 10 recipients with blood group "O" in this group developed pure red cell aplasia (PRCA), lasting 5 months. The acute GVHD occurred in 7 out of the 24 patients. The chronic GVHD occurred in 5 of 21 cases. Relapse was observed in 2 patients with acute leukemia. The actuarial probability of disease-free survival at 2 years was 63.3%. In conclusion, ABO-incompatible grafts for NAST have no adverse effect on engraftment, recovery of platelets, incidence of GVHD, relapse rate or survival. ABO-incompatible NAST is fairly safe if there is indication, however, the onset of erythropoiesis is delayed when major ABO mismatched.

Kinetic patterns of cytokine expressions in H-2 haploidentical nonmyeloablative bone marrow transplantation in mice / 中国实验血液学杂志

In order to explore what role relative cytokines play in acute GVHD (aGVHD) of mice transplanted with H-2 haploidentical nonmyeloablative bone marrow, a murine model was established by using C57BL/6J mouse as donor and (C57BL/6J x BALB/C) F1 mouse as the recipient. The recipient mice were given CsA and mycophenolate mofetile (MMF) for prophylaxis of aGVHD. The expression levels of IL-2, INFgamma, IL-4 and IL-10 in the spleen were detected by semi-quantitative RT-PCR at different time points after transplantation. The results showed that the expression levels of these cytokines increased slightly in the group only received nonmyeloablative conditioning. The expression levels of IL-2 and INF-gamma elevated significantly after transplantation in group 2 (without aGVHD prophylaxis), peaked at day 21 and day 14 respectively, then dropped rapidly, returned to the basal levels at day 35. The study on kinetic pattern of expression of IL-2 and INF-gamma in group 3 (with aGVHD prophylaxis) gave a similar result to group 2, but the peak value of cytokine expression in group 3 was much lower than that in group 2. The expression of IL-4 in Group 2 and group 3 all peaked at day 14, but the peak value in group 3 was higher than that in group 2, and decreased slowly in group 3. The expression of IL-10 increased gradually after transplantation, peaked at day 14, decreased after day 21, elevated again until day 35. It is concluded that the expression levels of these cytokines in the spleen all increase after H-2 haploidentical nonmyeloablative bone marrow transplantation. CsA and MMF can reduce the incidence and severity of aGVHD by down-regulating the expression levels of IL-2 and INF-gamma.

The clinical research of nonmyeloablative allogeneic peripheral blood hematopoietic stem cells transplantation for hematological diseases / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the significance of nonmyeloablative allogeneic peripheral blood hematopoietic stem cell transplantation in the treatment of hematological diseases.</p><p><b>METHODS</b>A nonmyeloablative conditioning regimen consisted of CD(3) monoclonal antibody, cyclosporine A, cyclophosphamide and cytarabine was used for allogeneic stem cell transplantation in 33 patients with hematological diseases. Of them, 11 were acute leukemia (AL) in first complete remission (CR(1)), 4 AL-CR(2) approximately 3, 3 refractory AL, 4 severe aplastic anemia (SAA), 7 chronic myeloid leukemia (CML), 2 myelodysplastic syndrome, 1 each of chronic lymphocytic leukemia (CLL) and myelofibrosis.</p><p><b>RESULTS</b>All 33 patients passed the hematopoietic suppression stage smoothly and achieved engraftment of the donor cells. There were 24 cases of full donor chimerism (13 cases converted from mixed chimerism), 4 mixed chimerism (MC) and 5 developed graft rejection. Of the 33 cases, 7 (21.2%) developed acute GVHD and chronic GVHD, 25 (75.8%) still live and 8 (24.2%) died.</p><p><b>CONCLUSIONS</b>Nonmyeloablative allogeneic peripheral blood stem cells transplantation is a safe, less toxic and curative approach for patients with hematological disease.</p>

Comparison of conditioning regimens containing or no fludarabine in nonmyeloablative allogeneic peripheral blood stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic effect of conditioning regimen containing fludarabine in nonmyeloablative allogeneic peripheral blood stem cells transplantation (NAST) in the treatment of hematological diseases.</p><p><b>METHODS</b>Thirty-six patients with acute leukaemia, severe aplastic anaemia, MDS and myelofibrosis received NAST from HLA matched donors' G-CSF mobilized peripheral blood stem cells after nonmyeloabalative conditioning. The conditioning regimen consisted of CTX, Ara-C, CsA, anti-CD(3) antibody or anti-thymocyte globulin and with or without fludarabine. GVHD prophylaxis was performed with cyclosporine combined methotrexate (no MMF group, n = 5) or mycophenolate mofetil (MMF group, n = 31).</p><p><b>RESULTS</b>All of the treatment was generally well tolerated and all cases achieved engrafted of the donor cells. In fludarabine group, engraftment was observed in 87.5% (14/16) patients with complete donor chimerism, graft failure was 12.5% (2/16) and in no fludarabine group, 80% (16/20) and 20% (4/20), respectively. The incidence of acute GVHD (grade I - IV) was 27.8% (10/36) and chronic GVHD 22.2% (8/36). In fludarabine group, grade I - II aGVHD was 37.5%, in no fludarabine group, 20%. cGVHD was 12.5% in fludarabine group and in no fludarabine group 30%, respectively. Interstitial pneumonia (IP) was observed in 16.7% (6/36) of the patients, being 18.7% (3/16) and 15% (3/20) in fludarabine and no fludarabine group, respectively. Overall survival rate was 80.5% (29/36) with a median follow-up of 13 months.</p><p><b>CONCLUSIONS</b>There was no significant difference between fludarabine based (n = 16) and non-fludarabine based conditioning regimen (n = 20) in NAST for the treatment of hematological diseases, regarding for incidence of GVHD, IP, engraftment and survival.</p>

Diagnosis and treatment of invasive pulmonary aspergillosis in malignant hematological diseases-3 cases reported / 中国实验血液学杂志

Invasive pulmonary aspergillosis is difficult to diagnose and a critical ill with high mortality. In this paper, the diagnosis and treatment of invasive pulmonary aspergillosis complicated in 3 cases of hematological malignancy (2 acute leukemias and 1 MDS-RA) were retrospectively analysed. All patients had histories of hypoimmunity and were received prophylactic antifungal treatment. Pulmonary aspergillosis infection still occurred and confirmedly diagnosed by sputum examination. After 7 to 14 days of combination treatment of liposomal amphotericin B, itraconazole and flucytosine, 2 cases were cured and another showed effective. In conclusion, early diagnosis and treatment of invasive pulmonary aspergillosis are very critical and the therapeutic effectiveness of combined scheme with liposomal amphotericin B, itraconazole and flucytosine is very effective for pulmonary aspergillosis.

Nonmyeloablative Allogeneic Peripheral Blood Hematopoietic Stem Cell Transplantation for Therapy of Hematologic Diseases / 中国实验血液学杂志

The objectve of this investigation was to explore the use of nonmyeloablative allogeneic peripheral blood stem cell tranplantation for treatment of hematologic diseases. Six patients were included: 3 cases with acute leukemia in first complete remission (2 AML and 1 ALL), 2 severe aplastic anemias and 1 chronic myeloid leukemia in chronic phase. All of the 6 cases were received HLA-identical siblings donor peripheral blood stem cell transplantation after a nonmyeloablative conditioning. The donor cells were engrafted in all patients (3 cases were full engrafment of donor cells and 3 were mixed chimerism). Hematopoietic recovery was appeared in all of the cases (ANC recovered to more than 0.5 x 10(9)/L and platelet count to more than 30 x 10(9)/L on day 9 to day 21 and day 14 to day 28 after transplantation, respectively). Two patients developed III or IV degree GVHD. Our preliminary results suggest that the procedure is more safe, efficient and less complications than myeloablative conditioning regimens and represents another new approach in the management of patients with hematologic diseases.

The Antitumor Activity of G-CSF-Mobilized Peripheral Blood Mononuclear Cells Activated by IL-2 Alone or in Combination with IL-12 / 中国实验血液学杂志

This study was undertaken to investigate the antitumor activity of rhG-CSF mobilized blood mononuclear cells (G-PBMC) from normal donor activated by rhIL-2 alone or in combination with rhIL-12 in vitro. LDH release assay was used to determine the cytotoxicity activity of G-PBMC against K562 (NK-sensitive) and Raji cells (NK-resistant). The phenotype of the activated G-PBMC was assayed by flow cytometry. Results suggested that the cytotoxicity of the fresh G-PBMC was low before IL-2/IL-12 stimulation. G-PBMC developed marked cytotoxicity after 24 hours of IL-2 activation, with more significant increase for further 48 hours induction. When G-PBMCs were exposed to combination of IL-2 and IL-12, the cytotoxicity was significantly enhanced. The proportions of CD3(+) T cells and CD8(+) T cells were increased when G-PBMC incubated with IL-2 for 72 hours. However, CD56(+) cells were significantly elevated when G-PBMC exposed to IL-2 and IL-12 for 7 days. It is concluded that G-PBMC activated by IL-2 had evident antitumor activity, which was further increased when IL-2 in combination with IL-12. These results demonstrate that IL-2 and IL-12 treatment in vitro might promote the graft-versus-leukemia (GVL) response of G-PBMC.