Objective To explore the diagnosis and treatment of Mycobacterium tuberculosis infection in a child with severe β Mediterranean anemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods The clinical data of a child with severe β mediterranean anemia who had Mycobacterium tuberculosis after allo-HSCT were retrospectively analyzed.The pertinent literatures were reviewed.Results Six-year-old girl with Mediterranean anemia was infected by Mycobacterium tuberculosis after allo-HSCT.After anti tuberculosis treatment by HRZE (isoniazid,rifampicin,pyrazinamide and ethambutol),the condition was improved.Conclusion It is rare of Mycobacterium tuberculosis infection after allo-HSCT,which needs timely diagnose and treatment.

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.

0.05).Conclusion ABO-incompatible allo-PBSCT is fairly safe if there is indication. There was no influence on engraftment, incidence of GVHD or prognosis. However, an O to A group transplantation contributes significantly to the post-transplant PRCA.

BACKGROUND: Currently the hematopoietic stem cells can be obtained from bone marrow, peripheral blood and cord blood, so it is expected to search a new source of stem cells in order to satisfy the clinical transplantation needs. From the 5th week of pregnancy, the blood sinusoid system develops completely in liver, and then hematopoietic stem cells can move with blood flow. OBJECTIVE: To observe the biological features of human fetal blood hematopoietic stem/progenitor cells (HS/PCs), and their transplantation into non-obese diabetic/severe combined immunodeficiency disease (NOD/ SCID) mice. DESIGN: Control trial. SETTING: Department of Hematology, First Affiliated Hospital of Guangxi Medical University. MATERIALS:①Cell resource: Twenty-one fetal blood samples were from dead fetus [gestational age of 18-29 weeks, mean (24.2±3.2) weeks] and twenty-one full-term cord blood samples were provided from the Department of Obstetrics, First Affiliated Hospital of Guangxi Medical University between October 2002 and February 2003, with the consent of their relatives.②Experimental animal: Twelve NOD/SCID female mice of 6-7 weeks old were bred in sterility and super-clean operation board. METHODS: Flow cytometer was used to assess cell surface markers of HS/PCs including CD34, CD38, HLA-DR and CD90 in 21 human fetal blood samples, and their expressions were compared with 21 human cord blood samples. Moreover, human fetal blood mononuclear cells (MNCs) were transplanted into 6 NOD/SCID mice irradiated sublethally. After 5 weeks, human leukocytic content was also detected in bone marrow of mice with flow cytometer while human Cart-1 gene in recipients' bone marrow was sensed with polymerase chain reaction (PCR).MAIN OUTCOME MEASURES: ① Expressions of HS/PCs surface markers in fetal blood and cord blood. ②Implantation of fetal blood cells into NOD/SCID mice.RESULTS: ①The percentage of CD34+ cells in fetal blood was significantly higher than that of full-term cord blood [(2.258 8±0.720 9)%,(1.572 9±0.478 3)%, P=0.000 4]. The percentages of CD34+CD38- cells and CD34+CD90+ cells in fetal blood were also higher than those of fullterm cord blood [(1.298 6±0.470 6)%, (0.871 0±0.409 5)%, P=0.001 6;(0.930 0±0.469 2)%, (0.560 0±0.365 8)%, P=0.032 4].②Four cases (4/6)of human fetal blood MNCs smoothly transplanted the hematopoiesis of sublethally irradiated NOD/SCID mice. Five weeks after the transplantation, human leukocyte and Cart-1 gene could still be detected in marrow cells of NOD/SCID mice.CONCLUSION: Human fetal blood contains more HS/PCs than cord blood. Human fetal blood MNCs can engraft bone marrow of NOD/SCID mice and reconstitute general hemopoiesis of marrow and lymph systems.Human fetal blood is a new possible source of pluripotential hematopoietic stem cell.

Objective To study the apoptosis mechanism of K562 cell lines induced by mangiferin. Methods The mRNA expression levels of apoptosis-related genes including bcl-2, bax, survivin of K562 cells treated by mangiferin (25—200 ?mol/L) for 24, 48, 72, and 96 h were determined by RT-PCR; the BCR/ABL protein P210 level was detected by Western blotting. Results Mangiferin up-regulated bax gene of K562 cells significantly and down-regulated bcl-2 gene slightly, resulting in an enhancement of the ratio of bax/bcl-2. Mangiferin down-regulated the expression levels of P210 in K562 cells in a time-and concentration-dependent manner and so is the expression level of survivin mRNA in K562 cells. ConclusionThe mechanism of mangiferin-induced apoptosis in K562 leukemic cells might be involved in up-regulating the gene expression of bax and down-regulating the mRNA expression of BCR/ABL protein P210, bcl-2, and survivin.

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.

OBJECTIVETo investigate the relationship between the expression of cyclin A and drug resistance in patients with adult acute leukemias.

METHODSThe mRNA expressions of cyclin A, mdr1, Topo II alpha and bcl-2 were measured in 64 patients with adult acute leukemia (AL) and 20 normal subjects by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR).

RESULTS(1) The cyclin A and Topo II alpha mRNA expression levels in the treatment resistant group were significantly lower than that in the sensitive group (P < 0.01). There was no cyclin A mRNA expression in the 20 normal subjects under the same experiment condition. (2) The mdr1 and bcl-2 mRNA expression levels in the resistant group were significantly higher than that in sensitive group (P < 0.01). (3) Cyclin A and Topo II alpha gene expression levels were positively correlated (r = 0.794, P = 0.000) in the 64 AL patients, and there was a negative correlation between the gene expression levels of cyclin A and mdr1 (r = -0.337, P = 0.029) in the drug resistant group. (4) Ten AL patients with low expressions of both cyclin A and Topo II alpha were all in the resistant group. Logistic regression of binary analysis showed a significant correlation between the low expression of cyclin A and drug resistance.

CONCLUSIONLow expression of cyclin A gene might be a unfavorable prognostic factor for patients with AL and measurement of both cyclin A and Topo II alpha gene expression would predict drug resistance for AL patients.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Acute Disease ; Adult ; Antigens, Neoplasm ; Cyclin A ; genetics ; metabolism ; DNA Topoisomerases, Type II ; genetics ; metabolism ; DNA-Binding Proteins ; Drug Resistance, Multiple ; genetics ; physiology ; Drug Resistance, Neoplasm ; physiology ; Female ; Humans ; Leukemia ; genetics ; metabolism ; Logistic Models ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; biosynthesis

In order to investigate the relationship between the expression of cyclin A and drug resistance in adult patients with acute leukemia (AL), the mRNA expression of cyclin A, mdr1, Top II alpha, bcl-2 was detected in 64 adult patients with AL and 20 normal controls by semi-reverse transcription polymerase chain reaction (semi-RT-PCR). It was found that the cyclin A and Top II alpha mRNA expression levels in drug resistant group were significantly lower than in sensitive group (P < 0.01). Under the same experimental condition no cyclin A mRNA expression was detectable in all normal controls. The mdr1 and bcl-2 mRNA expression levels in resistant group were significantly higher than in sensitive group (P < 0.01), cyclin A and Top II alpha gene expression levels were closely correlated (rs = +0.794, P = 0.000, n = 64) in all AL patients, but cyclin A was not correlated with mdr1 and bcl-2 gene expression levels. In drug resistant group there was a negative correlation between the gene expression levels of cyclin A and mdr1 (rs = -0.337, P = 0.029). The 10 AL patients with positive lower expression of both cyclin A and Top II alpha were all resistant to drugs. Logistic regression of Binary analysis showed the correlation between the lower expression of cyclin A and drug resistance. It was concluded that lower expression of cyclin A gene might be an unfavorable prognostic factor for patients with AL, and detection of both cyclin A and Top II alpha gene expression would predict drug resistance in AL patients.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Adolescent ; Adult ; Aged ; Cyclin A ; biosynthesis ; genetics ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Genes, MDR ; genetics ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Leukemia, Promyelocytic, Acute ; genetics ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; RNA, Messenger ; biosynthesis ; genetics

BACKGROUND:In recent years, the application of stem cel s to treat autoimmune diseases has become a hot spot. But, studies on umbilical cord blood mesenchymal stem cel s transplantation for the treatment of polymyositis/dermatomyositis are rarely reported. OBJECTIVE:To explore the immunologic mechanism of Th cytokines on the occurrence and development of polymyositis/dermatomyositis by observing the changes in serum interferon-γ, interleukin-4 and interleukin-17 in patients after umbilical cord blood mesenchymal stem cel s transplantation. METHODS:Eighty-one polymyositis/dermatomyositis patients were selected and divided into conventional therapy group (n=44) undergoing glucocorticoid and immunosuppressants therapy and cel transplantation group (n=37) undergoing intravenous infusion of umbilical cord blood mesenchymal stem cel s at a density of (3.5-5.2 )×107 . Dosing regimen was same in the two groups. After fol ow-up of 1, 3, 6 months, the changes of creatine kinase and myodynamia were evaluated;after fol ow-up of 3 and 6 months, lung imaging was evaluated;in the cel transplantation group, interferon-γ, interleukin-4 and interleukin-17 levels were detected before treatment and at 3 and 6 months after treatment. RESULTS AND CONCLUSION:At 1, 3, 6 months after treatment, the creatine kinase level was significantly decreased, and the muscle force grade was significantly increased in both groups (both P<0.001). Compared with the conventional therapy group, the creatine kinase level was lower and the muscle force grade was higher in the cel transplantation group (both P<0.001). Results from lung imaging test showed a remarkable improvement after cel transplantation, and it indicated that umbilical cord blood mesenchymal stem cel s transplantation had good stability. At 6 months after transplantation, the level of interferon-γwas significantly increased, while the interleukin-4 level was decreased significantly (both P<0.01);at 3, 6 months after cel transplantation, the levels of interleukin-17 were significantly decreased (P<0.01). Levels of interleukin-4 and interleukin-17 were positively correlated with the level of creatine kinase at 6 months after cel transplantation (r=0.467, 0.488, both P<0.05), but there was no obvious correlation between the levels of interferon-γand creatine kinase (r=0.213, P>0.05). These findings indicate umbilical cord blood mesenchymal stem cel s transplantation combined with glucocorticoid and immunosuppressants therapy can adjust immune network effects and improve the immune tolerance in polymyositis/dermatomyositis patients, which is safe and effective.