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【Objective】 To evaluate the safety and efficacy of the collection of peripheral blood stem cells (PBSCs) in pediatric patients with thalassemia major (TM) weighing 20 kg or less. 【Methods】 PBSCs collection data of 170 pediatric patients with TM weighing 20 kg or less from January 2013 to December 2020 in our center were reviewed. Safety was assessed by the occurrence of adverse events during apheresis procedures, and efficacy was evaluated by the number of CD34+ cells collected. 【Results】 A total of 171 PBSCs procedures were performed on 170 patients with TM weighing 20 kg or less, with a median age of (4.98±1.53) years and a median weight of (17.30±2.18) kg. The probability of collecting at least 1×106 CD34+ cells/kg during a single course of apheresis was 99.41% (169/170), with a median (5.88±4.23) ×106 CD34+ cells collected per kg of weight of the recipient. A minimum pre-apheresis hemoglobin (Hb) of 60 g/L in patients with TM weighing 20 kg or less was safe and feasible. The most common adverse event of G-CSF mobilization in TM patients is bone pain, with the incidence of 7.65% (13/170), which was higher than that of healthy children donors in our center. The most common adverse events during the collection were pain at the puncture site of the femoral vein (6.47%, 11/170) and low pressure of the fluid (2.92%, 5/170). And no serious complications related to PBSCs mobilization, central venous catheter(CVC)placement or the apheresis procedure occurred. 【Conclusion】 PBSCs collection by COM.TEC blood cell separator in children weighing 20 kg or less is safe and efficacious.
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Objective@#To analyze the risk factors of varicella zoster virus(VZV) infection after allogeneic hematopoietic stem cell transplantation(allo-HSCT) in children, and to provide reference for the diagnosis, monitoring and prophylaxis of VZV infection after allo-HSCT.@*Methods@#A total of 367 patients, who underwent allo-HSCT in Pediatric Transplantation Center of Nanfang Hospital Affiliated to Southern Medical University from January 2012 to June 2015 were collected.Clinical characteristics and risk factors of the patients complicated with VZV after allo-HSCT were retrospectively analyzed.@*Results@#Thirty-four patients (9.26%) were complicated with VZV infection after allo-HSCT.The median onset time was 96.5 d(19-326 d). Two of 34 patients relapsed 3 times, 1 case of them relapsed twice, 3 cases of them relapsed once, and 4 cases of 34 patients were complicated with VZV encephalitis.All cases were treated with antivirus drugs, infusion of immunoglobulin, reduction of immunosuppressant dosages, with external use of Acyclovir ointment.The median therapy time was 13 days (7-28 days). All of their herpes subsided, and neurological symptoms such as headache, vomiting and convulsion disappeared.VZV-DNA both in blood and cerebrospinal fluid turned negative.No patient had herpetic dissemination and visceral involvement, and no one died directly of VZV infection.Results indicated that age(χ2=6.863, P=0.009), underlying disease(χ2=14.793, P=0.022), type of HSCT(χ2=14.459, P=0.001) and resource of stem cell (χ2=20.585, P=0.002) were significant risk factors for VZV infection after allo-HSCT, while sex (χ2=0.106, P=0.745) and antithymocyte globulin in conditioning regimen(χ2=0.010, P=0.921) had no relation to it.@*Conclusions@#VZV infection mainly occur within 12 months after allo-HSCT and is prone to be complicated with VZV encephalitis.The effect of Acyclovir is good.Monitoring and prophylaxis of VZV infection after allo-HSCT should be strengthened in children with high risk factors.
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Objective To analyze the risk factors of varicella zoster virus( VZV)infection after allogeneic hematopoietic stem cell transplantation(allo－HSCT)in children,and to provide reference for the diagnosis,monitoring and prophylaxis of VZV infection after allo－HSCT. Methods A total of 367 patients,who underwent allo－HSCT in Pediatric Transplantation Center of Nanfang Hospital Affiliated to Southern Medical University from January 2012 to June 2015 were collected. Clinical characteristics and risk factors of the patients complicated with VZV after allo －HSCT were retrospectively analyzed. Results Thirty－four patients(9. 26﹪)were complicated with VZV infection after allo－HSCT. The median onset time was 96. 5 d(19－326 d). Two of 34 patients relapsed 3 times,1 case of them relapsed twice,3 cases of them relapsed once,and 4 cases of 34 patients were complicated with VZV encephalitis. All cases were treated with antivirus drugs,infusion of immunoglobulin,reduction of immunosuppressant dosages,with exter﹣nal use of Acyclovir ointment. The median therapy time was 13 days(7－28 days). All of their herpes subsided,and neurological symptoms such as headache,vomiting and convulsion disappeared. VZV－DNA both in blood and cerebro﹣spinal fluid turned negative. No patient had herpetic dissemination and visceral involvement,and no one died directly of VZV infection. Results indicated that age(χ2 ﹦6. 863,P﹦0. 009),underlying disease( χ2 ﹦14. 793,P﹦0. 022),type of HSCT(χ2 ﹦14. 459,P﹦0. 001)and resource of stem cell(χ2 ﹦20. 585,P﹦0. 002)were significant risk factors for VZV infection after allo－HSCT,while sex(χ2 ﹦0. 106,P﹦0. 745)and antithymocyte globulin in conditioning regimen ( χ2 ﹦0. 010,P﹦0. 921)had no relation to it. Conclusions VZV infection mainly occur within 12 months after allo－HSCT and is prone to be complicated with VZV encephalitis. The effect of Acyclovir is good. Monitoring and prophylaxis of VZV infection after allo－HSCT should be strengthened in children with high risk factors.
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Objective The research about the effect of different chemotherapeutic drugs on CD19?CAR?T cells with CCK8 test to provide the theoretical support about the development of chemotherapy for clinical support. Methods Extract T cells from a normal adult peripheral blood and synthesize CD19?CAR?T cell. CD19?CAR?T cells were treated with different doses of chemotherapeutic drugs for 24,48,72 h and(or)96 h,and inhibition rate was calculated. Results First,we observed that the inhibition rates of fludarabine and Mafosfamide for CD19?CAR?T cells were increasing with the time and concentration (P 0.05). Finally ,Cyclophosphamide had no effect in CD19?CAR?T cells in vitro (P > 0.05). Conclusion Mafosfamide and Fludarabine can inhibit the CD19?CAR?T cells. Cyclophosphamide have no activity in vitro.
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Objective To investigate the autophagy and apoptosis in acute myelogenous leukemia U937 cell induced by Sirolimus.Methods U937 cells were subcultured, and blank control group(normal) and Sirolimus treated groups(12 h, 24 h,48 h) were established.The Sirolimus treated groups were treated by 2 μmol/L concentration of Sirolimus for 12 h,24 h and 48 h, respectively.The cell morphology of U937 cells treated by Sirolimus was observed after 12 h,24 h and 48 h.The survival rate of cells was detected by cell counting kit-8 method.Cell apoptosis was detected by flow cytometry using Annexin V-FITC/PI double labeled.Real-time PCR was used to detect the level of mRNA expression in autophagy specific protein maker mictotubule-associated protein light chain 3 (LC3)-Ⅱ in different treated times by Sirolimus.Sirolimus LC3 protein expression levels after treatment were detected by Western blot method.Results Under inverted microscope, the cell number of Sirolimus treatment group reduced gradually after 12 h ,24 h and 48 h culture, volume of cells became smaller, cells got ruptured, and the nucleus pycnosis and cellular debris increased.With the extension of time, U937 cells survival rate was falling, and there was statistical differences compared with those of the control group(P =0.031).With Sirolimus treatment, U937 cells after 12 h,24 h and 48 h, U937 cell apoptosis rate increased, and there were statistically significances, compared with those of the control group (P =0.027).With Sirolimus treatment U937 cells after 12 h,24 h and 48 h,LC3-Ⅱ mRNA expression and protein expression were down-regulated compared with those of the control group, and there were statistically significances (P =0.029).Conclusions Sirolimus can induce autophagy and apoptosis in U937 cells.Autophagy protein LC3-Ⅱ in gene and protein expression levels were lowered, and LC3-Ⅱ may play an important role in regulating the leukemia cell autophagy.
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BACKGROUND:Cytokines play an important role in the occurrence and development of graft-versus-host disease, but there is a current lack of reports on the association between cytokines and graft-versus-host disease after al ogeneic hematopoietic stem cel transplantation for treatment ofβ-thalassemia major. OBJECTIVE:To investigate the association between cytokines and graft-versus-host disease after al ogeneic hematopoietic stem cel transplantation forβ-thalassemia major. METHODS:We observed the dynamic variation of interleukin 6, interleukin 8, interleukin 12, tumor necrosis factor-αand macrophage migration inhibitory factor in 11 children withβ-thalassemia major before onset of graft-versus-host disease, when graft-versus-host disease occurred, at days 4 and 7 after onset of graft-versus-host disease, and when graft-versus-host disease disappeared. RESULTS AND CONCLUSION:There was a significant difference in serum levels of interleukin-6, interleukin-12, tumor necrosis factor-α, macrophage migration inhibitory factor in different time points, and the highest levels of different cytokines appeared when graft-versus-host disease occurred, fol owed by those at 7 days after graft-versus-host disease. There was a significant difference in serum levels of interleukin-8 in different time points, and the highest level appeared at 4 days after graft-versus-host disease. The dynamic expression of interleukin-6, interleukin-8, interleukin-12, tumor necrosis factor-α, macrophage migration inhibitory factor can estimate the immune function ofβ-thalassemia major patients who develops graft-versus-host disease after al ogeneic hematopoietic stem cel transplantation, and can be used as the immunobiology indicators for the early diagnosis of graft-versus-host disease.
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<p><b>OBJECTIVE</b>To assess the value of magnetic resonance imaging T2* tests in the detection of myocardial and liver iron overload in patients with β-thalassemia major (β-TM).</p><p><b>METHODS</b>From 2010 to 2011, 28 β-TM patients over 10 years old under blood transfusion therapy and chelation care with serum ferritin (SF)>1000 µg/L underwent myocardial and liver MRI T2* tests on a voluntary basis. The results were analyzed in relation with age, SF, and left ventricular ejection fraction (LVEF).</p><p><b>RESULTS</b>Fourteen out of the 28 cases (50%) were found to have myocardial iron overload, including 7 severe cases, 2 moderate cases, and 5 mild cases. All the 28 cases had liver iron overload, including 2 mild cases, 7 moderate cases, and 19 severe cases. Two out of the 28 cases had lowered LVEF (7.14%), and one of them had severe myocardial iron overload. There was a negative correlation between myocardial MRI T2* and SF (r=-0.479, P=0.01). Myocardial MRI T2* was positively correlated with liver MRI T2* (r=0.378, P=0.047). Age was not significantly correlated with SF, LVEF, or liver MRI T2*.</p><p><b>CONCLUSION</b>Magnetic resonance imaging (T2*) detection is an effective and non-invasive means for detecting myocardial and liver iron overload in patients with β-thalassemia major receiving blood transfusion. T2* combined with SF is the main diagnostic indicator to assess iron overload in the vital organs.</p>
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Young Adult , Ferritins , Blood , Iron , Metabolism , Iron Overload , Diagnosis , Metabolism , Pathology , Liver , Metabolism , Magnetic Resonance Imaging , Myocardium , Metabolism , beta-Thalassemia , Diagnosis , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of different doses of antithymocyte globubin-fresenius (ATG-F) for allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with beta-thalassemia Major.</p><p><b>METHODS</b>Sixty-four children with beta-thalassemia major undergoing allo-HSCT were divided into two equal groups to receive ATG-F pretreatments at high (30 mg/kg) or low (15 mg/kg) doses as part of the conditioning regimen including mainly cyclophosphamide, busulfan, fludarabine, and thiotepa. The outcomes of the patients were compared between the two groups.</p><p><b>RESULTS</b>No obvious difference were noted in the time to leukocyte and platelet engraftment between the two groups. The incidence of grade II-IV acute graft-versus-host disease (aGVHD) appeared to be higher in the low-dose group than in the high-dose group (12.5% vs 9.4%). The incidence of grade III-IV aGVHD was also higher in the low dose group (12.5% vs 6.3%), but the difference was not statistically significant. Application of high-dose ATG-F was associated with a higher rate of probable and possible fungal infection (P<0.05).</p><p><b>CONCLUSION</b>The two doses of ATG-F is feasible as a part of the conditioning regimen for allo-HSCT in children with beta-thalassemia major.</p>
Subject(s)
Adolescent , Animals , Child , Child, Preschool , Female , Humans , Male , Rabbits , Antilymphocyte Serum , Allergy and Immunology , Hematopoietic Stem Cell Transplantation , Methods , Lymphocytes , Allergy and Immunology , Transplantation Conditioning , Methods , beta-Thalassemia , Allergy and Immunology , General SurgeryABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of human cytomegalovirus (HCMV) infection on T lymphocyte subsets in children with β-thalassemia major (TM) during the initial 6 months after allogeneic hematopoietic stem cell transplantation (Allo-HSCT).</p><p><b>METHODS</b>From January, 2010 to January, 2011, 35 children with TM underwent Allo-HSCT. Peripheral blood samples were obtained from the children 6 month after the transplantation to examine the changes of T lymphocytes subsets in relation to HCMV seropositivity.</p><p><b>RESULTS</b>Thirteen children were found seropositive and 22 were seronegative for HCMV. The HCMV-seropositive children had a higher CD8⁺ cell percentage but a lower CD4⁺ cell percentage than those without HCMV infection. Compared with those seronegative for HCMV, the children with HCMV seropositivity showed increased percentages of CD8⁺ cells and CD8⁺CD28⁻ cells with a decreased percentage of CD8⁺CD28⁺ cells. A positive linear correlation was found between the percentages of CD8⁺CD28⁻ cells and CD8⁺ cells.</p><p><b>CONCLUSION</b>HCMV infection can lead to the accumulation of CD8⁺CD28 cells to cause increased CD8⁺ T cells in the peripheral blood in TM children after Allo-HSCT. The percentages of CD8⁺CD28⁻ cells has a positive linear correlation to that of CD8⁺ cells.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , CD8-Positive T-Lymphocytes , Allergy and Immunology , Cytomegalovirus , Cytomegalovirus Infections , Allergy and Immunology , Hematopoietic Stem Cell Transplantation , Postoperative Period , T-Lymphocyte Subsets , beta-Thalassemia , Allergy and Immunology , General Surgery , VirologyABSTRACT
OBJECTIVE: To investigate the curative effect of combined transplantation of bone marrow and umbilical cord blood of same sibling in children with β-thalassemia major.METHODS: Eight thalassemia major patients undergoing combined transplantation of bone marrow and umbilical cord blood of same sibling aged from 4.0 to 7.5 years, 5 boys and 3 girls, were recruited at the Department of Pediatrics, Nanfang Hospital,Southem Medical University from January 2005 to March 2009. The patients were classified into three classes according to Pesarothalassamia classification, class Ⅰ to class Ⅱ 7 cases and class Ⅲ 1 case. Donors ranged 1-4 years received 10 μg/kg per day of subcutaneous granulocyte colony-stimulating factor (G-CSF) for 5 consecutive days. Bone marrow was harvested on the fifth day. Bone marrow and umbilical cord blood of the same sibling then were transfused into the patient.RESULTS: Recovery of hematopoiesis was gained in all patients 4 weeks following transplantation. Seven patients suffered from infection of different degree. Four patients developed mild venous occlusive disease. Two patients developed grade Ⅰ acute graft-versus-host disease (GVHD), and one developed grade Ⅰ chronic GVHD. Seven patients were alive and one died of pulmonary infection and heart failure 32 days following transplantation.CONCLUSION: Combined transplantation of granulocyte colony-stimulating factor primed bone marrow and umbilical cord blood of same sibling in children with β-thalassemia major is safe and effective with promising results. However, complications should be paid high attention following transplantation.