BACKGROUND:Allogeneic hematopoietic stem cell transplantation is currently the most effective method for the radical treatment of thalassemia major,but only half of patients can find compatible bone marrow or peripheral blood stem cells.Sib-derived umbilical cord blood stem cells have different characteristics from bone marrow and peripheral blood stem cells,and are a potential alternative source of hematopoietic stem cells for transplantation in patients with thalassemia major.OBJECTIVE:To investigate the therapeutic effect of human leukocyte antigen matched sibling fresh umbilical cord blood transplantation in the treatment of β-thalassemia major in children.METHODS:Forty-eight children with β-thalassemia major,including 28 males and 20 females,with a median age of 4 years old,were selected from Nanfang Hospital of Southern Medical University from June 2010 to June 2020.All of them received fresh cord blood transplantation from human leukocyte antigen matched sibling.Transplantation conditioning adopted a myeloablative regiment without anti-thymocyte globulin.A combination of cyclosporine A and mycophenolate mofetil with or without short-range methotrexate was administered for graft-versus-host disease.RESULTS AND CONCLUSION:(1)The median infused doses of total nucleated cells and CD34+cells were 8.17×107/kg and 2.40×105/kg,respectively in 48 children.The median follow-up time after cord blood transplantation was 98 months,and 44 cases were successfully engrafted.The median time to neutrophil and platelet engraftment was 28 and 31 days,respectively.Among them,37 cases were found to be donor-type complete chimerism detected as evidence of implantation after transplantation,7 cases were found to be stable mixed chimerism.(2)Among the 44 children with successful implantation,four patients developed acute graft-versus-host disease,and were scored as grade Ⅰ(n=2)and grade Ⅱ(n=2).All the affected organs were skin,and no chronic graft-versus-host disease occurred.(3)After umbilical cord blood transplantation,cytomegalovirus infection and activation occurred in 5 of the 48 cases,sepsis in 12 cases,invasive fungal disease in 3 cases,stomatitis in 21 cases,hemorrhagic cystitis in 8 cases,and hepatic vein occlusion in 1 case.(4)Among 48 children,47 patients survived;1 died of severe pneumonia combined with acute heart failure 28 days after transplantation;43 survived without disease;3 had primary implantation failure,and 1 had pancytopenia after transplantation.The 5-year probabilities of overall survival and disease-free survival were 98％and 89％,respectively.The cumulative incidence of transplant-related deaths at 1 year was 2.1％.(5)The above results indicate that human leukocyte antigen matched sibling fresh umbilical cord blood transplantation is effective in the treatment of β-thalassemia major in children with a low incidence of graft-versus-host disease.

BACKGROUND:Allogeneic hematopoietic stem cell transplantation is currently the most effective method for the radical treatment of thalassemia major,but only half of patients can find compatible bone marrow or peripheral blood stem cells.Sib-derived umbilical cord blood stem cells have different characteristics from bone marrow and peripheral blood stem cells,and are a potential alternative source of hematopoietic stem cells for transplantation in patients with thalassemia major.OBJECTIVE:To investigate the therapeutic effect of human leukocyte antigen matched sibling fresh umbilical cord blood transplantation in the treatment of β-thalassemia major in children.METHODS:Forty-eight children with β-thalassemia major,including 28 males and 20 females,with a median age of 4 years old,were selected from Nanfang Hospital of Southern Medical University from June 2010 to June 2020.All of them received fresh cord blood transplantation from human leukocyte antigen matched sibling.Transplantation conditioning adopted a myeloablative regiment without anti-thymocyte globulin.A combination of cyclosporine A and mycophenolate mofetil with or without short-range methotrexate was administered for graft-versus-host disease.RESULTS AND CONCLUSION:(1)The median infused doses of total nucleated cells and CD34+cells were 8.17×107/kg and 2.40×105/kg,respectively in 48 children.The median follow-up time after cord blood transplantation was 98 months,and 44 cases were successfully engrafted.The median time to neutrophil and platelet engraftment was 28 and 31 days,respectively.Among them,37 cases were found to be donor-type complete chimerism detected as evidence of implantation after transplantation,7 cases were found to be stable mixed chimerism.(2)Among the 44 children with successful implantation,four patients developed acute graft-versus-host disease,and were scored as grade Ⅰ(n=2)and grade Ⅱ(n=2).All the affected organs were skin,and no chronic graft-versus-host disease occurred.(3)After umbilical cord blood transplantation,cytomegalovirus infection and activation occurred in 5 of the 48 cases,sepsis in 12 cases,invasive fungal disease in 3 cases,stomatitis in 21 cases,hemorrhagic cystitis in 8 cases,and hepatic vein occlusion in 1 case.(4)Among 48 children,47 patients survived;1 died of severe pneumonia combined with acute heart failure 28 days after transplantation;43 survived without disease;3 had primary implantation failure,and 1 had pancytopenia after transplantation.The 5-year probabilities of overall survival and disease-free survival were 98％and 89％,respectively.The cumulative incidence of transplant-related deaths at 1 year was 2.1％.(5)The above results indicate that human leukocyte antigen matched sibling fresh umbilical cord blood transplantation is effective in the treatment of β-thalassemia major in children with a low incidence of graft-versus-host disease.

Diabetes mellitus (DM) is a major metabolic disease endangering global health, with diabetic nephropathy (DN) as a primary complication lacking curative therapy. Sporoderm-broken spores of Ganoderma lucidum (GLP), an herbal medicine, has been used for the treatment of metabolic disorders. In this study, DN was induced in Sprague-Dawley rats using streptozotocin (STZ) and a high-fat diet (HFD), and the protective mechanisms of GLP were investigated through transcriptomic, metabolomic, and network pharmacology (NP) analyses. Our results demonstrated that GLP intervention ameliorated renal damage and inflammation levels in DN rats. Integrative metabolomic and transcriptomic analysis revealed that GLP treatment modulated glucose and cellular energy metabolisms by regulating relevant genes. GLP significantly suppressed the inflammations by impacting glucose and energy metabolism-related gene expression (Igfbp1 and Angptl4) and enhanced metabolic biomarkers of 4-Aminocatechol. In addition, NP analysis further indicated that GLP may efficiently alleviate DN via immune-related pathways. In conclusion, this study provides supportive evidence of the anti-inflammatory effects of GLP supplements, highlighting their potential for promising clinical applications in treating DN.

Diabetes mellitus(DM)is a major metabolic disease endangering global health,with diabetic ne-phropathy(DN)as a primary complication lacking curative therapy.Sporoderm-broken spores of Ganoderma lucidum(GLP),an herbal medicine,has been used for the treatment of metabolic disorders.In this study,DN was induced in Sprague-Dawley rats using streptozotocin(STZ)and a high-fat diet(HFD),and the protective mechanisms of GLP were investigated through transcriptomic,metabolomic,and network pharmacology(NP)analyses.0ur results demonstrated that GLP intervention ameliorated renal damage and inflammation levels in DN rats.Integrative metabolomic and transcriptomic analysis revealed that GLP treatment modulated glucose and cellular energy metabolisms by regulating relevant genes.GLP significantly suppressed the inflammations by impacting glucose and energy metabolism-related gene expression(Igfbp1 and Angptl4)and enhanced metabolic biomarkers of 4-Aminocatechol.In addition,NP analysis further indicated that GLP may efficiently alleviate DN via immune-related pathways.In conclusion,this study provides supportive evidence of the anti-inflammatory effects of GLP supplements,highlighting their potential for promising clinical applications in treating DN.

【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.

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.

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). Secondly , Bus ulfan had no effect in CD19?CAR?T cells in vitro(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.

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.

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.

OBJECTIVETo 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).

METHODSFrom 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).

RESULTSFourteen 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*.

CONCLUSIONMagnetic 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.

Adolescent ; Adult ; Child ; Female ; Ferritins ; blood ; Humans ; Iron ; metabolism ; Iron Overload ; diagnosis ; metabolism ; pathology ; Liver ; metabolism ; Magnetic Resonance Imaging ; Male ; Myocardium ; metabolism ; Young Adult ; beta-Thalassemia ; diagnosis ; metabolism ; pathology