Adolescent ; Anemia, Aplastic ; blood ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Lymphocyte Subsets ; immunology ; Male ; Retrospective Studies ; Severity of Illness Index ; T-Lymphocytes, Cytotoxic ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

Objective To investigate the effects of in vivo electroporation on plasmid mediated reporter gene expression and immunogenicity of DNA vaccine. Methods Luciferase expression plasmid was administered intramuscularly to BALB/c mice at 8μg and 40μg dosage level through injection with or without eletroporation Luciferase expression level in murine muscle was detected by IVIS imaging system 24 h after injection. DNA vaccine plasmid p1.0-gp1455m carrying codon-optimized env gene of CN54 strain ( HIV-1 CRF07_BC) was administered to mice at dosages of 8μg and 40μg through the two approaches mentioned above. Mice were immunized at week 0,2 and 4. Env-specific immune responses were detected at two weeks post the second and the third vaccinations. Env-specific antibody immune responses were determined by ELISA. Euv-specific cellular immune responses were determined by IFN-γ ELISPOT. Results Luciferase expression level in murine muscle was significantly increased as much as 35 folds through in vivo eletroporation. Results of ELISA and ELISPOT revealed that in vivo eletroporation could significantly enhance both the humoral and cellular immune responses induced by DNA vaccination. The responses induced by electrodelivered p1.0-gp1455m at 8 μg dosage were better than those induced by simple intramuscular injection with 40 μg of plasmid DNA. On the other hand, 2 injections followed by electroporation elicited comparable level of humoral and cellular immune responses with those induced by 3 injections without electroporation. Conclusion In vivo electroporation was capable of enhancing both the plasmid-mediated gene expression and immunogenicity of DNA vaccine.

Objective To explore the changes of CD40 and CD40 ligand(CD40L) levels and investigate their significances in children with graft-versus-host disease(GVHD)after related-donor human leukocyte antigen(HLA) matching allogeneic hematopoietic stem cell transplantation(HSCT).Methods Nineteen patients with ?-thalassemia major and 1 patient with congenital inherent hemolytic anemia accepted umbilical cord blood transplantation(UCBT) and allogeneic peripheral blood stem cell transplantation(allo-PBSCT),respectively,and all cases were received successfully related-donor human leukocyte antigen matching allogeneic HSCT.Peripheral blood samples were obtained before and after transplantation,the time when GVHD happened,the expressions of CD40 and CD40L were measured by using immunofluresence asssy.Results Four UCBT children and 3 allo-PBSCT children had no acute GVHD.Thirteen children had acute GVHD(degreeⅠ-Ⅳ),the expressions of CD40L on CD4+ and CD8+ T cells in the patients with acute GVHD increased,especially in allo-PBSCT.Five cases of UCBT and 12 cases of allo-PBSCT patients had chronic GVHD,the expressions of CD40L+,CD25+ and CD69+ on CD4+ and CD8+ T cells in patients with chronic GVHD increased obviously.The expression of CD19+CD40+ was lower than normal within 3 months after transplantation.Conclusions The high expression of CD40L+T cells in periferal blood after HSCT was related to the activation and proliferation of T cells in the development of GVHD in HSCT.

The title of a book is, generally, the high concentration of the writer's intention and the theme of content. Translate the title of an ancient Chinese medical book or document accurately and plainly is meaningful for exhibiting the style of the book, also for promoting the international communication of TCM. The principle should be followed is to choose the translating terms accurately to reveal the theme of content and express the cultural connotation of the book perfectly.
Books ; history ; China ; History, Ancient ; Medicine in Literature ; Medicine, Chinese Traditional ; history ; Terminology as Topic ; Translating

Adult ; Dermatitis, Occupational ; etiology ; Drug Eruptions ; etiology ; Humans ; Male ; Phthalimides ; adverse effects

Adult ; Aged ; Aged, 80 and over ; Benign Paroxysmal Positional Vertigo ; diagnosis ; drug therapy ; economics ; Female ; Humans ; Male ; Middle Aged ; Surveys and Questionnaires ; Treatment Outcome

This study was aimed to investigate the effect of various cytokines on megakeryocytes expansion in vitro from human cord blood CD34(+) cells in order to establish an optimal culture system for MK expansion. Mononuclear cells were obtained by Ficoll-Hapaque density gradient separation. CD34(+) cells were positively isolated using a CD34 progenitor cell isolation kit. CD34(+) cells were placed into 24 well plates at a concentration of 2 x 10(4) per well. Each well contained 1 ml of IMDM with the present of effective MK cells growth cytokines. Clonogenic potentials of MK progenitor were assayed using a methylcellulose cultures system. The results suggested that four cytokines (IL-3 + IL-6 + TPO + FLT3L) culture system could effectively induce and expand cord blood CD41(+) MK cells. The number of CD41(+) cells expanded 154.67 +/- 32.21-fold on day 7, and 193.23 +/- 25.24-fold on day 14. In conclusion, established expansion system in vitro for MK cells provides experimental foundation for recovery of platelets after cord blood transplantation.
Antigens, CD34 ; analysis ; Blood Platelets ; cytology ; immunology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Fetal Blood ; cytology ; immunology ; Humans ; Interleukin-3 ; pharmacology ; Interleukin-6 ; pharmacology ; Megakaryocytes ; cytology ; immunology ; Membrane Proteins ; pharmacology ; Platelet Membrane Glycoprotein IIb ; analysis ; Stem Cells ; cytology ; immunology ; Thrombopoietin ; pharmacology

OBJECTIVEChronic graft versus host disease (cGVHD) is the most common late complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and it represents the major cause of mortality in long-term survivors. Over the past decade, although conventional therapy has achieved complete responses in approximately 50% of patients, the prophylaxis and treatment of cGVHD are still not satisfactory. In the late years, utilization of new immunosuppressant such as tacrolimus (FK506), mycophenolate mofetil (MMF) on cGVHD improved the curative effects. This study tried to analyze the results of combination of methylprednisolone (MP), MMF and FK506 or cyclosporine A (CSA) as immunosuppressive therapies for cGVHD and to explore the effective regimen for children.

METHODSForty-five patients received allo-HSCT. Among them 32 received UCBT and 13 received PBSCT. The conditional regimen mainly consisted of busalphan, cyclophosphamide, antihuman thymocyte globulin, fludarabin, melphalan, thiotepa and total lymph node irradiation. Prophylaxis of GVHD consisted of CSA, MP and MMF. Patients with cGVHD received a regimen with combination of MP, MMF and FK506 or CSA.

RESULTSSeventeen out of 32 patients who received UCBT were engrafted. while 9 out of 13 patients who received PBSCT were engrafted. Nine cases of the 30 engrafted patients developed cGVHD (morbidity 30%). Among the 17 patients who received UCBT, 3 developed cGVHD (18%). Among the 13 patients who received PBSCT, 6 developed cGVHD (46%). Six cGVHD continued from aGVHD (6/9). One patient was given CSA plus MMF, and 8 were given three-drug regimen with MP, MMF and FK506. The overall response rate was 100%. Two patients died of CMV-IP or septicemia (mortality 20%). Seven (78%) patients survived (event free survival, EFS) longer than 3 years. The side effects included hepatotoxicity, nephrotoxicity, hypertension, articular capsulitis and arrhythmia. The main complication and the major causes of death were infection.

CONCLUSIONThe incidence of cGVHD is low in children. The incidence of cGVHD after PBSCT is higher than that after UCBT. aGVHD is a highly dangerous factor. Combined therapy of MP plus MMF and FK506 or CSA is safe and effective for the treatment of cGVHD in children.

Child ; Child, Preschool ; Chronic Disease ; Drug Therapy, Combination ; Female ; Graft vs Host Disease ; drug therapy ; epidemiology ; prevention & control ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Incidence ; Male ; Methylprednisolone ; administration & dosage ; Mycophenolic Acid ; administration & dosage ; analogs & derivatives ; Tacrolimus ; administration & dosage

To construct pcDNA3.1(+)/A2E eukaryotic expression vector and obtain a stable expression on HLA-I negative human K562 cell, PCR technique was employed to amplify A2E cDNA from the multi-cistron expression vector pG/A2E carrying HLA-E and HLA-A2 cDNA through internal ribozyme entry site (IRES), the cDNA was subcloned into vector pcDNA3.1(+), thus a eukaryotic expression was constructed and named pcDNA3.1(+)/A2E; then, the recombinant plasmid was transferred into the target cells, followed by screening with G418 and limiting dilution; finally, flow cytometry was adopted to detect HLA-E expression on the target cells. The results showed that HLA-E molecules were successfully expressed on K562 cells transfected with pcDNA3.1(+)/A2E (27.76%) and the expression of HLA-E molecules was not detected on K562 cells transfected with pcDNA3.1(+). It is concluded that the pcDNA 3.1(+)/A2E eukaryotic expression vector was successfully constructed and the HLA-E molecules were expressed on K562 cells. The data presented here would be expected to lay a good basis for the research of the molecular mechanism of HLA-E function and the interaction between HLA-E and the receptor on NK cells, as well as the influence of the expression of HLA-E in vitro on NK cells.
Cloning, Molecular ; DNA, Complementary ; genetics ; Eukaryotic Cells ; metabolism ; Flow Cytometry ; Gene Expression ; Genetic Vectors ; genetics ; HLA Antigens ; biosynthesis ; genetics ; HLA-A2 Antigen ; biosynthesis ; genetics ; Histocompatibility Antigens Class I ; biosynthesis ; genetics ; Humans ; K562 Cells ; Polymerase Chain Reaction ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVEAplastic anemia is characterized by bone marrow failure and marked reduction of white blood cells, red blood cells and platelets in peripheral blood. Clinical studies have shown that immunosuppressive therapy greatly prolonged the long-term survival of some patients with aplastic anemia. But in severe aplastic anemia (SAA) patients whose ANC was < 0.5 x 10(9)/L, platelets were < 20 x 10(9)/L, very low bone marrow proliferation and high death rate were observed. The present study aimed to evaluate the efficacy of immunosuppressive treatments with cyclosporine A (CSA) alone or CSA combined with antithymocyte globin (ATG) in children with acquired SAA.

METHODSFifty-four cases with SAA were treated with immunosuppressive agents mentioned above in our department from Jan. 1997 to June 2003, 31 of the cases had treated with CSA combined with ATG. There were 18 cases with SAA type I and 13 cases with SAA type II in CSA combined with ATG group, and 13 cases had very severe aplastic anemia. The other 23 cases were treated with CSA alone (CSA group), 10 of these cases had SAA-I and 13 had SAA-II, and 5 cases had very severe aplastic anemia. The responsive rate, relapse, adverse reactions and event free survival (EFS) were compared between CSA combined with ATG group and CSA group.

RESULTSThe proportions of patients with different types of the disease and severity were comparable between the two groups. The responsive time of the CSA combined with ATG group and CSA group was 2.5 months and 3.5 months, respectively (P < 0.05), the responsive rate in two groups was 81% (25/31) and 52% (12/23), respectively (chi(2) = 4.962, P < 0.05). In 37 cases who were responsive to therapy, the relapse rate was 8% (2/25) and 50% (6/12) respectively (chi(C)(2) = 6.143, P < 0.05). There were no significant differences in adverse reactions to the immunosuppressive agents. All cases were followed-up for more than 1 year, and the event-free survival over one year in these two groups was 81% (25/31) and 52% (12/23), respectively. Forty-seven cases were followed-up for more than two years, and the event-free survival was 74% (20/27) and 50% (10/20), respectively (P < 0.01). Twelve cases were followed-up for over 5 years. There were no secondary tumor, myelodysplastic syndrome and other colony diseases.

CONCLUSIONThe immunosuppressive therapies for acquired severe aplastic anemia in childhood were effective. The effect of CSA combined with ATG was better than that of CSA alone, and the relapse rate was lower with the combined treatment. However, the long-term effect needs longer follow-up studies to evaluate.

Adolescent ; Anemia, Aplastic ; drug therapy ; Antilymphocyte Serum ; administration & dosage ; therapeutic use ; Child ; Child, Preschool ; Cyclosporine ; administration & dosage ; therapeutic use ; Female ; Humans ; Immunosuppressive Agents ; administration & dosage ; therapeutic use ; Male