Adolescent ; Adult ; Cause of Death ; Child ; Female ; Hematologic Neoplasms ; mortality ; surgery ; Hematopoietic Stem Cell Transplantation ; mortality ; Humans ; Male ; Middle Aged ; Tissue Donors ; Young Adult

Objective To investigate the efficacy of HLA-matched ABO-incompatible sibling al- logeneic peripheral blood stern cell transplantation(allo-PBSCT)in treatment of hematologic malig- nancies.Methods Between June 2001 and September 2005,68 patients with hematologic malignancies received allo-PBSCT from HLA-identical sibling donors.Among them,38 were ABO-compatible (ABO-compatible group),30 ABO-incompatible(ABO-incompatible group).Conditioning regimen of all patients:BU/CY used as conditioning regimen in AML,MDS and CML patients;TBI/CY used in ALL and NHL patients.TBI/CY/Melphan used in MM patients.GVHD prophylaxis:A combination of cyclosporine(CsA)and mycophenolate mofetil(MMF)and methotrexate(MTX)was administered for GVHD prophylaxis.Results (1)Sixty-seven patients were all engrafted,and only one patient was graft failure.The median time(range)to neutrophils≥0.5?10~9/L and platelets≥20?10~9/L was +12(+9～+15)and+21(+15～+40)days post-transplant respectively.The time to neutrophil and platelet engraftment had no significant difference between ABO-incompatible group and ABO-com- patible group(P＞0.05);(2)Thirty patients with ABO-incompatible allo-PBSCT had no evidence of hemolysis.In ABO-incompatible group,the onset of erythropoiesis after allo-PBSCT was delayed. Three of 7 patients with blood group“O”receiving a group“A”developed pure red cell aplasia (PRCA).The blood type of 30 patients would transfer to the type of donors in 60 days(24～153 days)after transplantation.(3)Following up to 30th,Sep.2005,the incidence of aGVHD was high- er in ABO-incompatible group(20.0%)than that in ABO-compatible group(2.6%,P=0.019). The incidence of cGVHD,VOD,HC,CMV infection,disease relapsed rates and mortality had no sig- nificant difference between ABO-incompatible and ABO-compatible recipients(P＞0.05).(4)Kaplan- Meier survival analysis revealed the rates of survival had no significant difference between ABO-incom- patible and ABO-compatible recipients(P＞0.05).Conclusions ABO-incompatible allo-PBSCT is fairly safe if there is indication.Although the incidence of aGVHD was higher in ABO-incompatible group than that in ABO-compatible group,but there was no effect on engraftment,incidence of cGVHD or prognosis.

The objective of study was to investigate the effect of liposomal transfection of antisense oligodeoxynucleotide (ASON) on alpha-globin gene expression and proliferation of K562 cells, to explore the new way of gene therapy in beta-thalassemia. Targeted ASON of alpha-globin was designed and synthesized, and compared with positive control [sense oligodeoxynucleotide (SON) group] and blank control. By liposomal transfection, ASON, SON was co-cultured with K562. The efficiency of transfection was assayed by fluorescence microscopy and flow cytometry (FCM), the alpha-globin gene expression of K562 was measured by real-time PCR, and the proliferation of K562 was determined by Cell Count Kit-8 assay. The results indicated that the highest efficiency was at 24 hours after liposomal transfection, the gene expression level of alpha-globin in ASON group was significantly lower than that in SON group and blank control (p < 0.01). The proliferation of K562 cells was obviously inhibited, meanwhile the above effect showed the dose-dependent manner. It is concluded that the liposomal transfection of ASON inhibits the alpha-globin gene expression of K562 cells, which may be the new target for gene therapy in beta-thalassemia.
Cell Proliferation ; drug effects ; Gene Expression ; Humans ; K562 Cells ; Liposomes ; metabolism ; Oligonucleotides, Antisense ; pharmacology ; Transfection ; alpha-Globins ; metabolism

This study was aimed to detect and identify the promoter CpG island methylation of γ-globin gene in peripheral blood mononuclear cells from patients with β-thalassemia major and healthy adult in Guangxi province, as well as to analyze the difference of promoter methylation rate of each CpG sites between them, and then to screen the promoter CpG island main methylation sites which maybe influence γ-globin expression. The template DNA was modified by bisulfite genomic sequencing PCR; the promoter sequences of γ-globin gene were amplified by technique Touchdown PCR, and then the PCR products were cloned and sequenced for obtaining methylation status of each CpG sites in target fragments, and then the accurate methylation sites and levels were detected quantitatively. The results indicated that the 4 CpG methylation sites locating at 28, 122, 231 and 234 bp in sequences were hypermethylated. As compared with healthy adults, the DNA methylation rate of 122 and 231 bp CpG sites in patients with β-thalassemia major was obviously lower, however, methylation rates of 28 and 234 bp sites were not significantly different between patients and healthy adults. It is concluded that the methylation sites 28, 122, 231 and 234 bp of γ-globin gene promoter are found both in patients with β-thalassemia major and healthy adults. The 122 and 231 bp sites are identified preliminarily to be involved in the regulation of γ-globin expression. This study provides the experimental evidence for alleviating the clinical symptoms of β-thalassemia major and targeting gene treatment through the regulation of γ-globin.
Adolescent ; Adult ; Case-Control Studies ; Child ; CpG Islands ; DNA Methylation ; Female ; Humans ; Male ; Promoter Regions, Genetic ; Young Adult ; beta-Thalassemia ; genetics ; gamma-Globins ; genetics

OBJECTIVETo study the effect of liposomal transfection of antisense oligonucleotide (ASON) on the erythroid cell alpha-globin gene in the patients with severe beta-thalassemia, and provide a new idea for beta-thalassemia gene therapy.

METHODSA highly effective ASON targeting alpha-globin gene was transfected into severe beta-thalassemic erythroid cells cultured in vitro by liposomal at an optimal concentration. The expression level of alpha, beta, gamma-globin gene, the level of hemoglobin, and the excess alpha-globin chains precipitates in ASON group and control group were carefully analyzed by quantitative real-time PCR(Q-RT-PCR), high performance liquid chromatography (HPLC), and electron microscope, respectively.

RESULTSThe mRNA expression of alpha-globin gene was significantly lower in ASON group (9.04 +/- 0.29) than in control group (24.23 +/- 0.29) (P<0.01). Simultaneously, the disequilibrium between alpha- and beta-, gamma-globin gene expression was partly modified by ASON, the ratios of ASON group and control group being 0.79 +/- 0.02 and 2.26 +/- 0.06 respectively (P<0.01). HPLC demonstrated that the levels of HbA2 and HbF increased with downregulation of alpha-globin gene in beta-thalassemic erythroid cells, particularly HbF. The precipitates of alpha-globin chains in ASON group were lessened under electron microscope, particularly in early erythroblast while no change in the control group.

CONCLUSIONThe high effective ASON contributes to inhibit the alpha-globin gene expression of severe beta-thalassemic erythroid cells, partly modify the disequilibrium between alpha-, beta- and gamma-globin gene expression and obviously reduce the precipitates of alpha-globin chains in erythroid cells. It might provide a new idea for gene therapy of beta-thalassemia.

Cells, Cultured ; Child ; Genetic Therapy ; Humans ; Liposomes ; Oligonucleotides, Antisense ; genetics ; Transfection ; alpha-Globins ; genetics ; metabolism ; beta-Globins ; metabolism ; beta-Thalassemia ; genetics ; metabolism ; therapy ; gamma-Globins ; metabolism

Objective To evaluate the effect of the intermittent deferomamine(DF) therapy on relieving iron overload caused by transfusion in children with ? thalassemia.Methods Sixteen children who were finally diagnosed as ? thalassemia major were treated with deferomamine for 124 times totally to low the iron overload. The serum iron(SI), serum ferritin(SF) and urine ferritin were detected each time with radio-immunity technique and difference was compared before and after treatment. Meanwhile, weather DF involved children′s liver and renal function was observed in whole procedure.Results Iron overload exists in 16 cases of ? thalassemia major children by a long- term hypertransfusion therapy, with average level SI 33.69?6.72 mmol/L,SF 441.19? 54.70 ?g/L,urine ferritin 8.64?6.79 ?g/L. The difference was significant (paired-samples t test,t =6.173 P 0.05).Conclusion The study suggest that intermittent low-dose DF therapy is effective for iron overload caused by transfusion in ? thalassemia children, without apparent side effects.

Objective To describe the prevalence of deliberate self-harm (DSH) and suicidal behaviors (SIB) as well as the relationship between them in a college student population.Methods A total of 4063 medical students were selected under the cluster sampling method in Anhui province.Data were analyzed by Pearson Chi-square and logistic regression.Results A total of 4063 ( 13.4％ )students reported that they had deliberately harmed themselves during the past 12 months.The acts of DSH with 1 and more than or equal to 2 times occurrence accounted for 3.7％ and 9.6％ among all the respondents.The act of DSH was significantly higher among boys than that among girls.Rates of suicide ideation,suicide plan,attempted suicide and SIB in the last year were 4.5％,1.4％,0.6％ and 4.9％,respectively.However no statistically significant difference was found in different sex.Students in the DSH group that with SIB were found to have lower positive coping levels (OR=0.5,95％CI:0.3-0.8),getting less support from fricnds (3-5 vs.less than or equal to 2:OR=0.6,95％CI:0.3-0.9;more than or equal to 6 vs.less than or equal to 2:OR=0A,95％CI:0.2-0.8),with higher negative coping levels (OR=2.1,95％CI:1.2-3.7),having more serious depressive symptoms (OR=2.9,95％ CI:1.6-5.2) and anxiety symptoms (OR=2.2,95％ CI:1.2-3.8),having more serious sleeping problems (OR=1.7,95％CI:1.1-2.8 ) and perceived fat (fat vs.moderate:OR=2.0,95％CI:1.1-3.6) than the DSH group without SIB.The rates of SIB in students with DSH behavior were significantly higher than those in students without those behaviors (OR=4.7,95％ CI:3.5-6.4).Psyehosocial variables could attenuate the relationship between the DSH status and suicidal events (OR=3.3,95％CI:2.4-4.5).The DSH frequency exhibited a curvilinear relationship to SIB (OR=3.1-10.0) and psychosocial variables also attenuated this relationship (OR=2.4-5.9).Conclusion It was well known that SIB was not a suicidal gesture but our findings suggested that the presence of DSH might trigger the suicidal attempts.

OBJECTIVETo explore the effect of liposomal transfection of cyclin A antisense oligodeoxynucleotide (ASON) on HL-60 cell proliferation and apoptosis.

METHODSBy liposomal transfection, cyclin A ASON was co-cultured with HL-60 cells, the cell growth curve was determined by MTT assay and cell apoptosis electron-microscopy in situ cell apoptosis detection kit (POD), the protein and mRNA of cyclin A and bcl-2 were measured by FACS and RT-PCR, the role of cyclin A ASON in the development of leukemia was tested by the tumor formation in nude mice.

RESULTS(1) In the cyclin A ASON liposomal transfection group (group A), the proliferation of HL-60 cell was significantly inhibited as compared to those in cyclin A ASON group (group B) (68.9% vs 24.8%) (P < 0.01). (2) The expressions of cyclin A and bcl-2 of group A were significantly lower than those in the control group (1.1% vs 38.8%, P < 0.01; 21.9% vs 65.0%, P < 0.01, respectively), and the DNA ladder and apoptosis body was displayed. (3) In group A, the rate of tumor formation in nude mice was lower, the time for tumor formation was longer and the volume of tumor was smaller than those in control group.

CONCLUSIONLiposomal transfection of cyclin A ASON can inhibit in vitro proliferation of leukemia cells and induce in vivo apoptosis of the tumor cell, which might provide a new target for gene therapy.

Animals ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Cyclin A ; genetics ; physiology ; Genetic Therapy ; HL-60 Cells ; Humans ; Leukemia ; therapy ; Liposomes ; Mice ; Mice, Inbred BALB C ; Oligonucleotides, Antisense ; pharmacology ; Transfection

OBJECTIVETo evaluate the efficacy and safety of deferasirox in heavily iron-overloaded patients with beta-thalassemia major.

METHODSA single arm, open-label clinical trial was conducted to evaluate the efficacy and safety of deferasirox in the treatment for 23 patients with beta-thalassemia major and heavily iron-overloaded in 3 years follow-up.

RESULTSThe 23 patients never received regular chelation before enrolling this trial [the mean baseline of serum ferritin was (5433.96 ± 2873.90) µg/L]. In this trial, a deferasirox dose of 20 mg×kg(-1)×d(-1) could stabilize serum ferritin levels, while of ≥ 30 mg×kg(-1)×d(-1) reduced the levels and achieved negative iron balance. There were no serious adverse events related to the drug. Most common adverse events were mild increases of liver enzyme and serum creatinine levels. Overall, 23 patients could tolerate the drug on schedule and all completed the trial.

CONCLUSIONAs a new oral iron chelator, deferasirox has a significant efficacy for the treatment of iron overload. The effectiveness is dependent on the courses of treatment and the dose of deferasirox. The single-dose used is safe and tolerated, so deferasirox can remarkably improve life quality of patients.

Humans ; Iron ; blood ; Iron Overload ; Quality of Life ; Treatment Outcome ; beta-Thalassemia ; blood