0.31 and

Objective To establish a method to induce dendritic cells (DC) from peripheral blood mononuclear cells of healthy people in normal human AB serum in vitro and to identify the phenotype and the function of DC. Methods Peripheral blood mononuclear cells (PBMNC) of healthy people were cultured in RPMI 1640 media including human AB serum, GM-CSF, rhIL-4, and/no rhCD40 for 7 days to generate DC,which were identified by morphological features, surface antigen expression and the ability to stimulate T cells.Results After cultured and induced, DC displayed typical morphology with elongated dendritic process viewed by inverse light microscope as well as Wright-Gimsa stain. Mature DC express CD83 and the costimulatory molecules CD40 CD80, and CD86 to effectively activate T cells. In the five time points of 0 day, 1st day, 3rd day, 5th day and 7th day, the expression of CD83, CD40, CD80, CD86 and CD14 were significantly different (F= 50.253, 243.769, 248.181, 191.267 and 226.339, respectively, P＜ 0.05). The ability to stimulate T cells in GM-CSF, rhIL-4, and rhCD40L group was also stronger than that in GM-CSF and rhIL-4 group. DC started to secrete IL-12 from 5th day, the values were (42.92±1.54) pg/ml and (136.18±5.27) pg/ml in group of plus CD40L and of non plus CD40L, respectively. The secretion of the two groups of IL-12 were (60.09±2.27) pg/ml and (322.30±30.60) pg/ml (t = -44.941, -22.611, bath P ＜ 0.05). There are significant differences between the two groups. Conclusion DCs can be cultured from the peripheral blood of healthy people in normal human AB and rhCD40L serum.

Objective To explore the expression of thioredoxin reductase (TrxR) mRNA and protein,as well as the relationship between TrxR and myeloid leukemia.Methods Bone marrow samples from 20 acute myeloid leukemia (AML),20 chronic myeloid leukemia (CML),and 20 healthy persons as normal control group were collected.The human non small cell lung cancer A549 cells were selected as the positive control group.The expression of TrxR mRNA was detected by real-time quantitative polymerase chain reaction.The expression of TrxR protein level was detected by Western blot.Results The relative quantitation expressions of TrxR mRNA were 6.751 (13.459),4.321 (11.389),18.477 (2.089),1.045 (0.467) in AML,CML,positive control and normal control group,were 17.814±3.979 and 4.860±1.550 in the primary diagnosed/relapse group and complete remission (CR) group of AML patients,and were 19.552 (5.758) and 3.459 (2.047) in the primary diagnosed and treatment group of CML patients.The expression levels of TrxR mRNA in AML,CML and positive control group were significantly higher than that in normal control group (H =43.978,P ＜ 0.001).For AML patients,the expression levels of TrxR mRNA in the primary diagnosed/relapse group,CR,positive control group were significantly higher than that in normal control group (F=246.793,P ＜ 0.001),and the difference between the primary diagnosed/relapse group and positive control group was no significant (P ＞ 0.05).The expression of TrxR mRNA were higher in the primary diagnosed/relapse group than that in CR group,and that in CR group than that in normal control group (both P ＜ 0.05).For CML patients,the expression levels of TrxR mRNA in the primary diagnosed,treatment,positive control group were significantly higher than that in normal controls (H =38.222,P ＜ 0.001),the difference between that in the primary diagnosed and that in positive control group was no significant difference (P ＞ 0.05),but the expression of TrxR were significantly higher in the primary diagnosed group than that in treatment group,and that in treatment group than that in normal controls (both P ＜ 0.05).The expression of TrxR mRNA in the primary diagnosed/relapse group of AML and the primary diagnosed group of CML showed no significant difference (P ＞ 0.05).The positive levels of TrxR protein were 0.679,0.606,0.877 and 0.095 in AML,CML,positive control and normal control group.TrxR protein was highly expressed in myeloid leukemia patients.Conclusion The expression levels of TrxR mRNA and protein are increased in myeloid leukemia.The low expression of TrxR in normal hematopoietic stem cells is highly expressed in myeloid leukemia,and is downgraded after treatment,which may be used as one of the parameters to diagnosis,effect and prognosis of myeloid leukemia.

Objective To investigate the personality traits, anxiety and depression in patients with leukemia. Methods A cross-sectional survey was performed in 120 patients with leukemia and 118 age and gender-matched controls. Personality was assessed using the Type A Behavior Pattern Questionnaire (TABPQ). Emotion was ascertained using the self-rating anxiety scale (SAS) and self-rating depression scale (SDS). The relationship between personality type A and anxiety and depression was evaluated by Pearson correlation analysis. Results The prevalence of personality type A was significantly higher in patients with leukemia than healthy controls [71.67 % (86/120) vs. 30.51 % (36/118), P< 0.01]. Patients with leukemia also had higher prevalence of anxiety and depression compared with healthy controls [36.67%(44/120) vs. 9.32%(11/118), P< 0.01; 29.17 % (35/120) vs. 5.93 % (7/118), P< 0.01]. Pearson correlation analysis showed that time-hurry (TH) and competition-hostility (CH) of personality type A correlated positively with anxiety (r=0.292, r= 0.277, respectively; both P< 0.05). Conclusion TH and CH of personality type A are associated with anxiety in leukemia patients.

Objective To explore the expression level of thioredoxin (Trx) in acute myeloid leukemia (AML) cells,and its association with clinical features and prognosis,as well as the effects of diamide,an inhibitor of Trx,in inducing AML cells apoptosis.Methods Expression of Trx on AML cells and fresh bone marrow cells from healthy adults were analyzed by Western-blotting. The inhibition of proliferation was measured by MTT assay.The anti-leukemia effect of diamide was observed by morphology and agarose gel electrophoresis.Results 75 ％ (15/20) of AML patients expressed Trx,while no expression was observed in control group.Higher expression level of Trx was associated with higher WBC counts (x2 =9.375,P ＜ 0.05),which suggested that overexpression was associated with leukemogenesis.The inhibition of diamide on AML cells showed time and dose dependent by MTT assay.The IC50 values of diamide at 24 h,48 h and 72 h were 98.26 mg/ml,47.53 mg/ml and 8.34 mg/ml,respectively.After AML cells were treated with diamide,the apoptotic body appeared by morphology,and the typical DNA “ladder” bands were confirmed by agarose gel electrophoresis.Conclusion Trx expression level in AML cells is significantly higher than that of control group.Diamide inhibites the proliferation of AML cells by inducing apoptosis,which might be a potential agent for AML.

ObjectiveTo induce monocyte-derived dendritic cells(MoDC)from acute myeloid leukemia (AML) and healthy persons by rhCD40L in normal human AB serum system in vitro and to identify the morphology and phenotype of MoDC. MethodsPeripheral blood mononuclear cells(PBMNC)of AML and healthy persons were cultured in RPMI 1640 media including human AB serum, GM-CSF, rhIL-4 and rhCD40L, respectively. MoDC were identified by morphological features, surface antigen expression and the ability to stimulate T cells. ResultsAfter cultured for 7 days, MoDC displayed typical morphology with elongated dendritic process,and upregulation of the costimulatory molecules CD40,CD80,CD86 and CD83.The morphology and expression of costimulatory molecules were not significantly different between AML and healthy persons (P＞0.05),but were significantly different between rhCD40L group and without rhCD40L group (P＜0.05). MoDC had the ability to activate T cells, and there were no statistical differences between AML and healthy persons(P ＞0.05), but were significant differences between rhCD40L group and without rhCD40L group (P＜0.05). MoDC started to secrete IL-12 on day 5, and there was no statistical differences between AML and healthy persons(P＞0.05),and had differences between rhCD40L group and without rhCD40L group (P＜0.05).ConclusionMoDC can be cultured from the peripheral blood of AML and healthy persons.There were no significant differences in morphology and phenotype.Monocyted-derived DC can be used as an alternative to generate leukemia-specific cytotoxic T cells,especially in the presence of rhCD40L.

Objective To develop a simple model for the noninvasive diagnosis of liver fibrosis in patients with chronic hepatitis B and to testify its diagnostic value. Methods One hundred and ninety patients with chronic hepatitis B who had undergone liver biopsy were divided into 2 groups:one for developing the model(n=110) and one for validation(n=80). Histological staging of liver fibrosis,assessed blindly and independently by 2 pathologists,was determined according to Scheuer fibrosis score.Twenty markers involved in the study were analyzed initially in the estimation group to derive a predictive model to discriminate the stages of fibrosis.The model created was then assessed with receiver operating characteristic curve(ROC)analysis. It was also applied to the validation group to test its accuracy. Results Haptoglobin(HPT),γ-glutamyl transpeptidase(GGT)and platelet were identified by logistic regression analysis as independent factors of fibrosis. A model developed from the above three markers was established to predict the stage of fibrosis(S). In ROC analysis,the area under curve(AUC) for identifying S≥1,S≥2,S≥3 and S=4 was 0.832,0.835,0.820 and 0.843 respectively. The model had a similar AUC in the vatidation group without statistically significant difierence. Using a cut-off of ＜0.18, significant fibrosis (S≥2)could be excluded in 27 patients of the total patient population(negative predictive value 90%).Similarly,applying a cut-off ≥0.70,significant fibrosis could be identified correctly in 67 patients of the total patient population(positive predictive value 82.7%).The model had a high level of diagnostic value in patients with HBeAg-positive chronic hepatitis B as well as in patients with HBeAg-negative chronic hepatitis B(AUC for identifying S≥2,0.857 vs 0.802). Restricting biopsy to patients with intermediate scores(≥0.70 and ＜0.18) may prevent liver biopsies in 58.4% of the patients while maintaining 84.7% accuracy. Conclusions A model including HPT,GGT and platelet is a simple and reliable index for predicting significant fibrosis in patients with HBeAg-positive chronic hepatitis B as well as in patients with HBeAg-negative chronic hepatitis B.

Objective To investigate the expression of nuclear factor erythroid-2 related factor 2 (Nrf2) gene in chronic myeloid leukemia (CML) and explore its relationship with clinical characteristics and thioredoxin reductase (TrxR).Methods The expressions of Nrf2 and TrxR genes in bone marrow cells and K562 cells were analyzed in 30 bone marrow preparations of CML patients in different phases,including 20 in chronic phase,3 in accelerated phase,7 in blastic phase by real-time quantitative polymerase chain reaction.Ten health subjects were served as normal controls.Results The relative quantitation expression of Nrf2 and TrxR mRNA were 5.601±1.069 and 9.017±2.398 in chronic phase,1.698±0.349 and 5.590±1.015 in accelerated phase,1.252±0.807 and 5.050±1.469 in blastic phase,1.377± 0.344 and 1.867±0.977 in normal controls.The expressions of both Nrf2 and TrxR mRNA in CML had significant differences from those of the normal controls (x2 =14.680,P =0.002,x2 =8.271,P =0.041).The expression of Nrf2 mRNA in accelerated phase,blastic phase group showed no significant difference (Z =0.011,P =0.496),but lower than that in chronic phase group (Z =2.155,P =0.016,Z =2.534,P =0.006).The difference between the first visit and post-treated group was significant (Z =2.015,P =0.022).The expression in K562 cells and normal controls had significant difference (Z =1.898,P =0.029).In CML patients,the expression of Nrf2 was positively correlated with that of TrxR (r =0.738,P =0.037).Conclusion The expression of Nrf2 gene is higher in the first visit group of CML than that in the other groups,and is decreased after therapy,which may be the molecular marker predicting the progress of CML.Nrf2 mRNA expression level is correlated with TrxR.

Objective To explore the activity of thioredoxin reductase (TrxR) in chronic myeloid leukemia cell line K562 and the anti-leukemia effect of BBSKE (a novel inhibitor of TrxR) in vitro. Methods The activity of TrxR on K562 cell lineage and fresh bone marrow cell from healthy adult was analyzed by insulin reduction assay. The inhibition of proliferation was measured by CCK-8 assay. The anti-leukemia effect of BBSKE was detected by laser scanning confocal microscope,agarose gel electrophoresis and flow cytometry with Annexin V -FITC/PI staining. Results TrxR activity of K562 cell lineage was significantly higher than that of normal bone marrow mononuclear cells. The apoptosis of K562 cells could be induced at concentrations of 10 μmol/L BBSKE after treated for 24 hours. The typical DNA ladder bans were observed by agarose gel electrophoresis. The apoptotic rates of K562 cells were (10.28±2.74) %. Application of 10 μmol/L BBSKE for 48 hours could also induce apoptosis of fresh bone marrow cell from chronic myeloid leukemia patients, and the apoptotic rates were (5.70±0.48) %. Conclusion TrxR activity in chronic myeloid leukemia cells was significantly higher than that of normal cells. BBSKE inhibits the TrxR activity and the proliferation of K562 by inducing apoptosis.It might be a potential medication for chronic myeloid leukemia.