Objective To explore the resistance and molecular mechanisms underlying the anticancer activity of daunorubicin in CD34 +acute myeloid leukemia(AML) cells. Methods CD34 +AML cell lines(KG1a and Kasu-mi-1)were used as objectives, and CD34 -AML cell line U937 was used as positive control. Western blot analysis was used to examine the protein expression of Bcl-2 and Bax in CD34 +AML and CD34 -AML cell lines incubated with/without daunorubicin to compare the sensitivity of CD34 +AML and CD34 -AML cells to daunorubicin. SiRNA against Bcl-2 was used in KG1a and Kasumi-1 cells and examined the effect on cell viability by MTT assay. Results Western blot analysis showed that Bcl-2 protein levels in CD34 +AML cells appeared to be significantly higher than in CD34 -AML cells. Western blot analysis showed that treatment with 0.4 μg/ml daunorubicin for 48 h caused down-regulation of Bcl-2 only in CD34 -AML cells,but not in CD34 +AML cells. Suppression of Bcl-2 with siRNA increased the susceptibility of KG1a and Kasumi-1 to daunorubicin. Conclusion CD34 +AML cell lines ex-press higher levels of Bcl-2 protein. Daunorubicin fails to down-regulate the high Bcl-2 protein levels in CD34 +AML cells. Suppression of Bcl-2 with siRNA increases the susceptibility of KG1a and Kasumi-1 to daunorubicin. The high Bcl-2 protein levels in CD34 +AML cells may be involved in the insensitivity to daunorubicin.

AIM:To investigate the anti-tumor effects of special cytotoxic T lymphocytes(CTLs)activated by dendritic cells(DCs)loaded with antigens and CD40L in vitro.METHODS:Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation from normal human heparinized blood.The adherent cells were cultured with granulocyte-macrophage colony stimulating factor(GM-CSF),interleukin-4(IL-4),alpha tumor necrosis factor(TNF-?),DCs were co-cultured with frozen-thawed antigen of K562 cells and CD40L,then triggered T cells into specific CTLs.RESULTS:Most suspended cells exhibited distinctive morphological features of DCs which expressed CD40 96%,CD86 97%,CD80 77%,CD1a 69%,and gained the powerful capacity to stimulate proliferation of allogenic lymphocytes.Under the effector∶target ratio of 20∶1,CTLs derived from cultures with DCs and frozen-thawed antigen of K562 cells were showed 71.3% cytotoxicities against K562 cells.CTLs derived from cultures with DCs loaded with frozen-thawed antigen and CD40L were showed 86.9% cytotoxicities against K562 cells.Cytotoxicities by CTLs derived from cultures with unloaded DCs against K562 cells were 37.6% and cytotoxicities by monocytes were 21.1%.Cytotoxicities by CTLs derived from experiment groups were stronger than control groups(P

AIM: To investigate the effect of granulocyte colony stimulating factor (G-CSF) on arabinosyl cytosine (Ara-C)-induced apoptosis in HL-60 leukemia cells. METHODS: The proliferation of HL-60 leukemia cell was observed by hemopoietic cell culture. Apoptosis was measured by the morphology of apoptosis cell , the quantitation of DNA fragmentation with the diphenylamine reaction. The change in drug sensitivity was measured by “MTT”. RESULTS: G-CSF could stimulate the proliferation of HL-60 leukemia cell and colonies of cell increased to 76.5?18.0, compared to the control group (46.5?13.5. P

BACKGROUND: Al ogeneic hematopoietic stem cel transplantation is the main method to cure leukemia, but the patients receiving al ogeneic hematopoietic stem cel transplantation stil have to face the risk of recurrence. Myeloid sarcoma is a rare extramedul ary relapse mode with worse clinical outcomes, so it is necessary to understand the characteristics of myeloid sarcoma and relative treatment methods. OBJECTIVE: To analyze the clinical characteristics and treatment methods of myeliod sarcomas in cardio-phrenic angle after al ogeneic peripheral blood stem cel transplantation. METHODS: One case was diagnosed as single myeloid sacoma in the cardio-phrenic angle after al ogeneic peripheral blood stem cel transplantation. The patient underwent surgical resection of the mass, chemotherapy and radiotherapy. The clinical effect, complications and survival situation were observed. RESULTS AND CONCLUSION: The patient suffered from bacteremia, fungal pneumonia and even life-threatening sepsis shock during two courses chemotherapies. Then, the patient received radiotherapy for mediastinum and the myeloid sacoma never relapsed. However, the patient suffered central nervous system leukemia after free of the disease for 25 months. Myeliod sarcoma after al ogeneic hematopoietic stem cel transplantation is rare and its manifestation is changeful. The diagnosis mainly depends on histopathology and immunohistochemistry. Surgery, chemotherapy, radiotherapy, second transplantation and molecular targeted drugs are the choices of treatment strategy. However, the optimal treatment strategy for individual patients remains to be determined.

Objective To evaluate and compare the value of predicting diseases such as hypertension and arteriosclerosis of the body mass index(BMI) and percent body fat(PBF),so as to provide the basis for evaluation of obesity.Methods Totally 3149 common residents in Jiangsu province were surveyed using the method of stratified and cluster sampling.Effective data included height,weight,PBF,blood pressure and brachial-ankle pulse wave velocity(baPWV).Obesity was defined by percent body fat with the evaluation of WHO and ASBP.Areas under ROC curves(AUC)of BMI and PBF were estimated by the nonparametric test and then the two diagnostic tests were compared by predictive value of related diseases.Results(1)Predictive value of BMI to diagnose obesity(defined by PBF):AUC are respectively 0.949(for WHO standard)、0.906(for ASBP standard)in women and 0.864 in men.In age group,the 20-39 years have the highest AUC of BMI to predict obesity.P＜0.01 for above nonparametric tests.(2)According to ROC curves we got the adjusted cut-off points of BMI are respectively 26 kg/m2 in men and 25 kg/m2 in women for WHO standard or 26 kg/m2 in men and 23 kg/m2 in women for ASBP standard,while the specificity of predicting obesity decreased from 90％ ～ 99％ to 76％ ～87％,but the sensitivity significant increased from 17％ ～ 43％ to 78％ ～ 89％;After adjusting the cutoff points of BMI,the value of kappa of prevalence of obesity increased from 0.475 to 0.537 in men and 0.115 to 0.655 in women.(3)To predict hypertension,areas of BMI were 0.688(95％CI:0.656-0.720) in men and 0.745 (95％CI:0.708-0.782) in women,similarly,of PBF were 0.687 (95％CI:0.655-0.718) in men and 0.723(95％CI:0.681-0.764)in women;To predict arteriosclerosis there showed highly consistency that the areas of BMI were 0.613(0.586-0.641)in men and 0.692(95％CI:0.659-0.726)in women,meanwhile,for PBF they were 0.635 (95％CI:0.608-0.663) in men and 0.683 (95％CI:0.648-0.718) in women (P＜0.01).(4) Paired test of the two areas under ROC curves showed that PBF had higher diagnostic value than BMI in men to predict arteriosclerosis(u=2.05,P＜0.05),however,no statistical difference in women(u=0.75,P＞0.05)and in predicting hypertension(u=0.75 to men and u=1.26 to women,P＞ 0.05).Conclusions Using BMI and PBF can all predict hypertension and arteriosclerosis effectively,thus,obesity can be evaluated suitably by both BMI and PBF in large-scale population study,especially in women and youth.Relatively,WHO standard of PBF is more suitable for Chinese population to evaluate obesity than ASBP standard.

AIM: To study the effects of WT1 peptide-loaded dendritic cells (DC) stimulating the cytotoxic T lymphocytes (CTL) on K562 cells in vitro. METHODS: DC were generated from normal human peripheral blood mononuclear cells (PBMC) in the presence of granulocyte-macrophage colony stimulating factor(GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-?) , DC were cultured with WT1 peptides , and then triggered T cells into specific CTL. RESULTS: Most suspended cells exhibited distinctive morphological features of DCs and they stimulated proliferation of allogenic lymphocytes. Under the effector : target ratio of ~20∶1 , CTLs derived from cultures with DC and WT1 peptides were showed 86.1%?26.8% cytotoxicity against K562 cells, cytotoxicity by CTLs derived from cultures with unloaded DC against K562 cells were 47.1%?20.8% and cytotoxicity by lymphocytes were 27.7%?15.3%. Cytotoxicity by CTLs derived from culture with WT1 peptides-loaded DC were the strongest among three groups (P

AIM: To investigate the changes of the expression of Bcl-2 and Fas protein and the apoptosis in HL-60 cells induced by cyclosporine A. METHODS: The expression of Bcl-2 and Fas protein and apoptosis in (HL-60) cells were measured by immunohistochemistry analysis and flow cytometric analysis. RESULTS: There was strong expression of Bcl-2 in HL-60 cells, treatment with cyclosporine A (CsA) for 8-10 h down-regulated the expression of Bcl-2. Fas protein expression in HL-60 cells was very low, CsA induced apoptosis of HL-60 cells, but didn't induce Fas protein expression. CONCLUSION: CsA induces apoptosis in HL-60 cells by down-regulating Bcl-2 expression. [

Abstract： Objective　To establish a screening-diagnosis-evaluation system of neonatal congenital heart disease (CHD) suitable for Hainan Province based on the application of percutaneous blood oxygen saturation measurement and cardiac auscultation screening, and to provide a basis for further promotion of the evaluation effect. Methods　Screening agencies (all midwifery institutions) used the "double index method" to screen newborns for congenital heart disease. Those who were screened positive for any index were transferred to diagnostic institutions (the people's hospitals of all cities and counties and some tertiary hospitals) for echocardiography examination within 1 week. Those with abnormal results were transferred to the treatment institution (two tertiary hospitals) for reconfirmation and evaluation. All data were managed online. Results　In this study, 96 913 newborns born in midwifery institutions in 19 cities and counties were screened, with a response rate of 99.44%. The ratio of male infants (50 836) to female infants (46 077) was 1.10∶1. A total of 2 284 positive patients were screened by dual index method, and ultrasound diagnosis was performed, with the rate of cardiac ultrasonography of 98.07% (2 240/2 284) and a high response rate. A total of 238 cases of congenital heart disease were diagnosed (incidence rate of 0.25%), with the top three main types were ventricular septal defect, atrial septal defect, and patent ductus arteriosus. The sensitivity (88.24%) and Youden index (0.86) of dual-indicator screening for CHD detection were significantly higher than the other separate indicators. Through Kappa consistency test analysis, the consistency of dual-indicator screening with simple heart murmur screening results was excellent, with a Kappa coefficient value of 0.835 (>0.75); the consistency of dual-indicator screening and simple POX screening results was good, with a Kappa coefficient value of 0.429 (between 0.40-0.75), and the differences were statistically significant (P<0.001). By the end of the study, 136 children had undergone open-chest/occlusion surgery, with good postoperative recovery, and the rest were followed up. Conclusions　The neonatal CHD screening-diagnosis-assessment technology system established in this study, with close connections between various blocks, high screening response rate and echocardiography examination rate, is beneficial for the early diagnosis and treatment of CHD children, and has certain application value in institutions at all levels in Hainan Province, and is worth further promotion.

Objective To investigate the mechanisms of oridonin inducing apoptosis on acute leukeamia NB4 cells and its mechanism. Methods NB4 cells in culture medium in vitro were given with different concentrations (8, 16, 24, and 32 μmol/L) of oridonin. The inhibitory rate of the cells was measured by MTT assay, cell apoptotic rate was detected by flow cytometry (FCM), morphology of apoptosis was observed by Hoechst 33258 fluorescence staining, DNA fragmentation was assayed by agarose gel electrophoresis, caspase-3 expression was detected by Western blotting, and caspase-3 activity was assayed with colorimetric assay kit before and after apoptosis occurred. Results Oridonin (over 16 μmol/L) could inhibit the growth of NB4 cells and cause apoptosis significantly, the suppression was both in a timeand dose-dependent manner. Marked changes of apoptosis including condensation of chromatin and nuclear fragmentation were observed very clearly by Hoechst 33258 fluorescence staining and a characteristic "ladder" of DNA fragments was elicited by agarose gel electrophoresis; Western blot analysis revealed that caspase-3 was activated by the loss of caspase-3 proenzyme (32 kDa) and the appearance of its 20 kDa subunit, and that along with the apoptotic process caspase-3 activity was increased concurrently. Conclusion Oridonin can induce apoptosis in NB4 cells via activation of caspase-3. These results will provide laboratory evidence for the clinical treatment of acute leukemia with oridonin.

ObjectiveTo unify the definitions of colonoscopic characteristics of Crohn disease (CD) and intestinal tuberculosis ( ITB),and to evaluate colonoscopic and clinical features in the differential diagnosis of CD and ITB.MethodsA collaborative group composed of 10 experts from 5 hospitals voted to identify and confirm the colonoscopic characteristics.Clinical and colonoscopic characteristics were analyzed,thereafter,characteristics were scored based on different diagnostic specificity.ROC curve was used for determining the cutoff point to differentiate CD from ITB.ResultsFirstly,standard endoscopic images and descriptions were determined.Secondly,colonoscopic parameters which were significantly different between the CD and ITB patients included the follows:involvement of more than four intestinal segments,anorectal involvement,longitudinal ulcers,cobblestone appearance and transverse ulcers.Clinical findings which were significantly different between the CD and ITB patients included active pulmonary tuberculosis,PPD-test strong positive,anal fistula/perianal abscess and extra-intestinal manifestations in CD.4.4％(6/136) patients were confirmed by histological evidence of caseating granulomas.By using our scoring system,39.7％ (54/136) confirmed diagnoses and 18.4％ (25/136) suspected diagnoses were made in patients without histological evidence.ConclusionIdentification of colonoscopic characteristics and unification of the colonscopic diagnostic criteria were helpful in the differential diagnosis between CD and ITB.The differential diagnosis rate could he improved by using the scoring system.Half cases could not be confirmed even with combined pathology and the scoring system,so a more comprhensive scoring system would be warranted.