Objective:To determine whether the bcl-2 antisense oligonucleotide (ASODN) can increase the sensitivity of NCI-H460 cell lines to docetaxel or radiation. Methods: Docetaxel in combination with bcl-2 ASODN or non-sense oligonucleotide (NSODN) was used to treat NCI-H460 cells, and then IC_~50 of docetaxel against NCI-H460 cells was determined with MTT method. In radiation group, NCI-H460 cells were divided into pure radiation, radiation plus bcl-2 ASODN, and radiation plus NSODN groups. The inhibition rates of cell growth were assayed by MTT, and the expression levels of Bcl-2 protein were assayed by immunofluorescence. Results: IC_~50 in bcl-2 ASODN plus docetaxel, pure docetaxel and NSODN plus docetaxel groups was (0.101?0.009) ?mol/L,(0.183?0.018)?mol/L, and (0.179?0.016)?mol/L, respectively. The differences between the first group and the latter 2 groups were significant (P

AIM:To explore the effects of the combined use of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN) and cisplatin on apoptosis of HL-60 cells. METHODS:PCR-ELISA was used to determine telomerase activity in HL-60 cells untreated or treated with ASODN, the expression levels of hTERT protein were assayed by immunofluorescence using fluoresce isothiocyanate (FITC) label. Apoptosis was detected by DNA electrophoresis and flow cytomertric cell cycle analysis. RESULTS:The expression levels of hTERT protein and telomerase activity in HL-60 cells treated with ASODN were shown to decrease with time, the DNA fragments were obviously found and the percentage of apoptosis cells was significantly enhanced in HL-60 cells with the combined use of hTERT ASODN and cisplatin compared with the combined use of hTERT sense oligodeoxynucleotide and cisplatin or cisplatin alone, respectively. CONCLUSION:Inhibition of telomerase activity by hTERT ASODN increases the cisplatin-induced apoptosis of HL-60 cells.

AIM: To explore the effects of the combined use of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN) and cisplatin on apoptosis of HL-60 cells. METHODS: PCR-ELISA was used to determine telomerase activity in HL-60 cells untreated or treated with ASODN, the expression levels of hTERT protein were assayed by immunofluorescence using fluoresce isothiocyanate (FITC) label. Apoptosis was detected by DNA electrophoresis and flow cytomertric cell cycle analysis. RESULTS: The expression levels of hTERT protein and telomerase activity in HL-60 cells treated with ASODN were shown to decrease with time, the DNA fragments were obviously found and the percentage of apoptosis cells was significantly enhanced in HL-60 cells with the combined use of hTERT ASODN and cisplatin compared with the combined use of hTERT sense oligodeoxynucleotide and cisplatin or cisplatin alone, respectively. CONCLUSION: Inhibition of telomerase activity by hTERT ASODN increases the cisplatin-induced apoptosis of HL-60 cells.

Objective:To investigate the effects of Bcl-2 shRNA on the growth of liver carcinoma cell line BEL-7402 and colorectal carcinoma cell line Caco2.Methods: Bcl-2 shRNA was cloned into Pgenesil-1 plasmid labeled with fluorescent protein and the product was transfected into BEL-7402 and Caco2 cells with Lipofectamine 2000.The study also included shRNA as negative control,Pgenesil-1,Lipofectamine and blank control groups.Transfected cells were visualized by inverted fluorescent microscope and then assayed by flow cytometry.The expression levels of Bcl-2 protein were assayed by Western-blot and cell proliferation was measured by MTT method.Results: There was no difference in transfection rate among cells in Bcl-2 shRNA,shRNA and Pgenesil-1 vector groups.Western-blot assay showed that the expression levels of Bcl-2 protein in BEL-7402 and Caco2 cells were significantly decreased in Bcl-2 shRNA group compared with those in other 4 groups(P

Objective:To study the mechanism of tretinoin-induced neural differentiation in fetal liver CD34+ cells in vitro. Methods :CD34+ cells from fetal liver were isolated with a magnetic cell sorting kit and were cultured. Cells pretreated with or without protein kinase C(PKC) inhibitor chelerythrine chloride(3 ?mol/L) were induced by 5?10-7mol/L tretinoin for 24 h, and then incubated in serum-free medium. Expressions of genes in treated cells were assayed by Western blotting and RT-PCR. Results: Tretinoin significantly promoted expression of neural specific genes such as Nestin, neuron-specific nuclear protein (NeuN) , neuron-specific neuronfilament-M (NF-M) , and MAP-2 in fetal liver CD34+ cells, with Nestin increased by 4. 09 folds, NeuN by 5. 12 folds, and NF-M by 7. 27 folds (all P

Objective:To isolate mesenchymal stem cells (MSCs) from mouse fetal liver and induce them differentiate into islet B-like cells. Methods:MSCs were isolated from C57BL/6J mouse fetal liver and were induced with high concentration of glucose, basic fibroblast growth factor(bFGF) ,and nicotamine medium. The gene expressions related to islet B cells such as pancreatic duodenal homeobox-1 (PDX-1), proinsulin-1 (INS-1) ,and glucose transporter-2 (GLUT-2) were detected by RT-PCR. Insulin in the treated cells was examined by immunocytochemistry. The insulin clusters were stained with dithizone (DTZ), a zinc-chelating agent known to selectively stain pancreatic B cells. Results: RT-PCR showed that the treated cells expressed PDX-1, INS-1 and GLUT-2, while the undifferentiated cells did not. After approximately 10 d of treatment, the fetal liver cells formed DTZ-stained cell clusters in flasks (80-120 clusters in a flask 25 cm2 in area). Immunocytochemistry also confirmed that these aggregates were strongly positive for insulin. Conclusion: MSCs derived from fetal liver can be induced into islet B-like cells in vitro.

Objective　 To investigate the indication and timing of surgery for acute necrotizing pancreatitis.　 Methods　 82 cases of acute necrotizing pancreatitis were analyzed retrospectively. 10 cases were treated non-operatively. Early operation was performed on 44 cases, while 28 cases underwent surgery on late stage.　 Results　 The overall morbidity and mortality was 24% and 18%, respectively. All 10 patients in the non-operative group were cured with a morbidity of 10%. The morbidity and mortality in the early operation group was 14% and 11%, respectively, compared with 46% and 36% of those receiving surgery on late stage（P＜0.01, P＜0.05）. Among those in late surgery group, patients not complicating infection had significant lower morbidity and mortality rate than those suffering from severe infection or organ dysfunction（P＜0.01, P＜0.05）.　 Conclusions　 Early operation is necessary for some severe cases.In patients with severe pancreatic necrosis surgery should be performed before severe infection occurs.

Objective To discuss Extracorporeal Membrane Oxygenation(ECMO) management method and effect during inter-hospital transport of potential cardiac death donors after cardiac death (DCD).Methods 8 potential donors after cardiac death with brain injury were supported by ECMO for inter-hospital transport.All donors were inserted Medtronic overall cannula into one side femoral artery and venous.The position of catheters were guided by ultrasound.The front-end of venous catheter located in the junction of atrium and inferior vena cava,meanwhile the front-end of artery catheter was below renal artery.100 IU/kg heparin was injected before inserting cannulas.Flow of ECMO maintained at 2.0～3.0 L/min,and oxygen flow was 2～3 L/min during ECMO supporting.When hemodynamics of potential donors were stable,patients were moved into ambulance with ECMO for inter-hospital transport.Results A total of 8 ECMO transports were performed for central circulatory collapse caused brain injury.Patients were previously cannulated and on ECMO prior to transport and transported a distance of more than 100 kilometer from our institution by ambulance.ECMO running times were 120 min,and operation process circulatory stable.Conclusion ECMO can ensure inter-hospital transport of potential donors after cardiac death safety.

Objective To evaluate the impact of sleep-disordered breathing(SDB)on the quality of life(QOL)in children with SDB and investigate the value of tonsillectomy and adenoidectomy in improving their QOL.Methods Questionnaire of disease-specific quality of life for children with obstructive sleep apnea with 18 items(OSA-18)was used to evaluate QOL in 168 children with SDB within four weeks before and six to 12 months after surgical operation,and correlation between OSA-18 scores to apnea hypopnea index(AHI)and minimum oxygen saturation(MiO2)in 62 of them was evaluated with polysomnography(PSG),with 50 healthy children without snoring as controls.Results Overall scores of OSA-18 in children of SDB group before surgical operation were significantly higher than those in children of control group(t=15.12,P＜0.01).and QOL was seriously affected in 30.4 percent of the children with SDB.Scores of OSA-18 correlated to AHI and MiO2 before surgical operation.QOL was improved significantly in 73.8 percent of children with SDB after tonsillectomy and adenoidectomy.Conclusions SDB has obvious impact on pediatric QOL,which can be improved by tonsillectomy and adenoidectomy.There exists significant correlation between OSA-18 scores and objective indicators of PSG,and the former can be used as clinical diagnosis for SDB in children and as quantitative evaluation for the effectiveness of intervention.

AIM: To investigate whether the bcl-2 antisense oligonucleotide increases the sensitivity of HL60 and K562 cell lines to daunorubicin. METHODS: IC50 for HL60 and K562 was determined with MTT method, the expression levels of Bcl-2 protein were assayed by immunofluorescence using fluoresce isothiocyanate labeling. In addition, apoptosis was detected by morphological observation and flow cytometric analysis of DNA fragmentation. RESULTS: It was found that the two oligonucleotides directed against the coding region and the translation initiation of bcl-2 mRNA, combined respectively with daunorubicin, inhibited expression of bcl-2 protein, increased apoptosis in HL60 and K562 cells, and decreased IC50 of daunorubicin significantly ( P