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 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

Matrine is an effective natural product isolated from the traditional herb,Sophora flavescens,with wide range of pharmacological and clinical effects and treating as the treatment for chronic viral hepatitis in clinic.Currently,the mainstream clinical preparations of matrine involved injections,tablets,capsules and suppositories,while preparations under development included targeting preparations,sustained-release preparations and transdermal drug delivery preparations.Admittedly,It can significantly improve the biological utilization of matrine through the optimization of its clinical preparations for with scientific researches and the development of new dosage forms.While the clinical applications will be expanded and the clinical efficacy of the preparations of matrine enhanced with the development of researches for new dosage forms and drugs and the application of new materials.It may be also benefical to the study of monomer compositions of Chinese material medica,including the development of the preparation of matrine.Rested on the recent studies,pharmacological activities,preparation methods and researches for the preparations of matrine was summarized in this text for providing a reference for developing its preparations in the future.

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.

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

AIM: To explore the effect of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (ASODN) on chemotherapeutic durges-induced apoptosis in CEM cells. METHODS: Cell viability was determined by trypan blue dye exclusion assay. Apoptosis was detected by morphological observation and flow cytomertric cell cycle analysis. RESULTS: The survival rates of CEM cells cultured with daunorubicin, vincristin and (etoposide) respectively were similar with that cultured with those chemotherapic durgs plus hTERT ASODN. The survival rates of CEM cells cultured with cis-diamminedichicloroplatinum (DDP) added 24 h later were higher than that cultured with hTERT ASODN and DDP added 24 h later. The survival rates of CEM cells cultured with DDP were similar with that cultured with hTERT SOND and DDP. In morphological observation of apoptotic cells using Giemsa staining, cells treated with DDP or DDP combined with hTERT ASODN ro SODN at 48 h, displayed classic apoptotic changes. Apoptosis rates of CEM cells treated with DDP for 48 h after 24 h of exposure to ASODN significantly increased. There was significant difference in the percentage of apoptotic cells of CEM cells between hTERT ASODN plus DDP and SODN plus DDP or DDP alone, respectively. CONCLUSION: hTERT ASODN enhances DDP-induced apoptosis of CEM cells.

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 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