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 investgate the expression of Survivin and Caspase-3 in primary ovarian carcinoma, and to explore their relationship. Methods The expression of Survivin and Caspase-3 was detected by flow cytometry (FCM) . Results FCM showed that the expression of Survivin in ovarian serous carcinoma was higher than that in serous cystadenoma and normal ovarian tissues (F=19.80;P=0.00) and correlated with the clinical stage( t=2.882,P=0.009).The expression of Caspase-3 in ovarian serous carcinoma was higher than that in serous cystadenoma and normal ovarian tissues (F=16.32,P=0.00)and has no correlativity with clinical stage,histological grade,and lymph metastasis(P＞0.05).Survivin expression was negatively correlated with Caspase-3 expression in ovarian serous tumors (r=0.43,P=0.029).Conclusion Survivin was negatively correlated with Caspase-3 in ovarian serous tumors.Combination of detecting the two protein expressions might be valuable in diagnosis and determining the malignant degree in ovarian serous carcinoma.

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 construct an eukaryotic expression vector of pre-miR-15a,and to investigate the inhibitory effect of pre-miR-15a to Raji cells. Methods The pGCSIL-GFP vector encoding pre-miR-15a nucleotides was transfected into the bacterial competent cells,and then confirmed by PCR and sequencing analysis. The identified vector was transfected into Raji cells with oligofectamine 2000. The cells were divided into 3 groups,blank,negative control and pre-miR-15a group. Semi-quantitative RT-PCR was used to detect the expression of Bcl-2 mRNA,and immunofluorescence indirect for Bcl-2 protein expression. The growth of Raji cells was assayed by trypan blue dye exclusion method. Results PCR and sequences analysis indicated that the recombinant clones was identical with target sequences. Many green fluorescent cells were observed under fluorescent microscopy. The levels of Bcl-2 mRNA at every group had no obviously difference. Bcl-2 protein expression was obviously decreased at pre-miR-15a group compared with the other groups. Trypan blue dye exclusion method showed the cell growth was inhibited at 48 h and 72 h post-transfection. Conclusion We successfully construct the eukaryotic expression vector of pre-miR-15a,and it can inhibit the growth of Raji 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

Objective It was designed to investigate the relationship between the productions of p16 and Rb genes expression and gastric carcinogenesis and evaluate the correlation between the expression of p16 and Rb genes in gastric carcinoma.Methods The expression of P16 and Rb proteins was examined in gastric carcinoma and precancerous lesion and normal gastric mucosa by streptavidin-peroxidase immunohistochemical method (S-P).Results The positive rates of P16 and Rb proteins expression showed as below: There were 96 25%(77/80) and 98 75%(79/80) in normal gastric mucosa, 92 00%(46/50) and 80 00%(40/50) in atypical hyperplasia gastric mucosa and 47 54%(58/122) and 59 84%(73/122) in gastric carcinoma respectively. The positive rates of P16 and Rb proteins expression in gastric carcinoma were significantly lower than that in normal gastric mucosa and atypical hyperplasia gastric mucosa (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.

AIM: To study the mechanism of butylated hydroxyanisole-induced neural differentiation of fetal liver Sca-1~+ cells in vitro. METHODS: Sca-1~+ cells from 14.5-day-old mouse fetal liver were isolated with a magnetic cell sorting kit. Cultured cells pretreated with or without extracellular signal-regulated kinase (MEK1) inhibitor, PD98059, were induced by 200 ?mol/L butylated hydroxyanisole (BHA) for 24 hours, and then incubated in serum-free medium. Expression of genes in treated or untreated cells were assayed by Western blotting and RT-PCR. RESULTS: There was low level of neuronfilament-L (NF-L) and brain factor-1 (BF-1) in fetal liver Sca-1~+ cells, but no neuronfilament-H (NF-H) and tyrosine hydroxylase (TH) was observed. BHA significantly promoted the expression of neuron-specific NF-L, NF-H, BF-1, and TH in fetal liver Sca-1~+ cells. NF-L, NF-H, BF-1 and TH increased by 6.32 fold, 2.73 fold, 3.37 fold and 2.68 fold, respectively (all P

Recent researches have showed that neural stem cells be isolated not only in the embryonic nervous system but also in adult nervous system. Neural stem cells have some capacity for self-renewal, migration and can differentiate into neurons, astrocyte and oligodendrocyte. This provides a basis in therapy of degenerative diseases and functional recovery of the central nervous system injury. Also helps in the study of determining the postmortem interval ( PMI) and postmortem injury. Distribution, biological characteristic, identification and the factors effecting the proliferation, differentiation and migration with reference to therapy of neural stem cells and applications in forensic pathology are reviewed.

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