AIM: To explore whether human telomerase reverse transcriptase (hTERT) antisense oligodeoxynucleotide (ASODN) could enhance the sensitivity of acute lymphoblastic leukemia cells from relapse patients to cisplatin. METHODS: The expression levels of hTERT protein were detected by immunofluorescence using fluorescence isothiocyanate (FITC), the number of viable cells was determined using the trypan blue dye exclusion assay, and apoptosis was detected by morphological observation and flow cytometric cell cycle analysis. RESULTS: The expression of hTERT protein was inhibited after treatment with hTERT ASODN. Treatment with cisplatin combined with hTERT ASODN had significantly reduced the number of viable acute lymphoblastic leukemic (ALL) cells (P<0.05). In morphological observation of apoptotic cells using Hoechst33258 and PI double staining techniques, cells displayed classic apoptotic changes in the presence of cisplatin or cisplatin combined with hTERT ASODN or ASODN at 48 h. Apoptotic rates of cells treated with cisplatin and ASODN were higher than that of cells treated with cisplatin alone (P<0.05). CONCLUSION: hTERT ASODN could increase sensitivity of cultured primary acute lymphoblastic leukemia cells from relapse patients to cisplatin.

Objective To explore the effect of carnosic acid combined with tamoxifen on breast cancer cells and the mechanism.Methods Breast cancer cell lines MCF-7,T47D were divided into 4 groups:PBS was given to control group,carnosic acid(CA)was added to CA group,tamoxifen(TAM)was added to TAM group,and CA+TAM group was treated with carnosic acid and tamoxifen.The proliferation ability was measured by MTT assay,the colony formation capacity was measured by colony formation assay,the invasion ability was measured by Transwell chamber assay,the migration ability was measured by wound-healing assay,the expression of Caspase-8,Caspase-3 and PARP protein were measured by Western blotting.Results CA,TAM and CA+TAM suppressed breast cancer cell proliferation with a dose-dependent manner.Compared with CA group and TAM group,the proliferation inhibitory rate of CA+TAM group was significantly increased(P<0.05).In MCF-7 and T47D cell lines,compared to control group,the rate of colony fromation,number of invasive cells and cell migration rate of CA,TAM,CA+TAM groups were significantly decreased(all P<0.05).These indexes mentioned above in CA+TAM group were significantly lower than those in CA group and TAM group(all P<0.05),but there were no significant differences between CA group and TAM group (all P>0.05).In two breast cancer cell strains,the expression of cleaved Caspase-8 protein between control,CA,TAM and CA+TAM groups was not significantly different(P>0.05).Compared to control group,the expression levels of cleaved caspase-3,cleaved PARP protein in CA,TAM and CA+TAM groups were significantly increased(P<0.05),and these protein levels were significantly higher in CA+TAM group than in CA and TAM group(all P<0.05).But there were no significant differences between CA group and TAM group(all P>0.05).Conclusion Carnosic acid combined with tamoxifen can suppress the proliferation,invasion and migration of breast cancer cells,and its mechanism may be related to the activation of Caspase-3/PARP signaling pathway.

A Review] The oncogene expression and growth of leukemic cells could be inhibited by antisense oligonucleotide. The selection of target genes is the key step in the research of antisense oligonucleotide on leukemia. The article would review the status and prospect of some target genes of leukemia in the investigation of antisense oligonucleotide.

AIM: To investigate whether antisense oligodeoxynucleotides of hTERT?bcl-2 and c-myc could enhance the sensitivity of leukemia cell K562 to cisplatin. METHODS: The inhibiting effects of cisplatin and cisplatin plus antisense oligodeoxynucleotide on K562 cells were determined by MTT. RESULTS: The inhibiting rate of 20 ?mol/L cisplatin to K562 cell is 17.17%?1.36% and it becomes 25.41%?1.77% ,26.18%?1.43% and 28.29%?1、05%, respectively, as combinated with antisense oligodeoxynucleotide of hTERT?bcl-2 or c-myc. CONCLUSION: These results indicated that antisense oligodeoxynucleotides of hTERT?bcl-2 and c-myc enhanced efficacy of cisplatin in K562 leukemic cells.

AIM: To observe the effect of antisense oligodeoxynucleotide of human telomerase reverse transcriptase(hTERT) on the telomerase activity in Jurkat cells and its mechanism. METHODS: The telomerase activity was detected by TRAP. hTERT protein and mRNA expressions were detected by flowcytometry and RT-PCR. RESULTS: The absorbency of the cells ( A values) was used to represent the telomerase activity. At 48 h and 72 h, the A values of the Jurkat cells treated with antisense oligodeoxynucleotide of hTERT decreased to 0.351?0.051 and 0.238?0.024, respectively, compared to the A value of 0.492?0.051 in the control cells without treatment ( P

AIM: To explore the effect of human telomerase reverse transcriptase (hTERT) gene antisense phosphorothioate oligodeoxynucleotide (AS PS-ODN) on cisplatin-induced apoptosis in cultured primary acute lymphoblastic leukemia (ALL) cells. METHODS: The expression levels of hTERT protein were detected by immunofluorescence using fluoresce isothiocyanate (FITC) lable. Cell surviving fraction was determined using the trypan blue dye exclusion assay. Apoptosis was detected by morphological observation and flow cytomertric cell cycle analysis. RESULTS: The expression of hTERT protein was inhibited after treated by hTERT AS PS-ODN. Treatment with cisplatin after 24 h of exposure to AS PS-ODN had significantly reduced the number of viable ALL cells. However,there was no difference on ALL cells survival between sense oligodeoxynucleotide (S PS-ODN) /CDDP combination and CDDP-treated cells alone. In morphological observation of apoptotic cells using Hoechst 33258 and PI double staining techniques,cells displayed classic apoptotic changes treated with CDDP or CDDP combined with hTERT AS PS-ODN or S PS-ODN at 48 h. Apoptotic rates of cells added CDDP and AS PS-ODN were higher than that of cells added CDDP only ( P 0.05). CONCLUSION: hTERT AS PS-ODN inhibited the expression of hTERT protein and increased the CDDP-induced apoptosis in primer acute lymphoblastic leukemic cells.

AIM: To explore the effect of hTERT antisense phosphorothioate oligodeoxynucleotide (ASODN) on apoptosis induced by chemotherapeutic drugs in Jurkat cell lines. METHODS: Cell viability was determined using the trypan blue dye exclusion assay. Apoptosis was detected by morphological observation, DNA gel electrophoresis and flow cytometry analysis. RESULTS: The survival rates of Jurkat cells cultured with daunorubicin, vincristin, and etoposide, respectively were similar with that cultured with those chemotherapic drugs plus hTERT ASODN. The survival rates of Jurkat cells cultured with cis-diamminedichicloroplatinum(DDP) added 24 hours later were higher than that cultured with hTERT ASODN and DDP added 24 hours later. The survival rates of Jurkat cells cultured with DDP were similar with that cultured with hTERT SODN and DDP. In morphological observation of apoptotic cells using Giemsa staining, cells displayed classic apoptotic changes treated with DDP or DDP combined with hTERT ASODN or SODN at 48 hours. Agarose gel electrophoresis of genomic DNA from Jurkat cells treated with ASODN and DDP combination for 48 hours showed typical DNA “ladder”. Neither the DNA from Jurkat cells treated with SODN plus DDP nor the DNA from the cells treated with DDP alone showed ladder pattern. Apoptosis rates of Jurkat cells treated with DDP for 48 hours after 24 hours of exposure to ASODN significantly increased. There was significant difference in the percentage of apoptotic Jurkat cells between hTERT ASODN plus DDP and SODN plus DDP or DDP alone, respectively. CONCLUSION: The hTERT ASODN complementary to the translation initiation region of hTERT mRNA enhanced DDP-induced apoptosis in Jurkat cells.

AIM: To explore the effect of hTERT gene antisense phosphorothioate oligodeoxynucleotide (ASODN) on telomerase activity in Raji cells. METHODS: Polymerase chain reaction enzyme-linked immuonassay (PCR-ELISA) was used to determine telomerase activity. The expression levels of hTERT mRNA and protein were assayed by reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence using fluoresce isothiocyanate (FITC) label,respectively. RESULTS: RT-PCR and immunofluorescence assay showed that the expression levels of hTERT mRNA and protein from Raji cells decreased with time after hTERT ASODN treatment. There was no difference in hTERT mRNA and protein levels between control and SODN-treated cells. Telomerase activity decreased when Raji cells were treated with ASODN for 48 h. Telomerase activity of Raji cells was significantly inhibited when treated with ASODN for 72 h. There was no difference in telomerase activity levels between control and hTERT SODN-treated cells. CONCLUSION: hTERT ASODN could inhibit telomerase activity in Raji cells.

Objective To compare the clinical efficacy of intralesional betamethasone versus triamcinolone acetonide acetate in the treatment of active alopecia areata. Methods A total of 160 patients with active alopecia areata were divided into two groups, test group (n = 100) treated with intralesional betamethasone, and control group (n = 60) treated with intralesional triamcinolone acetonide. Both injections were given once every 3 weeks for 12 consecutive weeks. Results After 12-week treatment, the cure rate, response rate, and total response rate were 60.0%, 32.0% and 92.0% in the test group, respectively, compared to 41.7%, 31.67% and 73.3% in the control group, respectively. A significant increase was observed in the cure rate and response rate in the test group compared with the control group (χ2 = 10.25, 5.06, P < 0.01 and 0.05). During the treatment course, 8 (8%) patients in the test group and 9 (15%) patients in the control group developed localized atrophy of the scalp; 8 (8%) patients in the test group and 3 (5%) patients in the control group developed localized folliculitis; no significant difference was observed between the two groups in the occurrence of adverse reactions (P> 0.05). Conclusion Intralesional use of compound betamethasone injection has a notable therapeutic effect on alopecia areata.

Objective To set up the isolated perfused rat liver model. Methods Rat livers were harvested after the cannulation of the portal vein and bile duct. The reperfusion solution was Krebs-Henseleit solution containing bovine albumin serum and sodium taurocholate. The original CZ-1 isolated perfused rat liver system contained two subsystems: recirculating perfusion system and heat-exchange system. Then we modified the original CZ-1 system and omitted the heat-exchange system.The modified CZ-1 system consisted of a thermostatically regulated water bath,a peristaltic pump,a 4-neck round-bottom flask, a flow meter, an in-line manometer, a glass organ chamber, an iron support and a set of recirculating pipe line. Then the livers were connected via the portal vein to the modified CZ-1 system for 120 min. After 120 min reperfusion, bile production was evaluated. Routine HE staining and electron microscopic examination of hepatic tissues were also performed. Results The was not significantly different from that reported by references. Hepatic tissues in reperfusion group were also morphologically normal Conclusion The CZ-1 isolated perfused rat liver system was cost-effective and reliable to use. It was easy to run and is the ideal model for investigation of organ preservation solution.