AIM To study the effect of antisense oligonucleotide s of telomerase on the telomerase activity and the mechanisms of its inhibiting cervical carcinoma HeLa cell line proliferation. METHODS The cytotoxic effect in vitro was determined using the trypan blue dye exclusion assay. The telomerase activity was performed by PCR-TRAP and the flow cytometry analysis was employed to investigate the effects of ASON on the cell cycle and apoptosis. RESULTS There was no significant decrease in cell viability after treated with ASON for 5 days and the ASON with FuGENE6 reduced the telmerase activity compared with the tandem primer and the empty controls. The efficancy depended on the dose of ASON and the time of treatment. ASON did not induce cell apoptisis but increased G 2/M-phase cells. CONCLUSION Antisense oligonucletides of telomerase inhibited the telomerase activity but did not induce apoptosis in HeLa cell lines .Retardation of G 2/M-phase may be the mechanism of inhibiting proliferation in HeLa cell lines.

Aim To investigate the effects of human telomerase reverse transcriptase(hTERT)antisense phosphorothioate oligodeoxynucleotide(ASODN)on telomerase activity and apoptosis induced by crisplatin(DDP)in HeLa cell lines.Methods HeLa cells were collected after transfection of 0.05,0.1,0.2 ?mol?L-1of ASODN and the total DNA was extracted.Telomeric repeat amplification protocol(TRAP)-AgNO3 stain was used to detect the activity of telomerase.Apoptosis of HeLa cells was detected by AO staining morphological observation and flow cytometry(FCM)analysis technology.Results The telomerase activity of HeLa cells was expressed in high level,which was significantly reduced dose-dependently and time-dependently after treatment with ASODN of hTERT.Two days after transfection of ASODN of 0.05 ?mol?L-1,0.1 ?mol?L-1,0.2 ?mol?L-1,the telomerase activity decreased by 21.77%,52.42% and 71.10% respectively.After treatment with ASODN for 24,48 h and 72 h,the telomerase activity decreased by 12.48%,38.27% and 71.10% respectively.In morphological observation using acridine orange(AO)staining method,HeLa cells displayed typical apoptotic figure after treated with DDP plus ASODN of hTERT.The percentage of apoptosis of HeLa cells treated with DDP of 1.5 mg?L-1,3.0 mg?L-1,for 48 hours after 24 hours exposure to ASODN of 0.2 ?mol?L-1(50.35%,29.67%)was significantly higher than that treated with DDP alone(19.67%,11.38%,P

Aim To investigate the apoptosis of cervical cancer HeLa cells cultured in vitro transfectead by combining telomerase anti-hTR and anti-hTERT. Methods The activity of telomerase was tested by polymerase chain reaction-telomeric repeat amplification protocolenzyme-linked immunosorbent assay (PCR- TRAP- ELISA). Cell morphologies were observed by fluorescence microscope stained with acridine orange. Apoptosis and cell cycle were examined by Flow Cytometer method (FCM). Results The telomerase activity of HeLa cells transfected by antisense oligonucleotides were significantly decreased compared with those of nontreated control,sense oligonucleotides and oligofectamine TM(P

Objective To investigate the inhibitory effect of combination of antisense human telomerase RNA (anti hTR) and antisense human telomerase catalytic subunit (anti hTERT) on proliferation of cervical cancer Hela cell line cultured in vitro Methods Cervical cancer Hela cells were transfected by anti hTR, anti hTERT, anti hTR + anti hTERT, sense hTR, sense hTERT with Oligofectamin TM reagent The proliferation activity of cervical cancer Hela cells was determined using methyl thiazolyl tetrazolium (MTT) The activity of telomerase was tested by polymerase chain reaction telomeric repeat amplification protocol enzyme linked immunosorbent assay (PCR TRAP ELISA) Cell morphologies were observed under fluorescence microscope with acridine orange staining Apoptosis and cell cycle were examined by flow cytometer method (FCM) Results The inhibitory rates on proliferation and apoptotic rates of Hela cells transfected by antisense oligonucleotides were significantly higher compared with those of control, sense oligonucleotides and Oligofectamin TM (all P

Objective:To analyze the curative effect of suppression expression of URG4 on cervical cancer mice and the possible mechanisms.Methods:The expression of URG4 were analyzed in cervical cancer tissues and para-carcinoma tissues.16 male BALB/c nude mice were divided into Group A ( inoculated with URG4-siRNA SiHa cells) and Group B ( inoculated with SiHa cells) . After 4 weeks,mice were executed and analyzed.Results:The expression of URG4 were higher in human cervical cancer tissues than that in para-carcinoma tissues ( P<0.05).After experiment,Group A had a lower tumor weight,but a higher body weight than these of Group B,the difference was statistically significant (P<0.05).Group A mice had a lower levels of URG4 in tumor than that of Group B mice (P<0.05).Group A mice had a lower levels of Ki67 in tumor than that of Group B mice,the difference was statistically significant (P<0.05).Group A mice had lower levels of CyclinD1,β-catenin and C-myc in tumor than these of Group B mice (P<0.05). Conclusion:The expression of URG4 in cervical cancer tissue is up-regulation.Suppression expression of URG4 can suppress tumor growth in vivo,which may though suppress the Wnt/β-catenin signaling pathway.

Aim To study the effect of Vitamin C on growth and apoptosis of human cervical cancer cell line HeLa and its mechanisms. Methods HeLa cell line was treated with VitC. Cell proliferation was evaluated with MTT assay. The distribution of cell cycle,the rate of apoptosis and the expression of HPV18 E6,p53,Bax and Bcl-2 protein of the heLa cells, were observed using flow cytometer respectively. TRAP-PCR-ELISA assay was used to evaluate telomerase activity. Results Vitamin C arrested the HeLa cell line in G_0/G_1 phase in a dose and time-dependent manner, and significantly inhibited HeLa cells growth and increased the rate of apoptosis.Expression of HPV18 E6 and Bcl-2 proteins, was down regulated expression of p53 and Bax proteins was upregulated, and the telomerase activity was reduced. Conclusion Vitamin C can significantly inhibit HeLa cells growth and proliferation,arrest them in G_0/G_1 phase and induce their apoptosis.The reason of these changes is the decrease of E6 gene′s expression, and the succedent changes of proteins′ expression in cancer cells,including the downregulated expression of HPV18 E6 and Bcl-2 proteins, upregulated expression of p53 and Bax proteins and reduced telomerae activity.

As the amount of PET/CT increases,the amount of cyclotron also increases.Hospitals are confronted with some similar problems in cyclotrons purchase.The article is about the stability,after service,yields of nuclide and the particle energy of cyclotron.The stability of cyclotron is the most important factor,and after service is in the next place,while the yields of nuclide are also very important.

Objective To explore the changes of estrogen receptor (ER) and progesterone receptor (PR) in the endometria of pro-liferative phase in polycystic ovary syndrome (PCOS) patients. Methods Thirty-two patients who suffered PCOS combined with infertilitas feminis were enrolled in this study, and 20 cases of tubal infertilitas feminis having the corresponding time period were selected as controls. The expressions of estrogen receptor (ER) and progesterone receptor (PR) in the endometria were observed by pathological examination and immunohistochemical staining. Results The expressions of ER and PR in the PCOS group in the glands and interstitium of endometrial a-mong three different periods were not significantly different. The expressions of ER and PR in the glands and interstitium of endometrial a-mong three different periods in the control group were significantly different. There was no statistical difference in the expressions of ER, PR in the glands and interstitium of the early endometria between PCOS group and control group. The expressions of ER in the glands and inter-stitium of the middle endometrial in PCOS group was significantly lower than that of control group. The expressions of ER and PR in the glands and interstitium of the late proliferative endometrial in PCOS group was significantly lower than that of control group. Conclusion ER and PR of endometrial in the PCOS patients decreased. The cyclical of ER and PR in the PCOS patient were irregular.

Objective To explore the clinical value of expression of CEA and CK-20 detected by flow cytometry (FCM) in peritoneal washes on predicting peritoneal metastasis and prognosis of gastric carcinama.Methods Clinicopathological characteristics and follow up data of 105 patients with gastric carcinoma who were underwent D2 radical resection (R0) were collected.Peritoneal washes was collected.Peritoneal lavage cytology examination (PLC) was used to find intraperitoneal free cancer cells (IFCC).FCM was used to determine the CEA and CK-20 expressions.Meanwhile,13 patients with benign lesion on the stomach and the gastric carcinama cell line SGC-7901 were served as the negative and positive control,respectively.Results Positive expression of CEA was in 48 (45.7 ％) patients,of CK-20 was found in 67 (63.8 ％) patients,and of CEA and CK-20 was in 85 (81.0 ％) patients by FCM.However,positive expression of IFCCs was found in 31 patients(29.5 ％) by PLC.The expression of CEA and CK-20 were related to serosa invasion,lymphnode metastasis and pTNM stage (P ＜ 0.05).The median survival of patients with negative expression of both CEA and CK-20 (n =20) was significantly longer those with positive expression (n =30)(52 vs 18 months,P ＜ 0.05).Conclusions Combined Detection of CEA and CK-20 in peritoneal washes by FCM can be used to predict peritoneal micrometastasis and may predict the prognosis of gastric carcinoma.

ObjectiveTo investigate the effect of neoadjuvant chemotherapy in locally advanced cervical cancer on cell cycle,proliferation,and evaluate the feasibility of proportion of cell in different phase 　and proliferating cell nuclear antigen(PCNA) as sensitive indices to assess chemotherapeutical sensitivity and therapeutical effect.MethodsForty-nine cases of locally advanced cervical cancer were divided into response group and no-response group according to the clinical efficacy of neoadjuvant chemotherapy.Compared the proportion of cell in different phase and proliferation index of PCNA between two groups.The clinical therapeutic effect was evaluated after 4 weeks in the second neoadjuvant chemotherapy regimen.Results In 49 patients with locally advanced cervical cancer,clinical effective of 39 cases (response group),no effective of 10 cases(no-response group).The S-phase proportion of cell in the pre-neoadjuvant chemotherapy and post-neoadjuvant chemotherapy of response group[ (21.47 ± 5.21 )％ and(18.32 ±5.07)％] were higher than those of no-response group [ (9.63 ± 2.58)％ and ( 10.14 ± 2.32)％ ] (P ＜ 0.05 ).The proliferation index of PCNA of cervical cancer in the pre-neoadjuvant chemotherapy of response group [ ( 81.67 ± 7.14)％ ] was higher than that of no-response group [ (66.99 ± 2.29 )％ ] (P ＜ 0.05 ).Conclusion The S-phase proportion of cell and proliferation index of PCNA in the pre-neoadjuvant chemotherapy are important indexes to assess chemotherapeutical sensitivity and therapeutical effect.