Objective:To study the inhibitory effect of E1A gene on the growth of tumors in nude mice implanted with nasopharygeal carcinoma CNE2 cells and its promotion effect on the racliosensitivity of CNE2-implanted tumors, and to investigate the related mechanism. Methods: E1A gene was transfected into CNE2 cells using adenovirus system, and sta-ble E1A positive clones were established. The inhibitory effect of E1A on tumor formation-ability of CNE2 cells was ob-served in nude mice. The efficacy of E1A gene therapy with or without radiotherapy against CNE2 cell-implanted tumors was evaluated. The effect of E1A gene therapy on the expression of P53 was detected by RT-PCR. Results: CNE2 cells stably transfected with E1A gene (CNE2-Ad-E1A) were successfully established. The tumor formation time was later and tumor size was smaller in CNE2-Ad-E1A cell-implanted mice compared with those in CNE2 cell- and CNE2-Ad-β-gal cell-implanted mice (CNE2 cells stably transfected with Ad-β-gal). Radiotherapy, E1A gene therapy and E1A gene + radio-therapy all suppressed the growth of implanted tumors, with the tumor suppression rates being (60.32±5.34) %, (70.53±6.12) %, and (97.15±4.87) % , respectively. E1A gene therapy significantly increased the expression of P53 gene in tumor tissues. Conclusion: E1A can inhibit the growth of tumors in mice implanted with nasopharygeal carcinoma cells, and enhance its sensitivity to radiotherapy, which may be related to the increased expression of P53 gene in tumor tissues.

Inhibitor of growth 2 (INC2 ) is a member of INC family, and cooperates with p53 in regulating cell cycle, apoptosis, and aging. It can also facilitate chromatin remodeling and transcription regulating through histone modifications. Researches suggest that INC2 protein is an important factor in the genesis, metastasis and prognosis of many kinds of tumors. Resent studies indicate that INC2 can not be simply classified as oncogene or tumor suppressor. At present, the function of INC2 in tumorigenesis and tumor development involves interaction with varied molecules, but the mechanisms remain largely unknown, which still require further investigation.

Objective To discuss the clinical effect and safety of transurethral holmium laser resection of bladder tumors.Methods A total of 20 patients with bladder tumors(stage Ta～T2a)was treated by holmium laser resection transurethrally.There were 17 patients with primary tumor and 3 patients with recurrent tumor.The laser power was set at 15～40 W.Small lesions were vaporized directly,while large ones(more than 1.0 cm in diameter and with broad pedicels)were incised from the pedicel,with neighboring tissues 1～2 cm in extent vaporized and cauterized.Results The tumors were removed on one session in all the 20 patients.The operation time was 10～70 min(mean,30 min).No complications such as obturator nerve reflex,bladder perforation,or overhydration occurred.No blood transfusion was required.The postoperative catheterization time was 1～5 days(mean,3 days).No recurrence was found during follow-up examinations for 3 months in 16 patients and 6 months in 4 patients(mean,3.6 months).Conclusions Transurethral holmium laser resection of bladder tumors is a safe and effective procedure for the treatment of bladder tumors.

Objective:To study the inhibitory effect of E1A gene on the growth of tumors in nude mice implanted with nasopharygeal carcinoma CNE2 cells and its promotion effect on the radiosensitivity of CNE2-implanted tumors,and to investigate the related mechanism.Methods: E1A gene was transfected into CNE2 cells using adenovirus system,and stable E1A positive clones were established.The inhibitory effect of E1A on tumor formation-ability of CNE2 cells was observed in nude mice.The efficacy of E1A gene therapy with or without radiotherapy against CNE2 cell-implanted tumors was evaluated.The effect of E1A gene therapy on the expression of P53 was detected by RT-PCR.Results: CNE2 cells stably transfected with E1A gene(CNE2-Ad-E1A)were successfully established.The tumor formation time was later and tumor size was smaller in CNE2-Ad-E1A cell-implanted mice compared with those in CNE2 cell-and CNE2-Ad-?-gal cell-implanted mice(CNE2 cells stably transfected with Ad-?-gal).Radiotherapy,E1A gene therapy and E1A gene+radiotherapy all suppressed the growth of implanted tumors,with the tumor suppression rates being(60.32?5.34)%,(70.53?6.12)%,and(97.15?4.87)%,respectively.E1A gene therapy significantly increased the expression of P53 gene in tumor tissues.Conclusion: E1A can inhibit the growth of tumors in mice implanted with nasopharygeal carcinoma cells,and enhance its sensitivity to radiotherapy,which may be related to the increased expression of P53 gene in tumor tissues.

Objective To investigate the proteomics differences between the high-sensitivity(HS) and the low-sensitivity(LS)groups of cervical carcinoma treated by concurrent chemoradiotherapy,and to confirm the sensitivity associated proteins in intermediate stage and advanced cervical carcinoma.Methods Fresh carcinoma tissues were collected from 10 untreated cervical carcinoma patients.According to the response to concurrent chemoradiotherapy,the tissues were classified into HS group and LS group.In the first part of our experiment,protein separation was performed using two-dimensional gel electrophoresis(2-DE)with Amersham 18 am linear pH 3-10 immobilized pH gradient(IPG)strips.The images of the gels were analyzed by PD-quest 7.0 software to find the differentially expressed protein-spots in each group.Then the differentially expressed protein-spots were incised from the gels and digested by trypsin.The peptide mass flngerprintings(PMF)was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS).The proteins were identified by data searched in the Mascot-database.Two differentially expressed proteins were assayed by western blot and immunohistochemical methods.Results Most of the gels were clear and successfully analyzed by PD-quest 7.0 software.Most of the protein-spots concentrated on the area of 20-100 KDa(Mw)and pH4-8.The average number of the protein-spots was 781±74 in HS group and 766±52 in LS group.The match rate was 87.6%between the two groups.Eight proteins highly in HS group but lowly expressed in LS group included hemoglobin subunit beta,caspase-14 precursor,calmodulin-like,S100-A9 protein(MRP-14),galectin-7,HSKERC4,keratin 19 and actin.Ten proteins highly in LS group but lowly expression in HS group included anti HBs antibody light-chain Fab,laminB1,WARS protein,flavin reductase,glutamate dehydrogenase 1,nuclear matrix protein 238,retinal dehydrogenase 1,AFl 65172,subunit of replicative DNA polymerase and HSP70.The higher expression of HSP70 in LS group and galectin7 in HS groups were further confirmed by western blot and immunohistochemical method.Conclusions The 2-DE gels images are successfully acquired from high-sensitivity group and low-sensitivity group of intermediate stage and advanced cervical carcinoma tissues treated by concurrent chemoradiotherapy.Some differentially expressed proteins between the two groups can be further confirmed by western blot and immunohistochemical method.

Objective To investigate the relationship between carotid stiffness and coronary angiography in elderly patients with coronary heart disease(CHD). Methods Thirty-five elderly patients with CHD who underwent coronary angiography were enrolled in the study. The carotid stiffness was measured by ultrasound, the results were compared with those in hypertension group,hyperlipemia group and healthy elderly group. Results There were significant differences in carotid tensity (8.15 ± 1.54), arteriectasis ( 0.34 ± 0.07 ) and carotid stiffness ( 640.51 ± 150.98 ) of elderly patients with CHD compared to other groups (all P＜0.05 or P ＜ 0. 01 ). There was significant correlation between coronary angiography score and carotid stiffness in elderly patients of CHD(P＜0.05 or P＜0.01). Conclusions There is close relationship between carotid atherosclerosis and coronary atherosclerosis in elderly patients with CHD.

Objective To study the effect of E1A gene on the radiosensitivity of nasopharyngeal carcinoma (NPC) cells and its mechanism. Methods Ad-E1A gene was transfected into human NPC cells (CNE2), then the positive clones (CNE2-Ad-E1A) were identified by RT-PCR. CNE2 cells, CNE2 cells transfected with Ad-β-gal (CNE2-Ad-β-gal) and CNE2-Ad-E1A cells were irradiated with 0 Gy,2 Gy,4 Gy,6 Gy and 8 Gy respectively using 6 MV X-ray. Clone forming assays were carried out, cell survival curves were drawn and the sensitivity enhancing ratio (SER) was calculated. The redistributions of cell cy-cle were analyzed by flow cytometry. RT-PCR was used to detect the expression of wtp53. Results RT-PCR confirmed that E1A gene had been integTated into positively transfected cells and stably expressed. Cell survival curves showed that the SER of D0,Dq and SF_2 value was 1.37, 1.95 and 1.46 in CNE2-Ad-E1A cells. The D_0,D_q and SF_2 value was 1.57 Gy,1.82 Gy, 0.89 in CNE-2 cells and 1.53 Gy,1.78 Gy,0.82 in CNE2-Ad-β-gal cells, respectively. The G_2/M arrest was shown in CNE2-Ad-E1A cells. Moreover, the expression of wtp53 gene was markedly enhanced in Ad-E1A-CNE2 cells. Conclusions E1A gene can ef-fectively enhance the radiosensitivity of human NPC cells, which may be associated the enhancement of wt-p53 expression and G_2/M arrest.

Objective To investigate the effect of El A gene on the radiosensitivity of human laryngeal carcinoma cells and its correlated mechanisms. Methods The Ad-E1A and Ad-β-gal were amplifieated in Hek293 cells, extracted by freezing (-80℃) and thawing(37℃) repeatedly (3 times) , purificated by the method of density gradient of CsC1 and titrated by plaque assay method. Then they were transfected into human laryngeal carcinoma cells (Hep-2) and authenticated by RT-PCR. The radiosensitivity of Hep-2 cells transfeeted with or without El A were studied by cell surviral curve. Finally we investigated the correlated mechanisms including cell apoptosis studied by flow cytometry and VEGF content studied by RT-PCR. Resuits The radiosensitivity of Hep-2 cells transfected with El A was intensified, Do and Dq were lowered and α was increased. Flow cytometry showed that the apoptosis rate of cells with E1A or with El A and radiotherapy was increased. The VEGF content of the cells transfeeted with E1 A or treated by radiotherapy was decreased, which reached the lowest level when the cells were treated with the both mathods. Conclusions E1 A gene can intensify the radiosensitivity and contribute to the apoptosis of human laryngeal carcinoma cells. El A gene and radiotherapy can markedly decrease the VEGF content.

Objective To investigate whether folate deficiency cause high expression level of interferon gamma (IFN-γ) resulted from IFN-γ gene ( IFNG) hypomethylation and then promote the pathogenesis and development of ulcerative colitis (UC ) in a dextran sulfate sodium (DSS )-induced experimental colitis model in mice .Methods A total of 24 female BALB/c mice were divided into four groups ,six mice in each group , including folate deficient/DSS+ group , standard diet/DSS+ group , standard diet/DSS - group and folate deficient/DSS- group .At the beginning of the sixth week since fed , the mice of model groups were treated with 5% DSS to establish experimental colitis .By the end of the sixth week ,disease activity index (DAI) of colitis and histological changes were evaluated .The folate level of peripheral blood serum of mice were detected by enzyme-linked immunosorbent assay (ELISA ) . The expression of IFN-γ in colonic mucosa of mice was examined by immunohistochemistry . The methylation level of CpG island in the promoter region of IFNG was determined by methylation specific polymerase chain reaction (MSP) .The t test was used for measurement data .Chi square test was performed for comparison between groups of count data . Spearman correlation analysis was used for correlation analysis .Results The folate levels of peripheral blood serum of folate deficiency/DSS+ group and folate deficiency/DSS- group ((2 .70 ± 0 .19) and (2 .80 ± 0 .25)μg/L) were significantly lower than those of standard diet/DSS+ group and standard diet/DSS- group ((13 .62 ± 0 .38 ) and (13 .52 ± 0 .77)μg/L ,t= -63 .33、32 .27 ,both P< 0 .05) ,resepectively .The expression of IFN-γ in colonic mucosa of folate deficiency/DSS+ group and standard diet/DSS+ group were significantly higher than those of folate deficiency/DSS- group and standard diet/DSS- group (χ2 = 22 .18 ,P< 0 .05 ) . And the expression of IFN-γ in colonic mucosa of folate deficiency/DSS+ group was also higher than that of standard diet/DSS+ group (χ2 = 12 .00 ,P< 0 .05) .The expression level of IFN-γ of folate deficiency/DSS+ group and standard diet/DSS+ group was positively correlated with (r=0 .998、0 .953 ,both P<0 .01) .The folate levels of peripheral blood serum of folate deficiency/DSS+ group was negatively correlated with IFN-γexpression level and DAI (r= -0 .880 and -0 .926 ,both P<0 .05) .No abnormal methylation was detected in IFNG promoter CpG island in colonic mucosa tissues of mice of each group . Conclusion In the mice model of DSS induced acute experimental colitis ,folate deficiency may increace the expression of inflammatory factor IFN-γand enhance the inflammation activity of colonic mucosa .

To study the effect of chitosan on delayed outward potassium current(IK) in single ventricular myocytes of guinea pig and investigate its antiarrhythmic mechanism from ion channel view. Patch clamp technique with whole-cell configuration. Holding potential was -40mV,commanding potential was -60- + 70mV ,step pulse +10mV,stimulating frequency 1 Hz,duration 300 ms and stimulating interval 6s. The result showed that Chitosan inhibited IK in a dose -dependent manner. Conclusion :Chitosan can inhibite IK in single ventricular myocytes of guinea pig.