Objective To study the effect of pluripotency factor Nanog on the expression of the cell cy-cle related proteins,and then to explore its effect on the proliferative ability of HepG 2 cells.Methods TALENs gene editing tool was employed to induce mutation and downregulation expression of Nanog .T7 endonuclease 1 and genomic sequencing was used to analyze the mutation efficiency of Nanog .RT-PCR and western blot were used to determine the expression of mRNA and protein of Nanog ,respectively .Real-time cell based assay system was used to measure the proliferative ability of wild -type HepG2 cells and monoclonal HepG 2 cells with Nanog mutation.Results TALENs successfully induced mutation of Nanog gene .The targeting efficiency of mixed cells was analyzed by T7 endonuclease 1 approached 40%after two transfection with plasmid of Nanog -TALENs.Ad-ditionally,the Nanog mRNA expression level of monoclonal HepG 2 with Nanog mutation was downregulated by 3.4 times compared to the wild type HepG 2 cells,and the Nanog protein expression level was downregulated by 3.6 times.The cell cycle related proteins CyclinD1/D3,CyclinE1 and CDK2 expression were downregulated in monoclonal HepG2 with Nanog mutation in comparison to the wild type HepG 2 cells.Conclusion Nanog plays a role in influencing the proliferative ability of HepG 2 cells through modulating the expression of the cell cycle re-lated proteins CyclinD1/D3,CyclinE1 and CDK2.The downregulation expression of Nanog can inhibit the prolif-erative capacity of HepG 2 cells via the regulation of the cell cycle related proteins .

Purpose:To determine the effect of doses of three-dimensional conformal radiotherapy(3D-CRT) for unresectable biliary cancer. Methods:48 patients with localized,unresectable cholangiocarcinoma were treated with 3D-CRT.Patients were grouped according to the radiotherapy dose: 12 patients received a total dose of 33-39Gy(low dose group),23 patients received 40-58Gy(medium dose group),and 13 patients received 59.4-68Gy(high dose group). The low dose group received 3Gy/fraction,medium dose group and high dose group received either 1.8 or 2Gy/fraction,5 fraction/week. Results:The median time to local progression for all patients was 10 months. The median overall survival was 12 months. 24(50%)had radiographic evidence of disease progression. 21 of those patients(87.5%) experienced local recurrence as the first radiographic sign of disease progression. The remaining 3 presented with metastatic disease as the initial sign of progression. The time of local failure(P=0.220) and the median survival(P=0.232) were not statistically different among the three groups. The rate of acute gastrointestinal side effects was not statistically different among the three groups(P= 0.485) . Conclusions:Local progresion was the predominant pattern of disease recurrence in this group of patients with unresectable biliary cancer. Because the power to detect even large difference with small numbers of patients is limited,a benefit from high dose radiotherapy cannot be excluded.

Objective To provide a powerful and conclusive result for the association between the GGN polymorphic repeats in androgen receptor (AR)gene and prostate cancer (PCa)risk.Methods CNKI,VIP,Wanfang,PubMed/Medline,Embase and The Cochrance Library electronic database were used to retrieve the eligible publications addressing the association between the AR gene GGN polymorphic repeats and prostate cancer risk.16 GGN polymorphism repeats were used as cut off value,meta-analysis was ap-plied to the study on the association between the length of polymorphism repeats and prostate cancer risk.Results 9 cases of con-trol studies were included in this meta-analysis and a total of 2 438 cases and 1 968 controls were included.People with ≤16 GGN polymorphism repeats displayed a higher risk of prostate cancer(OR =1.15,95%CI :1.00 -1.31,P =0.04).Conclusion ≤16 GGN polymorphism repeats polymorphism associated with increased risk of prostate cancer.

Objective To investigate the effect of autophagy on the proliferation and migration of cervi-cal cancer cells ,as well as the underlining mechanisms .Methods Rapamycin was used to induce the autophagy in HeLa cells,formation of autophagosomes was observed by staining with acridine orange under fluorescence mi -croscope.Western blot was used to detect the expression of LC 3 and PI3K/Akt/mTOR in HeLa cells.The LC3 plasmid was transfected into HeLa cells .The distribution of LC3 in cells and the expression of LC3 was identified by fluorescence microscope and Western blot ,respectively.The autophagy was inhibited with 3-methyladenine(3-MA )in HeLa cells.The cell proliferation was monitored by RTCA real -time instrument.Transwell chamber was carried out to assess cell migration .Results After 6h of rapamycin treatment ,the expression of LC3B was in-creased in HeLa cells ( P<0 .05 ) .The proliferative and migration ability were weakened compared to wild type HeLa cells(P<0.05).The same results in the presence of 3-MA.The expression of PI3K/AKT/mTOR path-way proteins were activated by rapamycin treatment and LC 3 overexpression(P<0.05).Conclusion Autophagy can suppress the proliferation and migration in cervical cancer cells ,which may relate to PI3K/Akt/mTOR path-way.

It is to discuss the method for studying uveoscleral pathway and the influence of uveoscleral outflow on aqueous humor drainage. Seventeen rabbits were randomly divided into the flow group that included 5 rabbits and the morphological group that included 12. Fluorescein-isothiocyanate bovine serum albumin (FITC-BSA) was perfused into anterior chambers of flow group rabbit eyes as the tracer for measuring uveoscleral outflow. This tracer was infused into anterior chambers of morphological group rabbit eyes for observing fluorescent intensity of all tissue of the uveoscleral pathway with fluorescence microscopy. Fluorescence was seen in all sites of uveoscleral pathway. The fluorescent intensity of ciliary body and suprachoroidal space was significantly stronger than that of anterior sclera (P＜0.05). The intensity of choroid and posterior sclera was significantly weaker than that of other tissues (P＜0.05). The average value of uveoscleral outflow was (0.18±0.031) μl/min. This study demonstrates that uveoscleral pathway plays an important role in aqueous humor drainage. Anterior sclera is the main site where aqueous humor flows out. FITC-BSA is an effective tracer for studying uveoscleral pathway from both morphological and functional aspects.

Objective To explore the indication and the methodology of transcatheter closure of coronary artery fistula.Methods Altogether 17 cases for intervention,included 13 using the Cook corp coil(2 detachable and another 11 un-detachable);1 with the Pfm corp detachable coil;3 with the AGA corp Amplatzer-Duct.The other 26 cases underwent surgery. Results Intervention group,showed 3 failure cases because of selecting too small and un-detachable coil in early stage with final displacement.Fourteen cases were successful.Follow-up 3 years showed no residue shunt and other complications.All patients were successful with surgery.Conclusions Transcatheter closure of the coronary artery fistula is mild invasive,short hospitalization,safe,and effective under proper indications.

Objective To evaluate the effect and complication of inductio n chemot herapy combined with three-dimensional conformal radiation therapy (3DCRT) for l ocally advanced non small cell lung cancer (NSCLC). Methods Ninety-two such pa t ients were randomized into radiation therapy alone group(RT-, 50 patients) and i nduction chemotherapy combined radiotherapy group (CMT-, 42 patients). The indu c tion chemotherapy consisted of 2-4 cycles of platinum-based regimen. Results Th e overall median survival time was 15 months with 12 months in the RT group and 18 months in the CMT group(P=0.014)respectively. The 1-year o verall survival rates were 48.6% and 71.2% in RT and CMT group,respectively (P=0.004). The 2-year survival rates w ere 20.8% and 37.6% in RT and CMT group, respectively (P=0.0 41). Treatment was w ell tolerated and the toxicities were similar in either group. C onclusion The ad dition of induction chemotherapy to 3DCRT takes a survival advantage over 3DCRT alone for Stage Ⅲ NSCLC without increasing toxicities.

ObjectiveTo analyze the association of human urate transporter 1 ( hURAT1 ) gene promoter single-nucleotide polymorphisms(SNPs) with primary hyperuricemia ( HUA ) in Chinese Han people.MethodsA total of 215 patients with HUA and 323 healthy subjects were chosen to be investigated of SNP of hURAT1 promoter by PCR and sequencing.ResultsFive SNPs were identified,including-454A/T,-434T/C,-382C/T,-87C/T,and + 118G/A.Pairwise linkage disequilibrium analysis displayed a high linkage disequilibrium between the five SNPs ( r2 =0.99).In HUA group,the heterozygous genotypos ( AT,CT,CT,CT,AG ) frequencies were significantly lower than those in control group ( P＜0.05 ).Logistic regression analysis showed that the heterozygosis genotypes ( AT,CT,CT,CT,AG) were protective factors of HUA ( OR 0.68-0.75 ).The minor allele ( T,C,T,T,A ) frequencies for both SNPs were significantly different between two groups ( P =0.022,P =0.038 ).ConclusionThese findings indicate that -454A/T,-434T/C,-382C/T,-87C/T,and + 118G/A SNPs of hURAT1 gene promoter area are associated with HUA in Chinese Han population.

Objective To analyze the level of cardiac troponin I (cTnI) in children with left-to-right shunt congenital heart disease (CHD). Methods In this study, 146 children with secundum atrial septal (ASD) defect, 132 children with ventricular septal defect (VSD) and 300 healthy children were recruited. The levels of cTnI and N-terminal pro-brain natriuretic peptide (NT-proBNP) were measured and their correlation with clinical data was analyzed. Results The serum cTnI and NT-proBNP levels in both ASD and VSD patients were signiifcantly higher than those in normal children (H=3.89 and 5.27, P<0.01). The serum cTnI and NT-proBNP levels in VSD patients were signiifcantly higher than those in ASD patients (P<0.05). The ratio of pulmonary to systemic arterial pressure (Pp/Ps), pulmonary vascular resistance index (PVRI) and standardized left ventricular end diastolic volume in VSD patients were signiifcantly higher than those in ASD patients (P<0.05). Multiple regression analysis showed that Pp/Ps was signiifcantly correlated with cTnI in VSD patients. (β=0.81, SE=0.03, P=0.000). Conclusions Signiifcant volume and pressure overload due to a left-to-right shunt induce myocardial injury and could lead to irreversible myocardial remodeling in children with CHD. The serum cTnI level is a sensitive biomarker for myocardial damage in VSD patients.

Objective To explore the indications and clinical assessment for catherization in congenital heart diseased patients with stage operation. Methods Sixty five patients were selected from Apr. 1999 to Dec. 2002 undergoing second catherization. 47 male, 18 females with age ranging from 5 months to 22 years old on average of 6.8 years. EKG, chest X-ray, ECHO, cardiovascular pressure and blood oxygen saturation were taken in all patients during the procedure. Results According to the physiologic data recorded during the procedure of catherization, 39 cases needed one more surgery, 9 cases for intervention, 4 cases would be placed with endovascular stent 9 cases should be follow-up and 4 cases couldn t be operated again. Conclusions Catherization is not only offering the accurate physiologic data, but also revealing the details about anatomy with important information to the diagnosis and treatment for the congenital heart disease, especially for the srage-operation.