Objective To construct pGL3-EPO-HRE reporter gene vector and to detect its function.Methods To synthesize two single-strand DNA fragments which had overlap sequence at the 3'end,then mixed them to construct the double-strand DNA fragment which include three copies of(57 bp) human erythropoietin(EPO) HRE.There was MluⅠor BglⅡ digestive site at the end of this DNA fragment.After inserting the(87 bp) fragment into pMD18-T for sequencing,the correct sequence was inserted into pGL3-promoter to construct pGL3-EPO-HRE.The recombinant vector was co-transfected into MCF-7 cells with pcDNA3-HIF-1?or pcDNA3 respectively,(12 h) later cells were treated with(100 ?mol/L) cobalt chloride(CoCl_(2)) for(24 h), then the expression of pGL3-EPO-HRE was observed by dual-luciferase reporter gene assay method.Results The sequence of pGL3-EPO-HRE was identical with human EPO HRE sequence.pGL3-EPO-HRE luciferase activity increased about 10 fold(P

Objective To investigate the ultrasound diagnostic value for nipple discharge disease by comparison with fiberoptic ductoscopy. Methods Fiberoptic ductoscopy and ultrasonic inspection were performed on fifty-seven patients with spontaneous nipple discharge,and the results were analysed. Results In all patients, 55 cases (96. 49 % ) were abnormal in high-frequency ultrasound inspection:27 mammary duct expansion(47.36%),19 occupying lesion with mammary duct expansion or cyst (33.33%),6 cystic echo ( 10. 52% ) ,3 only solid echopattern (5.26%) ,and 2 nothing found (3.51% ). In ultrasonic inspection,color and pulse Doppler signal were found in 7 cases. On the other hand, 26 galactophoritis (45.61%), 12mammary duct ectasia (21.05 %), 17 intraductal papilloma (29.82%), 2 breast carcinoma (3.50%) were diagnosed by fiberoptic ductoscopy. In all 19 cases with occupying lesion,ultrasonography's sensitivity was 52. 63% ,specificity was 89. 47%, misdiagnosis rate was 10. 50%, rate of missed diagnosis was 47.36%.Conclusions To nipple discharge disease, high-frequency ultrasound has high sensitivity, easy to find abnormal indication,but is not accurate enough in the diagnosis of smaller occupying lesion.

Objective To construct pcDNA3-HIF-1? eukaryotic expression vector and to investigate its function in breast cancer MCF-7 cells.Methods RT-PCR was applied to amplify human HIF-1? cDNA from MCF-7 cells with a pair of sequence specific primers carrying a restriction enzyme site XbaⅠ or HindⅢ on each 5′end.HIF-1?(cDNA) was inserted into pMD18-T after sequencing,and then inserted into pcDNA3 vector.The recombinant vector was transfected into MCF-7 cells to observe its expression and function by Western blot and dual-luciferase reporter gene assay.Realtime-PCR was performed to detect the inducible expression of C-MET mRNA by HIF-1?,the anti-apoptotic effect of HIF-1 under hypoxia was analysed by detecting the Caspase3/7 activity.Results The sequence of pcDNA3-HIF-1? is identical with the gene bank.It enhances the expression of HRE reporter gene and C-MET mRNA,but decreases the Caspase3/7 activity in MCF-7 cells under hypoxia.Conclusion The pcDNA3-HIF-1? eukaryotic expression vector was successfully constructed,it not only has strong DNA binding and inducing activity,but also has anti-apoptotic effect in hypoxia MCF-7 cells.

Objective: To determine the distribution characteristics of waist circumference (WC), waist height ratio (WHtR) of 6–18 years olds in Guangzhou, and to put forward the WC and WHtR appropriate boundary values for 6–18 years olds on the basis of cardiovascular disease (CVD) risk factor assessment. Methods: We analyzed the height, weight, WC and its metabolic indication data (blood pressure, fasting blood glucose, and blood lipids) of 15 000 children in Guangzhou, aged 6–18, with the receiver-operating characteristic curve (ROC), and explored the best value point of WC and WHtRfor the prediction of cardiovascular diseases. Results: When the WC percent reached P85, and WHtR reached 0.48, the cardiovascular risk factors of fasting blood-glucose, blood pressure, and blood fat were signiifcantly higher. Conclusion: The 85th percentile value of WC and 0.48 of WHtR are the appropriate boundary values in increasing the cardiovascular disease risk factors in Chinese children and teenagers. WC and WHtR as a relatively simple inspection method, can well predict cardiovascular diseases, and be used in the conventional measuring items among students.

Objective To evaluate the different change in atherosclerosis between apolipoprotein E knockout ( ApoE-/-) mice and low density lipoprotein receptor knockout ( LDL-R-/-) mice in different diet by vivo ultrasound biomicroscopy(UBM) . Methods Thirty-two 8 weeks age ApoE-/- mice and twenty-eight 20 weeks age LDL-R-/- mice were selected ,and each mouse model were divided into four groups randomly according to weight . Two groups of each model were fed with normal diet for 8 weeks and 16 weeks respectively as general groups ,and two other groups were fed with west diet as high-fat groups . Intima-media thickness ( IM T ) in the aortic root of all groups were assessed by Vevo 770 UBM , and all measurementsofUBMwerecomparedwithhistopathologyandblood-fat.Results ①Thickenwallor plaque in aortic root showed by UBM and histopathology could be found in ApoE-/- mice in all diet conditions ,but the changes in LDL-R-/- mice only could be found in high-fat diet conditions;② IM T of ApoE-/- mice measured in histopathology were significantly higher than LDL-R-/- mice in all diet condition ( P<0 .05) ,but in UBM ,the IMT of ApoE-/- mice showed significantly higher than that of LDL-R-/- mice only in general diet groups and 8 weeks high-fat diet group ( P <0 .05) ;③In blood-fat ,the total cholesterol ( TC) in ApoE-/- mice were significantly higher than that of LDL-R-/- mice in all diet conditions ( P <0 .05) . But triglyceride ( TG) was higher only in general diet when it was compared between ApoE-/- mice and LDL-R-/- mice . Conclusions It is recommend to select ApoE-/- mice as animal model in atherosclerosis study rather than LDL-R-/- mice ,because the higher serum TC and higher susceptible of atherosclerosis in ApoE-/- mice . The west diet is necessary if LDL-R-/- mice was used to atherosclerosis study .

To evaluate the relationship between atherosclerosis and hemodynamic of coronary artery in mice detecting by ultrasound bio‐microscopy flow imaging . Methods Double 14 20‐week‐old LDL‐R‐/‐and C57BL/6 male mice were selected ,and randomly divided into two groups in each genotype according to weight . Each two groups were fed to 28 weeks or 36 weeks age respectively with west diet . Coronary artery hemodynamics in these mice were assessed in vivo by Vevo ?2100 ultrasound imaging system ,then the intima‐media thickness( IM T ) of aorta in histopathology were analyzed . T he differences of coronary artery hemodynamic parameters such as maximum velocity ( Vmax ) ,mean velocity ( Vmean) and velocity time integral ( V T I) were compared between mice of different genotypes of the same week and mice of different weeks of the same genotype . And the relationship between coronary artery hemodynamic in ultrasound and aortic IM T in histopathology were analyzed . Results ① All coronary hemodynamic parameters in LDL‐R‐/‐ mice were significantly lower than those of wild‐type mice except the Vmax between two 28‐week‐old genotypes group at the same weeks of age of different genotypes ( all P ＜0 .05) . But there was no significant difference in coronary artery hemodynamic parameters between mice of the same genotype at different weeks of age( P ＞0 .05) . ②T he histopathological measurements of aortic IM T in LDL‐R‐/‐mice were significantly higher than those of wild type mice ( all P ＜ 0 .05 ) ,and those of 36‐week‐old mice were significantly higher than those of 28‐week‐old mice ( all P ＜ 0 .05 ) . ③ All coronary hemodynamic parameters such as Vmax ,Vmean and V TI were negatively correlated with pathological measurements of aortic IM T ( r ＝ －0 .532 , －0 .423 , －0 .524 ; all P ＜ 0 .05 ) . Conclusions The parameters of coronary artery hemodynamics obtained by ultrasound bio‐microscopy are well correlated with the pathological results of atherosclerosis . Ultrasound bio‐microscopic flow imaging can be used as a new method to evaluate the degree of atherosclerosis in mice by detecting the hemodynamic parameters of coronary artery .