AIM:To investigate the effects of rhTGF-?1 and TGF-?1 gene transfection on the proliferation of cultured rabbit corneal endothelial cells in vitro.METHODS:Cell growth induced by various concentrations of rhTGF-?1 was determined by MTT proliferation assay.Under the induction of liposomes,recombinant pSecTag2-TGF-?1MP vectors were transferred into the corneal endothelial cells.Morphological changes of transfected cells were observed by HE staining.The expression levels of TGF-?1 were assessed by ELISA.Cell cycle analysis was assessed by flow cytometry.DNA fragment analysis was used to confirm the presence of apoptosis.RESULTS:rhTGF-?1 in concentrations of 5-20 ?g/L showed a significant suppressive effect on the proliferation of corneal endothelial cells,0.5-1 ?g/L had no effect,0.05-0.1 ?g/L facilitated cell growth,as compared with negative controls.The morphous of transfected corneal cells had no significant abnormality compared with normal cells.According to the result of ELISA,the concentration of TGF-?1 in the supernatant was calculated to be(98?3)ng/L.Flow cytometry assay showed that S and G2/M phase of transfected cells decreased significantly compared with that of control group,but the cell cycle recovered normally after adding 10 ?g/L EGF into the culture medium.Agarose electrophoresis didn't show marked ladders in transfected group.CONCLUSION:Effects of rhTGF-?1 on the proliferation of corneal endothelial cells are different with various concentrations.TGF-?1 gene transfection shows suppressive effect on the proliferation of cultured corneal endothelial cells,but does not induce cell apoptosis.EGF is the antagonist of this suppressive effect.

Objective:To investigate the effect of the new nanomaterials,polyamidoamine dendrimers PAMAM-D,mediated foreign gene human decay accelerating factor(hDAF) for xenotransplantation transfected pig sperm cells.Methods:PAMAM-D/hDAF cDNA compounds were made.The compounds were divided into 0.2 μg,0.4μg,0.6μg,0.8 μg and 1.0μg groups (each group adding corresponding dose of PAMAM-D in accordance with the N/P ratio 10:1,20:1,40:1),then were digested by restriction enzymes.The compounds were incubated with washed pig sperm cells.Then the transfection efficiency was detected by in situ hybridization in the different groups.Results:The PAMAM-D molecule can prevent electrophoretic migration of DNA in the compound.After digested the compounds by restriction enzymes,DNA can not be degraded.The transfection efficiency was different in different groups.Among the total,the efficiency was higher in both groups of 0.4 μg and 0.6μg than that of others.The top was the group of 0.4 μg linear plasmid plus PAMAM-D when the N/P ratio was 20:1(47.5%±O.2%,167% vs control group).Conclusion:PAMAM-D can improve the efficiency of exogenous gene transfeeted pig sperm cells,which can reinforce the stable binding of exogenous DNA to sperm cells.

Objective: To investigate the possibility of heart rate as an optimum index for evaluating physical labor intensity. Methods: From September to October, 2015, a total of 111 male young adults were selected as study subjects. A cycle ergometer was used for exercise at different intensities (20, 40, 60, 80, 100, 120, 140, 160, and 180 W) , and the exercise at each intensity lasted for 6 minutes. The subjects wore the K4b2 portable cardiopulmonary function testing instrument for the real-time monitoring of heart rate (HR) , pulmonary ventilation volume (Ve) , oxygen consumption (VO₂) , and energy metabolic rate (E) . The origin software was used for correlation and regression analyses of HR, Ve, VO₂, and E collected at different intensities to establish Ve-HR, VO₂-HR, and E-HR regression equations. Results: At different intensities, HR was positively correlated with Ve, VO₂, and E (r=0.8985, 0.8733, and 0.8973, all P<0.01) . There was a significant difference in the R² value of Ve-HR, VO₂-HR, and E-HR regression equations (R²=0.8070, 0.7625, and 0.8049, all P <0.01) . The validation results showed that there was no significant difference between the calculated values and the measured ones (P>0.05) . Conclusion HR can replace Ve, VO₂, and E and be used as an optimum index for evaluating physical labor intensity.