Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.

The purpose of this study was to fabricate decelluarized valve scaffold modified with polyethylene glycol nanoparticles loaded with transforming growth factor-β1 (TGF-β1), by which to improve the extracellular matrix microenvironment for heart valve tissue engineering in vitro. Polyethylene glycol nanoparticles were obtained by an emulsion-crosslinking method, and their morphology was observed under a scanning electron microscope. Decelluarized valve scaffolds, prepared by using trypsinase and TritonX-100, were modified with nanoparticles by carbodiimide, and then TGF-β1 was loaded into them by adsorption. The TGF-β1 delivery of the fabricated scaffold was measured by asing enzyme-linked immunosorbent assay. Whether unseeded or reseeded with myofibroblast from rats, the morphologic, biochemical and biomechanical characteristics of hybrid scaffolds were tested and compared with decelluarized scaffolds under the same conditions. The enzyme-linked immunosorbent assay revealed a typical delivery of nanoparticles. The morphologic observations and biological data analysis indicated that fabricated scaffolds possessed advantageous biocompatibility and biomechanical property beyond decelluarized scaffolds. Altogether this study proved that it was feasible to fabricate the hybrid scaffold and effective to improve extracellular matrix microenvironment, which is beneficial for an application in heart valve tissue engineering.

The purpose of this study was to fabricate decelluarized valve scaffold modified with polyethylene glycol nanoparticles loaded with transforming growth factor-β1 (TGF-β1),by which to improve the extracellular matrix microenvironment for heart valve tissue engineering in vitro.Polyethylene glycol nanoparticles were obtained by an emulsion-crosslinking method,and their morphology was observed under a scanning electron microscope.Decelluarized valve scaffolds,prepared by using trypsinase and TritonX-100,were modified with nanoparticles by carbodiimide,and then TGF-β1 was loaded into them by adsorption.The TGF-β1 delivery of the fabricated scaffold was measured by asing enzyme-linked immunosorbent assay.Whether unseeded or reseeded with myofibroblast from rats,the morphologic,biochemical and biomechanical characteristics of hybrid scaffolds were tested and compared with decelluarized scaffolds under the same conditions.The enzyme-linked immunosorbent assay revealed a typical delivery of nanoparticles.The morphologic observations and biological data analysis indicated that fabricated scaffolds possessed advantageous biocompatibility and biomechanical property beyond decelluarized scaffolds.Altogether this study proved that it was feasible to fabricate the hybrid scaffold and effective to improve extracellular matrix microenvironment,which is beneficial for an application in heart valve tissue engineering.

Objective:To study on the effects of micro-plasma radio frequency technology on guinea pig skin pigmentation animal model induced by narrow band ultraviolet B (NB-UVB).Methods:We selected ten healthy guinea pigs (common grade), removed the brown hair of the guinea pig and established UVB to induce the skin pigmentation model of guinea pig. The pigmentation zone was randomly divided into four groups: Group A was used as blank control with no treatment and Groups B , C and D were treated with micro-plasma radio frequency technology using different energy parameters. Group B treatment parameters were as follows: output power was 10 watts, and treatment energy was 0.06J. Group C treatment parameters were as follows: output power was 20 watts, and treatment energy was 0.12J. Group D treatment parameters were as follows: output power was 30 watts, and treatment energy was 0.18J. They were compared before treatment and at 2, 4, and 6 weeks after treatment, based on melanin cell staining (epidermis separation-dopa staining) and melanin granules dyeing method (Masson-Fontana). Immunohistochemical staining (c-kit protein) was used to observe the tyrosinase activity in the melanocytes, melanin granule percentage area of the skin area, and the change in melanin cell activity. SPSS13.0 statistical software was used for statistical analysis of the experimental data. The significance of the difference ( P< 0.05) was measured. Results:The results showed that micro-plasma radio frequency (RF) technology treatment could reduce the tyrosinase activity in the melanocytes, 2, 4 and 6 weeks after the end of treatment: Groups B, C and D were lower than group A, and the difference was statistically significant ( P<0.05), and as the treatment energy increased, the speed of reduction was increased. Micro-plasma radio frequency technology could reduce skin pigmentation and decrease the amount of epidermal melanin granules, 2 and 4 weeks after the end of treatment: Groups B, C and D were lower than Group A; 6 weeks after the end of treatment, only Group D was lower than Group A, with statistically significant differences ( P< 0.05). An increase in the energy of the micro-plasma radio frequency sped up the decline in the amount of epidermal melanin granules.The number of c-kit positive cells was not increased in this experiment. Two weeks after the end of treatment, the number of c-kit positive cells was higher than those of Groups B, C and D. There were no statistically significant differences in Groups B, C and D ( P<0.05). No c-kit positive cells were found in Groups A, B, C and D at 4 and 6 weeks after the irradiation. Conclusions:Micro-plasma radio frequency technology could accelerate the fading of guinea pig skin pigmentation. It is effective to improve pigmentation of guinea pig skin after NB-UVB irradiation.

Objective:To study on the effects of micro-plasma radio frequency technology on guinea pig skin pigmentation animal model induced by narrow band ultraviolet B (NB-UVB).Methods:We selected ten healthy guinea pigs (common grade), removed the brown hair of the guinea pig and established UVB to induce the skin pigmentation model of guinea pig. The pigmentation zone was randomly divided into four groups: Group A was used as blank control with no treatment and Groups B , C and D were treated with micro-plasma radio frequency technology using different energy parameters. Group B treatment parameters were as follows: output power was 10 watts, and treatment energy was 0.06J. Group C treatment parameters were as follows: output power was 20 watts, and treatment energy was 0.12J. Group D treatment parameters were as follows: output power was 30 watts, and treatment energy was 0.18J. They were compared before treatment and at 2, 4, and 6 weeks after treatment, based on melanin cell staining (epidermis separation-dopa staining) and melanin granules dyeing method (Masson-Fontana). Immunohistochemical staining (c-kit protein) was used to observe the tyrosinase activity in the melanocytes, melanin granule percentage area of the skin area, and the change in melanin cell activity. SPSS13.0 statistical software was used for statistical analysis of the experimental data. The significance of the difference ( P< 0.05) was measured. Results:The results showed that micro-plasma radio frequency (RF) technology treatment could reduce the tyrosinase activity in the melanocytes, 2, 4 and 6 weeks after the end of treatment: Groups B, C and D were lower than group A, and the difference was statistically significant ( P<0.05), and as the treatment energy increased, the speed of reduction was increased. Micro-plasma radio frequency technology could reduce skin pigmentation and decrease the amount of epidermal melanin granules, 2 and 4 weeks after the end of treatment: Groups B, C and D were lower than Group A; 6 weeks after the end of treatment, only Group D was lower than Group A, with statistically significant differences ( P< 0.05). An increase in the energy of the micro-plasma radio frequency sped up the decline in the amount of epidermal melanin granules.The number of c-kit positive cells was not increased in this experiment. Two weeks after the end of treatment, the number of c-kit positive cells was higher than those of Groups B, C and D. There were no statistically significant differences in Groups B, C and D ( P<0.05). No c-kit positive cells were found in Groups A, B, C and D at 4 and 6 weeks after the irradiation. Conclusions:Micro-plasma radio frequency technology could accelerate the fading of guinea pig skin pigmentation. It is effective to improve pigmentation of guinea pig skin after NB-UVB irradiation.

Objective: Microtia is a relatively common birth defect, and its etiology is still unclear. In this paper, Phenolyzer, a kind of phenotype-based gene analyzer, was used to prioritize candidate genes involved in microtia. Methods: The search term " Microtia" was entered in the Phenolyzer interface. Through the steps of disease matching, gene query, gene scoring, seed gene expansion, the genetic information results of microtia were finally generated. This article traced 10 candidate genes in detail. Results: There were 13485 genes associated with microtia or related syndromes, of which 130 were seed genes. Each gene had a corresponding score, and the higher the score, the greater the correlation with the microtia. Among them, the top 10 genes included HOXA2, CHD7, CDT1, CDC6, ORC1, ORC4, ORC6, SMAD4, GLI3 and MED12. Conclusions Phenolyzer provides a detailed summary of candidate genes for microtia, providing accurate research directions for screening high-throughput sequencing results and subsequent studies.