This study examined the construction of eukaryotic expression plasmid of human transforming growth factor-β3 (hTGF-β3) and its inducing effect on the differentiation of precartilaginous stem cells (PSCs) into chondroblasts. hTGF-β3 gene was amplified by using polymerase chain reaction (PCR) and then inserted into the eukaryotic expression plasmid pcDNA3.1 to construct the eukaryotic expression plasmid pcDNA3.1(+)-hTGF-β3. Rat PSCs were isolated and purified by employing an immunomagnetic cell sorting system. pcDNA3.1(+)-hTGF-β3 was transfected into purified PSCs with the use of linear polyamines. The expression of TGF-β3 and cartilage-specific extracellular matrix (ECM) components was detected after transfection by real-time quantitative PCR, ELISA, immunochemistry and Western blotting, respectively. The results showed that the eukaryotic expression plasmid pcDNA3.1(+)-hTGF-β3 was successfully established as identified by enzyme digestion and DNA sequencing. Real-time quantitative PCR and ELISA revealed that hTGF-β3 was strongly expressed in pcDNA3.1(+)-hTGF-β3-transfected PSCs. Real-time quantitative PCR, immunochemistry and Western blotting showed that the cartilage-specific ECM markers, i.e., cartilage oligomeric matrix protein (COMP), Aggrecan, collagen type X and II were intensely expressed in the pcDNA3.1(+)-hTGF-β3-transfected cells. It was concluded that hTGF-β3 could be stably expressed in pcDNA3.1(+)-hTGF-β3-transfected PSCs and induce the differentiation of PSCs into chondroblasts.

This study examined the construction of eukaryotic expression plasmid of human transforming growth factor-β3 (hTGF-β3) and its inducing effect on the differentiation of precartilaginous stem cells (PSCs) into chondroblasts.hTGF-β3 gene was amplified by using polymerase chain reaction (PCR) and then inserted into the eukaryotic expression plasmid pcDNA3.1 to construct the eukaryotic expression plasmid pcDNA3.1(+)-hTGF-β3.Rat PSCs were isolated and purified by employing an immunomagnetic cell sorting system.pcDNA3.1(+)-hTGF-β3 was transfected into purified PSCs with the use of linear polyamines.The expression of TGF-β3 and cartilage-specific extracellular matrix (ECM)components was detected after transfection by real-time quantitative PCR,ELISA,immunochemistry and Western blotting,respectively.The results showed that the eukaryotic expression plasmid pcDNA3.1(+)-hTGF-β3 was successfully established as identified by enzyme digestion and DNA sequencing.Real-time quantitative PCR and ELISA revealed that hTGF-β3 was strongly expressed in pcDNA3.1(+)-hTGF-β3-transfected PSCs.Real-time quantitative PCR,immunochemistry and Western blotting showed that the cartilage-specific ECM markers,i.e.,cartilage oligomeric matrix protein (COMP),Aggrecan,collagen type Ⅹ and Ⅱ were intensely expressed in the pcDNA3.1(+)-hTGF-β3-transfected cells.It was concluded that hTGF-β3 could be stably expressed in pcDNA3.1(+)-hTGF-β3-transfected PSCs and induce the differentiation of PSCs into chondroblasts.

Objective: To compare the clinical effect of the Yu flap and the Karapandzic flap in repairing greater than 2/3 defects of the lower lip and to provide a reference for clinical application. Methods: Ten patients with greater than 2/3 lower lip defects after surgical resection of lower lip tumors and vascular malformations were enrolled: 5 patients were repaired with the Yu flap (Yu flap group) and 5 patients were repaired with the Karapandzic flap (Karapandzic flap group). Follow-up for at least 1 year was conducted to evaluate the morphology (symmetry, stoma, exposure of vermilion) and function (sensory function, motor function) of the reconstructed lower lip. Results : All the flaps survived, and all wounds showed primary healing. The lower lips reconstructed with the Yu flap or the Karapandzic flap obtained similar satisfactory oral function. The sensory function was essentially restored. There were no obvious obstacles in speech and expression, and no saliva leakage occurred. In the Yu flap group, only 1 patient had slight microstomia. In the Karapandzic flap group, 2 patients had slight microstomia and 3 patients had moderate microstomia. 90% (9/10) of the patients were very satisfied with the postoperative outcome, and 1 patient in the Karapandzic flap group was basically satisfied. Conclusion Both the Yu flap and the Karapandzic flap can be used to repair greater than 2/3 lower lip defects and reliable outcomes can be achieved. These two methods can achieve similar oral functions, but the effect of the Karapandzic flap is inferior to that of the Yu flap in terms of aesthetic appearance, and microstomia often occurs, while the Yu flap can generally maintain the original size of the mouth cleft.