BACKGROUND: Thoracodorsal artery perforator flap can relieve damage to donor site and avoid bulk in the recipient site,but dissociation of perforating branch took time.Some one believed that it should be done by very experienced physicians and some muscle tissues should be reserved.OBJECTIVE: To investigate the method,effectiveness and clinical application of improved latissimus dorsi flap based on perforator flap conception for reconstruction of soft tissue defects of lower extremity.METHODS: A total of 17 patients needing skin flap transplantation were selected.12 latissimus dorsi musculocutaneous/muscle flaps,3 latissimus dorsi flaps with a few muscle and 2 double-leaf segmental latissimus dorsi compound flaps were designed based on perforator flap conception.According to the territory of latissimus dorsi musculocutaneous flap,a skin paddle in which anterior underlying muscle and main perforator was designed,extend about to the anterior edge of the latissimus dorsi muscle.An additional latissimus dorsi muscle flap was selected for soft tissue enlargement if necessary.Sometimes,double-leaf segmental latissimus dorsi musculocutaneous/muscle flap,including one muscle-sparing latissimus dorsi musculocutaneous flap and the other segmental latissimus dorsi muscle flap nourished by the lateral branches of the thoracodorsal vessels was selected to repair two adjacent defects.The harvested tissue area ranged from 12 cm×8 cm to 28 cm×17 cm.Survival state of skin flap,together with shape and function of donor site and recipient site of skin flap were observed.RESULTS AND CONCLUSION: Following skin flap transplantation,one case developed vascular crisis that was relieved following re-exploration for vessel anastomosis.All skin flap survived.Second-stage skin grafting was done on one muscle flap wound.All donor sites were sutured directly.After a follow-up of 3 to 18 months in 15 cases,only two cases received two-stage plastic operation because bulky flaps brought some trouble in wearing shoes.Improved latissimus dorsi flap based on perforator flap conception can reduce damage to the donor site and the receipt area bulk.Double-leaf segmental latissimus dorsi compound flaps can repair both heel and toe wound.The versatile latissimus dorsi flap designed using thoracodorsal artery perforator flap conception is an ideal flap for repairing widespread soft tissue defects in the lower extremity.

We constructed inducible and constitutive heterotrophic expression vectors of Dunaliella salina (D. salina) and identified heterotrophic transformants. A gene encoding a glucose transporter (Glut1) was cloned from human placenta tissues by RT-PCR and sequenced. Inducible heterotrophic expression vector pMDDGN-Bar of D. salina, which included a duplicated carbonic anhydrase (DCA) promotor and a Bar selectable marker that could drive expression of the Glut1 gene in D. salina, was constructed by molecular biology methods. In addition, we constructed another vector G5Glut1-Bar that contained a constitutive ubiquitin promotor, Glut1 and Bar box. The two expression vectors were introduced into D. salina by electroporation method, and then screened the transformants with phosphinothricin (PPT). Total RNA of the transformants extracted was used to analyze the integration of the target gene (Glut1) by RT-PCR. The cloned Glut1 sequence was 1479 bp and encoded 493 amino acids. The results of all enzymes digesting showed that two expression vectors were successfully constructed. After screening by PPT for several weeks, the transfomants grew well whereas wild-type cells died completely. The result of RT-PCR indicated that two transformants both had an about 250 bp specific band and the sequence homology was 100% compared with the human Glut1 sequence by Blast analysis. Taken altogether, inducible and constitutive heterotrophic expression vectors of D. salina was constructed successfully and the Glut1 gene was integrated into the genome of D. salina. Expression vectors above-mentioned may be used for the expression of the foreign Glut1 gene in D. salina.
Base Sequence ; Chlorophyta ; genetics ; metabolism ; Cloning, Molecular ; Electroporation ; Genetic Vectors ; genetics ; Glucose Transporter Type 1 ; biosynthesis ; genetics ; Industrial Microbiology ; Molecular Sequence Data ; Transformation, Genetic

To optimize the electroporation system in Dunaliella salina (D. salina), we studied the effects of growth phase of cells, electric parameters, electroporation buffer and concentration of plasmid on transformation efficiency. The results showed that a transformation efficiency of 1.81 per thousand was achieved in D. salina cells at mid-log growth phase electroporated with plasmid (DCA-bar) 10 microg/mL, voltage 0.8 kV and capacitance 25 microF. However, when glycerol was added to electroporation buffer at a final concentration of 0.4 mol/L, the transformation efficiency was increased up to 2.03 per thousand. Additionally, transformation was done with plasmids DCA-bar, NR-bar, pUomega-bar respectively, under above optimum conditions, and similar transformation efficiencies were obtained. The findings indicate that an efficient and stable system of electroporation in D. salina has been developed, providing a powerful tool for the transgenic research of D. salina.
Chlorophyta ; cytology ; genetics ; Culture Media ; Electroporation ; Organisms, Genetically Modified ; genetics ; Transformation, Genetic ; genetics

In order to build an "Internet +" innovation and entrepreneurship education practice platform, 3,752 undergraduates from 5 medical colleges and universities were investigated by questionnaire. The results showed that there was a gap between the expectation of the students and the setting of entrepreneurship and innovation courses, project guidance and so on. In view of the contradiction between the supply of educational resources and the needs of students in medical colleges and universities, we have developed the "Internet +" innovation and entrepreneurship education practice platform, that including five main functions and three layers of framework based on cloud computing and open source technology. The platform integrates various resources of medical colleges and universities to make useful exploration for cultivating students' innovation and entrepreneurship ability, accelerating the implementation of projects, and promoting the connotation development of innovation.