1.Differentially expressed long non-coding RNAs in adipogenic differentiation of human adipose-derived stem cells
Wenjun YUE ; Xiangmei WANG ; Qin ZHANG ; Haihua HUANG ; Youwan WEI ; Zhi PENG
Chinese Journal of Tissue Engineering Research 2014;(28):4491-4497
BACKGROUND:The obesity has led to a plenty of diseases including hypertension, coronary heart disease, fatty liver, hyperlipidemia, and type 2 diabetes. Therefore, understanding the mechanism of adipocyte differentiation is of far-reaching significance to the prevention and treatment of obesity. For the current studies of the mechanism of adipocyte differentiation pay more attention to microRNA, rather than long non-coding RNAs (lncRNAs). OBJECTIVE:To obtain the lncRNAs whose fold change was apparent during adipogenic differentiation, and to further screen the lncRNAs that possibly play a crucial role in adipogenic differentiation for verification. METHODS:Subcutaneous fat was obtained from human abdomen. Adipose-derived stem cells were col ected using tissue culture method. The third passage of adipose-derived stem cells was used for adipogenic differentiation. Through microarray technology, the expression levels of lncRNAs and mRNA were analyzed at 0, 5 and 12 days in adipogenic differentiation. Combining with bioinformatics report, lncRNAs apparently presented fold change were screened and verified by qRT-PCR. RESULTS AND CONCLUSION:Fold change 1.5 (P<0.05) was considered as a criterion during adipogenic differentiation. The number of up-regulated lncRNAs was 748 for 5 days versus 0 day, 847 for 12 days versus 0 days, 593 for 12 days versus 5 days. At the same time, the down-regulated number was 828 for 5 days versus 0 day, 1 113 for 12 days versus 0 day, 750 for 12 days versus 5 days during adipogenic differentiation. In combination with bioinformatics analysis results, 3 of 28 lncRNAs were related to lipid metabolism:AK304548, BP216319 and DA852857, according to the standard that fold change in 0, 5 and 12 days was higher, and the target genes were known to be associated with adipogenesis-related genes. PCR results showed that the expression of AK304548 and BP216319 and its target gene presented an up-down trend, which is consistent with the microarray sequencing results. These results indicated that lncRNA plays a critical regulatory role in the adipogenic differentiation.
2.Treatment of plantar cutaneous deficiency with retrograde medial pedicled skin flap of the leg
Zhi PENG ; Zhenhua JIA ; Haihua HUANG ; Xiaorui GUO ; Youwan WEI ; Jing LI ; Zhiyuan WU ; Peihua ZHANG
Chinese Journal of Microsurgery 2011;34(1):31-33
Objective To introduce clinical application of retrograde medial pedicled skin flap of the leg to repair the plantar cutaneous deficiency. Methods From January 2002 to May 2008, in our hospital we adopted retrograde medial pedicled skin flap to renovate 12 cases of plantar cutaneous deficiency. The size of the skin flaps ranged from 10 cm× 7 cm to 13 cm × 12 cm. Results Ten cases of retrograde medial pedicled skin flap were survived, the rest had partial necrosis because of distal blood flow obstacle. One was cured by changing dressings; the other was repaired by secondary operation. Morphology and function of soft tissue defects were renovated well with 6-18 months follow-up. Conclusion This operation is an effective and reliable technique for plantar cutaneous deficiency.