1.Effects of ursolic acid from loquat leaves on proliferation inhibition andexpression of PPAR-γ, TGF-β1 in rat hepatic stellate cells
Yangwu ZHANG ; Weisheng LUO ; Shan CHEN ; Rui HUANG ; Quanxiao TAN ; Shiyan WANG ; Xia ZHANG ; Chuanfeng XUAN
Chinese Pharmacological Bulletin 2017;33(4):517-521
Aim To investigate the effects of ursolic acid from loquat leaves on proliferation inhibition and expression of PPAR-γ 、TGF-β1 in rat hepatic stellate cells, so as to explore the mechanism of anti-hepatic fibrosis of UA.Methods HSC-T6 cells were randomly divided into blank group, rosiglitazone control group, the low, medium and high concentration of UA group to detect the cell proliferation inhibition by CCK-8 after 24,48,72 h.The content of type collagen Ⅰ in cell culture supernatant of each group was examined by ELISA.The expression of PPAR-γ mRNA, TGF-β1 mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR.Effects on HSC-T6 PPAR-γ and TGF-β1 protein from each group were detected by immunocytochemical method.Results CCK-8 results showed that the inhibitory rate of UA on cell proliferation increased with the prolongation of drug action time(P<0.01).ELISA results showed that with the increase of the concentration of UA, the content of type Ⅰ collagen content decreased(P<0.01).Real-time PCR results showed that with the increase of the concentration of UA, and the expression of PPAR-γ mRNA increased, the expression of TGF-β1 mRNA decreased(P<0.01).The results of immunocytochemistry showed that the expression of PPAR-γ protein was increased, and the expression of TGF-β1 protein was decreased with the increase of the concentration of UA(P<0.01).All effects mentioned above were dose-dependent.Moreover, the effects in the high concentration groups were stronger than those in control group.Conclusion UA can inhibit the proliferation of HSC-T6 cells, Which may be associated with the up-regulation of PPAR-γ expression and the down-regulation of TGF-β1 expression.
2.Repair of forefoot soft tissue defect by free ultra-thin superficial peroneal artery perforator flap
Xiaodong YANG ; Yangwu LIU ; Jing YANG ; Genfu ZHANG ; Maochao DING ; Jin MEI
Chinese Journal of Microsurgery 2013;(2):123-125
Objective To investigate the clinical effect of free ultra-thin superficial peroneal artery perforator flap for repairing the wound in forefoot.Methods From January 2010 to June 2012,six cases were treated with free ultra-thin superficial peroneal artery perforator flap for repairing the wound in forefoot,including 4 cases of dorsal defect,two cases of wound on the toe.The size of the wound ranged from 3.0 cm × 5.0 cm 5.5 cm × 8.0 cm.Four cases for direct suture,two cases of donor site repairing with skin graft.Results All of 6 cases were repaired successfully and no vascular crisis occurred,and clinical follow ups were performed after 3-12 months,the results was satisfactory.Conclusion It is an effective method of repairing the wound in forefoot by free ultra-thin superficial peroneal artery perforator flap.
3.Differentiated expression of VvSUC12 and VvSUC27 in embryogenic and non-embryogenic calli of Vitis vinifera L.
Si CHEN ; Lei ZENG ; Shangwu CHEN ; Yangwu SUN ; Wen ZHANG ; Haiying XU ; Huiqin MA
Chinese Journal of Biotechnology 2010;26(4):530-537
We induced embryogenic calli (EC) and non-embryogenic calli (NEC) from flower filaments of Vitis vinifera L. cv. Chardonnay about 10 days before full bloom. The callus were sub-cultured, observed and verified by somatic embryo induction. PCR primers for VvSUC12 and VvSCU27 were designed according to the corresponding sequences in GenBank. After RNA extraction with RNAplant for EC and NEC cell lines, we synthesized the 1st strand DNA for semi quantitative RT-PCR, and normalized the density of the bands against house-keeping gene Actin. The results of 31 cycles semi-quantitative RT-PCR showed that VvSUC12 was highly expressed in both EC and NEC, with higher expression intensity in NEC than in EC, but not reached the significant level; while the expression of VvSUC27 was only detected in EC, and the expression level was significantly lower than that of VvSUC12. We increased the semi-quantitative RT-PCR cycle number to 35 and found that VvSUC27 gene was weakly expressed in NEC, in EC the intensity of the band was increased comparing with 31 cycles, and the expression level was higher than that of NEC. The paper discussed the differential expression of the two sucrose transporters and their relationship with the sucrose in the tissue culture medium.
Culture Techniques
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methods
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Gene Expression Regulation, Developmental
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Gene Expression Regulation, Plant
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Genes, Plant
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Membrane Transport Proteins
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biosynthesis
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genetics
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Plant Proteins
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biosynthesis
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genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Vitis
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embryology
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genetics
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growth & development
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metabolism