BACKGROUND:hIL-24, a tumor suppressor gene, can stimulate immune responses, inhibit the growth of tumor cel s, and the formation of tumor vessels, and induce cel apoptosis. OBJECTIVE:To explore the effects of hIL-24 gene on the proliferation and apoptosis of fibroblasts in the keloid and the underlying mechanisms. METHODS:Al the keloid specimens col ected from 13 patients were used for fibroblast culture and indentification. Fibroblast of the keloid was transfected with or without hlL-24 lentivirus. Subsequently, mRNA expressions of transforming growth factor-β, Smad3, proliferating cel nuclear antigen, matrix metal oproteinase-2,-9, and metal opeptidase inhibitor 1 were determined. RESULTS AND CONCLUSION:Immunofluorescent staining and flow cytometry showed that vimentin antibody was expressed positively in cytoplasma of fibroblast cultures, and the purity was more than 97.8%. Western blot assay showed that hIL-24 expression was significantly increased in the transfected fibroblasts. Quantitative PCR showed that the overexpression rate of hIL-24 in fibroblasts was 81.7%and mRNA expressions of transforming growth factor-β, Smad3, proliferating cel nuclear antigen, matrix metal oproteinase-2, and-9 were significantly decreased, while metal opeptidase inhibitor 1 mRNA expression was significantly increased in hIL-24 transfection group compared with control group (P<0.05). These findings suggest that hIL-24 gene inhibits the expressions of proliferating cel nuclear antigen, matrix metal oproteinase-2, and-9 in fibroblasts, and the underlying mechanism may involves TGF-β/Smad3 pathway.

OBJECTIVETo investigate the effect of hIL-24 gene on proliferation, migration and invasion activity of human keloid fibroblasts (KFs).

METHODShIL-24 gene was cloned into lentivirus vector, then the lentivirus particles expressing hlL-24 were infected into KF cells. Real-time PCR and Western blot were performed to examine the expression of hIL-24 in lentivirus infected cells. The growth ability was detected by MTT assay. The cell cycle was analyzed by flow cytometry, The invasion and migration were detected by matrigel invasion assay and wound healing assay.

RESULTSComparing to controls group and KF-NC group, the expression levels of hIL-24 mRNA and protein were both significantly up-regulated after 4 days of hIL-24 lentivims infection. Comparing with the KF-NC group, MTT assay showed that the A490 of KF-hlL-24 group was down-regulated after lentivims infection ( P < 0. 05 ). Comparing with the KF-NC group, Cell cycle test revealed hlL-24 gene could block KF cells in G1 [(75. 40 ±2. 10)% ] , the proportion of KF cells was decreased in S phase [(4. 96 ± 1. 60)% ] and G2 phase [(0.01 ± 0.01)% ]. After KF cells were infected(P <0.01). Transfection of hlL-24 lentivirus inhibited the migration and invasion activity of KF cells.

CONCLUSIONLentivirus-mediated hlL-24 gene efficiently inhibits proliferation, cell cycle progression, migration and invasion activity of KF cells.

Cell Cycle ; Cell Line, Tumor ; Cell Movement ; genetics ; Cell Proliferation ; genetics ; Down-Regulation ; Fibroblasts ; physiology ; virology ; Genetic Vectors ; Humans ; Interleukins ; genetics ; physiology ; Keloid ; genetics ; pathology ; Lentivirus ; RNA, Messenger ; metabolism ; Transfection ; methods

Objective To observe the anatomy of the radial proper digital arteries and their dorsal vessels of index fingers, and the relative position and orientation of them were summarized. To explore the surgical method and clinical appilication of the stepladder advancement flap pedicled with the radial proper digital artery of index finger in the finger tip defects. Methods From June, 2013 to June, 2016, 6 hand specimens were injected into the brachial artery with red latex to carry out the microanatomy of the index finger’s radial proper digital arteries and their dorsal vessels. And 1 vascular cast of hand specimen were observed the origin, number and oriention of the artery and its dorsal vessel. Ten cases with soft tissue defects of index finger in finger tip, were repaired with stepladder advance-ment flap pedicled with the radial proper digital artery. The evaluations and analysis were made in survival rate and finger's function by the postoperative regular consultations. Results There were 2 (4 hands) or 3 (2 hands) dorsal vessels in the proximal, and 2(6 hands) in middle segments of the radial proper digital arteries of index fingers in 6 hand specimens respectively. While the vascular cast of hand specimen showed that 3 dorsal vessels in the proximal, and 2 in the middle segments of the radial proper digital artery. Ten patients were performed the operation. The blood flow after the surgery were good and all flaps survived well. Followed-up time was 10-14 months. The color, feeling, contour and texture of flaps was good. The function of flexion and extension of the finger was good too, and no defor-mity of the purlicue. The resolution of static two points was 5.5-9.0 mm, averaged of 7.2 mm. Conclusion The stepladder advancement flap pedicled with the radial proper digital artery of index finger can extend the donor site. It is safe, reliable and effective, providing a alternation for the repair of the soft tissue defects of the index finger tip.

Objective:To investigate the effect of hsa-miR-199a-5p on keloid fibroblasts (KFs) fibrosis and its relationship with TGF-β/Smad signaling pathway in vitro. Methods:KFs were infected with the lentiviral vector carrying hsa-miR-199a-5p (hsa-miR-199a-5p group) or the blank vector (rLv-NC group), and a blank control group was set. The proliferation status of KFs were detected by CCK-8 assay. At 48 hours after infection, the expression of hsa-miR-199a-5p in KFs were detected by quantitative real-time PCR (qRT-PCR); cell apoptosis was detected by flow cytometry analysis; cell invasion ability was detected by Transwell; The mRNA and protein levels of collagen Ⅰ, collagen Ⅲ, α-smooth muscle actin(α-SMA), Smad3 and TGF-β1 were detected by qRT-PCR and Western blotting, respectively.Results:At 48 hours after infection with RLV-hsa-miR-199a-5p lentiviral vector, the mRNA expression level of hsa-miR-199a-5p in hsa-miR-199a-5p group KFs was significantly higher than that in rLv-NC group and blank control group, and the difference was statistically significant ( P<0.01). Compared with the rLv-NC and blank control groups, the proliferation rate and invasiveness of the hsa-miR-199a-5p group decreased, the apoptosis rate increased, with statistical significance ( P<0.01); the mRNA and protein expression levels of collagen Ⅰ, collagen Ⅲ, α-SMA, Smad3 and TGF-β1 in hsa-miR-199a-5p group were significantly lower than those in rLv-NC group and blank control group, with statistical significance ( P<0.01). Conclusions:hsa-miR-199a-5p may inhibit the proliferation and invasion of KFs by inhibiting TGF-β/Smad pathway, and promote the apoptosis of KFs, thus inhibiting keloid fibrosis.

Objective:To investigate the effect of hsa-miR-199a-5p on keloid fibroblasts (KFs) fibrosis and its relationship with TGF-β/Smad signaling pathway in vitro. Methods:KFs were infected with the lentiviral vector carrying hsa-miR-199a-5p (hsa-miR-199a-5p group) or the blank vector (rLv-NC group), and a blank control group was set. The proliferation status of KFs were detected by CCK-8 assay. At 48 hours after infection, the expression of hsa-miR-199a-5p in KFs were detected by quantitative real-time PCR (qRT-PCR); cell apoptosis was detected by flow cytometry analysis; cell invasion ability was detected by Transwell; The mRNA and protein levels of collagen Ⅰ, collagen Ⅲ, α-smooth muscle actin(α-SMA), Smad3 and TGF-β1 were detected by qRT-PCR and Western blotting, respectively.Results:At 48 hours after infection with RLV-hsa-miR-199a-5p lentiviral vector, the mRNA expression level of hsa-miR-199a-5p in hsa-miR-199a-5p group KFs was significantly higher than that in rLv-NC group and blank control group, and the difference was statistically significant ( P<0.01). Compared with the rLv-NC and blank control groups, the proliferation rate and invasiveness of the hsa-miR-199a-5p group decreased, the apoptosis rate increased, with statistical significance ( P<0.01); the mRNA and protein expression levels of collagen Ⅰ, collagen Ⅲ, α-SMA, Smad3 and TGF-β1 in hsa-miR-199a-5p group were significantly lower than those in rLv-NC group and blank control group, with statistical significance ( P<0.01). Conclusions:hsa-miR-199a-5p may inhibit the proliferation and invasion of KFs by inhibiting TGF-β/Smad pathway, and promote the apoptosis of KFs, thus inhibiting keloid fibrosis.

Objective: To explore the anatomy and clinical application of stepladder V-Y advanced flaps based on the lateral calcaneal artery perforator. Methods: Twenty-four lower extremities from 12 adult cadavers were dissected to investigate the perforators emerging along the lateral edge of the achilles tendon, in terms of amount and distribution, relative to the tip of the lateral malleolus. Based on the anatomical findings, stepladder V-Y advanced flaps, based on the perforator(s) of the lateral calcaneal artery (LCA) or the peroneal artery (PA), were used for reconstruction of lower-posterior heel defects in 22 cases. Results: Twenty-four lower extremities from 12 adult cadavers were successfully dissected. The peroneal artery continued to be the lateral calcaneal artery at the level of (6.0±1.4) cm above the tip of the lateral malleolus. Both of them have the perforators distributed to the skin in the achilles tendon area. In the space from the tip of the lateral malleolus to 8.0 cm and above, a total of 71 perforators with diameter larger than 0.5 mm were found in all extremities. Among them, 9%(7/71) were originated from peroneal artery, and 91%(64/71) were the lateral calcaneal artery. In the 22 cases of clinical application, the pedicle of flaps were used, based on the perforator of lateral calcaneal artery and (or) peroneal artery. The size of flaps ranged from 3.5 cm×9.0 cm to 5.0 cm×10.0 cm.There were 3 cases of skin necrosis at the tip of the flap, which were healed after dressing change. The other flaps had completely survived, and the wound healed in the first stage. Follow-up last for 8-24 months after operation. All patients had well recovery of ankle flexion, extension and appearance. They were able to weight-bearing walk, with natural gait. The texture and sensation of the flaps were similar to that of the surrounding skin. There were few obvious scar or contracture deformity. The functional recovery was satisfied. Conclusions The blood supply in the achilles tendon region is mainly from the lateral calcaneal artery, and partly from the peroneal artery. Stepladder V-Y advanced flap based on the lateral calcaneal artery perforator can be used with reliable blood supply and sensation reconstruction, which is a good choice to repair small to medial wounds in heel area.