ABSTRACT
To explore the effects of miR-101-3p on IL-1β-induced chondrocyte injury and its underlying mechanisms. Methods: Chondrocytes were divided into 4 groups: a control group (NC group), a IL-1β group, a negative control group (IL-1β+miR-NC group), and a miR-101-3p group (IL-1β+miR-101-3p group), which were treated with IL-1β after transfecting with miR-101-3p mimic or negative mimic. The expressions of miR-101-3p-5p and stanniocalcin 1 (STC1) at different concentrations of IL-1β (1, 5, 10 ng/mL)-induced chondrocytes were detected by Western blotting and real-time PCR. MTT assay was used to detect cell proliferation rate, while caspases assay kits and flow cytometry were used to measure the cell caspase and apoptosis level. Western blotting assay was used to detect the expression levels of pro-inflammatory and ECM-related protein, such as matrix metalloproteinase 9 (MMP9) and collagen Type II. In addition, 3'-untranslated regions (UTR) of wild-type STC1 (STC1-3'-UTR-WT) or 3'-UTR of mutant STC1 (STC1-3'-UTR-MUT) were co-transfected with miR-101-3p mimic or miR-NC, respectively, while luciferase reporter assay was used to examine the regulative role of miR-101-3p in STC1. In order to detect whether STC1 was involved in the effect of miR-101-3p on chondrocytes, miR-NC (miR-NC group), miR-101-3p (miR-101-3p group), anti-NC (anti-NC group) and anti-miR-101-3p (anti-miR-101-3p group) were respectively transfected into the cells, and the expression of STC1 protein was detected by Western blotting. Subsequently, the cells were randomly divided into a miR-101-3P group (IL-1β+miR-101-3p group), an over-expression control group (IL-1β+miR-101-3p+ad-GFP group), and an over-expression STC1 group (IL-1β+miR-101-3p+ad-STC1 group) to investigate whether STC1 was involved in the role of miR-101-3p in chondrocyte. Similarly, MTT assay was used to detect cell proliferation rate, caspases assay kits and flow cytometry were used to measure the cell caspase and apoptosis level. Western blotting assay was used to detect the expression levels of pro-inflammatory and ECM-related protein MMP9 and collagen Type II. Results: Compared with the 0 ng/mL IL-1β, the expression of miR-101-3p was decreased in chondrocyte at different concentration of IL-1β (1, 5, 10 ng/mL) (all P<0.05), while the level of STC1 was increased (P<0.05). Compared with the NC group, the chondrocyte proliferation rate was down-regulated (P<0.05), while the apoptosis rate, the levels of caspases, IL-6 and TNF-α were increased in the IL-1β group (P<0.05). Moreover, the MMP9 levels were increased obviously, and the protein levels of collagen Type II were decreased in the IL-1β group compared with the NC group (both P<0.05). Compared with the IL-1β+miR-NC group, the proliferation rate was increased (P<0.05), whereas the apoptosis rates, the caspase-3/9 levels, the IL-6 and TNF-α levels were increased in the IL-1β+miR-101-3p group (all P<0.05). Then MMP9 levels were decreased obviously (P<0.05), and the protein levels of collagen Type II were increased in IL-1β+miR-101-3p group compared with the IL-1β+miR-NC group (both P<0.05). In addition, the double luciferase assay showed that the STC1 levels could be inhibited in the miR-101-3p group compared with the miR-NC group (P<0.05). STC1 levels were decreased in the miR-101-3p group compared with the miR-NC group (P<0.05), and the STC1 levels were increased in the anti-miR-101-3p group compared with those in the anti-NC group (P<0.05). The results of miR-101-3p+ad-STC1 group showed that compared with the miR-101-3p+ad-GFP group, the STC1 could reverse the effects of miR-101-3p on IL-1β-induced proliferation, apoptosis, inflammatory responses and ECM protein of chondrocytes. Conclusion: The regulation of miR-101-3p/STC1 signal pathway may have a role in reducing the IL-1β-induced chondrocyte injury.
Subject(s)
Cell Proliferation , Chondrocytes , Glycoproteins , Metabolism , Interleukin-1beta , Metabolism , MicroRNAsABSTRACT
Objective: To discuss the effectiveness of using dorsal two wing-shaped advancement flap to reconstruct finger web for treatment of congenital syndactyly. Methods: Between August 2014 and August 2017, 30 cases of congenital syndactyly were treated, including 18 males and 12 females with an average age of 2.5 years (range, 1.5-5 years). Eight cases were of bilateral hands syndactyly and 22 cases of single hand syndactyly. There were 39 webs of syndactyly (including 1 case of syndactyly of middle finger, ring finger, and little finger). Among them, 11 webs were complete and 28 webs were incomplete. At the dorsum, a flap with V-shaped tip and two wing-shaped pedicle was designed and was just sewed up with an anchor-shaped incision at the palm. Distal end of fingers were separated by serrated flap and were sutured after removal of fatty tissue. In 11 cases with tight skin connection, the defect area at lateral and distal end of fingers was repaired by small pieces of full-thickness skin graft. Results: All the flaps survived completely after operation, and no flap necrosis occurred. The skin grafts on the distal side of the finger survived and the wound healed by first intension. All 30 cases were followed up 6-12 months, with an average of 9 months. Postoperative flexion and extension function of fingers were good, and the web depth and width were normal. At last follow-up, according to the Swanson et al. standard, 20 fingers were graded as excellent, 8 as good, and 2 as fair, with an excellent and good rate of 93.3%. Conclusion: The effectiveness of using dorsal two wing-shaped advancement flap to reconstruction finger web for treatment of congenital syndactyly is satisfactory.
ABSTRACT
Osteosarcoma is the most common maligant in skeletal system.Its treatment is always the focus of most clinical research workers.This article is a brief overview of the research progress of the therapy for osteosarcoma.In addition to neoadjuvant chemotherapy,surgical treatment,this article also includes molecular targeted therapy,gene therapy,and immune therapy.These new advances will contribute to the clinical work.