Mechanism of miR-26a-5p/cAMP response element binding protein 1 molecular axis regulating osteogenic differentiation of adipose-derived mesenchymal stem cells / 中国修复重建外科杂志

OBJECTIVE@#To investigate the regulatory effects of miR-26a-5p on the osteogenic differentiation of adipose-derived mesenchymal stem cells (ADSCs) by regulating cAMP response element binding protein 1 (CREB1).@*METHODS@#The adipose tissues of four 3-4 weeks old female C57BL/6 mice were collected and the cells were isolated and cultured by digestion separation method. After morphological observation and identification by flow cytometry, the 3rd-generation cells were subjected to osteogenic differentiation induction. At 0, 3, 7, and 14 days after osteogenic differentiation induction, the calcium deposition was observed by alizarin red staining, ALP activity was detected, miR- 26a-5p and CREB1 mRNA expressions were examined by real-time fluorescence quantitative PCR, and CREB1 protein and its phosphorylation (phospho-CREB1, p-CREB1) level were measured by Western blot. After the binding sites between miR-26a-5p and CREB1 was predicted by the starBase database, HEK-293T cells were used to conduct a dual-luciferase reporter gene experiment to verify the targeting relationship (represented as luciferase activity after 48 hours of culture). Finally, miR-26a-p inhibitor (experimental group) and the corresponding negative control (control group) were transfected into ADSCs. Alizarin red staining, ALP activity, real-time fluorescent quantitative PCR (miR-26a-5p) and Western blot [CREB1, p-CREB1, Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN)] were performed at 7 and 14 days after osteogenic induction culture.@*RESULTS@#The cultured cells were identified as ADSCs. With the prolongation of osteogenic induction culture, the number of calcified nodules and ALP activity significantly increased ( P<0.05). The relative expression of miR-26a-5p in the cells gradually decreased, while the relative expressions of CREB1 mRNA and protein, as well as the relative expression of p-CREB1 protein were increased. The differences were significant between 7, 14 days and 0 day ( P<0.05). There was no significant difference in p-CREB1/CREB1 between different time points ( P>0.05). The starBase database predicted that miR-26a-5p and CREB1 had targeted binding sequences, and the dual-luciferase reporter gene experiment revealed that overexpression of miR-26a-5p significantly suppressed CREB1 wild-type luciferase activity ( P<0.05). After 7 and 14 days of osteogenic induction, compared with the control group, the number of calcified nodules, ALP activity, and relative expressions of CREB1, p-CREB1, OCN, and RUNX2 proteins in the experimental group significantly increased ( P<0.05). There was no significant difference in p-CREB1/CREB1 between the two groups ( P>0.05).@*CONCLUSION@#Knocking down miR-26a-5p promoted the osteogenic differentiation of ADSCs by up-regulating CREB1 and its phosphorylation.

Preliminary effectiveness of carpal arthroscopic adjuvant treatment of intra-articular fractures of distal radius / 中国修复重建外科杂志

Objective: To investigate the early-term effectiveness of carpal arthroscopy in the treatment of intra-articular fractures of distal radius. Methods: The clinical data of 50 cases of intra-articular fractures of distal radius between January 2015 and December 2017 were retrospectively analyzed. According to the different methods of intraoperative assisted treatment, the patients were divided into the trial group (11 cases with carpal arthroscopy assisted treatment) and the control group (39 cases with traditional open reduction). There was no significant difference between the two groups in general data such as gender, age, affected side, cause of injury, time from injury to operation, and preoperative displacement ( P>0.05), which were comparable. Six patients in the trial group had triangular fibrocartilage complex (TFCC) injury and received one-stage repair. Postoperative X-ray films were taken to estimate the fracture reduction. Patient-Rated Wrist Evaluation (PRWE) wrist function score and modified Mayo score were used at 3 months after operation to evaluate the function of the wrist. The range of wrist flexion, extension, pronation, and supination motion of the two groups were recorded and compared at 3 months after operation. Patients in the trial group were further divided into the reduction group after arthroscopic exploration (group A, 6 cases) and the simple cleaning group after arthroscopic exploration (group B, 5 cases), and their wrist motions were compared. Results: The operation time of the trial group was greater than that of the control group ( t=11.08, P=0.00). There was no significant difference in intraoperative blood loss and fracture reduction between the two group ( P>0.05). X-ray film at 1 day after operation showed that the degree of fracture displacement was significantly decreased when compared with preoperative one in each group ( P0.05). Conclusion: Carpal arthroscope assisted treatment of intra-articular fractures of distal radius can achieve good reduction and postoperative function. Meanwhile, TFCC, ligament, articular cartilage, and other injuries can be repaired in one stage.

Clinical comparative study of thoracoscopic assisted reduction and traditional manual reduction with percutaneous intramedullary nail internal fixation for mid-clavicular fractures / 中国修复重建外科杂志

Objective: To compare the effectiveness of thoracoscopic assisted reduction and traditional manual reduction with percutaneous intramedullary nail internal fixation in the treatment of mid-clavicular fractures. Methods: A prospective randomized controlled trial was conducted. Twenty-two patients with mid-clavicular fractures who met the selection criteria between March 2012 and March 2017 were recruited and randomly divided into trial group (7 cases, thoracoscopic assisted reduction and percutaneous intramedullary nail fixation) and control group (15 cases, traditional manual reduction and percutaneous intramedullary nail fixation). There was no significant difference in gender, age, side, cause of injury, fracture classification, interval between injury and operation between the two groups ( P>0.05). The operation time and fracture healing time were recorded and compared between the two groups. The effectiveness was evaluated by Constant-Murley scale at 6 months after operation, which included subjective evaluation indexes (functional activity and pain) and objective evaluation indexes (range of motion of shoulder joint and muscle strength). Results: The operation time of the trial group was significantly longer than that of the control group ( t=5.881, P=0.000). Patients in both groups were followed up 7-20 months, with an average of 11 months. Satisfactory anatomical reduction achieved in all patients, and all incisions healed by first intension. In the control group, 1 patient had difficulty in removing the intramedullary nail, and 1 patient had fracture nonunion. No fracture nonunion or intramedullary nail rupture in the other patients of two groups. There was no significant difference in fracture healing time between the two groups ( t=0.764, P=0.453). At 6 months after operation, there was no significant difference in Constant-Murley scale between the two groups ( P>0.05). Conclusion: The treatment of the mid-clavicular fracture by using thoracoscopic assisted reduction with intramedullary nail internal fixation requires longer operation time, but does not require fluoroscopy. The effectiveness is comparable to that of traditional surgery.

Effectiveness analysis of modified tarsal sinus approach for Sanders Ⅱ- Ⅲtype calcaneal fractures / 中国修复重建外科杂志

Objective: To investigate the short-term effectiveness of modified tarsal sinus approach and traditional tarsal sinus approach in the treatment of Sanders Ⅱ-Ⅲ type calcaneal fractures. Methods: Between January 2015 and August 2017, 53 patients with Sanders Ⅱ-Ⅲ type calcaneal fractures were selected and divided into observation group (21 cases, using modified tarsal sinus approach for fracture reduction after exposure of the subtalar joint below the long and short fibular tendon) and control group (32 cases, using traditional tarsal sinus approach) by random number method. There was no significant difference between the two groups in terms of gender, age, side, cause of injury, fracture type, injury to operation time, and preoperative Böhler angle, Gissane angle, visual analogue scale (VAS) core ( P>0.05), which were comparable. The operation time, postoperative drainage volume, postoperative Böhler angle, Gissane angle, and postoperative angle improvement values of the two groups were recorded and compared. VAS score, American Orthopaedic Foot and Ankle Society (AOFAS) score, and short-form 36 health survey scale (SF-36) score were used to evaluate the effectiveness. Results: All the 53 patients successfully completed the operation without serious complications such as vascular and nerve injury and perioperative death. There was no significant difference in operation time and postoperative drainage volume between the two groups ( P>0.05). Patients in both groups were followed up 12-36 months (mean, 17 months). No infection, fracture displacement, failure of internal fixation, and malunion of fracture occurred after operation. None of the patients underwent secondary joint fusion. There was no significant difference in fracture healing time between the two groups ( t=0.30, P=0.77). The postoperative Böhler angle and Gissane angle at 2 days in the two groups were significantly improved when compared with those before operation ( P0.05). VAS scores at 24 hours and 1 year after operation were significantly improved when compared with that before operation in both groups ( P0.05). There was no significant difference in AOFAS scores between the two groups at 1 year after operation ( t=1.46, P=0.15). However, the SF-36 scale score at 1 year after operation was significantly higher than that of the control group ( t=2.08, P=0.04). At last follow-up, 2 patients in the observation group and 8 patients in the control group presented subtalar joint stiffness or pain, and there was no significant difference in the incidence between the two groups ( χ2=1.98, P=0.16). Conclusion: The modified tarsal sinus approach for the treatment of Sanders Ⅱ-Ⅲ type calcaneal fractures has the advantages of minimal invasion, clear reduction under direct vision, reliable reduction and fixation, and low incision complications.