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 ( P<0.05); however, there was no significant difference in Böhler angle, Gissane angle, and improvement value between the observation group and the control group at 2 days after operation ( P>0.05). VAS scores at 24 hours and 1 year after operation were significantly improved when compared with that before operation in both groups ( P<0.05). There was no significant difference in VAS scores between the two groups at 24 hours and 1 year after operation ( P>0.05). 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.

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.

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 ( P<0.05), but no significant difference was found between the two groups at 1 day after operation ( t=0.19, P=0.85). Patients in both groups were followed up 8-20 months, with an average of 12 months. There was no significant difference in fracture healing time between the two groups ( t=0.52, P=0.60). At 3 months after operation, the PRWE score, modified Mayo score, and wrist motions in the trial group were all better than those in the control group ( P<0.05). There was no significant difference in wrist motions between group A and group B ( P>0.05). 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.

Objective To explore the targeting homing capacity of bone-marrow derived mesenchymal stem eells(MSCs) on rabbit myocardial ischemia by intravenous injection of MSCs under the mediation of diagnostic ultrasound and microbubble. Methods Density gradient centrifugation and adherent culture method were used in the isolation and cultivation of MSCs. MSCs were labeled with DAPI. Rabbit myocardial infarction(MI) models were builded by totally ligation of left anterior descending branch of coronary artery. DAPI labeled MSCs were implanted by intravenous injection with or without the mediation of diagnostic ultrasound and microbubbles. Forty-eight hours after cell transplantation, the hearts of MI rabbits were made of frozen section and observed under fluorescent microscope. The DAPI positive cells were counted in the MI and border area of rabbit heart and compared between two groups. Pathological changes of MI area were observed with HE staining under light microscope and transmission electronic microscope. Results The number of DAPI positive cells in MI and border area of rabbit in both groups were counted under fluorescent microscope. There were more DAPI positive cells in the MI area in ultrasound + microbubble + MSCs group (213.2±26.5) than that in the intravenous injection group (146.8±18.78, P<0.01). There were erythrocytes leaking out of the vessels in MI area in HE staining section under light microscope in ultrasound + microbubble + MSCs group while there were nearly none in the intravenous injection MSCs group. The intercellular space of endothelial cells of the vessels wall was increased and serum component leaked out of the vessel wall in ultrasound + microbubble + MSCs under transmission electronic microscope. Conclusions The targeted homing capacity of BM-MSCs in the MI area of rabbit heart can be enhanced under the mediation of diagnostic ultrasound and microbubbles.

Objective To explore the value of diagnostic ultrasound mediated microbubble destruction in improving the myocardial perfusion and left ventricular systolic function when cooperated with the mecsenchymal stem cells(MSCs) transplantation in rabbit myocardial ischemia. Methods One week after myocardial ischemia (MI) modeling,36 rabbits were divided into 3 groups,the control group(group Ⅰ) ,intravenous injection of MSCs group(group Ⅱ) and ultrasound + microbubble + MSCs group (group Ⅲ). Myocardial contrast enhancement (MCE) was performed and quantification analysis of anterior wall was assessed with Photoshop. Left ventrieular systolic function was assessed with M-mode echocardiography and bi-plane Simpson's method. CD34 expression in heart was detected with immunohistochemisty(IHC). Western blotting was applied to detect the level of VEGF in three groups. Results The differences of gray scale analyzed with histogram of Photoshop in anterior wall of ischemia myocardium between the group Ⅰ and group Ⅱ or group Ⅲ were significant,and P value was 0. 032 and 0. 000 , respectively. There were significant differences of FS between group Ⅲ (30. 43±4.09)% and group Ⅱ (26.29±2.93)%, P<0.01, and similar to group Ⅰ (19.28 ± 2.84)%. The difference of EF(%) between group Ⅲ and group Ⅱ was significant [(61.5±5.8 vs 53.6±4. 71), P<0. 05] ,or markedly significant between group Ⅲ and group Ⅰ [(61.5±5.8 vs 42.6± 5.0), P <0.01]. EF(%) assessed with bi-plane Simpson's method was significantly increased from (34.64 ± 4.59) in group Ⅰ to (41.78 ± 4.21) in group Ⅱ and (48.6±3.96) in group Ⅲ. The expression of CD34 assessed with immunohistochemistry was the highest in group Ⅲ. The level of VEGF with western blotting in group Ⅲ was significantly higher than other two groups. Conclusions It is an efficacious transplantation means of MSCs infusion under the ultrasound mediated microbubles destruction in improving the myocardial perfusion and cardiac systolic function.

Objective To establish the methodology of combining BOLD and 1H-MRS for investigating correlation between the deactivation in medial prefrontal cortex (MPFC) and gamma-aminobutyric acid (GABA) concentration by acupuncture at LI4 (Point Hegu),and to optimize the experimental technique and procedure.Methods Twenty healthy adult volunteers were enrolled.During fMRI-BOLD scanning,each subject received acupuncture at right LI4 (Point Hegu).MRS scanning was based on MEGA-PRESS sequence,and ROIs were located at bilateral MPFC.The task BOLD fMRI was block design,including 3 stimulations (30 s) with 2 intervals (2 min).Then MRS scanning was performed before and after BOLD.The quantitative values of the BOLD positive and negative activations (Pm) and GABA concentrations were calculated.Results All 20 subjects completed BOLD fMRI scanning,and met the postprocessing requirements.MRS images of 9 subjects with good image quality were included in analysis.Among all 20 subjects,positive activation (Pm=1.17± 0.16) was observed in 9,while negative activation (Pm =-1.31 ± 0.17) was observed in 11 subjects.The GABA average values before and after the acupuncture were (19.93 ±1.04) nmol/L and (20.04±0.81)nmol/L,respectively,and the average amplitude between post-and pre-acupuncture was (0.11 ± 1.60)nmol/L.Conclusion The success rate of this method for quantitative study of brain function established multimodal-functional (BOLD-fMRI and MRS) was acceptable,and the multimodal brain function changes as well as the quantitative values were observed in the brain region during acupuncture.Combined BOLD and MRS quantitative method is feasible for testing acupuncture response in the brain.

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.
Animals ; Female ; Mice ; Cell Differentiation ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Cyclic AMP Response Element-Binding Protein/metabolism* ; Mesenchymal Stem Cells ; Mice, Inbred C57BL ; MicroRNAs/metabolism* ; Osteocalcin/metabolism* ; Osteogenesis/genetics* ; RNA, Messenger/genetics*