Objective To probe into the optimal concentration of Wnt-11 to induce the differentiation of bone marrow mesenchymal stem cells (BMMSCs)into cardiomyocyte-like cells in vitro.Methods BMMSCs were isolated from the bone marrow of SD rats using whole bone marrow culture method.After cultured for 48 h, BMMSCs of the second generation were utilized for directed induction.Based on the final concentration of Wnt-11 , BMMSCs were divided up into Group A (100 ng/mL),Group B (200 ng/mL),Group C (400 ng/mL)and Group D (blank control).After 72-hour induction,the cells were cultured in complete medium for 4 weeks while cells in Group D were cultured only in the complete medium.The morphological changes were observed under the phase contrast microscope.Surface antigen expression of BMMSCs was identified by flow cytometry.When cells were cultured for 4 weeks,the expressions of Desmin,Connexin43 and cTnI were detected by immunocytochemistry. Meanwhile, the ultrastructural changes were observed using transmission electron microscope. The mRNA expressions of cardiac transcription factors GATA-4,Nkx2.5 andα-MHC in BMMSCs were detected by RT-qPCR at 1,2 and 4 weeks after induction.Results Primary BMMSCs formed cell colonies at 2 weeks;the cells were mainly fusiform or star-shape,and a few irregularly-shaped ones were also visible.The passaged cells were larger than those of primary culture.After induction,the cells exhibited long shuttle-shape and were aligned in parallel. Flow cytometery displayed that the positive rate of the surface antigens of BMMSCs CD29,CD45,and CD90 was 97.9%,0.4% and 99.5%,respectively.When BMMSCs-induced via Wnt-11 were cultured for 4 weeks,Desmin, cTnI and Connexin43 were all positively expressed in induction groups.Whereas in the blank control group they were slightly positive or negative;the positive rate in Group B was the highest (P<0 .05 ).Transmission electron microscopy exhibited that organelles such as rough endoplasmic reticulum,mitochondria,as well as some ribosomes were visible in the cytoplasm of these cells in each induction group.In addition,myofilaments were arranged in parallel in the cytoplasm.The cells in induction groups could express GATA-4 and Nkx2 .5 in the first week,and then the expression of them decreased in the second week,but then increased in the fourth week;gene expression in induction Group B was significantly higher than in the other two induction groups (P<0 .05 ).The expression of GATA-4 and Nkx2 .5 in Group D was 1 ,α-MHC was not expressed in the four groups during the culture period. Conclusion Wnt-11 can induce the differentiation of BMMSCs into cardiomyocyte-like cells in vitro,and the optimal concentration of Wnt-11 is 200 ng/mL.

Objective To investigate the clinical features, operation procedures and clinical outcomes of cervical spondy-lotic myelopathy with different imageological characteristics. Methods Two hundred and twenty one cervical myelopathy with different imageological characteristics patients between November 2011 and October 2013 were involved in this retrospective study. Patients were distributed into three groups, namely mild group (A), moderate group (B) and severe group(C), based on severi-ty of complexity by imageological variables (severity of cervical spine degeneration, the number of spinal cord compression, severi-ty of spinal cord compression and difficulty in operation). Preoperative and postoperative Japanese Orthopaedic Association (JOA) score and mean recovery rate were collected to evaluate the postoperative clinical effects. Demographic data and imageological characteristics were recorded;Univariate analysis or analysis of variance was conducted to analyze the correlation between post-operative JOA recovery rate with gender, age, course of disease, severity of spinal cord compression, the number of segment with spinal cord compression, severity of disc degeneration, MRI T2 increased signal intensity, cervical instability, cervical kyphosis, developmental cervical spinal stenosis and ossification of posterior longitudinal ligament. Results There was no significant differ-ence in gender between three groups. There was significant difference in age, preoperative JOA score and improvement rate be-tween group A and group B (C). All patients received decompression and reconstruction by anterior or posterior approach. The ra-tio of anterior approach in group A, B and C was 91.1%(72/79), 79.8%(71/89), 35.8%(18/53). The last follow-up JOA recovery rate of group A, B and C was 75.4%±6.4%、67.7%±8.7%、62.8%±10.4%. The last follow-up JOA recovery rate was correlated with duration of disease, the number of spinal cord compression and severity of spinal cord compression. Age, duration of course, preop-erative JOA score, degree of cord compression, the number of segment with cord compression, degree of disc degeneration, MRI T 2 increased signal intensity, cervical instability, cervical kyphosis, developmental cervical spinal stenosis and OPLL. Conclusion Patients suffering from cervical myelopathy with different imageological have good prognosis by appropriate operation procedure.

ObjectiveTo discuss the selection of surgical approach,operative methods,and stability of reconstitution of affection of cervicothoracic junction.MethodsFrom January 2001 to February 2009,86cases with affection of cervicothoracic junction were treated surgically.The mean age of patients at the time of surgery was 43.1 years (range,17-70).Fifty-seven patients were treated with anterior approach (fixation with autologous bone grafts was done in 38 patients,Cage fixation in 5,titanium mesh in 14),21 with posterior approach(the fixation of lateral mass screw combined with pedicle screw was used in 12 patients,fixation with pedicle screw in 9),and 8 with anterior combined posterior approach.The neurological function of 53 cases of injury of cervicothoracic junction was assessed by American Spinal Injury Association (ASIA) criteria,and the rest was assessed by Japanese Orthopaedic Association(JOA) criteria; bone arthrodesis and restoring lordosis of cervical spine were assessed by Bohlman radiographic criteria.ResultsSixty-nine cases were followed up for an average of 12.4 months(range,3-45).ASIA score increased from 1.8 preoperatively to 2.3 postoperatively,and JOA score increased from 10.3 preoperatively to 12.8 postoperatively.Bone fusion reached in all patients,and lordosis of the cervical spine of 62 cases was restored.One case with respiratory dysfunction,1 case with cerebrospinal fluid leakage,2 cases with hoarseness,and 1 case with loose lateral mass screw were found after the surgery.ConclusionThe advantages of surgical reconstitution of the cervicothoracic junction included promoting recovery of neurological function,restoring the alignment and lordosis of the cervical spine,decreasing the rate of complication,which were dependent on the suitable surgical indications,surgical approach,and way of surgical reconstitution.The selection of reconstituted methods depends on disease,lesion site,type and degree of injury,experience of doctor,decompression and spinal stabilization synthetically.

Objective To explore the indications of cervical artificial disc replacement (ADR) based on radiographic evaluation and different anterior decompression methods. Methods From January 2008 to July 2009, 175 patients with cervical spondylosis or disc herniation who underwent anterior decompression were involved in this study. Patients were distributed to different operative groups based on the preoperative radiographic evaluation. One hundred and forty-five cases were treated with fusion operation, and the others received ADR. Operative methods were as follows: 1) Anterior cervical discectomy and fusion (ACDF); 2)Anterior cervical discectomy and subtotal vertebrectomy; 3) Anterior subtotal vertebreetomy and fusion; 4)ProDisc-C ADR. The patients with single-level of cervical spondylotic myelopathy were divided into ACDF and ADR groups according to different operative methods. Clinical outcomes of two groups were evaluated by Japanese Orthopaedic Association (JOA) score. The range of motion (ROM) of the segment was recorded in ADR group at the 1st month, 3rd month, 6th month and 12th month postoperatively. Results The indication of ADR was cervical spondylosis with slight disc calcification or small vertebral posterior osteophytes. Under this condition, decompression could be obtained thorough intervertebral space and ADR be implanted. If cervical spondylosis was associated with vertebral posterior huge osteophytes, serious intervertebral narrow or fusion, serious disc calcification ,ossification of the posterior longitudinal ligament and extensive cervical spinal stenosis, subtotal vertebrectomy was necessary. The mean improvement rates of ACDF and ADR were 66.05% and 67.13%. There was no difference between two groups (P ＞ 0.05). No difference of ROM was found before and after surgery in ADR group (P ＞0.05). Conclusion Only decompression can be achieved thorough through the intervertebral space, and ADR is suitable for cervical spondylosis. ACDF and ADR have similar outcomes in treatment of single-level of cervical spondylotic myelopathy. But ADR has the advantage of maintaining ROM of the operative segment.

Objective To study the roles of ethyl pyruvate ( EP ) in spinal cord edema after spinal cord injury ( SCI ) and in spinal cord astrocytic swelling after oxygen-glucose deprivation and reoxygenation ( OGD/R) in vitro in rats. Methods After SCI models were established in adult Sprague-Dawley ( SD ) rats, an intraperitoneal injection of EP was conducted to inhibit high mobility group box-1 ( HMGB1 ). Effects of EP on spinal cord edema, HMGB1 expression and astrocyte activation ( glial fibrillary acidic protein ( GFAP ) expression) in SCI rats were analyzed. Spinal cord astrocytes were cultured in post-natal SD rats and incubated under OGD/R procedure. Effects of EP on cell swelling, expression of HMGB1, aquaporin-4 ( AQP4 ) and toll-like receptor 4 ( TLR4 ) , and nuclear expression of nuclear factor-kappa B ( NF-κB ) in spinal cord astrocytes were observed. Results The water content in the spinal cord was increased significantly more at 1 d after SCI than at 12 h and 3 d ( P ＜0.05 ). Intraperitoneal injection of EP at 50 mg/kg reduced spinal cord water content, HMGB1 expression and astrocyte activation ( GFAP expression ) in SCI rats signif-icantly more than that at 25 mg/kg or 100 mg/kg ( P ＜0.05 ). The volume of spinal cord astrocytes cultured in vitro after OGD 6 h/R 24 h was significantly greater than that after OGD 6 h/R 6 h or OGD 6 h/R 12 h ( P ＜0.05 ). EP at 12 μmol/L reduced cell swelling, decreased expression of HMGB1, AQP4 and TLR4, and downgraded nuclear expression of NF-κB in spinal cord astrocytes after OGD/R significantly more than EP at 6 μmol/L( P ＜0.05). Conclusion EP may reduce early spinal cord edema after SCI, attenuate spinal cord astrocyte swelling and decrease AQP4 expression after OGD/R in vitro by inhibiting HMGB1 in rats.

Objective: To evaluate the difference of plasma myeloperoxidase (MPO) level in different types of acute coronary syndrome (ACS) patients, and the value of baseline MPO level in predicting short-term and long-term outcomes in patients with ACS. Methods: The study cohort was derived from "the 12th Five-Year" National Science and Technology Support Program Project "Study on Comprehensive Intervention and Prognosis of Acute Coronary Syndrome" . We enrolled all hospitalized ACS patients who were enrolled in "the 12th Five-Year" cohort from January 1, 2011 to December 31, 2013. A total of 630 patients from 20 centers were enrolled. According to the diagnosis, the patients were divided into two groups: ST-segment elevation myocardial infarction (STEMI) group and non-ST-elevation acute coronary syndrome (NSTE-ACS) group. Plasma levels of MPO were measured by ELISA method. Cardiovascular events in the hospital were recorded. All patients were followed-up by telephone, follow-up ended December 31, 2015. The occurrence of major adverse cardiovascular events (MACE, defined as cardiac death, recurrent myocardial infarction, unscheduled coronary revascularization procedure and stroke) and all-cause death were recorded. Logistic regression analysis and Cox regression analysis were used to evaluate the predictive value of baseline MPO levels obtained during hospitalization and the long-term outcomes of ACS patients. Results: A total of 597 ACS patients were enrolled in final analysis. Level of plasma MPO in STEMI patients was significantly higher than that of NSTE-ACS patients (34.02(19.31, 67.87) μg/L vs. 27.25(16.69, 52.92) μg/L, P=0.028) . MPO was not related to the in-hospital cardiovascular events (OR=0.797, 95%CI 0.366-1.737, P=0.569). Follow up was completed in 476 patients, median follow-up time was 796 (32, 1 816) days. There were 23 all-cause deaths and 51 MACE. Plasma MPO level was not an independent predictor for all-cause death (HR=1.434, 95%CI 0.502-4.100, P=0.501) and MACE (HR=1.271, 95%CI 0.662-2.442, P=0.471). Conclusion In hospitalized ACS patients, level of plasma MPO was significantly higher in STEMI patients than in NSTE-ACS patients, but MPO could not predict the short-term or long-term outcomes in patients with ACS.

Objective:To determine the relationship between tibial plateau stresses and malunion by exploring the changes in mechanical conduction in the knee joint after malunion of Hoffa fracture of the tibial plateau.Methods:This study selected 28 knee joint specimens treated with formalin for preservation, half of which were from male and half from female individuals with an age of (51.4±9.5) years. Their structures were intact, and flexion-extension activities normal. X-ray examinations excluded osteoporosis, tuberculosis, and diseases that could have potentially affected bone quality. The knee specimens were divided into a control group (intact tibia) ( n=4) and 6 groups of tibial plateau Hoffa fracture malunion model: 3 vertical malunion groups (groups V1, V2, and V3, with a vertical displacement of 1, 2, and 3 mm, respectively, n=4) and 3 separation malunion groups (groups S3, S5, and S7, with a separation displacement of 3, 5, and 7 mm, respectively), with half males and half females in each group. After a 600N vertical load was applied at passive knee flexions at 0°, 30°, 60°, 90°, and 120°, the stress levels in the medial and lateral compartments of the knee joint were measured using pressure-sensitive films. Results:Under a vertical load of 600 N, when the knee joint was in a neutral position (flexion of 0°), the differences in the medial and lateral tibial plateau stress values were not statistically significant between the malunion models groups and the control group ( P>0.05). When the knee flexion increased to 30°, the medial tibial plateau stress in the V3 and S7 groups was significantly greater than that in the control group ( P<0.05). At a knee flexion of 60°, the medial plateau stress was significantly greater in the V3, S5 and S7 groups than that in the control group, and the differences were significantly greater than the comparisons at a knee flexion of 30° (all P<0.05). When the knee flexion was 90°, the medial plateau stress in the V2, V3, S5 and S7 groups was significantly greater than that in the control group ( P<0.05), but the lateral tibial plateau stress in the V3 group was significantly smaller than that in the control group ( P<0.05). When the knee flexion was further increased to 120°, the differences in the medial and lateral plateau stress values were statistically significant between all the malunion groups and the control group ( P<0.05), and the differences significantly greater than the comparisons at a knee flexion of 90° (all P<0.05). Under a vertical load of 600 N, the differences in the stresses on the medial and lateral plateaus were not statistically significant between the control group and all the malunion groups at a knee flexion of 0° ( P>0.05). When the knee flexion increased to 30°, the difference between the medial and lateral stresses was not statistically significant in the control group ( P>0.05), but was statistically significant in the V3 and S7 groups ( P<0.05). When the knee flexion reached 60°, 90°, and 120°, the differences between the medial and lateral tibial plateau stresses in all the groups were statistically significant ( P<0.05). Conclusions:The peak knee stresses after malunion of Hoffa fracture of the tibial plateau correlate with the severity of malunion and knee flexion angles. The mechanical properties are not significantly different between a mild malunion knee and a normal knee, but a significant displacement (vertical displacement >2 mm and separation displacement ≥5 mm) may increase the peak knee stresses to increase the risk of knee osteoarthritis. When the severity of malunion is certain, an increase in knee flexion angle increases the difference in the peak stress between the medial and lateral tibial plateaus, thus increasing the risk of knee osteoarthritis.