Objective To detect the methylation status of CDH1 3 gene promoter in colon cancer by using methylation-specific polymerase chain reaction(MSP)technique.Method The tissue specimens from 32 cases of colon cancer(observation group),and 12 cases of normal colon tissues(control group)were examined,the methylation of CDH13 gene promoter was detected by MSP.Result The methylation of CDH13 gene promoter was detected in 19 cases(59.4％,19/32)in observation group,1 case(8.3 ％,1 / 12)in control group,there was statistical significance between two groups(P =0.002).Conclusion Frequency of the methylation of CDH13 gene promoter is apparently higher in colon cancer tissues than that in normal colon tissues,it reveals that CDH13 gene promoter may contribute significantly to the development of colon cancer.

Objective To optimize the enzymolysis technology of ginsenoside Rg1.Methods Taking ginsenoside Rg1 content as the index,orthogonal design method was used for optimization and HPLC for determination.Results Cellulase enzymolysis was the best extracting process,and enzyme amount,enzymolysis time,and enzymolysis temperature had obvious effect on the extraction of ginsenoside Rg1.The optimum extraction technologies were as follows: cellulase amount was 1.4%,enzymolysis time 60 min,the enzymolysis temperature 45 ℃.Conclusion The optimization extraction technology is simple,steady,and the extracting rate is high.

Objective To investigate the molecular mechanism of Loureirin A mediated anti-hepatic fibrosis by evaluting its effects on proliferation , secretion ofα-smooth muscle actin (α-SMA) and transforming growth factor-beta1 (TGF-β1), and expression of rat hepatic stellate cells in vitro . Methods Primary hepatic stellate cells were isolated and cultured from Sprague-Dawley rats. After activating and inducing primary hepatic stellate cells from qHSC to aHSC, the activated hepatic stellate cells model in vitro was established. Then we observed the morphological changes of static hepatic stellate cells and activated hepatic stellate cells with inverted phase contrast microscope. Cultured hepatic stellate cells were treated with different concentrations of loureirin A and the inhibitory rate of HSCs proliferation was measured by MTT assay. The expression of Frizzled-4 was measured by western blot analysis. The content ofα-SMA and TGF-β1 in the cultured HSCs'supernatant were measured by enzyme-linked immunosorbent assay (ELISA) . Results Loureirin A the proliferation of inhibited activated hepatic stellate cells in a time-dose-dependent manner compared with the control group,IC50=0.30 μg/μL. After loureirinA treatment of the HSCs, western blot analysis showed that Frizzled-4 expression level was obviously lower than control group. Loureirin A also inhibitedα-SMA and TGFβ1 (P<0.05) secretion in the cultured HSCs'supernatant in different degree by the assay of ELISA. Conclusions The molecular mechanism of Loureirin A and Wnt signaling pathway mediated anti-hepatic fibrosis and anti-angiogenesis may involve down-regulation the expression of Frizzled-4, inhibiting the synthesis and secretion ofα-SMA,TGF-β1and the proliferation of HSCs.

Objective To describe the pathological unit and octagonal en bloc resection for the treatment of ossification ligamentum flavum(OLF)in thoracic spine with spondylotic myelopathy.Methods Ninety-five patients from January 2002 to January 2007 were diagnosed as thoracic OLF,61 males and 34 females with an average age of 53.9 years(range,31-78 years).There were upper thoracic spine OLF in 32 cases,middle thoracic spine OLF in 24 cases and lower thoracic spine OLF in 39 cases.Single-segment OLF was found in 53 cases,double segments OLF was found in 38 cases and three segments OLF was found in 4 cases.CT scan multiplanar co-localized reconstruction was employed to detect the structure of spine with OLF.The Japanese Orthopaedic Association(JOA)lower limb motor function score,sphincter function score and motor function improvement rate were used to evaluate the outcomes.Results CT scan was engaged to observe 141 OLF pathological unite.The OLF pathology unit was defined as all the spine structures between the extension lines of the lower margin of the OLF two adjacent pedicles.Each OLF associates with an OLF pathology unit.The mean follow up duration was 38.3 months(range,24-60 months).Among 86 patients with sensations disturbance before operation,67 totally recovered and 19 relieved after operation.Trunk restrictions in 69 cases before operation were completely recovered after operation.Postoperative JOA sphincter function score was 2.651±0.334,comparing with preoperation score(2.262±0.561),and the difference was statistically significant.Postoperative JOA motor function score was 3.694±0.429,which was significantly increased than preoperative score 1.539±0.873,and motor function recovery rate was 87.57%.There was excellent in 71 cases,good in 17 cases and fair in 5 cases.The excellent and good rate was 94.74%.Conclusion The octagonal en block resection is relative safe for treatment thoracic OLF with myelopathy.Pathological unit of OLF in thoracic spine is more accurate to summarize the pathological contents and features of the OLF and its adjacent structure.

Objective To investigate the efficacy of transcatheter closure of coronary artery fistula (CAF) by transthoracic echocardiography(TTE) and the role of TTE in this therapy. Methods CAF were occluded with transcatheter closure techniques in 17 patients. TTE was performed before and after the treatments. The key points were retrospectively analyzed including: the sites of CAF, the position and diameter of CAF,the shape and position of the devices after the intervention,the residual shunt,and cardiac chamber size. Results Before the therapy,TTE made definite diagnosis of CAF. All patients had lesions in single coronary artery, and the diameter of CAF was 2-14 (6.4 ± 3.5)mm. All patients underwent transcatheter closure successfully. TTE revealed the shunts disappear 1-4 days after the occlusion. During a follow-up period of 1- 29 (11.7± 7.9)months,no residual shunts, no complications, and normal size of thecardiac chambers were recognized by TTE. Conclusions The transcatheter closure of CAF has emerged as a less invasive, safe and effective strategy. Echocardiography has important role in primary screening of patients and the follow-up after the treatments.

Objective To evaluate the value of echocardiography in transcatheter closure of ruptured sinus of Valsalva aneurysm(TC-RSVA).Methods TC-RSVA was attempted in 11 patients.The location,shape,size of defects and its relationship with the neighbor structures were revealed before the procedure.Then the deployment of occluder was monitored during the procedure,and the effectiveness was observed in the follow-up.Results Eleven patients were diagnosed as the isolated RSVA by echocardiography.The size of defects was 2 - 13 mm estimated by echo.The procedures were successful in all patients.Usually the Amplatzer duct occluders were chosen to be 1 to 5 mm larger than the size of defects.Three patients had mild residual shunt during the procedure,which all dispeared in the first month of follow-up,but one of them demonstrated recurrent mild residual shunt in the 32nd month of follow-up.There was no aggravating aortic regurgitation in the follow-up.Conclusions TC-RSVA is relatively safe and effective.Observation of long-term effectiveness is still necessary.Echocardiagraphy plays an important role in TC-RSVA.

Objective To sum up a new surgical method to treat knee ankylosis with skeletal traction following arthrolysis. Methods Skeletal traction through supracondyle of femur or through patella was applied for 48 patients with knee ankylosis after dissection of both intra and extra articular adhesion and quadricepsplasty. Results The follow up period ranged from 3 to 36 months. The flexion angles of the knee joint increased 70?to 105?in 34 patients. Conclusion Skeletal traction following arthrolysis to treat knee ankylosis can prolong the length of quadriceps,increase the flexion angle of the knee joint step by step and facilitate early functional recovery after operation.

Objective To obtain and anlysis the diffuse idiopathic skeletal hyperostosis(DISH) related miRNAs under 3-D adhesion for cell culture.Methods From January 2012 to January 2014,4 ossific ligamenta flava tissues were obtained from DISH patients and 4 normal ligamenta flava tissues were obtained from trauma patients surgically.Fibroblasts were separated by using collagenase technique and then cultured on human acellular amniotic membrane (HAAM).Each sample was identified by immunofluorescence before harvested.Total RNA was extracted and then quantified by microfluidics analysis.The small RNAs (＜300 nt) were isolated by using a YM-100 Microcon centrifugal filter.μParafloTM MiRNA microarray assay was performed using a service provider to identify miRNAs whose expression was significantly different between the two groups.Part of differential expression miRNAs were verified by qRT-PCR.Targets of miRNAs were obtained using PicTar 2005,miRanda v5,TargetScan 5.1,their function were analyzed by using Gene Ontology.Functional pathway analysis of miRNAs was performed using KEGG Pathway Analysis.TRANSFAC 7.0 public and Patser were used to get the distribution of transcription factor binding sites.Results When grown on HAAM,fibroblasts kept their morphology,distributed in the way of cluster,lived in multi-level of HAAM,and established linkage.Collagen Ⅰ and Ⅲ were tested positive in normal group cells.Collagen Ⅰ,Ⅱ,Ⅲ and Osteocalcin were tested positive in DISH group cells by immunefluorescence.In total 15 miRNAs showed differential expression,12 were up-regulated and 3 were down-regulated.The result of qRT-PCR was consistent with MiRNA microarray assay.Totally 67 target genes were predicted which participated in cell differentiation,cell adhesion,mineralization et al,and had function in regulating MAPK,Wnt,TGF-β,Focal adhesion signal pathway et al.In total 10 transcription factors were predicted in differentially expressed miRNAs.Conclusion HAAM can provide fibroblasts with 3D adhesion growth,Some differentially expressed miRNAs may participate in the pathogenesis of DISH.

Objective To observe the clinical effects of double support plate via posteromedial approach in the treat?ment of tibial plateau fracture of posterior column. Methods A total of 21 cases of tibial plateau fracture in closed posterior column with an average age of 34.6 (range, 21 to 56) years old were treated using double support plate through posteromedial approach from January in 2010 to January in 2013(16 males and 5 females). Among all, 4 cases were combined with lateral column fracture and 17 cases were three column fracture. Wound healings were observed after operation. X-ray examina?tions were performed at 2 weeks, 3 months, 4 months, 6 months and 12 months after operation as well as at last follow up. The averaged healing time was calculated. Changes of tibial plateau angle and posterior slope angle were compared between time points at 2 weeks after operation and at last follow up. Rasmussen knee score criteria was employed to assess knee joint recovery. Results The mean follow-up was (18.2 ± 1.8) months. No infection was reported. The average fracture healing time was 3.8(3.8±0.6)months. There was no significant difference between tibia plateau angle and posterior slope angle be?tween time points at 2 weeks after operation and at last follow-up (P<0.05). According to Rasmussen knee score criteria at last follow-up, 13 cases were graded as excellent, 6 cases as good, and 2 case as fair, with an excellent and good rate of 90.5%. Conclusion With posteromedial approach, the fractures of medial column, posteromedial and posterolateral of pos?terial column can be exposed well. Using double support plates to fix posteromedial and posterolateral of posterior column fracture can effectively prevent loss of reduction and the function of knee recovered well postoperatively.

Objective To observe the clinical effect of arthrodesis in treating severe hallux rigidus. Methods Totally 30 patients (30 feet) with severe hallux rigidus treated by arthrodesis from January in 2010 to January in 2013 were retrospec?tively analyzed. AOFAS and VAS scoring system were employed to evaluate the clinical curative effect including acquiring the fusion rate, the time to fusion and the incidence rate of hallux valgus. Forefoot-plantar pressure values before and after operation were measured by Plantar pressure measurement system. Results The mean follow-up time was 18 months. AO?FAS score was raised from (48.2±7.6) to (72.8±6.2) after operation (P<0.01). VAS score was also increased from (1.7±0.7) to (8.3 ± 1.1) after operation (P<0.01). The average healing time of knitting was 12.1 weeks. None was non-union and devel?oped to hallux valgus. The forefoot-plantar pressure returned to normal postoperatively from abnormal laterally distribution.Conclusion The surgical strategy of arthrodesis is proved to be effective in treating severe hallux rigidus, in terms of capa?bility of relief from pain, recovery of forefoot-plantar pressure distribution, low incidence of complication.