Abstract:Intervertebral disc degeneration（IDD） is mainly seen in the lumbar disc herniation，spinal stenosis and other spinal dis－eases and may cause symptoms like muscle strain and back pain，seriously bothering the normal lives of patients. Bone morphogenetic protein（BMP） is a kind of protein proven to be closely related with bone formation and cartilage formation. The effect of BMP on the development and function maintaining of intervertebral disc has attracted a large number of researchers to explore its mechanism. The preventive treatment of IDD by gene therapy is one of the current research hotspots. The research progresses of mechanism of BMP in the repairing of intervertebral disc degeneration were reviewed in this article.

Abstract:Objective：To investigate the effects of recombinant adenovirus-mediated bone morphogenetic protein-7（Ad-BMP-7） over-expression on osteogenic differentiation of prenatal mouse intervertebral disc nucleus pulposus（NP） cell，and to explore whether BMP-7 regulates classic Notch signal pathway in this process. Methods：Recombinant Ad-BMP-7 and adenovirus-mediated green fluores－cent protein（Ad-GFP） were amplified by HEK293 cell and were used to infect NP cells. The experiment was designed into BMP-7 group（n=3），GFP group（n=3） and blank group（n=3）. Real-time PCR was performed to detect the mRNA expression level of BMP-7，real-time PCR was carried out to detect the mRNA expression level of osteogenesis index osteopontin（OPN），osteoprotegerin（OPG） as well as the classic Notch signal pathway members. Western blot was used to detect the protein expressions level of osteogenesis transcription factor Runx2 and OPN. Alkaline pho-sphatase（ALP） activity was measured by using ALP assay and staining. Results：The high titers Ad-BMP-7 and Ad-GFP were generated by adenovirus amplification in HEK293. The basic mRNA expression of BMP-7 was almost undetectable in NP cell. Compared with those in blank group，Ad-BMP-7 mediated over-expression of BMP-7 significantly increased the mRNA expressions of OPN，OPG and the protein expressions of osteogenesis transcrip－tion factor Runx2，OPN（OPN：F=613.815，P=0.000；BMP-7 group vs. GFP group and blank group，P=0.000；OPG：F=10.046，P=0.012；BMP-7 group vs. GFP group P=0.016；BMP-7 group vs.blank group，P=0.047）. The activity of ALP，a specific indicator for steogenesis in Ad-BMP-7 group was statistically higher than that in blank group（F=513.417，P=0.000；BMP-7 group vs. GFP group and blank group，P=0.000），with enhanced positive purple-blue stained cells. After 3 days of Ad-BMP-7 infection，the mRNA expression of classic Notch signal pathway members Notch-1，Notch-2，Jag-1，Hey-1 significantly increased（Notch-1：F=94.574，P=0.002；BMP-7 group vs. GFP group，P=0.003；BMP-7 group vs. blank group P=0.004）（Notch-2：F=318.962，P=0.001；BMP-7 group vs. GFP group and blank group，P=0.000）（Jag-1：F=166.476，P=0.001，BMP-7 group vs. GFP group and blank group P=0.000）（Hey-1：F=1 082.054，P=0.000；BMP-7 group vs. GFP group and blank group，P=0.000），while reversed to the baseline at day 7. There was no difference in others Notch signal pathway members among treated groups. Conclusion：Adenovirus-mediated BMP-7 over-expression can induce osteogenic dif－ferentiation of NP cell，which partly due to transient up-regulation of some classic Notch signal pathway members.

Abstract:Objective：To investigate the mechanism of using massage to promote muscle fiber regeneration by studying on the effect of massage on myogenic regulatory factor of rabbit quadriceps injury Myf5 mRNA and MyoD mRNA. Methods：The 42 healthy adult New Zealand white rabbits，weighting（2.0±0.5） kg，were randomly divided into four groups：normal group（A，n=3），control group before massage（B group，n=3），massage group（C group，n=18） and natural recovery group（D group，n=18）. No treatment was used in A group while hitting by a homemade device was used in B，C and D groups to build rabbits’ quadriceps injury model in right and hind limb. In group C，massage was provided in the first four days after modeling，with speed of 2 600 r/min and frequency of 1/d，for 15 min. In group D，no massage was given. Quadriceps specimens of nine rabbits in C group and D group were got respectively on the 5th d and 10th d after modeling and pathological changes were observed by HE staining. The relative expression of Myf5 mRNA and MyoD mRNA was detected by real-time quantitative PCR. Results：The myogenic regulatory factor Myf5 mRNA expression was （7.82 ±0.19） and （4.99±0.17） in A and B groups，（5.28±0.15） and （5.78 ± 0.04） in C and D groups on the 5th d after modeling，（5.95 ± 0.16） and （6.26±0.26） in C and D groups on the 10th d after modeling，with statistical differences（P<0.01）. MyoD mRNA expression was （14.94±0.12） and （12.12±0.03） in A and B groups，（12.52±0.12） and （12.88±0.14） in C and D groups on the 5th d after modeling，（13.28 ± 0.02） and （13.81±0.17） in C and D groups on the 10th d after modeling，with statistical differences（P<0.01）. HE staining showed that the damage was mitigated and the muscle fibers were more orderly with larger diameter in C groups than in D groups. Conclusion：Massage can promote the synthesis of collagen fibers of injured quadriceps muscles in rabbits，which will help re－pair damaged tissues；its mechanism may be through the pro－motion of myogenic regulatory factor Myf5 mRNA and MyoD mRNA expression.

Abstract:Objective：To compare the stability of expansive pedicle screw（EPS） and polymethylmethacrylate-augmented pedicle screw （PMMA-PS） in osteoporotic synthetic bone blocks. Methods：Thirty osteoporotic synthetic bone blocks were randomly divided into three groups（10 in each group）. A pilot hole was prepared in advance using the same method in all samples. Then，the conventional pedicle screw（CPS） was inserted into the hole in CPS group，the pilot hole was filled with PMMA（2.5 ml） followed by CPS insertion in PMMA-PS group，and EPS was inserted into blocks in EPS group. Twenty-four hours later，X-ray and CT examination and axial pullout tests were performed on all samples. The maximam pullout strength（Fmax） and energy absorbed value（EAV） were measured. Then，the destruction of blocks was observed and the diameter of hole（DOH） was measured. Results：In CPS group，the screw was surrounded directly by synthetic bone without any other materials. In PMMA-PS group，the screw was wrapped up by PMMA totally. The PMMA was evenly distributed in synthetic bone around screw，which obviously improved the local density around track. In EPS group，anterior part of EPS presented an obvious expansion in synthetic bone and formed an unguiform structure which pressed surrounding synthetic bone. Results in the ax－ial pull-out tests revealed that screw stability in both EPS group and PMMA-PS group were significantly enhanced compared with that in CPS group（P<0.05）. But screw stability in PMMA-PS group was significant higher than that in EPS group（P<0.05）. After pullout of screw，the destructions of blocks were the most severe in PMMA-PS group and the lightest in CPS group. The DOH in PMMA-PS group and EPS group were significant bigger than that in CPS group（P<0.05）. But the DOH in PMMA-PS group was significant bigger than that in EPS group（P<0.05）. Conclusion：EPS and PMMA-PS can both significantly increase screw fixation strength in osteoporotic synthetic bone blocks. In addition，EPS can avoid the shortcomings caused by PMMA. Though，EPS shows less effect on augmentation of screw stability compared with PMMA-PS in osteoporotic synthetic bone blocks，it also provides a new study direction to prevent screw loosening in osteoporosis in clinic and avoid risks caused by PMMA in traditional method.

Abstract:Objective：To research the effect of activator protein 2α on the process of bone morphogenetic protein 9（BMP9）-induced osteogenesis differentiation of mouse mesenchymal stem cells（C3H10） and its mechanism. Methods：C3H10 cells were infected with Ad-BMP9. Adenovirus-mediated dominant negative mutants Ad-dnAP2α-△bHLH and Ad-dnAP2α-△TAD were used to inhibit AP2α transcriptional activity. RT-PCR，real-time PCR and Western blot methods were used to detect the expression of AP2α，BMP9 and osteogenetic related markers. Alkaline phosphatase reading/staining and alizarin red staining were performed to detect the status of osteogenetic differentiation. Results：Over-expression of BMP9 could accelerate osteogenesis differentiation but did not affect the expression of AP2α. AP2α dominant negative mutants could inhibit the BMP9-induced increase of osteogenesis related markers osteocalcin（OC），osteopontin（OPN）and osteoprotegerin（OPG）expression（OC：F=56.929，P=0.000；OPN：F=54.789，P=0.000；OPG：F=159.451，P=0.000）and reduce the alkaline phosphatase activity（F=69.776，P=0.000） and red staining of calcium salt deposi－tion. Conclusion：AP2α transcription activity might regulate the process of BMP9-induced osteogenesis differentiation.

Abstract:Objective：To compare the results of bone regeneration condition in mouse calvarial defects measured by Mimics or Micro-CT scanner’s built-in software and to evaluate the differences of the two methods. Methods：Defect of 4 mm in diameter was prepared on the right cranium of mice，and in vivo Micro-CT scan was performed immediately after surgery，6 weeks and 12 weeks postopera－tively. Then new bone volume and bone healing rate within the defect were measured after reconstructing 3D images by Micro-CT scanner’s built-in software. The new bone volume and bone healing rate within the defect were quantitatively analyzed using Mimics software. The 2D data of Micro-CT were imported into Mimics software and 3D images were reconstructed by it as well. Results：The bone regeneration volume and bone healing rate at 12 weeks postoperatively were significantly higher than those at 6 weeks postoper－atively，no matter measured by Micro-CT scanner’s built-in software or Mimics software（P<0.05）. In addition，there was no signifi－cant difference in the new bone volume and bone healing rate between the results of Mimics software and the results of Micro-CT scanner’s built-in software at 6 weeks and 12 weeks postoperatively（P<0.05）. Conclusion：Similar results of new bone regeneration within the calvarial defect are obtained by the two softwares at 6 weeks and 12 weeks postoperatively . However，mimics software is simpler to operate than Micro-CT scanner’s built-in software.

Abstract:Objective：To investigate the osteogenic differentiation of bone marrow mesenchymal stem cells（MSCs） derived from Sprague Dawley rats on TiO2 nanotubes under different inducing conditions. Methods：Isolated and purified MSCs were cultured on the surface structures of plane titalium and TiO2 nanotubes anodized of 30，70，100 nm diameter. Four osteogenic differentiation methods were ap－proached：regular culture medium（control group），regular medium with L-Ascorbic acid and β-glycerophosphate，regular medium with L-ascorbic acid，β-glycerophosphate and 1α，25-dihydroxyvitamin D3 and regular medium with L-ascorbic acid，β-glycerophosphate and dexamethasone. Alkaline phosphatase（ALP） activities of all groups were analyzed on 3，7，14 d after the culture，respectively. Calcium deposition was detected on 21 d after the culture. Real-time PCR was applied to quantitate the expression of osteoblast related gene Runx2 and OSX on 14 d after the culture. Results：MSCs cultured on the same surface structure demonstrated higher ALP activity（F=338.542，P=0.000），calcium deposition（F=417.012，P=0.000） as well as Runx2（F=14.419，P=0.000） and OSX（F=42.011，P=0.000） gene expression in the L-ascorbic acid + β-glycerophosphate + dexamethasone group than in the others，with statistically significant differences. Within the same culture condition，ALP activity（F=53.170，P=0.000），calcium deposition（F=264.268，P=0.000） and Runx2（F=3.196，P=0.037） and OSX（F=5.895，P=0.003） gene expression were much higher on the surface structure of 30 nm diameter TiO2 nanotubes，with statistically significant differences. Culture condition and material structure had interactions（ALP activity（F=6.322，P=0.000），calcium deposition（F=33.330，P=0.000） and gene expression of OSX（F=2.825，P=0.015）），in which L-ascorbic acid + β-glycerophosphate + dexamethasone culture and 30 nm diameter TiO2 nanotubes was the best combination. Conclusion：The osteogenic differentiation of MSCs can be influenced by both the structure of material surface and chemical stimulation. On the same surface structure，MSCs in the L-ascorbic acid + β-glycerophosphate + dexamethasone culture group has greater potential of osteogenic differentiation than the others. Under the same chemical stimulation，MSCs on 30 nm diameter TiO2 nanotubes are more apt to differentiating towards osteoblasts than the way on other surface structures. The combination condition of L-ascorbic acid + β-glycerophosphate + dexamethasone culture and 30 nm diameter TiO2 nanotubes is most conductive to the os－teogenic differentiation of MSCs.

Abstract:Objective：To investigate the effects of recombinant adenovirus-mediated co-infection of bone morphogenetic protein 9（BMP9） and vascular endothelial growth factor（VEGF） gene on osteogenic differentiation of bone marrow mesenchymal stem cells（BMSCs）. Methods：C3H10T1/2 cells were infected by recombinant adenovirus-mediated of BMP9 and VEGF as follow five groups：Ad-BMP9 group，Ad-VEGF group，Ad-VEGF+Ad-BMP9 group，Ad-null group and control group. After infecting for 7 days，RT-PCR was performed to detect the mRNA expression level of BMP9 and VEGF，and real-time PCR was carried out to detect the mRNA ex－pression level of osteogenesis index osteocalcin（OC），osteoprotegerin（OPG），osteopontin（OPN）. Western blot was used to detect the protein expressions level of OPN. Alkaline phosphatase（ALP） activity was measured by quantitative and staining assay，and calcium deposition was determined by Alizarin Red S staining in each group. Results：BMP9 and VEGF co-expressed effectively in Ad-VEGF+Ad-BMP9 group. The mRNA level of OC，OPG，OPN and the protein level of OPN in Ad-BMP9 group and Ad-VEGF+Ad-BMP9 group were statistically higher than those of other groups（P<0.05），and were the highest in Ad-VEGF+Ad-BMP9 group（P<0.05）. The activity and staining assay of ALP，and the deposition of calcium in Ad-VEGF+Ad-BMP9 group were statistically higher than those in the other groups（P<0.05）. Conclusion：Combination of BMP9 and VEGF may promote osteogenic differentiation of C3H10T1/2 cells in vitro.

Abstract:Objective：To observe the effects of compound rhubarb powder on adjuvant arthritis（AA） rats. Methods：The experimental AA model was set up by intradermal injection of freund’s complete adjuvant（FCA）. Normal group，control group，voltaren-treated group and compound rhubarb powder-treated group were established with twelve in each group.The swollen volume of rats’ claw was calculated by plantar circumference method. The morphology of ankle joint was observed under light microscopy. The levels of tumor necrosis factor-α（TNF-α） and interleukin-10（IL-10） in synovial fluid and blood serum and ankle joint were separately measured by ELISA and immunohistochemistry. Results：After six weeks of treatment，compared with those of control group，the swollen volume of rats’ claw was significantly reduced（voltaren-treated group=3.32±0.10；compound rhubarb powder-treated group=3.35±0.07；F=491.02；P=0.00） in treatment group；the expression of TNF-α in blood serum（voltaren-treated group=77.32±2.65；compound rhubarb powder-treated group=75.90±1.95） and ankle joint（voltaren-treated group=4.83±0.30；compound rhubarb powder-treated group=4.83±0.30） was decreased（blood serum：F=1 115.61，P=0.00；ankle joint：F=122.39，P=0.00）；but the expression of IL-10（voltaren-treated group：blood serum=47.04±1.58，ankle joint=5.83±0.39；compound rhubarb powder-treated group：blood serum=46.06±2.03，ankle joint=5.50±0.36） was increased（blood serum：F=1 511.81，P=0.00；ankle joint：F=133.94，P=0.00）. There was no statisti－cal difference between voltaren-treated group and compound rhubarb powder-treated group in the swollen volume of rats’ claw and the ex－pression of TNF-α and IL-10（the swollen volume of claw：F=491.02，P=0.43；synovial fluid and blood serum：TNF-α：F=1 115.61，P=0.21；IL-10：F=1511.81，P=0.31；ankle joint：TNF-α：F=122.39，P=1.00；IL-10：F=133.94，P=0.50）. The inflammatory cells and pannus were reduced and articular cartilage was damaged less in each treatment group under light microscopy. Conclusion：Compound rhubarb powders can obviously decrease joint in－flammation of AA rats as voltaren.Its mechanism may be related with decreasing inflammatory factors（TNF-α） and increasing anti-inflammatory factors（IL-10） so as to eliminate joint swelling and reduce joint inflammation.

Abstract:Objective：To evaluate the prevention of bone cement leakage with gelatin sponge pre-injection during percutaneous verte－broplasty（PVP） operation. Methods：The thoracolumbar samples from elderly female corpse were selected to make compression frac－ture models. The models were randomized into low viscosity bone cement group（group A） and gelatin sponge pre-injection plus low viscosity bone cement group（group B）. The bone cement dynamic viscosity under the temperature of 19 ℃ in both groups was esti－mated. Bone cement leakage rate and leakage quantity of each group were detected by using imitated percutaneous vertebroplasty （PVP） method and by separately injecting 5 ml bone cement at constantly slow speed at points of 3，6，9，12 min after the bone cement reconciliation. Results：The viscosity of low viscosity bone cement increased with the time while adding gelatin sponge exerted great impact on the viscosity. The viscosity of bone cement viscosity in group B was higher than that of group A，with statistical significant differences（P=0.000）. The appropriate time point for adding gelatin sponge was 9 min after cement mixing. The leakage quantity of group B was significantly fewer than that of group A（P=0.035）. Conclusion：The traditional method of combining low viscosity bone cement with gelatin sponge pre-injection can obviously decrease percolation rate and percolation quantity of the bone cement. This method is more economical，easier to operate and has a good prospect of clinical promotion.

Abstract:Objective：To evaluate the clinical outcome of percutaneous vertebroplasty（PVP） in treating osteoporotic vertebral fracture pseudarthrosis. Methods：Fifty-four patients underwent PVP were involved. Visual analogue scale（VAS） and MacNab scale were recorded before the operation and on 1 d and at 1 month，3 months，12 months after the operation. In addition，X ray and CT exami－nation were conducted on 1 d postoperatively. After data collection，SAS software was used to do ANOVA for repeated measurement. Results：Follow-up（12 months） was achieved in 48 patients out of 54 patients. VAS（F=142.481，P=0.000） and MacNab（F=144.207，P=0.000） scores significantly improved on 1 d（VAS=3.120±1.076；MacNab=2.940±0.598） and at 1 month（VAS=2.620±0.874；MacNab=3.060±0.697），3 months（VAS=2.320±0.726；MacNab=3.210±0.683），12 months（VAS=1.920±0.686；MacNab=3.440±0.501） after the operation compared with those before the operation（VAS=8.490±0.689，MacNab=1.270±0.449）. Conclusion：Per－cutaneous vertebroplasty is effective in the treatment of osteoporotic vertebral fracture pseudarthrosis.

Abstract:Objective：To assess the vertebral fracture by dual-energy X-ray absorptiometry（DXA） and to analyze the risk factors of fragility fracture in postmenopausal women. Methods：Totally 203 postmenopausal women underwent vertebral fracture assessment （VFA） by DXA. Bone mineral density（BMD）（g/m2） at different positions was detected and the corresponding T-score（SD） was attained. All patients were grouped based on whether having vertebral fracture and the different parts of fracture. Results：Low BMD（g/m2） and T-score（SD） in patients with both thoracic fracture and thoracic and lumbar fracture were significantly lower than those without vertebral fracture（P<0.05）. The occurrence of vertebral fracture was affected by age（yr）. Conclusion：Lower BMD（g/m2） and T-score（SD） may be important predictors for thoracic fracture and thoracic and lumbar fracture，while exerts little effect on fracture of lum－bar vertebra. Age（yr），BMD（g/m２） of total hip and T-score（SD） are risk factors for vertebral fracture. BMD（g/m2） com－bined with VFA is recommended in the assessment of vertebral fracture.

Abstract:Objective：To evaluate the efficacy of single-level osteotomy in the treatment of malformation typed congenital kyphoscol－iosis（CKS）. Methods：Nineteen consecutive cases of malformation typed CKS patients from July 2006 to May 2012 with single-level osteotomy and posterrior pedicle screw fixation were investigated retrospectively，in which 9 cases were conducted by pedicle subtrac－tion osteotomy（PSO） and 10 cases were conducted by vertebra column resection（VCR）. The average age was 19.2 years and the av－erage follow-up time was 4.2 years. The surgical and radiographic information were comparatively analyzed. Visual Analogue Scale（VAS），the American Spinal Injury Association （ASIA） classification and the complications along the follow-up time were reviewed. Results：Mean operative time was （302.1±43.2） min with average blood loss of （589.5±151.5） ml and average fusion segment of 6.0±1.5. The correction rates of scoliosis Cobb’s angle，segmental kyphosis Cobb’s angle and distance of vertical line between the 7th vertebra and central sacra（C7-CSL） were （79.7±9.2）%，（81.1±21.9）%，（82.7±12.5）%，respectively. The parameters above were not significantly different between PSO group and VCR group except the preoperative segmental kyphosis Cobb’s angle（P >0.05），of which，PSO group was 22.7°±22.0°，VCR group was 62.2°±21.0°（t=4.009，P=0.001）. At last follow-up，no significant progress was observed in scoliosis Cobb’s angle，segmental kyphosis Cobb’s angle and C7-CSL. VAS improvement rate was （85.9±15.9）%. Three cases improved from D to E in ASIA classification. One case of cerebrospinal fluid leakage，two cases of unilateral numbness and one case of back pain were observed without any sign of wound infection or implant failure. Conclusion：Single-level osteotomy with posterior pedicle screw fixation is safe and feasible with ex－cellent correction. For sagittal correction，VCR procedure shows significant advantage.

Abstract:Objective：To analyze the characteristics and key diagnosis points of children humerus distal epiphysiolysis fracture；to study the surgical treatment methods，significance and related issues. Methods：From 2004-2012，197 children with distal humerus epiphysiolysis were received and cured. According to the Delee standards：there were 5 cases of type A，138 cases of type B and 54 cases of type C. Totally 115 cases were cured within 24 h after the injury，69 cases were cured within 1-7 d and 13 cases were cured over 7 d. All children were operated by open reduction and internal fixation through the anterior elbow approach. X-ray was performed after surgery within 24 h. Kirschner wire was removed and excises were started within 4-6 weeks. Results：Totally 197 children were followed up for 6 months to 2 years with an average of 15 months. Kirschner wire was removed within 4-5 weeks with an average of 5 weeks. According to the Flynn standards，elbow function was evaluated：179 cases were excellent，11 cases were good，4 cases were fair，3 cases were bad and the excellent and good rate was 96.4%. Children underwent surgery within 7 d had higher excellent and good rate than those underwent surgery over 7 d，with statistical difference between two groups（P<0.05）. Conclusion：Treating child humerus distal epiphysiolysis fracture through anterior elbow approach is easy to perform. The fracture can be easily restored and the restored fracture can be anatomized. It can reduce the incidence of cubitus varus and cubitus valgus and retain limb function to the greatest extent.

Abstract:Objective：To evaluate the results of single-frame external fixator in treating obsolete Monteggia’s fractures in children．Methods：From January 2008 to December 2011，a total of 22 children with obsolete Monteggia’s fracture were reviewed including 14 males and 7 females with an average age of 7.4-year-old（ranged from 2-year-old to 14-year-old）. According to the Boda classification system：there were 12 cases of type Ⅰ（extension type，54.5%），4 cases of type Ⅱ（flexion type，18.3%），5 cases of type Ⅲ（adduction type，22.7%），1 case of type Ⅵ（4.5%）. All patients were operated with single-frame external fixator after the reduction. The plaster was kept for 4-6 weeks postoperatively. Results：All patients were available at the final follow up with a mean of 2.8 years（ranged from 2 years to 4.8years）. Good union was achieved in all cases after 11.6 weeks（ranged from 10 weeks to 20 weeks）. The radio－humeral joints of 21 cases were returned to normal and subluxating of the radius was observed in one case. There was no ulna nonunion and malunion after the operation. According to the Mackay scoring system，the motion function of 18 cases（81.8%）was excellent，3 cases（13.6%） was good，1 case（4.6%） was poor，with the excellent and good rate of 95.5%. No case of atrogenic vascular injury，os－seous connection of ulna and radius，capitellum and radial head fracture was observed. Conclusion：Single-frame external fixation is effective in the treatment of obsolete Monteggia’s fractures in children. It can rectify the elbow malformation，adjust the operation and restore elbow function. Single-frame external fixation is simple，safe，minimally invasive and has good theraputic effects.

Abstract:Objective：to investigate the effects of combination treatment of hepaCAM and pirarubicin（THP） on the proliferation of BI－U-87 cells. Methods：BIU-87 cells were designed as blank treatment group，adenovirus-GFP-hepaCAM treatment group（Ad-GFP-hepaCAM treatment group），THP treatment group，Ad-GFP-hepaCAM and THP combination treatment group. MTT was used to detect cell proliferation；cell cycle was evaluated by flow cytometry；the mRNA expressions of cyclinD1 and CDK2 were examined by re－al-time PCR. Cells that were designed as blank treatment group，Ad-GFP treatment group，Ad-GFP-hepaCAM treatment group were subcutaneously injected in nude mice. The volume of nude mice was evaluated. After 4 weeks，all mice were sacrificed，transplanted tumors were excised and tumor tissues were used to perform HE staining and immunohistochemistry was performed to examine hep－aCAM expression. Results：Ad-GFP-hepaCAM combined with THP exhibited more intense inhibitory effects on proliferation of BIU-87 cells than THP alone（F=2.702，P=0.000）. Meanwhile，the cell in G0/G1 was significantly increased（F=10.989，P=0.000） and mR－NA levels of cyclinD1 and CDK2 were significantly lowered in Ad-GFP-hepaCAM and THP combination treatment group than in THP group（F=5.4299，P=0.000）. In addition，the volume of tumors induced by hepaCAM-expression BIU-87 cells was significantly decreased compared with that of blank treatment group（F=12.587，P=0.000）. Conclusion：HepaCAM could enhance the inhibitory ef－fects of THP on the proliferation of bladder cancer BIU-87 cells.

Abstract:Objective：To determine the expression of Ghrelin gene in ovarian granulosa cells（GCs） of patients with polycystic ovarian syndrome（PCOS） and to discuss the relationship between them． Methods：GCs were obtained at the time of oocyte retrieval from a total of 47 women undergoing in vitro fertilization and embryo transfer（IVF-ET） treatment due to male factors or tubal factors. Eleven PCOS women were taken as PCOS group and 36 non-PCOS women as control group. The expression of Ghrelin mRNA was measured by real-time PCR and Western blot was used to detect the expression of Ghrelin protein in GCs. Results：The expression of Ghrelin mRNA in GCs was significantly lower in PCOS group than in control group（t=2.73，P=0.015） and the expression of Ghrelin protein in GCs was also significantly lower in PCOS group than in control group（t=7.82，P=0.001）. Conclusion：The mRNA and protein expressions of Ghrelin in GCs are significantly lower in women with PCOS than in non-PCOS women，suggesting that Ghrelin may be involved in the disturbance of follicular development in PCOS.

Abstract:Objective：To observe the expression of the gene of TSO45W-4B of Taenia solium in bifidobacterium longum（B.longum） on the basis of successful construction of the escherichia coli（E.coil）-bifidobacterium shuttle plasmid pGEX-TSO45W-4B of Taenia solium. Methods：The shuttle expression plasmid pGEX-TSO45W-4B of Taenia solium was electroporated into B.longum. After induc－tion with isopropyl-β-d-thiogalactoside，the expression of the recombinant protein was identified by sodium dodecyl sulfatepolyacry－lamide gel electrophoresis（SDS-PAGE） and Western blot. Results：Restriction analysis，polymerase chain reaction and sequencing showed that the recombinant plasmid pGEX-TSO45W-4B was successfully transformed into B.longum. As demonstrated by SDS-PAGE，the relative molecular mass（Mr） of the expressed recombinant protein was approximately 40 kD，and the purity of the recom－binant protein was 85% after purification withglutathione S-transferase affinity chromatography. The rabbit antiserum of TSO45W-4B，cysticercosis swine serum and cysticercosis patients serum could bind to the recombinant protein in Western blot assay. Conclusion：The gene of TSO45W-4B of Taenia solium could be expressed in B.longum and the expressed recombinant protein shows specific antigenicity.

Abstract:Objective：To construct lentivirus vector targeting integrin β1（ITGβ1） gene and to study its effect on migration and inva－sion of Tamoxifen-resistant MCF-7（MCF-7R）. Methods：Four shRNA sequences targeting ITGβ1 gene and one negative control se－quence were designed，and were cloned into lentivirus vector，respectively. Recombinant lentivirus and control were extracted after transfecting 293T with the recombinant vector and helper vectors. The lentivirus with the best interfering effect was determined by real-time PCR and was chosen to infect MCF-7R. The expression of ITGβ1 gene was determined by Western blot. Transwell assay was used to detect the difference of migration and invasion. Results：Four recombinant lentivirus vectors were successfully constructed and the packaged lentivirus with the best interfering effect was used to infect MCF-7R. Western blot results showed that the protein expression of ITGβ1 was significantly reduced in MCF-7R（P=0.000） and its migration and invasion abilities were markedly suppressed（P=0.000）. Conclusion：Lentivirus vector targeting ITGβ1 gene can efficiently suppress the expression of ITGβ1 in MCF-7R，thereby leading to decreased migration and invasiveness abilities of MCF-7R

Abstract:Objective：To investigate the effect of high intensity focused ultrasound（HIFU） on the lysozyme molecular structure. Meth－ods：The lysozyme solution was exposed at HIFU with pulse frequency of 0.94 MHz，sound intensity of ３.25×104 W/cm2，pulse repetition period of 4 Hz and duty cycle of 10% for 10 s. The effect of HIFU on the lysozyme molecular structure was studied using CD spectra，fluorescence spectrum，1H nuclear magnetic resonance（1H-NMR） and sodium dodecyl sulfate-polyacrylamide gel（SDS-PAGE） method. Results：The Result showed that after HIFU treatment，there was no obvious change in CD spectrum，indicating that HIFU might cause little effect on lysozyme lower structure. Whereas a weaken fluorescence intensity appeared indicating lysozyme conformation change. Data of SDS-PAGE showed obvious degradation of lysozyme branches. 1H-NMR analysis demonstrated that tryptophan（Trp） residue of lysozyme displayed chemical shift after treatment. Conclusion：The conformation structure of lysozyme could be changed by HIFU.

Abstract:Objective：To identify the expression of TRIM28 in non-small cell lung cancer（NSCLC），to study the effect of TRIM28 gene on the growth of non-small cell lung cancer. Methods：Immunohistochemistry was used to detect the expression of TRIM28 in clinical tissue of non-small cell lung cancer patients. After TRIM28 gene was inhibited by RNA interference assay，soft agar assay and MTT cell proliferation assay were used to observe the growth and proliferation of lung adenocarcinoma PAa，and flow cytometry was used to an－alyze the cell cycle. Results：TRIM28 positive staining was observed in 24 of 48（50.0%）non-small cell lung cancer tissue samples，whereas no staining was observed in any of the normal portions of the same tissues（ ?字2=27.034，P=0.000）. TRIM28 knockdown resulted in a decrease in cell proliferation and a significant population in G1/S fraction. Conclusion：TRIM28 expresses highly in non-small cell lung cancer tissues. The growth of lung adenocarcinoma PAa transfected with TRIM28 siRNA is inhibited suggesting that TRIM28 may be new therapy target for NSCLC.

Abstract:Objective：To investigate the effect and the mechanisms of lentiviral-shRNA-mediated signal transducers and activators of transcription 3（STAT3） silencing on the chemosensitivity to 5-fluorouracil（5-Fu）， of human gastric cancer cells SGC-7901. Meth－ods：The shRNA sequences targeting STAT3 was constructed and lentiviral vectors was used to transfect human gastric cancer cells SGC-7901. The interference effect of STAT3-shRNA was detected by real-time PCR and Western blot. The chemosensitivity to 5-Fu was analyzed by MTT assay and the flow cytometry was used to examine cell apoptotic rate. Western blot was used to compare the ex－pressions of anti-apoptotic proteins Bcl-2，Mcl-1，survivin. Results：Lentiviral vectors containing shRNA targeting STAT3 gene were successfully established and transfected into SGC-7901 cells．Real-time PCR and Western blot analysis showed that the STAT3-shRNA significantly inhibited the mRNA expression by 54% and protein expression by 40%（P<0.05）． The inhibitory rate of cell pro－liferation and the apoptotic rate were obviously higher in the cells treated by STAT3-shRNA（P<0.05）. STAT3-shRNA also induced downregulation of the anti-apoptotic proteins Bcl-2，Mcl-1 and survivin（P<0.05）. Conclusion：In summary，STAT3-shRNA could effectively inhibit the proliferation and accelerate the apoptosis of SGC-7901 cells，leading to the enhancement of their sensitivity to 5-Fu. The mechanism may be related with block of STAT3 signaling pathway though inhibiting STAT3 activation，thus decreasing ex－pressions of its downstream anti-apoptotic proteins Bcl-2，Mcl-1 and survivin.

Abstract:Objective：To research the effect of c-Ski oncogene on the activation of breast cancer-associated fibroblasts（CAFs）. Methods：The c-Ski gene was cloned into the vector pBABE-puro to construct the expression vector pBABE-puro-Ski then was transfected into normal fibroblasts（NFs）；the stable transfected fibroblasts were selected by puromycin. Western blot was used to ana－lyze the expression of c-Ski，α-smooth muscle actin（α-SMA） and fibroblast activation protein（FAP） after the transfection. MTT as－say，flow cytometry，wound-healing experiment and Transwell assay were used to study the differences of cell proliferation，migration and invasion before and after the transfection. Conditioned media were collected and used to study the effect of fibroblasts on the in－vasion of MDA-MB-231 cells by Transwell assay. Results：The recombinant plasmid pBABE-puro-Ski was successfully constructed. Western blot showed that the expressions of c-Ski，α-SMA and FAP were higher in cells transfected with pBABE-puro-Ski than in those untransfected cells（P=0.000，P=0.001，P=0.002）. MTT showed that the proliferation of pBABE-puro-Ski transfected cells was higher than that of untransfected cells（P=0.011）. FCM showed that the S phase of pBABE-puro-Ski transfected cells（38.58±1.28） was significantly higher than that of empty vector transfected（25.05±0.52） or untransfected cells（26.01±0.86）（P=0.000）. Wound-healing experiment and Transwell assay revealed enhanced migration and invasion of the pBABE-puro-Ski transfected cells com－pared with those of the other two groups. Transwell assays showed that pBABE-puro-Ski transfected cells could promote the invasion of MDA-MB-231 cells. Conclusion：Overexpression of c-Ski could stimulate the activation of breast CAFs and activated CAFs further promote the invasion of MDA-MB-231 cells.

Abstract:Objective：To observe the mouse ovarian preantral granulose cells proliferation and expression of proliferating cell nuclear antigen（PCNA） in order to investigate the mechanism of nerve growth factor（NGF） and K252α and UO126. Methods：The mouse ovarian preantral granulose cells were treated with NGF and inhibitors K252α and UO126 in vitro. Cell proliferation was measured by cell counting Kit-8（CCK-8） assay and change in the expression of PCNA was detected by Western blot. Results：The data showed that the proliferation of granulose cells was significantly increased in NGF group（A value=1.58） than in control group（A value =1.31）（PNGF＝0.000）. The proliferation of granulose cells was significantly decreased in K252α group（A value=1.21），NGF+K252α group（A value=1.19），NGF+UO126 group（A value=1.14） than in control group（PK252α＝0.001，PNGF+K252α＝0.001，PNGF+UO126＝0.000） after applying K252α and UO126. Relative expression of PCNA was higher in NGF group（5.21） than in control group（1.65）（PNGF＝0.000）. Relative expression of PCNA was respectively 1.48，0.84，0.41 in K252α group，NGF+K252α group and NGF+UO126 group，with statistical differences compared with that in control group（PK252α＝0.02，PNGF+K252α＝0.000，PNGF+UO126＝0.000） and that in NGF group（PK252α＝0.000，PNGF+K252α＝0.000，PNGF+UO126＝0.000）. Conclusion：NGF could significantly promote the proliferation of granulose cell and at the same time induce the expression of PCNA. However，the inhibitors suppress the proliferation of granulose cell，which might be associated with the declining expression of PCNA.

Abstract:Objective：To establish protein expressing profile of human bone marrow mesenchymal stem cells（hMSCs） which were induced into osteoblasts through proteomic and bioinformatic techniques and to further elucidate the precise molecular mechanism of hMSCs osteoblast differentiation. Methods：The total protein extracts were obtained with cell lysis buffer from hMSCs which were induced into osteoblasts. Two-dimensional gel electrophoresis（2-DE） and 2-DE gel analysis software were used to recognize and test protein spots. Clearly expressed protein spots were selected and were identified and analyzed by matrix-assisted laser desorption ionization-time of flight mass spectromety（MALDI-TOF-MS） and bioinformatics． Results：Protein expressing maps of hMSCs induced into osteoblasts were set up. Seventy-seven proteins were identified by MALDI-TOF-MS and bioinformatics，which were classified by Gene Ontology according to molecular function and biological process. Conclusion：Protein expressing maps of hMSCs induced into osteoblasts are built，which will provide basal information to illuminate the molecular mechanism of hMSCs differentiation into osteoblasts and to build protein expression database of osteoblast differentiation from hMSCs.

Abstract:Objective：To construct recombinant adenovirus vector（pAdxsi-GFP-VP3） containing VP3 gene and to investigate the apoptosis-inducing effect of VP3 gene on human large cell lung cancer NCI-H460 cells. Methods：VP3 gene was gained by enzyme digestion with pcDNA3.0-VP3 recombined plasmid as a template and was diverted to shuttle vector. The pAdxsi-GFP-VP3 was ob－tained by connecting with pAdxsi vector. After enzymatic digestion and verification by sequencing，virus amplification and purification were conducted，and the titer of the recombinant adenovirus was determined by biological and physical assays finally. The pAdxsi-GFP-VP3 was used to transfect human large cell lung cancer NCI-H460 cells with optimum multiplicity of infection. Expression of Apoptin in large cell lung cancer cells was respectively detected by reverse transcription polymerase chain reaction（RT-PCR） and Western blot. Cell ultrastructure at different periods of time after the transfection was observed by transmission electron microscopy（TEM）. Ef－fect of Apoptin on cell proliferative vitality was determined by MTT assay. Apoptotic rate and cell cycle in NCI-H460 cells after transfection were measured by flow cytometry（FCM）. Results：The pAdxsi-GFP-VP3 was successfully constructed by enzyme diges－tion and sequencing identification and the virus titer was 1.6×109 PFU/ml. RT-PCR results showed the mRNA expression of Apoptin in each experimental group at 48 h after the transfection. Western blot confirmed that Apoptin protein was highly expressed in each experimental group at 72 h after the transfection. The typical morphological changes in early apoptosis such as cell shrinkage，chromatin condensation and apoptotic bodies were observed under TEM. MTT assay indicated that the proliferative activity of trans－fected NCI-H460 cells was remarkably decreased and present－ed in a time-dependent manner（F=93.384，P=0.000） and there were statistical significances among different groups（F=18.427，P=0.003）. FCM detection showed that the expression of Apoptin in NCI-H460 cells after the transfection could induce cell apoptosis with a gradual increase in the apoptotic rate over time（F=47.307，P=0.000；F=303.237，P=0.000）. Moreover，the proliferation of NCI-H460 cells was slowed down；the proportion of cells was decreased at S phase（F=80.240，P=0.000） and was blocked at G2/M phase after the transfection compared with that of control group（F=110.080，P=0.000）． Conclusion：Our data demonstrate that Apoptin can effectively induce apoptosis of NCI-H460 cells，which may lay an ex－perimental basis for applying Apoptin in human NSCLC gene therapy.

Abstract:Objective：To research the growth and development of lucilia cuprina larvae fed by different organization sources. Methods：Under the condition of constant temperature（26 ℃）and relative humidity（70%），muscle tissue，hepatic tissue，brain tissue，muscle and fat mixture（6∶4） were used to feed lucilia cuprina larvae. Length and weight of larvae and were measured at an interval of 12 h while length and weight of pupa were measured after eclosion. Development stages of larvae，mortality of larvae and pupa as well as sex ratio of adults were recorded. Results：Growth was retarded in 40% fat mixed group than in the other three groups，achieving maximum individual delay time of 24 h. Larvae body length of hepatic tissue group（15.02±1.39） mm and 40% fat mixed group（14.85±1.43） mm were statistically shorter than those of brain tissue group（17.53±2.08） mm and muscle tissue group（16.82±2.15） mm（F=5.440，P=0.003）. Larvae body weight of hepatic tissue group（72.8±14.2） mg and 40% fat mixed group（70.4±15.4） mg were statistically lighter than those of brain tissue group（89.6±19.2） mg and muscle tissue group（85.9±16.7） mg（F=3.320，P=0.030）. Pupa body weight and body length of 40% fat mixed group were statistically lighter and shorter than those of brain tissue group and muscle tissue group（F=7.890，P=0.001；F=9.280，P=0.001）. Larvae and pupa mortality of 40% fat mixed group were statistically higher than that in the other three groups（F=3.470，P=0.040；F=4.720，P=0.035）. Conclusion：Lucilia cuprina larvae fed by 40% fat mixed food have longer growth and development period. Body weight and body length of larvae and pupa are lighter and shorter in hepatic tissue group and 40% fat mixed group than in the other groups.

Abstract:Objective：To assess the safety and efficiency of metformin and glyburide in gestational diabetes mellitus by comparing with insulin. Methods：Relevant 16 randomized controlled trials published before December，2013 were searched from PubMed，EMBASE，the Cochrane Central Register of Controlled Trials（CENTRAL），and Chinese biomedicine（CBM） literature database. The safety and efficiency between metformin and glyburide was assessed using direct and indirect results through R 2.15.2. Results：A total of 2 665 participants were identified in this Meta analysis. There were more macrosomia in glyburide group and in insulin group（OR=3.299，95%CI=1.649 to 6.600，P=0.001），more macrosomia in glyburide group than in metformin group（OR=0.234，95%CI=0.061 to 0.893，P=0.033），however there was no difference in macrosomia between metformin group than insulin group（P=0.198）. Maternal weight gain was lower in metformin group than in insulin group（SMD=-0.539，95%CI=-0.716 to -0.362，P=0.000）. Gestational age was smaller in metformin group than insulin group（SMD=-0.142，95%CI=-0.246 to -0.039，P=0.007）. Conclusion：Metformin can better control blood glucose and maternal weight gain and lower glycosylated hemoglobin level without increasing maternal hypoglycaemia，but it may significantly shorten gestational age. Incidence of macrosomia is higher in glyburide group.

Abstract:Objective：To systematically review the clinical efficacy of aspiration thrombectomy during ST-segment elevation of acute myocardial infarction（STEMI）. Methods：Literature search was performed among Pubmed，CNKI，VIP，Wangfang databases to search for available randomized control trials comparing aspiration thrombectomy combined with percutaneous coronary intervention（PCI） and single PCI. The Meta-analysis was conducted by using RevMan 5.2 software. Results：Totally 13 trials concerning 11 823 patients were enrolled. There was no statistically significant difference in all-cause mortality between aspiration thrombectomy combined with PCI and single PCI（RR=0.85，95%CI=0.73 to 1.00，P=0.05）. Major adverse cardiac events（MACE） was significantly reduced in aspira－tion thrombectomy combined with PCI than in single PCI（RR=0.81，95%CI=0.68 to 0.97，P=0.02）. Conclusion：Aspiration thrombec－tomy is conducive to reducing MACE.

Abstract:Objective：To evaluate the effect of the probiotics as adjunctive therapy on the eradication rate and side effects of anti-He－licobacter pylori（H.pylori） eradication treatment in children. Methods：All randomized controlled trials（RCTs） comparing probiotics supplementation with placebo or no extra intervention during anti-H.pylori regimens in children were systematically searched from the databases of PubMed，Embase，Cochrane Controlled Trials Register（CCTR），Web of Science，CNKI，VIP and Wangfang（from their in－ception to October 2013）. The qualities of included RCTs were evaluated and the Meta analysis was conducted by means of RevMan 5.2. Results：Eleven randomized controlled trials with 769 cases were enrolled in the analysis. The eradication rates（RR=1.20，95%CI=1.10 to 1.30，P=0.000） and the total occurrence of side effects（RR=0.44，95%CI=0.26 to 0.73，P=0.001） were significantly better in the treatment group（using probiotics combinded with anti-H.pylori eradication therapy） than in control group（using anti-H.pylori eradication therapy alone）. Moreover，the incidences of diarrhea，nausea and vomiting were significantly lower in treatment group than in control group（RR=0.27，95%CI=0.14 to 0.52，P=0.000；RR=0.47，95%CI=0.24 to 0.93，P=0.030）. Conclusion：The probiotics sup－plementation is effective not only in increasing the eradication rate of anti-H.pylori eradication treatment in children but also in alle－viating anti-H.pylori therapy related side effects.

Abstract:Objective：To compare the pharmacokinetic characteristics between obese and normal patients. Methods：The databases of Medline，Sciencedirect，Cochrane，CEPS，CETD，CNKI and WanFang were systematically searched. RevMan4.2 was used for meta-analysis. Results：A total of 5 researches were included in the research. Meta analysis showed that vancomycin clearance（CL） and distribution volume at steady state（Vdss） were significantly higher in obese population than in normal population. Obese population had higher CL/IBW and Vd/IBW. However，there was no significant difference in CL/TBW between the two groups. Conclusion：CL and Vdss are significantly higher in obese population，but CL/TBW is similar in the two groups.

Abstract:Objective：To evaluate the efficacy of statins for preventing gallstone formation and reducing the risk of cholecystectomy. Methods：PubMed，EMBASE，Cochrane Central Register of Controlled Trials，CBM and VIP library were searched for all studies relat－ing to statins in the prevention of gallstone formation and reduction of cholecystectomy risk. Meta analysis was performed by using Rev Man 5.0 software and Stata11.0 software. Results：Fourteen studies were in enrolled including 5 RCTs，3 self pre-and post-control ob－servations，4 case-control studies，2 cohort studies with a total of 811 393 people（65 605 patients and 745 788 controls）. The result of RCTs and self-control studies showed that there were significantly statistical differences between statin group and control group in lowing biliary cholesterol concentration（SMD=0.41，95％CI＝0.08-0.73，P=0.009）. The cholesterol saturation index（CSI）（SMD=0.53，95％CI＝0.22-0.84，P=0.00１） in the statin group was significant lower than that in the control group. The Meta analysis results of case-control and cohort studies showed that there were statistical differences in reducing cholecystectomy risk among different exposure duration of statins（1-2 years：OR=0.89，95％CI＝0.84-0.94）；>2 years：OR=0.79，95％CI＝0.67-0.94）；<1 year：OR=1.13，95％CI＝1.04-1.22）. Conclusion：Statin can reduce the biliary cholesterol concentration and bile CSI and may have certain effect on inhibit－ing the formation of gallstones. The exposure duration of statins more than one year exerts effect on lowering the risk of gallstone dis－eases after cholecystectomy.