Abstract:Objective：To construct a eukaryotic expression plasmid carrying let-7a and to establish the Ewing’s sarcoma（ES） cell line A673 and SK-ES-1 stably expressing let-7a. Methods：Fragment of attB1-K- eGFP-let-7a-1-attB2 coding let-7a was amplified using Gateway protocols. The let-7a gene was recombined with donor vector pDONR221 to construct an entry plasmid pDONR221-eGFP-let-7a，which forwarded to recombine with pLV.Des2d.P/neo to reconstruct pLV.Des2d.P/neo-EF1A>eGFP/Let-7a. PCR and se－quencing were used to confirm the plasmid. Lenti-let-7a transfected ES cell lines A673/let-7a and SK-ES-1/let-7a which can sta－bly express let-7a were screened out. A673/let-7a and SK-ES-1/let-7a were confirmed by real-time PCR. Western blot analysis was performed to detect the expression of CDK6 in A673/let-7a and SK-ES-1/let-7a cells，which was the target gene of let-7a. Results：PCR results confirmed that the value of target fragment was consistent with the theoretic value. Sequencing analysis showed that let-7a was correctly inserted into the pLV.Des2d.P/neo-EF1A>eGFP/let-7a plasmid. Real-time PCR indicated that the expression of let-7a was increased remarkably in A673/let-7a and SK-ES-1/let-7a compared with that of control and blank cells；expressing values of let-7a were as high as 8.5 folds（P=0.000） and 6.5 folds（P=0.001） in A673/let-7a and SK-ES-1/let-7a respectively. Western blot analysis showed that ectopic expression of let-7a down-regulated the expression of CDK6. Conclusions：ES cell lines A673/let-7a and SK-ES-1/let-7a which can stably express let-7a are successfully established.

Abstract:Objective：To explore the clinical outcomes of interbody bone fusion combined with transpedicular screw internal fixation in selective treatment of thoracolumbar fracture-dislocations complicated with severe intervertebral disc injury through posterior ap－proach. Methods：From January 2005 to Marth 2011，26 patients with thoracolumbar fracture-dislocations complicated with severe in－tervertebral disc injury were treated by this method. Inclusion criteria：①thoracolumbar fracture dislocation in single segment，less than 1/3 upper parts of fractured vertebra being compressed or burst，middle and inferior parts of fractured vertebra being normal；②dam－aged segment complicated with severe intervertebral disc injury. Demogrophic data，AO classification of fracture，American Spinal In－jury Association grade of neurological status，visual analogue scale（VAS），Bridwell fusion grade，segmental height，segmental kyphosis and treatment-related complications were evaluated. Results：Based on AO classification，20 patients had type B facture and 6 had type C. Except 16 patients with grade A spinal cord function，spinal cord function of all patients improved by 1-2 degrees. VAS scores were （9.00±0.76） before operation and were （5.18±1.43） at one week post-operation and （0.74±0.57） at the last follow-up. Twenty-three patients had grade I fusion and 3 patients had grade Ⅱ fusion at the last follow-up. Height of the fractured vertebra was （84.27±5.27）% before the operation，（97.38±1.20）% at one week after the operation and （97.04±0.96）% at the last follow-up. Kyphosis angel was （22.50±6.20）° before the operation，（2.60±1.86）° at one week after the operation and （3.50±2.65）° at the last follow-up. Cerebrospinal fluid leakage occurred in 4 patients. There was no case of incision infection，cage subsidence and screw loosen or breakage. Conclusions：With the correct selection of operative indications，resection of damaged intervertebral disc through posterior approach，cage interbody bone reconstruction and transpedicular screw internal fixation are effective methods in treating thora－columbar fracture-dislocations complicated with intervertebral disc injury.

Abstract:Objective：To explore a method to prepare deproteinized osteo-articulation（DPOA） with hydrogen peroxide and ether as well as to discuss its physicochemical characteristics and biological safety. Methods：Distal femur semi-joint cancellous bones of adult hybrid canine were made into bio-derived osteo-articulation materials with hydrogen peroxide and ether by degreasing and deproteinizing；outsaide surface，pore texture of interal surface，pore size and traffic situation among pores of DPOA were observed by stereology microscope and scanning electron microscope；eliminated cell composition of DPOA was observed by HE staining；culture solution pH values of DPOA were detected by instrument；protein contents of DPOA were detect by micro Kjeldahl method；stress weight was detect by biomechanics test；biological safety in vitro was tested by cytotoxicity test and hemolysis test. Results：Results of stereology microscope and scanning electron microscope reveal that DPOA had three-dimensional and porus-grid structure system of original bone tissue. Results of computer picture analytical system reveal that porosity was （84.76±4.63）%，maximum aperture was （318.64±16.35） μm and minimum aperture was （112.13±15.42） μm. HE staining revealed that cell composition of DPOA was eliminated entirely. pH value of DPOA culture solution was 7.382±0.009. Protein content of DPOA was （0.017±0.003）%. Maximum stress weight of biomechanics test was （226.00±7.65） N，approaching mechanics loading of normal osteo-articulation.Cytotoxicity was 0 grade. Result of hemolysis test was < 5%. Conclusions：Allogeneic natural derived DPOA made by hydrogen peroxide and ether method retains pore net structure of original bone tissue and possesses the intrinsic anatomical configuration. Its physicochemical characteristics and biological safety are qualified and can be used as an osteo-articulation grafting trestle material.

Abstract:Objective：To establish three dimensional finite element analysis（FEA） model of single external fixation screw penetrating into tibia shaft. Methods：Softwares of Mimics，Pro/E，Geomagic Studio，Ansys Workbench13 were used to set up FEA model of tibia-screw assembly successfully；results after applying force and moment loads were observed. Results：The shaft of tibia and single exter－nal fixation screw generated 3 710 volume meshes，6 952 nodes. Von-Mises stress and Total Deformation revealed the maxium stress of the screw and total deformation were located at contact surface and screw tail respectively. Conclusions：Three dimensional tibia-screw assembly model can provide basis for quantitative studies of stresses of the screw and the tibia as well as for biomechanical study of screws made by different materials and with different loads.

Abstract:Objective：To quantify the possibility and efficacy of internal fixation with bioabsorbable screw and stick in the treatment of humerus epicondyle fracture. Methods：Totally 23 children with humerus epicondyle fracture who received internal fixation using bio-absorbable screw and stick from 2008 to 2009 were enrolled. Functional exercise was taken 3 weeks after the operation. Follow-up lasted for 6 months to 3 years，18 months in average. Results：All 23 cases were followed up and healed. Function of the elbow joint was excellent in 20 cases，good in 3 cases and bad in 0 case，with good and excellent rate being 100%. Conclusions：With satisfactory fracture reduction and firm internal fixation，bioabsorbable screw and stick is effective in the treatment of humerus epicondyle frac－ture. Bioabsorbable screw and stick will break down as time goes by，without harming human body influencing imaging examination. The second stage operation is needed to take out the materials. Early functional exercise is conductive to promoting the healing results. El－bow function development of the epiphysis will not be affected.

Abstract:Objective：To evaluate the preliminary efficacy of nano-hydroxyapatite/polyamide 66（n-HA/PA66） cage in reconstruction following anterior cervical intervertebral discectomy and fusion in cervical spondylosis. Methods：Clinical data of 46 patients with cer－vical spondylopathy and cervical disc herniation who underwent anterior cervical intervertebral discectomy and fusion between June 2008 and June 2011 were retrospectively analyzed. Cervical pathologies included cervial spondylotic myelopathy and cervical radicu－lopathy. All patients were assessed clinically and radiologically. Clinical outcome was evaluated using Odom’s criteria；radiology results included subsidence of the cages，angle of cervical lordosis，fusion rate and intervertebral height. Results：Based on Odom’s criteria，excellent rate was 80%. All cases achieved graft fusion at 6 months after the operation without displacement and subsidence. There were significant differences in angle of cervical lordosis between preoperation and 6 months after operation（（2.29±3.04）° vs. （8.04±3.06）°，P＜0.05）；last follow-up showed（6.58±2.35）° and the mean loss of angle of cervical lordosis was （1.45±1.96）°. There were significant differences in intervertebral height between preoperation and at 6 months after the operation（（7.76±0.55） mm vs. （4.40±1.36） mm，P＜0.05）；last follow-up showed（7.36±0.59） mm and the mean loss of intervertebral height was （0.41±0.28） mm. Conclusions：n-HA/PA66 cage is effectively in the reconstruction following anterior cervical intervertebral discectomy and fusion since it can effec－tively maintain the biological alignment and cervical intervertebral height with high fusion rate.

Abstract:Objective：To summarize the experiences of posterior osteotomy for spinal kyphosis. Methods：Five patients with ankylosing spondylitis and an average kyphosis angle of 79.0° underwent double-segment wedge osteotomy in L1 and L3. Five patients with trau－matic or tuberculosis kyphosis and kyphosis angle less than 50.0° underwent single-segment wedge closing osteotomy. Five patients with kyphosis angle more than 50.0° underwent osteotomy with cage inserting anteriorly and closing posteriorly. Their clinical data were retrospectively analyzed. Results：Fifteen patients had an average of 11 months’ follow-up. All had bone-fusion at six months after the operation. Average kyphosis angle was 62.3° preoperatively and 15.5° postoperatively. Average correction angle and rate of kyphosis were 46.8° and 75.1% respectively. Average height was increased by 4.4 cm. For five patients with nervous injury，one pa－tient with Frankle C recovered to Frankle D；one with Frankle C to Frankle E，and three with Frankle D to Frankle E. Back pain was released obviously. Conclusions：Long round-shaped deformity can be corrected through multi-vertebral osteotomy. Severe kyphosis can be corrected through osteotomy with cage inserting anteriorly and closing posteriorly. Osteotomy correction by posterior approach is an effective method for spinal kyphosis.

Abstract:Objective：To compare therapeutic effects between proximal femoral locking compression plate（LCP） and proximal femoral nail antirotation（PFNA） in the treatment of femoral intertrochanteric fractures. Methods：Totally 125 patients with intertrochanteric fractures from 2005 to 2011were enrolled and were randomly divided into two groups. Patients in the two groups were treated with PF－NA and proximal femoral LCP respectively. Follow-up was conducted for 12-24 months，averaged 18.2 months. Condition during the operation and after operation was compared between two groups. Results：There was no significant difference in general data between the two groups（P >0.05）. There were significant differences in length of incision，blood loss，bed-lying time and postoperative drainage between two groups（P<0.05）. There was no significant difference in operation time，fracture healing time，Harries score and compli－cations between two groups（P>0.05）. Conclusions：For the treatment of intertrochanteric femoral fracture，both proximal femoral LCP and PFNA fixation could achieve good outcome，however，PFNA has more advantages in treating unstable femoral intertrochanteric fractures.

Abstract:Objective：To summarize the therapeutic effect of single-stage anterior release and reduction through oral pharynx and pos－terior fixation and fusion for irreducible atlantoaxial dislocation. Methods：Seventeen cases of irreducible atlantoaxial dislocation were treated in our hospital from November 2006 to August 2011 including 10 males and 7 females aged from 31 years to 68 years（aver－aged 46.5 years）. The 17 patients included 9 patients with obsolete odontoid process fracture，2 with inborn dissociation odontoid pro－cess，4 with atlan-occipital fusion deformation，2 with rheumatoid disease. Japanese Orthopaedic Association（JOA） scores ranged from 4 to 11 with an average of 7.47±2.15. All dislocation could not be reduced through skull traction under anesthesia. Anterior release and reduction through oral pharynx was performed and posterior fixation and fusion was performed by one stage. Among posterior fix－ation and fusion，13 were performed through atlantoaxial pedicle，4 were performed through dentata pedicle and occipital. Results：All patients pulled through perioperative period securely. Complication including arteria vertebralis injury，spinal cord injury and incision infection were not occurred. All patients were followed up for 9 to 56 months，with an average of 32.6 months. All patients gained anatomic reduction and bone fusion. At latest time follow up，JOA scores ranged from 11 to 17 with an average of 14.70±2.22. Conclusions：With the merits of sufficient reduction，thorough decompression and good spinal column stability reconstruction，single-stage anterior release and reduction through oral pharynx and posterior fixation and fusion is effective in the treatment of irreducible atlantoaxial dislocation.

Abstract:Objective：To analyze the efficacy of posterior decompression，fusion，pedicle screw system fixation in the treatment of de－generative lumbar scoliosis with stenosis（DLSS）. Methods：Thirty-three patients with DLSS in our hospital from July 2008 to June 2011 were analyzed retrospectively，including 11 males and 22 females. All the patients were underwent posterior decompression，fusion，pedicle screw system fixation. Visual analog symptom rating scale（VAS），Oswestry disability index（ODI） and the Cobb angle on the lumbar coronal film were used for efficacy assessment. Results：All patients were followed up for 12-26 months，averaged（17.27±3.78） months. Mean preoperative VAS score，ODI score and Cobb angle were （7.00±0.90） points，（70.61±7.29）% and （20.58±9.96） degrees while those at the last follow-up were （3.00±0.97） points，（31.42±9.22）% and （6.12±3.22） degrees，with significant differences between preoperative and postoperative results（P<0.05）. Excellent rate of ODI was 72.73%. During the follow-up，the height of disc space was remained，fusion cage were not displaced，no case of internal fixations breakage was observed and rate of fusion for bone graft was 100%. Conclusions：Posterior decompression，fusion，pedicle screw system fixation is an effective surgical treatment for DLSS.

Abstract:Objective：To compare curative effect between anterior decompression and posterior decompression in the treatment of tho－racolumbar burst fracture with neurologic deficit. Methods：From March 2007 to March 2010，62 cases of thoracolumbar burst fracture with neurologic deficit were treated. There were 43 males and 19 females with an average age of 42.3 years（22-68 years）. All cases were randomly divided into 2 groups：group A（anterior spinal canal decompression） and group B （posterior spinal canal decompres－sion）. According to American Spinal Injury Association（ASIA） standard，group A had 19 cases of C and 12 cases of D；group B had 16 cases of C and 15 cases of D. Results：Totally 62 cases were followed up for 2.0-3.5 years（28 months in average）. All wounds achieved primary healing postoperatively and all cases achieved satisfactory results in vertebral canal decompression and reduction. No loose－ness，protrution and breakage of internal fixator was observed. Blood loss was （675.3±124.1） ml in group A and （381.3±77.8） ml in group B，with statistically significant differences（t=11.175，P=0.000）. Operative time was （193.68±22.01） min in group A and （120.51±19.83） min in group B，with statistically significant differences（t=13.748，P=0.000）. Preoperative vertebral canal volume in group A was （43.05±8.53）% and （45.29±7.60）% in group B，without statistically significant differences（t=-1.092，P=0.279）. Post－operative vertebral canal volume in group A was （91.50±2.91）% and （89.81±2.60） % in group B，with statistically significant dif－ferences（t=2.411，P=0.010）. Vertebral canal volume in the last follow-up in group A was （91.89±2.16）% and （90.90±1.86）% in group B，with statistically significant differences（t=1.934，P=0.029）. Preoperative，postoperative and final vertebral heights in group A were （43.97±8.26）%，（90.45±2.26）%，（89.67±2.09）% and （45.48±6.52）%，（89.63±2.16）%，（88.95±1.85）% in group B，with－out statistically significant differences（t=-0.799，t=1.460，t=1.436，P＞0.05）. At 2 years postoperatively，27 cases in group A partly re－covered（curative rate of 87.10%） and 24 cases in group B partly recovered（curative rate of 77.45%），showing no statistically signifi－cant differences（χ2=0.995，P=0.800）. There were statistically significant differences in the postoperative pain，medical cost and time in hospital between groups A and B（t=2.181，t=11.808，t=5.848，P＜0.05）. Conclusions：Posterior decompression can achieve the same effects as anterior decompression in the treatment of thoracolumbar burst fractures with neurologic deficit as long as indications are strictly controlled. Posterior decompression is easier and has more merits than anterior decompression：shorter operative time，less blood loss，less damage，lower medical cost. Posterior decom－pression is more suitable for hospitals at primary level.

Abstract:Objective：To explore the clinical effects of impacted bone graft combined with structural bone graft for the reconstruction of acetabular bone defects in adult developmental dysplasia of hip（DDH）. Methods：Totally 11 adult patients suffered from DDH were treated in our hospital from January 2007 to June 2012. According to Crowe classification，there were 5 cases of type Ⅱ and 6 cases of type Ⅲ dysplasia，respectively. All acetabular bone defects were type Ⅲ defects（combined defects） according to the American Academy of Orthopaedic Surgeons classification. Femoral head allograft and femoral head autograft were used in structural bone graft for 3 cases and 8 cases，restpectively. All defects were further treated with autologous bone acquired from the femur or ilium using impacted morselized bone grafting technique. Results：Patients were followed up for 6-48 months，with no complications such as infection，hip pain and cup loosening being observed. Average Harris hip score increased from 39.7（18-52） before the surgery to 85.7（52-96） at the last follow-up. Conclusions：Structural bone graft and appropriate cup can provide instant support for the acetabular bone defects，while impacted bone graft can promote the fusion of acetabular cup with surrounding bone.

Abstract:Objective：To study the in vitro immunosuppression of interleukin-10（IL-10） modified bone marrow mesenchymal stem cells（BMMSCsIL-10）. Methods：IL-10 was cloned by RT-PCR and was used to construct pLNCX-IL-10. The constructed retrovirus was transfected into rat BMMSCs. T lymphocyte subsets in vitro influenced by BMMSCsIL-10 were detected by flow cytometry. Effects of BMMSCsIL-10 and its culture supernatant on in vitro mixed heterologous T lymphocyte proliferation were detected by CCK-8. Results：BMMSCsIL-10 significantly reduced CD4+ T and lowered the ratio of CD4+/CD8+，more intense than BMMSCs（P=0.000）. BMMSCsIL-10 and its culture supernatant significantly inhibited T lymphocyte proliferation，more intense than BMMSCs（P=0.000）. Conclusions：IL-10 modification can enhance the in vitro immunosuppressive effect of BMMSCs.

Abstract:Objective：To investigate the effect of massage on quadriceps femoris repair after injury by external force with the histologi－cal observation and ultrasound imaging observation as well as the influences of basic fibroblast growth factor （bFGF） in New Zealand rabbits so as to explore the possible molecular mechanisms of massage in repair of muscle injury. Methods：Thirty-two New Zealand white rabbits，weighing （2.0±0.5） kg，were randomly divided into 4 groups：normal control（group A，n=4），control group before the massage （group B，n=4），self-repairing group（group C，n=12，further divided into 7，14，21 d groups） and massage group（group D，n=12，further divided into 7，14，21 d groups）. In group A，no treatment was made and rabbits were taken as normal controls；in groups B，C，D，rabbit models of quadriceps femoris injury were prepared by self-made beater；in group C，no massage therapy was given and rabbits were taken as natural recovery controls；in group D，massage therapy was given at the 5th after model was built. At 7，14，21 d after injury，4 different animal quadriceps samples were taken in groups C and D respectively. Immunohistochemical method was used to detect expression of bFGF protein. HE staining was employed to inspect pathological changes. Ultrasound imaging was employed to inspect the tissue necrosis situation at 7，14 d after model build. Results：After massage intervention，expression of bFGF was higher in group D than in group C at 7 d（P=0.016），higher in group C than in group D at 14，21 d（P=0.000，P=0.020）. Ultrasound of mi－cro-bubble angiography and HE staining showed that injury was lighter，blood supply was richer，necrotic area was significantly de－creased and vascular restored faster in group D compared with those in group C. Conclusions：Massage can effectively prevent muscle atrophy，degeneration and necrosis，provide a rich blood supply for the damage regions and promote the angiogenesis. Mech－anism of beneficial effect of massage on injury repair and neovascularization may be related with bFGF.

Abstract:Objective：To observe the effects of hydroxymethylglutaryl coenzyme A reductase inhibitor fluvastatin（Flu） on renal fibrosis induced by cyclosporine A（CsA） in rats． Methods：Totally 30 adult male SD rats were randomly divided into three groups．Controls were fed with olive oil，CsA treated rats were given CsA（30 mg/（kg·d）），and Flu treated rats were given CsA（30 mg/（kg·d）） and Flu（7 mg/（kg·d）） by gavage． Five rats of each group were sacrificed on the 14th d and the others were sacrificed at the end of the study（the 28th d）. Histological changes in renal tissue were observed． Sites and levels of expression of Smad protein 2/3，Smad protein 7 and transforming growth factor-β1（TGF-β1） were examined by immunohislochemistry staining． Renal function and lipid levels were measured in every week． Results：Serum creatinine（SCr） in Flu treated group on the 7th d was all equivalent to that of control group（P=0.564）． From the first week after treatment，levels of SCr in CsA treated group were significantly increased compared with those in control group（P＝0.000）． Compared with those of control group，Smad protein 2/3 and TGF-β1 expression on the 14th and 28th d was significantly increased in CsA treated rats（P＝0.000），while Smad protein 7 expression was markedly decreased（P＝0.000）． These lesions were significantly ameliorated by Flu. Levels of total cholesterol（P=0.865，0.997，0.505，0.871，0.540），triglyc－erides（P=0.894，0.663，0.826，0.635，0.928） had no statistical difference at same time point among all three groups. CsA valley con－centration had no statistical difference between Flu treated group and CsA treated group（P=0.183，P=0.123）． Conclusion：Flu could slow the progression of cyclosporine nephropathy.

Abstract:Objective：To study the effects of recombinant human acidic fibroblast growth factor（rhaFGF） and recombinant human epi－dermal growth factor（rhEGF） on wound healing of rabbits’ ears with full-thickness skin injury. Methods：Models of rabbits’ ears with full-thickness skin injury were made and randomly divided into four groups：group A（rhaFGF 60 U/cm2+rhEGF 20 U/cm2），group B（rhaFGF 60 U/cm2），group C（recombinant bovine basic fibroblast growth factor 60 U/cm2）and group D（equivalent normal saline）. The reagents in the brackets were given every two days. Wound healing rate and time were respectively recorded at different time points through hyaline membrane tracing. Wound tissues were obtained on the 3rd，7th and 14th d to evaluate wound healing state through HE staining. Results：Wound healing time was statistically different between group A（（13.33±0.12） d） and other groups（P＜0.05）. Wound healing rate was also statistically different between group A and other groups. HE staining showed that on the 3rd and 7th d after modeling，growth of fibroblasts and capillary was more vibrate in group A and C than that in group B and D. On the 14th d after modeling，wound was almost healed in group A and fibroblasts and capillary of group A were relatively rare compared with those in other groups. Conclusions：rhaFGF combined with hEGF can promote wound healing in rabbits’ ears. Combination of the two drugs can stimulate the growth of granulation tissue，capillary formation and re-epithelialization. In the later stage of wound healing，capillary and fibroblasts grow slower，which may reduce the possibility of scar formation.

Abstract:Objective：To evaluate influences of different blood purification treatments on expressions of ATP-binding cassette receptor A1（ABCA1） in patients with maintenance dialysis. Methods：Between August 2010 and March 2012，30 uremia patients treated by hemodialysis（HD），30 uremia patients treated by hemodiafiltration（HDF） and 20 healthy volunteers（control group） were investigated. Peripheral blood mononuclear cells were isolated and stimulated to transfer to macrophages. Macrophages were identified by Oil red O dyeing. Macrophages were collected to detect ABCA1 and nuclear factor-κB p65（NF-κB p65）expression by using real-time PCR and Western blot. Results：Expression of ABCA1 was significantly decreased in HD and HDF groups compared with that in control group（P<0.01）. Expression of ABCA1 was increased in HDF group compared with that in HD group（P<0.05）. Furthermore，expression of NF-κBp65 mRNA and protein in HD and HDF groups was higher than that in control group（P<0.01，P<0.05）. Expression of NF-κBp65 mRNA and protein in HDF group was significantly lower than that in HD group（P<0.01）. Conclusions：HDF may increase cholesterol efflux in human monocyte-macrophage derived foam cells，the mechanism of which may associate with inactivation of NF-κBp65 pathway and increase of the expression of ABCA1 mRNA and protein.

Abstract:Objective：To establish animal model of guinea pig anaphylactic shock，to explore the expression of interleukin-4（IL-4），interleukin-13（IL-13） and immunoglobulin E（IgE） in guinea pig serum and to provide scientific and objective evidences for clinical and medicolegal diagnosis of anaphylactic shock as well. Methods：Totally 30 guinea pigs were died of anaphylactic shock were divided into experiment group（n=20） and control group（n=10）. ELISA method was used to detect IL-4、IL-13 and IgE by extracting serum of guinea pigs in experiment group and control group. Results：Serum IL-4 level in experimental group was higher than that in control group，P=0.000；serum IL-13 level in experimental group was higher than that in control group，P=0.000；serum IgE level in experi－mental group was higher than that in control group，P=0.000. Conclusions：Serum levels of IL-4，IL-13 and IgE in experiment group are obviously higher those in control group，which provides evidences for anaphylactic shock forensic identification and clinical diag－nosis.

Abstract:Objective：To detect whether Panax notoginseng saponins（PNS） could improve the erectile function of diabetic mellitus （DM） rats with erectile dysfunction（ED），to find the possible ways by which PNS worked and to explone effects of PNS on Bax and Bcl-2 expressions. Methods：Model of DMED was established and the rats were randomly divided into four groups：PNS treated group（intraperitoneal injection with doses of 50 mg/（kg·d），100 mg/（kg·d），150 mg/（kg·d）） and DMED group（intraperitoneal in－jection with saline）；at the same time，normal rats injected with saline were taken as controls（normal control group）. Four weeks later，ratio of intracavernous pressure（ICP） and mean arterial blood pressure（MAP） was measured by electro-stimulating of cavernous nerve. Then cavernous bodies of penis of all experimental rats were harvested and their expressions of Bcl-2 and Bax were determined by immunohistochemistry. Apoptosis of penile tissue cell was detected by TUNEL. Changes of the cavernous bodies in ultrastructures were observed by electron microscopy. Results：ICP/MAP was elevated and value of Bcl-2/Bax was increased in middle and high dose group than in DMED group. Results of electron microscopy uncovered that there were ruptured endothelium and impaired smooth muscle cells in the low dose PNS group and DMED group and even thrombosis in DMED group；in the middle，high dose group and normal control group，there were no abnormal changes. Conclusions：PNS might elevate the value of Bcl-2/Bax to protect endothelium and smooth muscle cells from apoptosis as well as protect the function of penis cells，thereby to improve erectile function.

Abstract:Objective：To investigate the expression of B7 homolog 4（B7-H4） in human breast cancer and to analyze its relationship with number of tumor-infiltrating FOXP3+ Tregs and cancer lymphonode metastasis. Methods：B7-H4 expression was detected by im－munohistochemistry in 16 cases of normal breast tissues and 79 cases of breast cancer. Number of tumor-infiltrating CD3+ T cells and FOXP3+ Tregs was determined by indirect immunofluorescent double-staining in 64 cases of invasive breast cancer. Correlation be－tween B7-H4 expression and number of tumor-infiltrating CD3+ T cells and FOXP3+ Tregs as well as lymphonode metastasis was analyzed. Results：B7-H4 expressed in 6.3% normal breast tissues，40.0% ductal cancer in situ and 78.1% invasive breast cancer. B7-H4 expression in invasive breast cancer was significantly higher than that in normal breast tissues and ductal cancer in situ（P=0.000，P=0.009） without significantly difference between normal breast tissues and ductal cancer in situ（P=0.037）. Intensity of B7-H4 was positively correlated with number of infiltrating FOXP3+ Tregs（rs=0.368，P=0.003） but was negatively correlated with number of infiltrating CD3+ T cells（rs=-0.316，P=0.011） and had no relationship with lymphonode metastasis of breast cancer（P >0.05）. Conclu－sions：B7-H4 overly expresses in breast cancer. B7-H4 is in association with the local antitumor immune inhibition not lymphonode metastasis of tumors.

Abstract:Objective：To study the effects of cutting off the seventh and the eighth intercostal nerve in the right side on hyperplasia of mammary gland（HMG） in SD rats. Methods：Totally 60 SD rats were randomly divided into 4 groups：normal group，model group，in－tercostal nerve transection group and ovariectomized group. Intramuscular injection of estradiol benzoate combinng with progesterone was provided to establish a rat model HMG. Diameter and height of the third nipple by right side in each group were measured at 2，4，6 weeks，respectively. Moreover，morphological changes and Ki-67 expression of the breast tissues were observed. mRNA levels and protein levels of estradiol receptor（ER） and progestogen receptor（PR） in breast tissues were measured by real time PCR and West－ern blot; protein levels of proliferating cell nuclear antigen（PCNA） were measured by Western blot respectively. Results：Diameter and height of the third nipple at the right side were measured. Diameter and height were lower in intercostal nerve transection group than in model group（P<0.05）. Layers of epithelial cell，numbers of acinus，diameter of acinus and diameter of duct of mammary gland were markedly reduced in intercostal nerve transection group than in model group（P<0.05）. In addition，mRNA and protein levels of ER and PR and the protein levels of PCNA in breast tissues were lower in intercostal nerve transection group than in model group（P<0.05）. Conclusions：Intercostal nerve transection could reduce pathological changes of HMG and reduce mRNA and protein ex－pressions of ER and PR as well as protein expressions of PCNA in rat mammary gland.

Abstract:Objective：To investigate the correlation between QT dispersion（ＱＴd） and serum enzyme changes in patients with acute cerebrovascular disease and to evaluate the diagnosis value of QTd and serum enzyme changes. Methods：Totally 90 patients with acute cerebrovascular disease from February 2011 to January 2012 were served as observation group；90 healthy subjects from February 2011 to January 2012 were served as control group. All patients and healthy subjects received 12-lead electrocardiogram（ECG） and QTd was measured. Commercial kits were used to measure the activity concentration of transferase（AST），lactate dehydrogenase（LDH），α-hydroxybutyrate dehydrogenase（α-HBDH），creatine kinase（CK） and myocardial-type creatine kinase isoenzyme（CK-MB）. Corre－lation analysis of QTd and serum enzymes was performed. Results：Average activity concentrations of serum AST，LDH，α-HBDH and CK of observation group were significantly higher than those of control group（t=10.13，P=0.000；t=12.55，P=0.000；t=13.14，P=0.000；t=20.91，P=0.000），but average activity concentration of myocardial-type CK-MB has no significant difference between the two groups（t=0.88，P=0.378）. Average QTd of observation group was significantly higher than that of control group（t=9.09，P=0.000）. QTd and serum enzymes（AST，LDH，α-HBDH and CK） activity were positively correlated（r=0.913，P=0.001；r=0.923，P=0.002；r=0.727，P=0.021；r=0.823，P=0.017）. Conclusions：QTd and serum enzyme（AST and LDH） activity concentration combined detection has potential clinical value for diagnosis of acute vascular disease.

Abstract:Objective：To investigate the effect of telmisartan on Ras/Raf/ERK1/2 signal pathway in rats with myocardial hypertrophy. Methods：Totally 30 rats were randomly divided into 3 groups including control group，model group and intervention group. Each group has 10 rats. Hypodermic injection of normal saline was applied in control group；hypodermic injection of isoproterenol was ap－plied in model group；hypodermic injection of isoproterenol and telmisartan was applied in intervention group. Myocardial index and pathological detections were performed. RT-PCR assay was used to detect ANP mRNA expression. After the model was successfully made，rats’ left cardiac tissues were collected and p-Raf protein and p-ERK1/2 protein expression was detected by immunohis－tochemistry. Results：Average integral optical density in p-Raf positive area was the highest in model group（P=0.000） followed by intervention group and control group（P=0.041，P=0.031）. Average integral optical density in p-ERK1/2 positive area was the highest in model group（P=0.000） followed by intervention group and control group（P=0.0039，P=0.023）. Conclusions：Telmisartan resists my－ocardial hypertrophy since it can reduce expressions of Raf protein and ERK1/2 and inhibit the activation of Raf/MEK/ERK signal pathway

Abstract:Objective：To investigate the expression of P300/CBP-associated factor（PCAF） and its correlation with the clinicopatholog－ical features in hepatocellular carcinoma（HCC）. Methods：Forty samples of surgically resected HCC and adjacent noncancerous tis－sues were collected. PCAF expression in the samples was detected using RT-PCR and immunohistochemical staining. Correlation be－tween PCAF protein expression and clinicopathological feature was analyzed. PCAF protein expression in normal hepatic cell line （LO2） and HCC cell lines（HepG2 and SMMC-7721） was detected. Results：mRNA and protein expression of PCAF in HCC tissues was significantly down-regulated compared with that of adjacent noncancerous tissues（P<0.05，respectively）. Clinicopathological analysis suggested that PCAF protein low-expression was associated with tumor intrahepatic metastasis（rs=0.413，P=0.037），portal vein invasion（rs=0.589，P=0.011） and advanced TNM stage（rs=0.513，P=0.029）. Expression of PCAF protein in HCC cell lines HepG2 and SMMC-7721 was significantly lower compared with that in hepatic cell line LO2（P<0.05，respectively）. Conclusions：Expression of PCAF is down-regulated in HCC cell lines and HCC tissues and the low-expression of PCAF is correlated with poor clinico-pathological characteristics of HCC.

Abstract:Objective：To explore the effect of gardnerella vaginalis（G.vaginalis） and its culture supernatant on the injury of fallopian tube tissue. Methods：Two strains of G.vaginalis were cultured in vitro. Different concentrations of bacterial suspension and sterilie bac－terial filtrate were prepared. Bacterial filtrate was repeatedly frozen thawed and inactivated. Bovine oviduct was cultured with or with－out different concentrations of bacterial suspension and processed bacterial filtrate. Ultra-high-power microscope was used to observe the cilia beating intensity and amplitude at different time points in different groups. Effects of G.vaginalis and different treatment forms of its culture supernatant on fallopian tube were determined. CD59 function antibodies and mitogen-activated protein kinase p38（p38 MAPK） pathway inhibitor was used to determine the damage process. Results：Sterile bacterial filtrate，heat treated bacterial filtrate，repeatedly frozen thawed bacterial filtrate all can cause damage to athletic ability of oviduct cilia and weaken their beating intensity and amplitude（P<0.007）. Tissue sections failed to find evidence of damage，but scanning electron microscope examination found that the number and length of the oviduct mucosal layer cilia were decreased. CD59 functional antibody and p38 MAPK path－way inhibitor can alleviate such toxins damage. Conclusions：G.vaginalis affect the function of the oviduct by a soluble toxin via CD59 dependent p38 MAPK pathway.

Abstract:Objective：To evaluate the therapeutic effect of peripheral blood stem cell transplantation（PBSCT） in the treatment of acute leukemia（AL），to discuss advantages and disadvantages of allogeneic PBSCT（Allo-PBSCT） and autologous PBSCT（Auto-PBSCT） and to choose the appropriate preparative transplantation regimen. Methods：All fifty patients with AL underwent PBSCT from 2004 to 2012 in our hospital were retrospectively analyzed including 21 male and 29 female. Twenty－eight patients were treated with Allo-PBSCT and 22 with Auto-PBSCT. Effects of transplantation and complications were analyzed. Differences in transplantation relapse rate，disease-free survival（DFS） and transplant related mortality（TRM） were compared. Preparative regimen related death and suc－cess rate were analyzed. Results：No pretreatment scheme related death was occurred except 2 cases of MAC（melphalan+Cytarabine+cyclophosphamide） preparative regimen implanted failure. Forty－eight patients engrafted successfully with transplant success rate of 96%. Median time of white blood cell>0.5×109/L was 11.3 d（（7-24） d） and platelets>20×109/L was 14.3 d（（5-44） d）. Allo-PBSCT and Auto-PBSCT related death cases were 8 cases and 3 cases respectively with TRM of 28.6% and 15.0%，respectively. During the follow-up，accumulative DFS for Allo-PBSCT and Auto-PBSCT were 46.4% and 35.0% respectively；accumulative relapse rates were 25.0% and 55.0%，respectively. During follow-up，overall survival（OS） rates of Allo-PBSCT and Auto-PBSCT were 57.1% and 50.0%，respectively. Among patients under the condition of complete remission at first course（CR1） and complete remission after two or more course or non-remission before transplantation，OS rates were 61.5%，53.3% and 45.5%，55.6%；DFS were 46.2%，46.7% and 27.3%，44.4%；death rates were 38.5%，46.7% and 54.5%，44.4%，respectively after transplantation. Conclusions：PBSCT is effective in the treatment of AL. Effect is better for patients under the condition of CR1 before transplantation. Relapse rate is lower，long-term DFS is higher and transplant related complications are more in Allo-PBSCT than in Auto-PBSCT. Optimization of preparative regimen is the key to improve transplantation efficacy，preparative regimen based on busulfan/cyclophosphamide regimen has a better effect of pulp and tolerance.

Abstract:Objective：To develop a ergonomics situ intelligent artificial anal sphincter（IAAS） with bio-feedback function and to observe its fecal continence function. Methods：Function of the IAAS was assessed in vitro and in vivo in New Zealand rabbits. Physiological saline was infused into the bowel to simulate the formation of stool. Simultaneously，function of the pressure feedback system and the bowel-restrained function of L-style artificial anal sphincter were observed. Results：Pressure of the feedback cuff was positively correlated with the pressure and volume of the bowel within a certain range；the pressure feedback system can reflect the change of the intraintestinal pressure effectively. L-style artificial anal sphincter can control fecal incontinence under the control of chip micro－computer. Conclusion：IAAS could efficiently maintain continence and reflect the change of the intraintestinal pressure.

Abstract:Objective：To explore of the efficacy and safety of risperidone combined with small doses of chlorpromazine in the treatment of schizophrenia with bronchial asthma at acute stage of seizures. Methods：A total of 64 patients with schizophrenia with bronchial asthma at acute stage of seizures were randomly divided into study group（risperidone combined with a small does of chlorpromazine，n=32） and control group（risperidone，n=32） and continuous treatment was made for six weeks. Efficacy and adverse events were as－sessed with the positive and negative symptom scale（PANSS），clinical global impression scale and treatment emergent symptom scale（TESS） before and after the treatment. Assessment on asthma symptoms and sign disappearing time at the end of the 2nd week after the treatment was conducted. Results of arterial blood gas analysis and changes of respiratory and heart rate of two groups were mea－sured before treatment and at the end of the first week after the treatment. Results：PANSS total scores，positive symptom scores，nega－tive symptom scores and general psychopathology scores of the two groups were significantly decreased at the end of the 6th week af－ter the treatment and were statistically different compared with those before the treatment（P<0.01）. Reduction rate was significantly higher in study group than in control group at the end of the first week after the treatment with statistical differences（P<0.05）. There was no statistical difference in PANSS total scores between the two groups at the end of the 6th week after the treatment（P >0.05）. There was no statistical difference in total effective rate between the two groups（P >0.05）. Marked effective rate of study group was 62.50%（20/32），statistically higher than that of 40.63%（13/32） in control group（P<0.05）. Reducing time of wheezing sound，disap－pearing time of clinical symptoms and disappearing time of wheezing sound were shorter in study group than in control group at the end of the 2nd week after the treatment，with statistical differences（P<0.05 or P<0.01）. Arterial blood gas analysis of PaCO2 and improvement of respiratory frequency were faster in study group than in control group at the end of the first week after the treatment，with statistical differences（P<0.05）. There was no statistical difference in TESS total scores between the two groups at the first and the 6th week after the treatment（P >0.05）. Side effect rates in study and control group were 62.50%（20/32） and 53.13%（17/32） respectively，without statistical differences（P >0.05）. Conclusions：With mild side effect，risperidone combined with small doses of chlorpromazinen is more effective in the treatment of schizophrenia with bronchial asthma at acute stage of seizures than single risperidone treatment.

Abstract:Objective：To explore the electroencephalogram（EEG） features and pathological changes of brain tissue with EEG abnor－malities in patients with supratentorial tumors complicated with secondary epilepsy. Methods：Totally 36 patients with nonfunctional area supratentorial tumors complicated with secondary epilepsy were enrolled. Tumor and epileptiform discharges resection under the electrocorticography（ECoG） monitoring were performed. The resected brain cortex with epileptic discharges was as defined as experi－mental group and normal dumb organization（parenthesis of tumor resection） without epileptic discharges under the microscope in the tumor surrounding or decompression brain tissue was defined as control group. Characteristics of ECoG wave in experimental group were analyzed and postoperative pathologic results in experimental group and control group were compared. Results：（1）Characteristics of ECoG wave in patients with supratentorial tumors：14 cases（38.89%） showed intermittent polyspikes，followed by continuous spike and generalized sharp wave rhythm.（2）Pathological features：21 cases（58.33%） in experimental group showed irreversible glioma cell and simple glial cell hyperplasia and only 7 out of 30 cases（23.33%） in control group found glial cell hyperplasia. Glioma or glial cell hyperplasia was significantly higher in experimental group than in control group（P<0.05）. （3）Pathological results and characteristics of ECoG wave in experimental group：in nonreversible group：continuous single spike was mainly seen（P<0.05）. Intermittent polyspikes and generalized high amplitude sharp wave rhythm can seen both in two groups（P >0.05）. Conclusions：ECoG wave manifests mostly as continuous single spike in patients with supratentorial tumors complicated with secondary epilepsy. Pathological features of brain tissues with ECoG abnormalities are glioma cells and glial cell hyperplasia. Cerebral cortex around the tumor with recurring single high amplitude spikes，clusters multi-spikes and spike-sharp-wave rhythm of discharge in ECoG should be actively treated.

Abstract:Objective：Papillary thyroid carcinoma is often complicated with lymph node metastases（LNM） and extent of lymph node dissection（LND） is under disscussion. The objectives of this retrospective study were to evaluate the pattern and predictive factors of cervical LNM and to determine the optimal lymph node management strategy. Methods：Between February 2010 and February 2013，96 patients who underwent surgical treatment for PTC at our hospital were retrospectively analyzed. All patients underwent LND at central and lateral sides. Neck dissection specimens were marked and separated according to neck levels and sent for pathology examination. Results：Overall，63 patients（65.6%） had nodal metastases to level Ⅵ，57（59.4%） to level Ⅲ，42（43.8%） to level Ⅱ，35（36.5%） to level Ⅳ，8（8.2%） to level Ⅴ. Gender（P=0.019） and extracapsular invasion（P=0.026） were significant predictive fac－tors of cervical LNM，and central LNM was significantly related with lateral LNM（P=0.014）. Conclusions：Ipsilateral central LND is recommended for all PTC patients. Whether to perform lateral LND and the extent of lateral LND are determined by results of central LND.

Abstract:Objective：To analyze the disease constitution and death cause of hospitalized patients with cardiovascular diseases and to offer some epidemiology evidences for clinic and research of cardiology. Methods：Patients discharged from the cardiology department of the First Affiliated Hospital of Chongqing Medical University from January 2011 to December 2012 were retrospectively investigated and were divided into 4 groups：aged below 44，aged 45 to 59，aged 60 to 74 and aged over 75. Their disease constitution，death cause and proportion of heart failure and sudden cardiac death were analyzed. Results：There were 5 096 male and 5 212 female patients and the average age was （65.16±14.37） years. The top three diseases leading to hospitalization were coronary heart disease（CHD），hy－pertension and arrhythmia with constituent ratios of 41.29%，27.19% and 10.41% respectively. The top three diseases leading to hos－pitalization were similar between males and females，but different among different age groups：in 12-44 age group，the top three dis－eases were arrhythmia，hypertension and vital myocarditis；the top three diseases were the same between 45-49 age group and 60-74 age group；in 75-105 age group；the top three diseases were CHD，hypertension and sick sinus syndrome. There were 117 deceased among 10 308 inpatients and the mortality was 1.14%. The top three death cause diseases were CHD（71 cases，60.68%），hyperten－sion（17 cases，14.53%），rheumatic heart disease（15 cases，12.85%）. The direct death causes diseases were heart failure（62 cases，52.99%） and sudden cardiac death（33 cases，28.21%）. Conclusions：CHD and hypertension are the leading causes of hospitalization and death of patients with cardiovascular diseases，which should be better prevented and controlled.

Abstract:Objective：To study the safety and approach of laparoscopic biliary tract surgeries for patients with history of upper abdomi－nal laparotomy. Methods：From May 2008 to September 2012，103 patients with upper abdominal laparotomy history underwent la－paroscopic biliary tract surgeries were retrospectively analyzed. Results：Ninety-six patients out of 103 patients were successfully op－erated under laparoscope，7 patients were converted because of excessive adhesion. No complication occurred in all patients. Con－clusion：Laparoscopic biliary tract surgeries are safe and feasible for patients with history of upper abdominal laparotomy.

Abstract:Objective：To evaluate the prognosis of asymptomatic children with renal stones. Methods：Totally 75 asymptomatic children with renal stones were followed up for 28-45 moths averaged 41 months after the initial diagnosis，including 37 melamine-associated renal calculi（MARC） children and 38 non melamine-associated renal calculi（NMARC） children. Urinary system ultrasound and uri－nalysis were examined at the end of follow-up. Results：Follow-up was completed in 53 children（34 MARC children and 19 NMARC children） and renal calculi were excreted in 31 children（58.49%）. Spontaneous discharge occurred more often in infants and in chil－dren with stone of diameter no than 5 mm，while unlikely occurred in children aged older than 3-year-old and in children with stone of diameter larger than 10 mm（P=0.003，P=0.000）. At the end of follow-up，urinalysis showed that the total abnormal rate（15.38%） at 41 months after the initial diagnosis was lower than that（41.03%） at initial diagnosis，with statistically significant differences（P=0.010）. No MARC child had proteinuria，significantly better than that of NMARC children（P=0.030）. Conclusions：Most asymptomatic children with renal calculi discharged spontaneously at the end of follow-up. Stone size and suffering age are two risk factors of dis－charge. Overall prognosis is fine and prognosis of MARC children is superior to that of NMARC. Long term prognosis of NMARC children needs further follow-up.

Abstract:Objective：To explore the main risk factors of recurrent aphthous ulcer（RAV） by investigating its pathogenic factors. Meth－ods：Using the self-made questionnaire of RAU’s risk factors to do the investigation. The high-risk factors of RAU were screened out from all correlative factors by non-conditional logistic regression analysis. Results：The high-risk factors including chronic digestive system disease，mental pressure，liking spicy food，family history，constipation，calculus index，smoking index and liking acidic food were introduced in the regression equation. OR values and regression coefficient of smoking index factor and liking acidic food fac－tor were less than 1 and 0，respectiuely（P＜0.05）. RAU and smoking index showed a negative correlation（rs=－0.170，P=0.000） while RAU and calculus index showed a positive correlation（rs=0.133，P=0.000） based on Spearman rank correlation analysis. Conclusions：These pathogenic factors from high-risk to low-risk were chronic digestive system disease，mental pressure，liking spicy food，family history，constipation and calculus index. Smoking and liking for acidic food may be the protective factors of RAU.