[Objective]To introduce the technique important points of intramedullary fixation with bone cement add the plate fixation for treated the unstable proximal humeral fractures in elderly patients,and proceed the analysis to the principle of bone cement fix.[Method]Ten cases of the unstable proximal humeral fractures were treated with open reduction,intramedullary fixation with bone cement first,then fix with the plate.Three days after operation functional trainings were began.The radiographic outcome and shoulder joint functions were observed postoperatively at 1 month,3 months,6 months and 12 months.[Result]The results were excellent in 5 cases,good in 4 cases,fair in 1 case according to the JOA function standard.The rate of excellent and good clinical results was 90%.Radiography showed that the fractures dissects to reply in 7 patients,fractures still had 3～5 mm moving in 3 cases.All plates were fixed dependable,with no loosening.All fractures had bone fusion.[Conclusion]The intramedullary fixation with bone cement add fixation with plate for the unstable proximal humeral fractures in the elderly can proceed the functional training of the earlier period,acquire the satisfactory function instauration.

Background Studies showed that some members of matrix metalloproteinases (MMPs) play an important role in the pathogenesis of diabetic retinopathy (DR).However,whether MMPs inhibitor can deter blood retinal barrier (BRB) from damage is below understood.Objective This study was to investigate the effects of GM6001,a MMPs inhibitor,on BRB permeability.Methods Twenty-four adult clean SD rats were randomized to the control group,diabetic group and diabetes+GM6001 group according to randomized number table.Diabetic models were induced by the intraperitoneal injection of streptozotocin in the rats of the diabetic group and the diabetes + GM6001 group,and equal volume of citrate buffer was used in the same way in the rats of the control group.GM6001 10 μ1 (100 μ mol/L) was intravitreously injected in the third and fourteenth day after modeling in the diabetes+ GM6001 group,and equal volume of normal saline solution was injected in the same way in the control group and the diabetic group.The rats were sacrificed and eyeballs were extracted 1 month after injection,and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in rat retinas were detected by reverse transcription PCR (RT-PCR).Evens blue (EB) was infused via the right jugular vein,and paraformaldehyde solution 1％ was then infused via left ventricle at the perfusion pressure 120 mmHg.The eyeballs were extracted 2 minutes later,and the leakage of EB in rat retinas was examined.Results RT-PCR electrophoresis exhibited the response bands of MMP-2 mRNA,and MMP-9 mRNA and GAPDH,with the gene size of 436,536 and 484 bp,respectively.The difference of the MMP2 mRNA and MMP-9 mRNA was statisticaly significant (F =20.336,P =0.000 ; F =8.742,P =0.002) ; and the relative expressions of MMP-2 mRNA and MMP-9 mRNA were significantly higher in the diabetic group and diabetes +GM6001 group than those in the control group (all at P＜0.01),and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in the diabetes+GM6001 group was significantly reduced in comparison with the diabetic group(both P=0.01,P=0.02).The standardized EB content in the retinas of the control group,diabetic group and diabetes+ GM6001 group was (12.60±3.50) ng/mg,(26.52±7.14) ng/mg and (17.55±2.65) ng/mg,showing a significant difference (F=17.032,P＜0.01),and EB content in rat retinas in the diabetic group was higher than that in the control group (P=0.003),and that in the diabetes+GM6001 group was lower in comparison with the diabetic group (P=0.020).Conclusions Intravitreal injection of GM6001 can down-regulate the expression of MMP-2 mRNA and MMP-9 mRNA in diabetic rats and therefore protect BRB.

Objective To provide the experimental basis for the further research of the interacting proteins with Stathmin ,the Stathmin gene Pichia pastoris expression system was constructed ,the expressed Stathmin product was purified and identified .Meth‐ods Stathmin gene was amplified from tumor cell line of SKBR3 by PCR method and cloned into the yeast expression vector pPIC3 .5K .The recombinant vector pPIC3 .5K‐Stathmin was constructed and transformed into Pichia pastoris GS115 .The positive clones were screened by YPD medium containing Geneticin 600 μg/mL .Expression was induced with 0 .5% methanol and expres‐sion products were identified by SDS‐PAGE and Western Blotting .Results DNA sequencing result showed that the gene fragment was consistent with Stathmin gene sequence .pPIC3 .5K‐Stathmin was selected from YPD culture medium containing Geneticin ,and the positive clones were identified by PCR .SDS‐PAGE showed that a 37 × 103 protein band could be seen on the PAGE gel after Coomassie Blue staining ,which was further confirmed and identified as Stathmin protein by Western Blotting .Conclusion Stathmin yeast expression vector is successfully constructed and expressed in Pichia pastoris ,which laid the foundation for the study of inter‐acting proteins with Stathmin ,and for the preparation of the biological treatment drugs of Stahtmin target .

Objective To explore the relationship between the Stathmin gene expression in breast cancer cells MDA-MB-231, MCF-7 and the biological behaviours such as cell growth,adhesion and invasion,and provide experimental basis of breast cancer metastasis for further study.Methods Used RT-PCR and Western Blot methods to detect the Stathmin gene expres-sion levels in MDA-MB-231 and MCF-7 cells,and in the mean while to test the MDA-MB-231 and MCF-7 cell growth,adhe-sion,invasion ability by CCK-8 cell proliferation experiments,cell adhesion experiments,cell invasion experiments,then, analyed the relationship of Stathmin gene expression and cell growth,adhesion,invasion ability.Results Over-expression levels of Stathmin gene were observed both in the MDA-MB-231 and MCF-7 cells (F=10.173,P<0.05),and furthermore, the expression levels of Stathmin gene in MDA-MB-231 cells was higher than in MCF-7 cells (t=4.562,P<0.05).While, the growth,adhesion and invasion ability of the MDA-MB-231 cells was higher than that of MCF-7 cells(P<0.05).Conclu-sion The higher level of Stathmin gene expression,the stronger breast cancer cells had ability of growth,invasion,and ad-hesive.The Stathmin gene expression levels was closely correlated with breast cancer cell invasive.

Objective To explore the value of echocardiography in the diagnosis of the parachute mitral valve(PMV).Methods The echocardiographic characteristics of 12 patients with PMV between 2005 and 2012 were reviewed retrospectively,including the morphology,movement and blood flow of mitral valve apparatus,and other associated cardiac abnormalities.Results Of the mitral valve leaflets in these 12 cases,all were thickened and opened restricted in diastole.Axial view showed that in all cases mitral valves opened eccentrically and in 2 cases they didn't close tightly.Each patient only had single papillary muscle in left ventricle,which received all chords of mitral valves.Of papillary muscles in these patients,4 were located at posteromedial wall,3 at anterolateral wall,2 at middle posterior wall,2 at apical wall,and 1 at posterolateral wall.Of the 12 patients,only one were simple PMV,11 had associated cardiovascular anomalies,2 had mild regurgitation in systolic period and 11 had increased trans-valve peak flow velocity.Conclusions Echocardiography is reliable in the diagnosis of PMV,which could provide a comprehensive evaluation of valvular lesions and other associated cardiovascular anomalies.

Objective To evaluate left ventricular(LV) systolic function by two-dimensional speckle tracking imaging(2D-STI) in severe aortic stenosis(AS) patients.Methods Standard echocardiography and 2D-STI examinations were performed in a total of 54 subjects including 26 consecutive patients with severe AS with LV ejection fraction(LVEF) ≥50％ and 28 controls.2D-STI including systolic longitudinal strain (LS) and strain rate(LSr) were assessed from the apical 4-chamber,3-chamber and 2-chamber views,the circumferential strain(CS) and strain rate(CSr),radial strain(RS) and strain rate(RSr) were measured from the standard short axis views (averaging 6 segments per view).The above parameters of AS groups were compared with those of normals.The repeatability of LVEF,LS and RS was evaluated in 6 AS patients and 6 normal volunteers selected at random from the investigation.Results ① There was no significant difference between AS group and control group in LVEF,LV end-diastolic dimension(LVEDD),LV endsystolic dimension (LVESD) and midwall fractional shortening (mwFS) (P ＞ 0.05).② Significant differences were detected between the two groups.LS,RS,CS,LSr,RSr,and CSr were obviously decreased in AS group(P ＜0.05).③The repeatability of LVEF,LS and RS was good by consecutive measurement of identical and independent observers.Conclusions Despite the presence of normal LVEF,the LV systolic function is impaired in severe AS patients.

Objective To investigate the clinical characteristics of cervical spine injury associated with chest injury by contrast with simple cervical spine injury.Methods A retrospective analysis was performed on records of 116 patients with cervical spine injury hospitalized from March 2009 to September 2014.There were 65 patients with simple cervical spine injury (simple injury group) and 51 patients with associated chest injury (associated injury group).Data recorded were the causes of injury, injury segment, treatment choices (tracheotomy rate, mechanical ventilation use and non-operative treatment), treatment time (operation rate at different time, time from injury to operation and length of hospital stay) , complications (electrolyte disorder, respiratory infection, respiratory dysfunction or failure, urinary tract infection, gastrointestinal bleeding and multiple organ dysfunction syndrome), and treatment outcome.Results The main cause of injury for the two groups was high falling.Lower cervical segment was the most likely to be affected.Significant differences were detected between the simple injury group and associated injury group with regard to tracheotomy rate (63％ vs.42％), rate of mechanical ventilation (41％ vs.25％), rate of early surgery (29％ vs.58％), rate of delayed surgery (69％ vs.30％), time from injury to operation [(7.2 ± 3.7) d vs.(3.1 ± 1.3) d], length of hospital stay [(22.6-± 5.5) d vs.(17.3 ± 3.7)d], electrolyte disorder rate (35％ vs.17％), incidence of respiratory system infection (55％ vs.35％), and respiratory dysfunction (43％ vs.25％) (P ＜0.05).After treatment, American Spinal Injury Association (ASIA) scale for grade D was significantly lower in associated injury group than in simple injury group (25％ vs.39％ P ＜ 0.05).Conclusion Cervical spine injury associated with chest injury is severe injury, often requiring tracheotomy and mechanical ventilation, and demonstrates difficulties to be effectively treated in the early phase, long hospitalization, various complications and high morbidity.

Objective To explore the clinical effect and side-effect of oxaliplatin(L-OHP)-containing regimen and cisplatin (DDP)-containing regimen in TACE of advanced hepatocellular carcinoma (HCC). Methods 108 patients with advanced HCC were randomly divided into experimental group(n=55) and control group (n =53). The experimental group were treated with TACE using L-OHP. After dilute with glucose solution, L-OHP(130 mg/m2) and FT207 (500-750 mg/m2) were injected into blood vessel respectively. ADM (40 mg/m2) and LP (10～30 ml) were emulsified and then used for vessel embolism according to the size of focus. The control group received TACE with DDP, DDP (40 mg/m2) and F]'207(500～750 mg,/m2) were diluted with glucose solution, and also according to the size of tumor' s focus, ADM(40 mg/m2) and LP(10～30 ml) were emulsified for vessel embolism, and then diuretic. Results The total effective rate of experimental group was 67.3 % (37/55), and that of control group was 47.2 % (25/53), and the difference was with statistical significance (P <0.05). The descent rate of AFP of experimental group was 73.1%(31/43), and that of control group was 44.7 % (17/38), and the difference was with statistical significance (P <0.05). The main side effects were gastrointestinal reactions, the incidence rate of nausea and vomiting in experimental group were lower than control group, and the difference was with statistical significance. The incidence rate of leucopenia, damage of hepatic function and peripheral neuritis were not significant. Damage of heart and kidney were not found in the two sets. Conclusion L-OHP -containing regimen in TACE of advanced HCC is an efficient method, with good security and good tolerance to patients.

Objective To study the effects of monosialoganglioside combined with edaravone in the treatment of cerebral ischemic neuronal injury in rats.Methods Healthy SD rats were randomly divided into model group and experimental group (monosialin ganglioside 20 mg · kg-1 + edaravone 3 mg · kg-1),each group of 21 rats.The experimental group was injected one,twice a day respectively.The model group using the same amount of 1％ saline,the two groups were administered 14 days.Animal model of focal cerebral ischemia-reperfusion was made by middle cerebral artery occlusion.Using the image analysis system,according to the method of positioning the cerebral ischemic semi-dark bag in the same field of vision,the upper middle cerebral artery occlusion of the same side of the parietal lobe of the upper part of the cortex is defined as a semi-dark band.The silt was observed by immunity.Comparison of the average absorbance and positive area of 3-phosphoinositidedependent(PDK1),phosphatidylinositol-3-kinase (PI3 K),serine-threonine protein kinase (Akt) and glycogen synthase kinase-3β (GSK-3β) protein immunoreactive cells in histochemical method.Results The expression of the PI3K protein,Aktprotein,PDK1 protein were increased while expression of the GSK3β protein was decreased in the model group and the experimental group.After 7 days,positive unit area of PI3K protein,Aktprotein,PDK1 protein and GSK3β protein in the model group and the experimental group were (19.58 ± 1.13),(26.91 ± 0.90) mm2;(15.98 ± 0.48),(22.87 ± 1.20)mm2;(17.97 ± 0.58),(26.02 ± 0.54) mm2;(22.03 ± 0.92),(14.02 ± 0.45) mm2,the difference between two groups was significant (all P ＜ 0.05).The average absorbance density of the four protein in the model group and the experimental group were 0.19 ±0.01,0.28 ±0.02;0.16 ±0.01,0.25 ±0.01;0.16 ±0.01,0.25 ±0.01;0.22 ±0.01,0.14 ±0.01,the difference between two groups was statistically significant (all P ＜0.05).Conclusion Single sialic acid ganglioside combined with edaravone against cerebral ischemia injury of nerve cells in the region.

BACKGROUND:Transfusion of bone marrow mesenchymal stem cells may become a novel and effective biological therapy for inflammatory bowel disease in clinical practice. Nevertheless, the oncological safety of the treatment is worrisome, and is a key to determine whether mesenchymal stem cells can be widely used in treatment of inflammatory bowel disease, and deserves further investigation. OBJECTIVE:To evaluate the therapeutic effect of bone marrow mesenchymal stem celltransfusion against inflammatory bowel disease in mouse models, and to clarify the effects of mesenchymal stem cells on tumorigenesis of inflammatory bowel disease. METHODS:Mouse model of colitis was established using Balb/c (H-2d) mice exposed to dextran sulfate sodium. Syngeneic bone marrow mesenchymal stem cells were transfused into mouse model through caudal vein. The therapeutic effect of mesenchymal stem cells was compared and observed, and pathological remission of colitis was evaluated. Mouse model of colitis-driven colon carcinogenesis was established using Balb/c (H-2d) mice exposed to dextran sulfate sodium and azoxymethane. Tumor formation within the murine colon was compared and observed after transfusion of mesenchymal stem cells. RESULTS AND CONCLUSION:In models of dextran sulfate sodium-induced colitis, weight loss and fecal occult blood were lessened in the bone marrow mesenchymal stem cellgroup compared with the phosphate buffered saline group. Histological damage score of colitis was less in the bone marrow mesenchymal stem cellgroup:mucosal structure of distal colon was almost intact under microscope, and there was smal area of epithelial defects and cryptal defects. Inflammatory cellinfiltration, proliferation of capil ary and smal vessels could be observed in mucosa and submucosa. Homing and colonization of mesenchymal stem cells in submucosa of inflamed colon could also be observed by in vivo tracing. In the dextran sulfate sodium/azoxymethane model of colitis-driven colon carcinogenesis, the number of intestinal tumors and tumor load were obviously less in the bone marrow mesenchymal stem cellgroup than in the control group. Results indicated that transfusion of bone marrow mesenchymal stem cells can apparently improve colitis lesions of mice with inflammatory bowel disease and inhibit carcinogenesis of colitis, which may provide theoretical support for the biological safety of mesenchymal stem cells transplantation for inflammatory bowel disease.