Objective To study the effects and mechanisms of diet-induced hyperhomocysteinemia on intimal hyperplasia of balloon-injured carotid arteries in rats. Methods Wistar rats were bred with 1 g/(kg?d) L-methionine. After 4 weeks, all animals underwent left common carotid arteries balloon denudation and the left common carotid arteries was extracted at day 14 and 28. The areas of carotid arteries intima and media, the proliferative index of smooth muscle cell and the percentage of reendothelization were measured and calculated. Results Intimal hyperplasia developed in all balloon-injuried arteries. The neointimal hyperplasia in the HHCY group was more obvious than that of the control group. The intimal thickness, neointimal area, Intima/Media area ratio were increased by 36%, 41%, 36% at 14 day and 33%, 30%, 21% at 28 day, respectively. Luminal area was decreased markedly in HHCY group. The percentage of intimal and medial PCNA positive cells increased by 1.7-fold and 2.3-fold at 14day. The percentage of endothelial covering area was decreased by 52% at 14day and 31% at 28day in HHCY group. Conclusion Diet induced hyperhomocysteinemia can stimulate SMC proliferation and inhibit endothelial regeneration, which leads to exacerbated intimal hyperplasia. This study implicates that homocysteine may be a significant risk factor for restenosis following vascular intervention.

Background and purpose:The screening of the genes being related closely with the mechanism of osteosarcoma metastasis was a difficult point in the realm of orthopaedics.We screened differential expression gene of human osteosarcoma MG-63 cell sublines with different metastatic capabilities with cDNA microarray,and studied the molecular mechanism of osteosarcoma metastasis.Methods:Total RNA of human osteosarcoma MG-63 cell sublines A1 and A2 was extracted,purified to mRNA and then reversely transcripted to cDNA probe respectively.The cDNA probe of A1 was labelled with Cy3 and the cDNA probe of A2 was labelled with Cy5.The two samples were hybridized with the cDNA microarray.The hybridization signals were scanned by Agilent Scanner and obtained data were analyzed using Ima Gene 3.0 software and Genespring software.Results:222 differential expression genes were found between cell sublines A1 and A2 by analyzing gene expression profile.There were 119 upregulated genes and 103 downregulated genes in cell sublines A1.All differential expression genes belonged to six main function groups and 49 genes of these had very obvious differentce in expression.Conclusions:There were many differently expressed genes between A1 and A2 cell sublines and only part of them were closely associated with mechanism of osteosarcoma metastasis.The technology of cDNA microarray could analyze effectively gene expression profile of human osteosarcoma MG-63 cell sublines, and supply a new approach to study the mechanism of osteosarcoma metastasis

Objective To study the effectiveness and safety of intravenous recombinant tissue plasminogen activator (rt-PA) thrombolysis therapy for acute ischemic stroke in patients with atrial fibrillation (AF).Methods A total of 94 patients treated with intravenous rt-PA thrombolysis within 4.5 hours after cerebral stroke onset were analyzed and divided into two groups:a non-AF group (n =64) and an AF group (n =30).Another 30 acute ischemic stroke patients with AF without thrombolytic therapy were selected as a control group.The prognosis of the three groups was compared.The National Institute of Health Stroke Scale (NIHSS) was used for comparison among the three groups before therapy and 7 days after therapy.The incidences of intracerebral hemorrhage (ICH) and symptomatic ICH (SICH) were recorded.The patients were followed up for 90 days and their clinical outcomes were assessed by using the modified Rankin scale (mRS).Results There were no significant differences in the NIHSS scores among three groups before treatment (P ＞ 0.05).The NIHSS scores were significantly lower in the AF and non-AF groups 7 days after thrombolysis therapy than those before thrombolysis therapy (P ＜0.05 and P ＜0.01),and no significant difference in the NIHSS scores was found in the control group before and after therapy (P ＞ 0.05).The incidence of ICH was significantly higher in the AF group than in the non-AF group (26.7％ vs.9.4％,P ＜ 0.05).No significant difference in the incidence of SICH was found between the AF and non-AF groups (13.3％ vs.6.3％,P ＞ 0.05).The favorable prognosis rate was higher in the AF and nonAFgroups than in control group (40.0％ vs.16.7％,P＜0.05; 45.3％ vs.16.7％,P＜0.01).No significant difference in very unfavorable prognosis rate was found between the AF and non-AF groups (20.0％ vs.18.8％,P ＞0.05).Conclusions It is effective and safe of rt-PA thrombolysis therapy for acute ischemic stroke patients with AF.

Objective To investigate the relation between brain natriuretic peptide (BNP) rs198388 polymorphism and the susceptibility of essential hypertension in Han population of Hunan. Methods A total of 567 patients with hypertension (the hypertension group) and 555 healthy volunteers (the control group) were enrolled. Gender, age, smoking and drinking history of the 2 groups were not significantly different. Blood pressure was measured in the 2 groups. After fasting for 12 h or more, blood glucose, total cholesterol, triglycerides, high density lipoprotein cholesterol and low density lipoprotein cholesterol were measured. DNA polymorphism analysis was done by polymerase chain reaction-restriction fragment length polymorphism method, and genotype was determined by agarose gel electrophoresis. Results The GG, GA, and AA genotypeswere detected.The frequencies of GA and AA genotypes and A allele were significantly lower in the hypertension group (GA and AA:12.3%;A:6.9%) than those in the control group (GA and AA:18.4%; A:9.7%; P=0.009, and P=0.014, respectively). Conclusion BNP rs198388 polymorphism may be associated with essential hypertension in Han people in Hunan. Carrying rs198388 GA and AA genotypes and A allele may be the reason for low risk of hypertension.

Objective To study the clinicopathological and imaging features,key points of diagnosis and prognosis of endolymphatic sac tumor.Methods Seven cases of endolymphatic sac tumor were studied with routine histopathological,immunohisochemical staining and CT,MRI scanning.Patients' prognosis and life quality were aesessed by follow-up.Results The clinical symptoms of patients were progressive sensorineural deafness,tinnitus,vertigo and affection of cranial nerves.CT revealed reticular of moth-eaten bony destruction of the petrosal region,and MRI revealed a region of high-signal intensity void of vascular flowing signals.The histopathologic examination of the tumor presented a papillary-cystic pattern.The tumor cells were round or cubaid in shapen,with nuclei in similar shape and size.There were cyst-like structares filled mith pinkish gelatinous.Tumor cells were roundor cuboid in shape,and some of them contained clear cytofrlasm.There was materials,which were stained strongly with PAS.Hemorrhage,cholesterol crystal and foreign body giant cells were found in the tumor stroma with abundant small blood vessels and extravasation of blood,cells with intracellular hematin,and giant cells collagen.There was also proliferation of fibrous connective tissues.Necrosis and bone destruction were also found in some cases.Immunohistochemical results showed that CK and NSE were positive in all the tumors,and CD56 and S-100 were positive in part of cases,GFAP,EMA,CgA,TG and Syn were negative in all the specimens.One patient had received radiotherapy after partial resection.Relapse occurred only in one patient two years after the first surgery,and no metastasis or relapse occurred in the rest of patients.Conclusions Endolymphatic sac tumor is a rare neoplasm.The diagnosis relies mainly on histopathological and immunohisochemical findings.The patients often have good prognosis.

Objective To study the relationship between ultrasonography of arteria dorsalis pedis and Hemorheology in patient with diabetic foot.Methods According to with or without frank anabrosis,51 patients were devided into two sub-groups:anabrosis group (23 patients) and no-anabrosis group (28 patients).The correlativity was analysed between the parameters of ultrasonography in arteria dorsalis pedis and hemorheological indices on patients with diabetic foot.Results When compared to patients without ulcers,vascular inner diameter (D),peak systolic velocity (PSV),resistent index (RI) and pulsatility index (PI) on patients with frank anabrosis were decreased significantly,but whole blood viscosity at high shear rate (BVH),whole blood viscosity at low shear rate (BVL),plasma viscosity (PV),fibrinogen (Fib) were increased.Multiple regression analyses indicated that the relationship between BVH,BVL and D was presence,partial regression coefficient (r) was -0.004 and -0.009 ; the relationship between PSV and BVH,PV was intimate,r was -0.957 and -25.655 ; the relationship between RI and BVH,PI and BVH,PV,Fib were relatively intimate,r was -0.009 , -0.141 , -2.666 , -0.383 ,respectively.Conclusions The changes of hemorheology can aggravate the abnormality of hemodynamics and the degree of vascular lesion in small vessels of lower limb,so it is one of influencing factor leading to lower limb arterial ischaemia and diabetic foot deterioration.

Objective To study the clinical significance of end-tidal carbon dioxide partial pressure (PetCO2 ) during cardiopulmonary resuscitation (CPR) and seek the fixed value according which to decide if we should and when to give up.Methods This was a prospective,observational study.A total of 124 patients with cardiac arrest in or out-of-hospital from may 2003 to March 2009 in emergency department of our hospital were selected.All of them had definite etiological factors.Changes of PetCO2 in 124 cardiac arrest patients during CPR were tracked.Results The gender,age,rescue time in seventy-one patients with the return of spontaneous circulation (ROSC) after endotracheal intubation have a significant difference with that in fifty-three patients without ROSC (P ＜0.01 ).The PetCO2 of the survival were higher than that of patients without ROSC or with ROSC,but finally died (P ＜0.01 ).A fixed point 14.4 mmHg of PetCO2 after 20 minutes' CPR can be used as a reference value to guide CPR or predict prognosis.Conclusions Monitoring PetCO2 during CPR has a predictive value on the success of resuscitation.

Objective To evaluate the association of Smad2/3,Smad4 and P-Smad3 protein expressions in tissue with liver fibrosis in patients with chronic HBV infections.Methods Liver biopsy was performed to determine the liver fibrosis grades in 131 cases of chronic HBV infections.Immunohistochemistry and semiquantitative analysis were applied to detect the expression of Smad2/3,Smad4 and P-Smad3 proteins in liver tissues.Results Smad2/3,Smad4 and P-Smad3 were detected mainly in fibrous septum,portal areas,myofibroblasts,sinus and cytoplasm in the liver tissue.The expression of Smad2/3,Smad4 and P-Smad3 was increased with the development of fibrosis ( r =0.81,0.58and 0.68,P =0.000),and a strong positive correlation was observed among three proteins (r =0.75,0.87and 0.84,P =0.000).Conclusion The expression of Smad2/3,Smad4 and P-Smad3 is correlated with liver fibrosis in chronic HBV infection,which suggests that the up-regulation of Smad proteins may be involved in the progression of liver fibrosis.

Objective To investigate the effects of IL-1 β on proliferation and migration of gallbladder cancer cells.Methods The secretion of IL-1 β in tissues of gallbladder cancer, chronic cholecystitis and normal gallbladder as well as in supernatant of gallbladder cancer cell lines (GBC-SD, SGC996) and HIBEpic cells was determined by enzyme-linked immunosorbent assay (ELISA) method.The levels of IL-1 β mRNA in GBC-SD, SGC996 and HIBEpic cells were measured by RT-PCR assay.The effects of exogenous IL-1 β on the proliferation of GBC-SD and SGC996 cells in vitro and in vivo were evaluated using WST-1 assay and xenograft tumor model, respectively.The effects of exogenous IL-1 β on the migration of GBC-SD and SGC996 cells in vitro were measured by Tranwell assay.The levels of Twist protein in GBC-SD and SGC996 ceils were examined by western blot assay after treatment with exogenous IL-1 β.In addition, the proliferation and migration of GBC-SD and SGC996 cells after gene silencing of Twist by Twist-siRNA were also evaluated.Results The level of IL-1β protein in normal gallbladder was low (66.4 ± 35.0)pg/ml,while it was significantly increased in gallbladder cancer and chronic cholecystitis [(616.4 ± 95.7) pg/ml and (422.3 ± 48.9) pg/ml, P ＜ 0.05].The levels of IL-1 βin GBC-SD and SGC996 cell culture medium [(587.4 ± 99.8) pg/ml and (657.2 ± 76.6) pg/ml] were much higher than those in the HIBEpic cells [(38.4± 12.1)pg/ml, P ＜ 0.05].Exogenous IL-1β promoted the proliferation of GBC-SD and SGC996 cells both in vitro and in vivo as well as migration in vitro (P ＜ 0.05).The level of Twist protein was significantly increased after treatment with exogenous IL-1 β.In addition, gene silencing of Twist blocked IL-1 β-induced proliferation and migration of GBC-SD and SGC996 cells.Conclusion IL-1 β promoted proliferation and migration of gallbladder cancer cells via Twist activation.

Objective To analyze the image and histological results of the combined use of allograft/extracorporeally frozen tumor-bearing bone and vascularized fibular flap for the reconstruction of bony defects following tumor resection,guiding clinical practice.Methods From March 2007 to June 2013,we enrolled 63 patients who had combinative biological reconstruction after bone tumor resection (11 in humerus,22 in femur,21 in tibia,4 in calcaneus).There were 36 male and 27 female in this series.The average age at time of operation was 20 years,ranging from 9 to 48 years.The follow-up ranged from 16 to 102 months with average of 48 months.We investigated the X-ray and CT images for all patients and histological findings of two patients.Patients were assessed functionally with the Musculoskeletal Tumor Society 93 score.Results Three patients with local soft tissue recurrence and one patient with infection underwent amputation.The survival of construct was 93.6％.Bone union achieved in all cases with the average MSTS score of 92.8％.Bone union ranged from 11 to 28 months in allograft group and 9 to 14 months in devitalized tumor bearing bone group.Significant difference of bone union time was found between two groups (Z=-3.638,P=0.000).Viability of the fibular grafts was verified in 58 of 63 patients (92％).Three types of images were observed in complex.Osteopenia and spongy change in fibula were found in 51 patients (81％) with stable fixation of the complex.Five complexes with failed blood supply of fibula and stable fixation revealed no density change of fibula,small amount of callous formation and relative delayed union.In seven complexes (11％) with unstable complex due to patients' incompliance,fibula reacted with dense hypertrophy and microfracture.Fusion of grafts with amount of callus was obviously observed.Union at allograft-host bone junctions occurred by residual host bone-derived external callus and fibular-derived internal callus that bridged the junction and filled the gap between abutting cortices.Callus from fibular graft was mature than that from periosteum of residual host bone.Internal repair was observed at the internal surface of the allografts.Fibula showed significant spongy changes.Conclusion Recycled tumor-bearing bone in combined with fibular flap is a reliable reconstruction as an alternative to traditional Capanna technique.The survival of the fibula is a cornerstone in success of complex reconstruction.Sponginess of fibula and internal repair of allograft compromise the intensity of complex,necessitating the strong instrumentation during reconstruction.