Objective To investigate of human esophageal squamous carcinoma cells(Eca-109) in which cyclooxygenase (COX)-2 gene was knocked down by adenovirus delivered siRNA.Methods Based on the plasmid pSUPER cloned with RNA polymerase Ⅲ-dependent promoter HI,the interfering plasmid psiRNA/COX-2 targeting human COX-2 mRNA was constructed,The siRNA/COX-2 fragment was derived from psiRNA/COX-2 digested by Not I and Xho 1.and was cloned into the shuttle plasmid pAdTrack.Then pAdTrack/siRNA/COX-2 was obtained and co transfected into the E.coli strain BJ5183 with the bone plasmid pAdEasy-1,the recombinant adenovirus Ad/siRNA/COX-2 was generated by homologous recombination.Having been packaged and amplified in cells 293,Ad/siRNA/COX-2 was transfected into Eca-109 cells.The PGE2 concentration in the cells culture supernatant was determined by ELISA,and the level of COX-2 mRNA in the cells was tested by real time PCR.Moreover,cell cycle and apoptosis were determined by flow cytometry,and cells growth curve was protracted.Results The recombinant adenovirus Ad/siRNA/COX-2 was successfully constructed.Ninty six hours after Ad/ siRNA/COX 2 transfecting into Eca 109 cells,COX-2 mRNA was reduced by 71.7%,and PGE2 concentration in the cells culture supernatant was decreased by 62.0%.Correspondingly,the growth of cells slowed down.At the same time,the cells in G0-G1 phase was increased by 32.24%,and those in S phase and G2-M phase were reduced by 16.38% and 15.86%.respectively.And cells apoptosis index was increased by 9.19%.Conclusion The adenovirus based-RNAi was capable of knocking down remarkably COX-2 of human esopbageal carcinoma cells,which lead to growth of cells slowing down.

The purpose of the present study was to examine the effects of oxidative stress on ventricular arrhythmias in rabbits with adriamycin-induced cardiomyopathy and the relationship between oxidative stress and ventricular arrhythmia. Forty Japanese white rabbits were randomly divided into four groups (n=10 in each): control group, metoprolol (a selective β1 receptor blocker) group, carvedilol (a nonselective β blocker/α-1 blocker) group and adriamycin group. Models of adriamycin-induced cardiomyopathy were established by intravenously injecting adriamycin hydrochloride (1 mg/kg) to rabbits via the auri-edge vein twice a week for 8 weeks in the adriamycin, metoprolol and carvedilol groups. Rabbits in the control group were given equal volume of saline through the auri-edge vein. Rabbits in the metoprolol and carvedilol groups were then intragastrically administrated metoprolol (5 mg/kg/d) and carvedilol (5 mg/kg/d) respectively for 2 months, while those in the adriamycin and control groups were treated with equal volume of saline in the same manner as in the metroprolol and carvedilol groups. Left ventricular end diastolic diameter (LVEDd) and left ventricular ejection fraction (LVEF) were measured by echocardiography. Plasma levels of N-terminal pro B-type natriuretic peptide (NT-proBNP), malondialdehyde (MAD) and superoxide dismutase (SOD) were detected. The left ventricular wedge preparations were perfused with Tyrode's solution. The transmural electrocardiogram, transmural action potentials from epicardium (Epi) and endocardium (Endo), transmural repolarization dispersion (TDR) were recorded, and the incidences of triggered activity and ventricular arrhythmias were obtained at rapid cycle lengths. The results showed that TDR and the serum MDA and NT-proBNP levels were increased, and LVEF and the serum SOD level decreased in the adriamycin group compared with the control group. The incidences of triggered activity and ventricular arrhythmia were significantly higher in the adriamycin group than those in the control group (P<0.05). In the carvedilol group as compared with the adriamycin group, the serum SOD level and the LVEF were substantially increased; the TDR, and the serum MDA and NT-proBNP levels were significantly decreased; the incidences of triggered activity and ventricular arrhythmia were obviously reduced (P<0.05). There were no significant differences in the levels of MDA and SOD, LVEF, TDR and the incidences of triggered activity and ventricular arrhythmia between the adriamycin group and the metoprolol group. It was concluded that carvedilol may inhibit triggered activity and ventricular arrhythmias in rabbit with adriamycin-induced cardiomyopathy, which is related to the decrease in oxygen free radials.

ObjectiveTo explore the role of BTBD10 overexpression in the proliferation of insulinoma cell line INS-1and its mechanism. MethodsThe recombined expression plasmid of pcDNA4.0-BTBD10 was constructed by gene cloning technique and was transfected into INS-1 cell by lipofectamine 2000. The stable overexpression BTBD10 of INS-1cell was selected at 48th hour after transfection.INS-1 cell proliferation activity was measured by MTT method.The expression of BTBD10,protein kinase B(Akt),phospho-Akt(p-Akt),mammal target of rapamycin (mTOR) and phospho-mTOR (p-mTOR) were determined by Western blot.ResultsThe stable overexpression BTBD10 of INS-1 cell wassuccessfullyconstructed.OverproductionofBTBD10promotedbetacellproliferation.The phosphorylation of Akt and mTOR was increased and the ratio of p-Akt/Akt and p-mTOR/mTOR was enhanced in the INS-1 overexpressed by BTBD10.But the expression of total Akt and mTOR presented no obvious changes. Conclusion The overexpression BTBD10 of INS-1 cell could activate of Akt/mTOR signalling pathway via stimulating phospho-mTOR and Akt,and enhance overall cell protein translation,so as to promote proliferation of INS-1 cell.

The present is aimed at identifying the activation of TRAFs in n asopharyngeal carcinoma (NPC) in vitro. The differential expression of TRAF2\,TRAF3 was not detected in RN A and protein level, whereas the expression of TRAF1 in HNE2-LMP1 cell lines wa s much more abundant than that in HNE2 cell lines, suggesting that TRAF1 may be an inducible molecule; More importantly, TRAF1, TRAF2 or TRAF3 were activated in the HNE2-LMP1 cells, whereas TRAF1, TRAF2 or TRAF3 were not activated in HNE2 cells as detected by the immunoprecipitation-Western blotting assay. These data provide an experimental basis for our study beginning from the signal transduca tion pathway for the eluccidation of the mechanism of LMP1 carcinogenesis in NP C.

Background Choroidal neovascularization (CNV) is a main cause of visual impairment in many retinal diseases.To create an ideal CNV animal model is very important for the experimental and clinical study of CNV.The assessment method of repeatable and reliable for CNV model is still seldom.Objective This experiment was to explore the label value of fluorescent antibody for visualizing and quantifying the morphologic changes associated with laser-induced CNV.Methods Laser-induced CNV models were created in 30 eyes of 15 male SPF C57BL/6J mice by Krypton red laser irradiating fundus 2 spots around the optical disc with the wavelength 647.1nm,power 260 mW,spot diameter 50μm and exposure time 0.05 seconds.The CNV was evaluated at 5 minutes,4,7,14 and 28 days after laser injury by using fundus photography and fundus fluorescein angiography (FFA),and the successful models were identified as the rupture of Bruch's membrane.The mice were then immediately sacrificed and the eyeballs were enucleated to prepare the choroidal flatmounts.The posterior eye cups were fluorescently labeled with markers of cell nuclei (DAPI,4',6'-diamino-2-phenylindole),endothelial cells (isolectin-B4),and filamentous actin (phalloidin).The CNV areas from specimens were measured by Image pro plus 6.0.Two eyes from one matched mouse without receiving photocoagulation were used as the controlls.This study followed the Standard of Association for Research in Vision and Ophthalmology.Results No any CNV was seen in photocoagulated eyes in 5 minutes and 4 days after laser irradiation.The first sign of CNV appeared at 7 days following photocoagulation.The incidence of fluorescein leakage was 76.47% (26/34),81.81% (18/22),50.00% (5/10) at 7,14 and 28 days,respectively.The fluomicroscope examination showed that in unphotocoagulated areas,retinal pigment epithelial (RPE) cells were visualized with a uniform hexagonal array.Immediately after laser exposure,a circular area devoid of fluorescent labeling was observed,indicating disruption of the choroid-Bruch membrane-RPE complex.On the fourth day,cellular debris and fragmented nuclei were presented and an autofluorescent ring was visible at the site of Bruch's membrane disruption.The number of CNV vessels increased exponentially during the next 3 days.At 7 days,a well-defined isolectin-B4 labeled CNV network was exhibited and lasted for 28 days.The CNV areas were (7.99±0.42)×103μm2,(16.89±3.77)×103μm2,(14.37±4.02)×103μm2 at 7,14 and 48 days after photocoagulation respectively,showing a significant difference among these three groups (F=17.340,P=0.000),and the CNV area was significantly increased in the photocoagulating eyes in 14 days and 28 days compared with 7 days (q=16.46,q=15.54,P＜0.01).Conclusion Fluorescent antibody labeling allows the well identification and measurement of laser-induced CNV lesions in mouse choroid/RPE flatmounts.This technique offers excellent morphologic detail and facilitates the study of critical early events in CNV.CNV complexes are labeled at an early stage,providing a more accurate preclinical evaluation of antiangiogenic molecule.

Objective To investigate the clinical features and distribution of G types of rotavirus diarrhea in infants and children in Suzhou and Maanshan areas.Methods 1267 stool specimens were collected from children under 5 years with acute diarrhea. Enzyme-linked immunosorbant assay (ELISA) and nested polymeras chain reaction (Nested-PCR) were used to detect rotavirus and serotypes.Results In 1267 stool specimens, 378 were detected RV positive, and serotype G was the prevalent type in the two cities with number of 250 and the G3 and G1 types were the most prevalent which accounted for 40%(100/250) and 44.40 % (111/250), respectively. While the G2 ,G4, G9 types and mixed infections were 8,3,3,4 and the un-identified type was 8.40%(21/250). G1 type was the most prevalent in Maanshan are which accounted for 58.54% in all cases,while the G3 type accounted for the most of case(47.85%) in Suzhou area.Conclusions The results indicated that rotavirus was the most important etiologic agent in Maanshan and Suzhou areas,G1 and G3 were the most prevalent types,respectively. J Appl Clin Pediatr,2005,20(3):208-210

OBJECTIVETo explore the clinical outcomes of open reduction and internal fixation with calcaneal locking plates in treating Sanders type II and III calcaneal fractures.

METHODSFrom January 2010 and October 2012, 38 calcaneal fractures with Sanders type II or III were treated with open reduction and internal fixation with calcaneal locking plate. According to the Sanders classification, 15 fractures were classified as type II, 23 fractures as type III. The patients were divided into two groups (group A and B) according to the different fixed methods. Sustentaculum tali was fixed with one screw in group A, including 13 males and 5 females, with a mean age of (38.56±8.03) years old (ranged, 25 to 55). And sustentaculum tali was not fixed in group B, including 16 males and 4 females, with a mean age of (42.35±8.29) years old (ranged, 29 to 53). Clinical effects were evaluated according to the changes of Böhler's angle and the Maryland Foot Score and VAS score.

RESULTSAll patients were followed up from 12 to 20 months with a mean of 14 months. Böhler's angles and subtalar joints obtained satisfactory reconstruction in all patients. One year after operation, the mean Maryland Foot Score was 88.61±7.59 in group A; and was 82.40±9.24 in group B; Maryland Foot Score of group A was higher and foot functional rehabilitation was better than group B. The mean VAS score was 13.39±11.47 in group A; and was 22.50±13.10 in group B; VAS score of group A was lower and foot pain was less than group B.

CONCLUSIONSustentaculum tall screw fixation has advantages of strong fixed strength, high stability, less postoperative pain, rapid functional recovery in treating Sanders type II and III calcaneal fractures.

Adult ; Bone Plates ; Bone Screws ; Calcaneus ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Recovery of Function

Objective To investigate the clinical teaching situation by using developmental inspection of School of Medicine,Shanghai Jiaotong University(SJTU-SM),and to put forward some suggestions. Methods By checking questionnaires and informal discussions,the relevant information was collected and analyzed by using SPSS statistics sofware. Results The clinical teaching quality of SJTU-SM was basically satisfied.The satisfaction from internship of grade 2004 was better than that of grade 2003.However,some problems in clinical teaching must be improved.Conclusion The investigation showed that the clinical teaching quality of SJTU-SM is being improving.However,in order to achieve the international accreditation standards,the quality guarantee system of clinical teaching need to be further perfected.

Objective To study the protective effect and its mechanism of anti-tumor necrosis factor-?antibody (TNF-? Ab) on lung injury after cardiopulmanary bypass(CPB).Methods Twenty-eight healthy rabbits were selected and randomly divided into four groups:group Ⅰ only received open chest operation;groups Ⅱ-Ⅳ underwent CPB.In the group Ⅳ,rabbit TNF-? Ab (2 400 pg/kg) was dropped into the intracheal tube before operation and just after releasing the aortic clamp.Saline was given to the group Ⅲ in- stead.Blood neutrophils count,TNF-?,MDA from the right and left atrium in the four groups were determined perioperatively.Water volume,TNF-? mRNA,TNF-? protein,apoptosis and pathomorphological changes were measured in the lung tissues.Results TNF- ? Ab can restrain leukocyte accumulation,reduce releasing of TNF-? and MDA in the lung.It can also reduce the occurrence of apop- tosis and attenuate pathomorphological changes in the lung tissue.However,it cannot reduce the secretion of TNF-? at the transcrip- tion level and protein level.Conclusion Intratracheal TNF-? Ab administration has markedly protective effect on lung injury after CPB.

OBJECTIVETo explore the feasibility of using transpedicular balloon kyphoplasty for aged osteoporotic thoracolumbar burst fractures with an in vitro model.

METHODSSimulated osteoporotic thoracolumbar burst fractures were created in 11 vertebral bodies. The burst fractures without obvious canal occupation were confirmed by spiral CT before the procedure. This operation involved the percutaneous insertion of two inflatable bone tamps into a fractured vertebral body transpedicularly under fluoroscopic guidance. Inflation of the bone tamp elevated the endplates, restored the vertebral body height, while created a cavity to be filled with bone cement. Preoperative and postoperative heights were measured, preoperative and postoperative sagittal diameter of vertebral canals were measured, and the repaired vertebral bodies were compressed to determine strength and stiffness values. The cement distribution and extravasation was evaluated by spiral CT after the procedure.

RESULTSThe transpedicular balloon kyphoplasty resulted in the significant restoration of the vertebral body height. The mean vertebral body heights was (20.73 +/- 1.37) mm before operation and (24.82 +/- 1.61) mm after operation (P < 0.01). Preoperative and postoperative sagittal diameter of vertebral canals were similar (P > 0.05). The cement distribution was regular, and the extravasation into spinal cannula was found in one case. The preoperative strength was significantly larger than the postoperative strength, whereas the postoperative stiffness was significantly less than the preoperative stiffness (P < 0.05).

CONCLUSIONSTranspedicular balloon kyphoplasty is an effective way to treat aged osteoporotic thoracolumbar burst fractures without obvious canal occupations and neurologic deficit in vitro. The procedure can increase strength and restore height of aged osteoporotic thoracolumbar burst fractures. Cement distribution and vertebral canal stenosis can be avoided.

Cadaver ; Humans ; Kyphoplasty ; instrumentation ; methods ; Kyphosis ; diagnostic imaging ; etiology ; surgery ; Osteoporosis ; complications ; Spinal Fractures ; complications ; diagnostic imaging ; surgery ; Spinal Fusion ; methods ; Thoracic Vertebrae ; pathology ; surgery ; Tomography, X-Ray Computed