Human hepatocarcinoma cells, HepG2 were cultured onto biodegradable polyglycolic acid (PGA) polymer scaffolds, which were cultured in a rotating cell culture system (RCCS) to form a three-dimensional (3D) multicellular culture in vitro. The RCCS can simulate microgravity effects with low shear stress and well exchanging for gas. Then the growth characteristics and some mechanism of the cells in RCCS were detected by scanning electron microscopy (SEM), transmission electron microscopy (TEM), RT-PCR and flow cytometry (FCM). The results indicate that the cells grew well with polyhedron morphology and lots of microvilli, mitochondria and tight junctions in this system, which means that this system is useful for cells to form 3D structure to mimic cell status in vivo. The expression of some cell adhesion molecules (CAMs) were changed markedly, which are closely associated with cancer invasion and metastasis. The characters of increased expression of integrin ?1(CD29), CD44, intercellular adhesion molecule-1(CD54) and depressed expression of E-cadherin presumably show that the HepG2 cells cultured in RCCS could recur some characters of primary liver cancer in vivo, the capacity of invasion and metastasis. It is necessary for acquiring perfect and external results to select an appropriate research model for studying in vitro. This 3D culture in vitro under simulated microgravity can provide a useful and reasonable model for oncology, anticancer drugs research and other research.

Objective To observe the expression changes of peripheral endothelial progenitor cells (EPCs) and Ang-1/Tie2 in patients with pulmonary arterial hypertension (PAH).Methods From Jun 2011 to Dec 2012,45 patients with PAH charged in Affiliated Hospital of Hangzhou Normal University were divided into 3 groups according to mean pulmonary arterial blood pressure (15 per group):mild(Group L),moderate(Group M),and severe(Group S),with another 15 normal people as control group(Group C).The EPCs were isolated from peripheral blood of every patient,number counting using fluorescence activated cell sorter (FACS),function test using cell culture in vitro.Expression of Ang-1 and Tie2 in the peripheral EPCs were measured by RT-PCR or Western-blot.Non normal data was analyzed by non parametric statistical test.Results The statistical discrepancy existed among Group L,M,S and the control in the number of EPCs [32.0 (27.0,37.0),26.0 (19.0,31.0),24.0 (22.0,26.0) vs 40.0 (37.0,51.0),P ＜ 0.05].The ability of migration [32.1 (26.5,37.5),26.8 (22.4,35.4),21.0 (17.8,34.0) vs 39.0 (33.3,42.4),P＜0.05] and adhesion of the EPCs [57.1(50.9,61.8),51.8(45.2,58.7),46.0 (37.2,55.1) vs 64.1 (56.2,75.0),P ＜ 0.05] among study groups and control group was different in statistic,the same with the proliferation activity of EPCs in different groups [0.6 (0.5,0.7),0.5 (0.4,0.6),0.4(0.3,0.5) vs 0.7(0.6,0.8),P ＜0.05].The mRNA expression of Ang-1 and Tie2 in Group M & S were significantly reduced compared with control [4.33 (2.49,4.62) and 2.89 (2.39,3.44) vs 5.31(3.78,6.22),P＜0.05],Tie2 mRNA[1.32(1.23,1.34)and 1.23(1.08,1.42)vs 1.49(1.25,1.66),P ＜ 0.05],and the protein expression of the phosphorylated Tie 2 in Group M &S were decreased [0.16 (0.15,0.24) and 0.12 (0.08,0.18) vs 0.22 (0.19,0.28),P ＜ 0.05].No significant difference of Ang-1 and Tie2 expression was observed between Group L and control [5.42 (4.72,5.95),1.54 (1.43,1.66) and0.23(0.19,0.33),P=0.674,0.867 and 0.674].Conelusion With the severity of PAH,the number and function of circulating EPCs decreased,as consistent with Ang-1 and Tie2 expression changes,suggesting that function decrease of EPCs in patients with PAH may be associated with the decrease of Ang-1/Tie2 expression.

Objective To observe the effect of early non-invasive positive pressure ventilation(NIPPV)on the treatment of serious hypoxemia induced by acute left heart failure. Methods Forty patients with acute left heart failure( Grade Ⅳ heart function)were randomly divided into two groups. Patients in both groups accepted supportive treatment included cardiotonics, diuretics, vasodilators, in additional to these high concentrations of oxygen were given in conventional group, and non-invasive positive pressure ventilation were given in NIPPV group by biphasic positive airway pressure(BiPAP). Systolic blood pressure, heart rate, respiratory rate, blood-gas analysis( pH, PaO2, PaCO2, SaO2 )and clinical signs were observed at 2 hours after treatments. Results Compared to control,RR( [ 19.55 ± 1.88] vs [21.85 ±3.51 ] ) BPM] ,HR ( [96.40 ±2.80] vs[ 103.20 ±6.78 ] BPM), SBP ( [ 116.30 ± 8.95 ] mm Hg vs [ 122.50 ± 6.13 ] mm Hg), pH (7.404 ± 0.027 vs 7.358 ±0.05) ,SaO2 ( [93.57 ± 1.18]% vs [91.97 ± 1.85]% ) ,PaO2 ( [75.58 ±4.61 ]mm Hg vs [68.38 ±7.95]mm Hg), PaCO2 ( [ 38.69 ± 3.06 ] mm Hg vs [ 43.61 ± 2.65 ] mmHg) were significantly different in NIPPV group( t = 2.582,4.146,2.558,3.534,3.256,3.505,5.428, Ps ＜ 0.05 ). We found no significant differences in the comparisons before treatments. Hypoxia improved in NIPPV group,and the total effective rate was 95% in NIPPV group and 70% in control group,which showed significant difference( x2 =4.329 ,P ＜0.05 ) Conclusion BiPAP non-invasive positive pressure ventilation combined with routine treatment in treating heart failure, could promptly correct hypoxia, improve heart function and shortening disease course.

Objective To investigate the expression of a novel imprinted gene,PEG10,in human gastric adenocarcinoma,and the effect of PEG10 on cell growth and proliferation of gastric cancer cells.Methods The PEG10 mRNA expressions in 40 human gastric adenocarcinoma,the corresponding adjacent normal tissues and 6 nomal gastric tissues were detected by RT-PCR.The expression vectors of PEG10 were constructed and transfected into gastric cancer cell line MKN45 which had no endogenetic PEG10 expression.Cell growth ability was measured by MTT assay.Results High PEG10 mRNA expression level was detected in 9 of 20(45.0%) human gastric adenocarcinoma which was significantly higher than those of the matched normal tissues(10.0%)(P

Objective To investigate the influence of different doses and courses of cigarette smoking on the expression of ICAM-1 mRNA, ICAM-1 in rats' cerebral vascular endothelial cells and volumes of cerebral infartion.Methods 105 male Wistar rats(weight 210～220 g) were randomly grouped into long-term massive(20 cigarettes/d,75 d), short-term massive(20 cigarettes/d,30 d), long-term trifle (8 cigarettes/d, 75 d), anti-smoking(20 cigarettes/d, 30 d, denying 45 d)and normal control(no smoking). In situ hybridization, immunohistochemistry were used to determine the expression of ICAM-1 mRNA, ICAM-1 in rat's cerebral vascular endothelial cells (VECs).The VEC's structures were observed by electronic microscope. Left middle cerebral artery occlusion model was established and pathologic image analyzing system were used to determine the ratio of the volume of infarction in hemisphere of the same side. Results On contrast to normal controls, the expression of ICAM-1mRNA, ICAM-1 and the volume of infarction were increased in smoking(P

Objective To study the differentially expressed genes in the vascular tissue of diabetic gangrene toot and the possible mechanism of diabetic gangrene.Methods The differentially expressed genes in the vascular tissue of diabetic gangrene foots were screened by the functional classific gene chip.Reverse transcription polymerase chain reaction(RT-PCR) detection was used to verify the differentially expressed 3 genes.Results In the detected 113 genes,13 of the gene expression level increased more than 2 times,3 of the gene expression level decreased more than 2 times.Three genes were verified with RT-PCR detection,which were results similar to those with chip testing.Conclusions The abnormal expression in a variety of genes in tissues of vascular lesions plays an important role in the course of development and progression of diabetic gangrene.Analysis of differential gene expression can contribute to understanding of the possible mechanisms of diabetic gangrene.

Objective To evaluate the clinical efficacy of a self-designed novel n-shaped aiming device in aiding the percutaneous cannulated screwing for treatment of odontoid fractures of Anderson-D' Alonzo type Ⅱ and shallow type Ⅲ.Methods A retrospective analysis was conducted of the 17 patients who had been treated at our department for odontoid fracture of Anderson-D' Alonzo type Ⅱ and shallow type Ⅲ between January 2009 and December 2013.They were 13 males and 4 females,with an average age of 37.5 years (from 16 to 61 years).All received surgical treatment after skull traction or jaw pillow traction preoperatively for reduction.Intraoperatively,percutaneous cannulated screwing via the anterior cervical approach was performed with the help of a self-designed novel n-shaped aiming device to fix the odontoid.Postoperatively,X-ray and CT scan were used to check the screw locations and evaluate the fracture healing.Results The 17 cases were followed up for 6 to 15 months (average,10.5 months).No intraoperative injury to the esophagus,artery,spinal cord or nerve root happened.The operating time ranged from 60 to 125 min.The bleeding was from 10 to 40 mL.Intraoperative observation revealed fine locations of the screws.Bony union was achieved in 16 cases 3 months postoperatively while separation and hardening of the fracture ends was found in one ease.Conclusion Our self-designed novel n-shaped aiming device can facilitate the percutaneous cannulated screwing for treatment of odontoid fractures of Anderson-D'Alonzo type Ⅱ and shallow type Ⅲ,given that it has advantages of simple manipulation,accurate insertion,minimal invasion,quick recovery and reliable efficacy.

objective To investigate the effects of different sections of receptor associated protein (RAP) on the expression and distribution of TRPC6,synaptopodin and podocalyxin in passive Heymann nephritis(PHN). Methods Male Sprague-Dawley rats were injected with three kinds of antisera (anti-RAP full-length serum,anti-RAP N-terminal serum and anti-RAP C-terminal serum)to establish three kinds of PHN models.The control group was injected with normal rabbit serum.The quatitation of 24 h urinary protein,serum albumin and creatinine were taken before injection and one week after PHN model successfully induced.The histopathologic changes of renal tissues were observed by light microscopy.The expression and distribution of TRPC6,synaptopodin and podocalyxin in glomerular podocytes were observed by laser scanning confocal microscopy and analyzed by fluorescence quantitative software after indirect immunofluorescence double staining.Results The quantities of 24 h urinary protein in the three model groups were significantly higher than those of themselves before injection and control groups (P<0.01,respectively).The values of serum albumin and creatinine were not significantly different before and after injection (P>0.05).The expression of TRPC6 in podocytes was higher in the PHN model groups than that of control group.Fluorescence intensity of TRPC6 in RAP full-length group was stronger than that in RAP N-terminal or C-terminal groups.The expressions of synaptopodin and podocalyxin distributed along the glomerular basement membrane as spot,discontinuous short line and defect of some segments,and were lower in three PHN groups than those of control group.Fluorescence intensity of synaptopodin and podocalyxin among three PHN groups had no differences. Conclusions RAP full-length and N-terminal or C-terminal parts can increase the expression of podocyte TRPC6,but decrease the expressions of synaptopodin and podocalyxin,and alter their distribution,which may be associated with the proteinuria,however,their role in the PHN pathogenesis needs further study.

Objeetive To assess the study outcomes in a consecutive series of patients with thoracic disc herniation (TDH) who undergone posterior spinal canal decompression and discectomy with segmental instrumentation and fusion.Methods Between January 2005 and June 2012,the data of 17 patients (11 males and 6 females) was retrospectively reviewed and analyzed.Disc herniation was classified as central in 10 cases and paracentral in 7 cases.The average canal encroachment was 71.7％±9.6％ (range,52％ to 90％).Their mean age at surgery was 55.7 years (range,21 to 81 years).All patients underwent a transfacet decompression and segmental instrumentation with interbody fusion.The data of patients included clinical presentation,blood loss,operative time,complications,visual analogue scale (VAS),Japanese Orthopaedic Association (JOA) score,and Frankel grading system.Results The average follow-up period was 37±12 months.The average surgical time was 120±63 min.The mean blood loss was 471±198 ml.Mean preoperative VAS score was 8.01±0.21,which improved to a mean of 1.12±0.07 at final follow-up.Average pre-and post-operative at 12 months JOA scores were 3.17±0.83 and 8.78±0.94 points,respectively.The average recovery rate was 65.1％±23.4％.Overall JOA scores showed a significant postoperative improvement.All patients reported 1 or 2 grade improvement in Frankel grading compared with preoperative status except for 1 patient whose grade had not changed.There were three complications,including one patient developing postoperative wound infection,one experiencing a cerebrospinal fluid leakage,one developing further neurological deterioration.Conclusion The results suggested that the posterior approach using special shaped osteotomy is feasible for central calcified TDH.No major complications occurred for achieving adequate decompression for central calcified TDH.

Objective To prospectively compare cadmium-zinc-telluride (CZT) SPECT gated myocardial perfusion imaging (GMPI),conventional SPECT MPI and cardiac MRI for the assessment of left ventricular volume and ejection fraction in patients with heart failure.Methods From July 2016 to October 2016,a total of 35 patients (27 males,8 females,average age:(52.7±14.9) years) with heart failure were consecutively included.All patients underwent CZT SPECT GMPI,conventional SPECT GMPI and cardiac MRI within 7 d.LVEDV,LVESV and LVEF of three imaging modalities were calculated.One-way analysis of variance,Pearson correlation analysis and Bland-Altman analysis were used.Results CZT SPECT showed excellent correlation with conventional SPECT for LVEDV,LVESV and LVEF (r values:0.983,0.986 and 0.910,respectively;all P＜0.001).Bland-Altman analysis revealed good agreement between CZT SPECT and conventional SPECT for LVEDV,LVESV and LVEF.The correlation between CZT SPECT and cardiac MRI for LVEDV,LVESV and LVEF were all significant (r values:0.864,0.896 and 0.836,respectively;all P＜0.001).Compared with cardiac MRI,CZT SPECT showed systemic underestimation of LVEDV and LVESV and good agreement of LVEF by Bland-Altman analysis.Conclusions CZT SPECT has high clinical value for patients with heart failure.Despite underestimating LVEDV and LVESV,it correlated well with cardiac MRI.It also has a good agreement with conventional SPECT on left ventricular volume and LVEF.