ObjectiveTo study the change of autophagy of human pancreatic cancer cell MiaPaCa-2 before and after Ad5-KAI1 tranfection,and to investigate the possible mechanism.MethodsThe MiaPaCa-2 cells without KAI1 expression were infected with Ad5-KAI1 with KAI1 target gene,and Ad5-null was used as negative control,and parental cell was used as blank control.The formation of autophagosomes was observed by electromicroscopy.The green fluorescent protein-labeled light chain 3 (LC3) associations with autophagosome membranes was detected by confocal microscopy.PD98059,LY294002 were applied to pre-treat the cells.The expression levels of beclin 1,AKT,ERK,the phosphorylation of AKT and ERK protein and the ratio of LC3-Ⅱ to LC3- Ⅰ were detected by Western blotting.ResultsAfter 100 MOI Ad5-KAI1 infections for 24 h,the rate of cell expressing KAI1 protein reached (84.97 ±8.56)％,number of LC3 increased from 4 to 20; and swelling,degeneration of mitochondria was observed,and bilayer-like structure in cytoplasm was found.The expression of beclinl increased (1.4 ±0.3 ) folds,and the expression of LC3-Ⅱ/LC3- Ⅰ increased (8.00 ±2.78) folds.PI3K blockade LY294002 pretreatment significantly suppressed the phosphorylation of AKT of MiaPaCa-2 (2.756 vs 1.516),but it did not inhibit the increase of ratio of LC3-Ⅱ to LC3- Ⅰ(0.770 vs 1.403).ERK blockade PD98059 pretreatment not only significantly suppressed the phosphorylation of ERK of MiaPaCa-2 ( 1.637 vs 0.403 ),but also inhibit the up-regulation of beclin 1 protein expression ( 2.377 vs 1.150) and increase of ratio of LC3- Ⅱ to LC3- Ⅰ (2.225 vs 0.680).ConclusionsKAI1 can significantly induce autophagy of human pancreatic cell line MiaPaCa-2 through phosphorylation of ERK rather than AKT.

Objective To explore the pathway of KAI1 induced autophagy regulating apoptosis in human pancreatic cancer cell line MiaPaCa-2.Methods There were three groups in the experiment,which were extracellular regulated protein kinases (ERK) phosphorylation inhibitor PD98059 pretreated group,Caspase-3 activation inhibitor VAD-FMK pretreated group and no PD98059 or VAD-FMK pretreated groups.And each group was divided into three sub groups with different treatment,which were adenovirus AD5-null vector infected control group,the human KAI1 gene recombinant adenovirus vector AD5 KAI1 infected group and autophagy inhibitor 3-MA pretreated and AD5-KAI1 infected group.The cell apoptosis was observed by AnnexinV-FITC/PI double staining.Caspase-3 activation level was evaluated by flow cytometry.ERK phosphorylation and poly(ADPribose) polymerase (PARP) cleavage were determined by Western blot.Results After the cancer cells infected with AD5 KAI1,KAI1 protein was expressed and GFP-LC3 green particles increased.Caspase-3 activation,PARP cleavage,ERK phosphorylation and apoptosis increased obviously.After autophagy inhibitor 3-MA pretreated,the percentage of apoptosis increased from (63.0 ± 7.9)％ to (88.0±4.5) ％ and Caspase-3 activation increased from (34.0±2.8) ％ to (44.2±4.0) ％ and PARP cleavage more.The apoptosis induced by 3-MA could be totally inhibited by Caspase-3 activation inhibitor VAD-FMK pretreated but could not be inhibited by ERK phosphorylation inhibitor PD98059.Conclusion KAI1- induced autophagy inhibits apoptosis through the downregulation of Caspase-3activation and PARP cleavage instead of ERK phosphorylation.

Objective To investigate the relationship among early parenchymal enhancement,post- gadolinium(Gd)hyperintense middle cerebral artery(HMCA),and subsequent hemorrhagic transformation (HT)in patients with hyperacute ischemic stroke.Methods Twenty-four consecutive patients with ischemic stroke who underwent MRI within 6 h[(4.3?1.4)h]of symptom onset were retrospectively reviewed.All of these patients underwent at least one follow-up MRI or non-enhanced CT study at 2 to 7 days.Post-Gd T_1WI were analyzed for parenchymal enhancement and hyperintense MCA.Gradient echo MRI and CT were used for assessment of HT.Results Ten patients developed HT on follow-up imaging (hemorrhagic group).Early parenchymal enhancement was found in 6 patients with HT(P

[Objective]To investigate the regular pattern of references quoted in theses of Orthopedic Journal of China from year 2005 to 2006,and to study the characteristics and experiences of reference quoting in Chinese orthopedic academic sphere.[Method]Forty-eight issues of Orthopedic Journal of China from year 2005 to 2006 were downloaded from the website www.chinajournal.net.cn.There were 466 theses about clinical research.All references in these theses were collected as origin of data.Excel software was applied to analyze all references.[Result]References were found in all 466 papers with the reference rate of 100%.The average reference amount in each thesis was 7.15(3 334/466).A total of 3110 references were from journals(93.28%)and 214 references were from monographs(6.42%).The Price Index reached 52.13%.And the number of self-quoted references reached 614 with the rate of self-quoting of 18.42%(614/3 334).References from Chinese and foreign journals with the first 20 highest being-quoting rates were collected,and it reached 56.12% of all references.[Conclusion]The content of references in Orthopedic Journal of China ranges widely.The main quoting origin is from journals.The mainly-quoted language is English,and references from other languages are rarely quoted.Most references showed new and original ideas.

Objective To compare the treatment effect of internal fixation with retrograde interlocking intramedullary nails or condylar plates in the treatment of comminuted fracture of the femoral condyle. Methods 36 cases of comminuted fracture of femoral condyle admitted from 2000 to 2005 were treated with retrograde interlocking intramedullary nails (n=19) or condylar plates (n=17). Union time of fracture and the recovery of knee function were observed respectively. Results The average union time of fracture of the cases treated with retrograde interlocking intramedullary nails was 102 days, and the score of knee function was 84.59. Whereas in the cases treated with condylar plates the union time of fracture was 91.35 days, and the score of knee function was 91.10. Significant differences in both union time of fracture and knee function between the two therapeutic methods were observed (P

Aiming at motion artifacts and off-resonance artifacts in multi-shot diffusion magnetic resonance imaging (MRI), we proposed a joint correction method in this paper to correct the two kinds of artifacts simultaneously without additional acquisition of navigation data and field map. We utilized the proposed method using multi-shot variable density spiral sequence to acquire MRI data and used auto-focusing technique for image deblurring. We also used direct method or iterative method to correct motion induced phase errors in the process of deblurring. In vivo MRI experiments demonstrated that the proposed method could effectively suppress motion artifacts and off-resonance artifacts and achieve images with fine structures. In addition, the scan time was not increased in applying the proposed method.
Artifacts ; Diffusion Magnetic Resonance Imaging ; Humans ; Image Processing, Computer-Assisted ; Motion ; Phantoms, Imaging ; Vibration

Acetamides ; adverse effects ; Adult ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; Case-Control Studies ; Humans ; Liver ; drug effects ; metabolism ; pathology ; Liver Diseases ; epidemiology ; Middle Aged ; Occupational Exposure ; adverse effects ; Young Adult

Adult ; Bronchoalveolar Lavage ; nursing ; Female ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; nursing ; therapy ; Retrospective Studies

Objective To investigate the roles of adrenomedullin(ADM) and endothelin-1(ET-1) in left-to-right shunt congenital heart disease(CHD) complicated with congestive heart failure(CHF).Methods Plasma ADM and ET-1 concentrations were measured by radioimmunoassay in heart failure stage and recovery period of 18 infants with left-to-right shunt CHD.Also 20 healthy infants acted as control group were measured.Results Plasma concentrations of ADM and EF-1 were significantly higher in heart failure stage of left-to-right CHD infants than those in their recovery period,and both in their heart failure stage and in their recovery period were higher than that in control group(P

Objective To explore the clinical features and operative method of complicated spinal tuberculosis. Methods Totally 562 inpatients suffering from spinal tuberculosis during January 1997 to December 2006 were reviewed retrospectively. A new definition of complicated spinal tuberculosis was made. Complicated spinal tuberculosis was classified into different types. Every type had its special surgical operation approach. Results Among the 87 cases being followed up for 45 months, 75 patients were totally recovered, and 12 patients were obvious improved on the mend according to Dr. Fang Xian-zhi’s standard. The classification of function which above good was 97.1%. The fusion rate was 96.3% during 87 patients who received bone grafting treatment 9 months ago. Posterior convex angle was 24? to 57?, average 35.3?. Conclusion We consider that complicated spinal tuberculosis will not be surgical contraindication. Different types of complicated spinal tuberculosis by different operation approach, and careful preparation before operation will gain good curative outcome.