Objective To investigate flow patterns at carotid bifurcation in vivo by combining computational fluid dynamics(CFD)and MR angiography imaging.Methods Seven subjects underwent contrast-enhanced MR angiography of carotid artery in Siemens 3.0 T MR.Flow patterns of the carotid artery bifurcation were calculated and visualized by combining MR vascular imaging post-processing and CFD. Results The flow patterns of the carotid bifurcations in 7 subjects were varied with different phases of a cardiac cycle.The turbulent flow and back flow occurred at bifurcation and proximal of internal carotid artery (ICA)and external carotid artery(ECA),their occurrence and conformation were varied with different phase of a cardiac cycle.The turbulent flow and back flow faded out quickly when the blood flow to the distal of ICA and ECA.Conclusion CFD combined with MR angiography can be utilized to visualize the cyclical change of flow patterns of carotid bifurcation with different phases of a cardiac cycle.

Objective To explore the association between genetic polymorphisms and haplotypes of tumor necrosis factor-related apoptosis inducing ligand (Trail) and ulcerative colitis (UC).Methods A total of 331 patients with UC and 832 age and sex-matched healthy controls were collected.After Trail gene was amplified by PCR,the genetic polymorphisms of three single nucleotides (G1525A/G1588A/C1595T) in 3' non coding regions of Trail gene were examined by direct sequencing.The relation between Trail haplotype and UC was analyzed.Results Compared with control group,the frequencies of variant allele A and genotype GA+ AA in Trail G1525A were significantly lower in UC group (both P＜0.01).The frequencies of variant allele A and T in Trail G1588A and C1595T were also significantly lower in UC group than that of control group,and the difference was statistically significant (both P ＜ 0.01 ).In mild and moderate UC patients,the frequencies of variant allele T and CT+TT in Trail C1595T were 49.15％ and 64.51％,in severe UC patients were 72.37％ and 84.21％,and the differences were significant between the two groups (OR=2.710 and 2.935,95％CI:1.598～4.596 and 1.188～7.249,all P ＜0.05).In severe UC patients,the frequency of variant allele A in Trail G1525A was 48.69％,which was higher than that of mild and moderate patients (35.16％,OR=1.750,95％CI:1.082～2.830,P=0.021).In UC group,the frequency of AAT haplotype was significantly lower than that of controls (43.09％ vs 58.41％,P＜0.01).The frequency of GAT haplotype was significantly higher in UC group (10.15％vs 0.18％,95％ CI:0.005 ～ 0.051,P＜ 0.01).Conclusion The genetic polymorphisms and haplotypes of Trail (G1525A/G1588A/C1595T) gene may be closely correlated with the susceptibility to UC.

Objective To investigate the spatial distribution and clustering areas of Budd-Chiari syndrome in Heze City,Shandong Province,and to provide epidemiological information for further exploring the etiology and related risk factors of the disease.Methods Detailed residential addresses of 342 cases of patients (residents of Heze City) with diaphragm type Budd-Chiari syndrome diagnosed between 1995 and 2004 in Heze Municipal Hospital,Heze Shan County Central Hospital,Affiliated Hospital of Xuzhou Medical College,Shandong Provincial Hospital and Beijing Xuanwu Hospital were collected.Geographic information system (GIS) was used as a platform for data management and display.The nearest neighbor index,Ripley's K(d) function,Ripley's L(d) function and the nearest neighbor clustering method were applied to detect the spatial characters of Budd-Chiari syndrome in Heze City,Shandong Province.Crimestat 3.0 was used for spatial analysis.Results The nearest neighbor distance analysis showed that the nearest neighbor index was 0.6767 (Z =-11.4387,P ＜ 0.01).That was an aggregation at the first-order spatial scale.Within the study area,the first clustering radius of Budd-Chiari syndrome was 6.66 km,and the first clustering strength was 5.40; the average radius of the strongest clustering area was 126.61 km,and the clustering strength was 12.52,while the biggest clustering radius was larger than 222 km.After corrected by population,the gathering strength was slightly higher than that before the correction.Ten first-order hot spots were formed,and 95％ confidence interval aggregation number was 7,which meant the results were statistically significant(P ＜ 0.05),main clustering areas are in Mudan District,Shan County and Juancheng.One secondorder hot spot was gathered based on the first-order hot spot.Conclusions Spatial distribution of Budd-Chiari syndrome in Heze City,Shandong Province has showed spatial aggregation and heterogeneity.This study has a great epidemiological significance for further exploring the cause of Budd-Chiari syndrome.

Enterocolitis, Necrotizing ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Infant, Newborn ; Intestinal Mucosa ; metabolism ; pathology ; Male ; Platelet Activating Factor ; metabolism

0.05),the mean FA on the left was higher than the right(t=1.912,P

OBJECTIVE Toprepareapolyclonalantibodyforspindlin1protein,anovelcancer related protein,and to provide the data for a better understanding of its functions and screening tu mor. METHODS Purifiedspindlin1proteinwasinjectedintorabbitstoproducethepolyclonalantiserumafter removing glutathione S-transferase (GST)from the fusion protein spindlin 1-GST that was expressed in Escherichia coli..The antiserum was purified through the Hitrap Protein A system,and the titer of spin-dlin 1 polyclonal antibody was detected by ELISA.The specificity of the polyclonal antibody was deter-minedbyWesternblottingandimmunohistochemistry.RESULTS Thetiterofspindlin1polyclonalanti-body was 1∶2000.Western blotting detection demonstrated that the spindlin 1 polyclonal antibody recog-nized myc-spindlin 1 reco mbinant fusion protein in HeLa cells transfected with pAdeasy-myc-spindlin 1 , which also corresponded with Myc.antibody.The HeLa cells were transfected with enhanced green fluo-rescence protein (EGFP)and spindlin 1 vector(pEGFP-C3-spindlin 1 ),which was confirmed by the in-dependent GFP fluorescence assay.The results of immunohistochemistry detection with the spindlin 1 polyclonal antibody suggested that spindlin 1 was mainly expressed in the nuclei of HeLa cells.More i m-portantly,in i mmunohistoche mical assays,the spindlin 1 antibody recognized nuclear spindlin 1 expres-sioninclinicalovariancancertissues.CONCLUSION Thespecificspindlin1polyclonalantibodyispre-pared,which may be used to detect cancer-related protein spindlin 1 in HeLa cells and ovarian cancer tissues.

OBJECTIVETo investigate the effects of the combination of musk and olibanum on the tight junction protein expressions in prostatic epithelial cells of normal and chronic prostatitis (CP) rats.

METHODSEighty male SD rats were randomly divided into 8 groups of equal number: normal control, normal musk, normal olibanum, normal musk + olibanum, CP model control, CP model musk, CP model olibanum, and CP model musk + olibanum. At 60 days after modeling, the rats in the control, musk, olibanum, and musk + olibanum groups were treated intragastrically with normal saline, musk (0.021 g per kg body weight per day), olibanum (1.05 g per kg body weight per day), or musk + olibanum respectively, all for 3 days. Then, all the rats were sacrificed and their prostate tissues harvested for detection of the expressions of the tight junction proteins Claudin-1, Claudin-3, Occludin, and ZO-1 in the prostatic epithelial cells by immunohistochemical staining.

RESULTSIn the CP models, only the expression of Claudin-1 was significantly increased. In the normal rats, the expression of Claudin-1 was remarkably upregulated after treated with musk (824.6 ± 393.3, P < 0.05), olibanum (982.0 ± 334.0, P < 0.05), and musk + olibanum (1088.1 ± 640.2, P < 0.01); that of Claudin-3 was elevated markedly by olibanum (1 009.5 ± 243.6, P < 0.05) and insignificantly by musk (597.5 ± 80.7), but the increasing effect of olibanum was reduced by musk + olibanum (678.4 ± 255.1). No statistically significant differences were found in the expression of Occludin among the rats treated with musk (693.0 ± 424.8), olibanum (732.1 ± 302.0), and musk + olibanum (560.2 ± 202.3), or in that of ZO-1 in the animals treated with musk (290.0 ± 166.8) and olibanum (419.7 ± 108.1), but the latter was markedly decreased in the musk + olibanum group (197.7 ± 98.2, P < 0.05). In the CP rat models, both the expressions of Claudin-1 (823.0 ± 100.1, P < 0.01) and Occludin (1160.0 ± 32.2, P < 0.05) were significantly increased. The expression of Claudin-1 was remarkably down-regulated by musk (764.9 ± 179.0), olibanum (468.4 ± 220.4), and musk + olibanum (335.1 ± 204.0) (all P < 0.05), but that of Claudin-3 up-regulated by musk (744.6 ± 94.5) and olibanum (700.1 ± 223.7) (both P < 0.05). The expression of Occludin was reduced by musk (615.0 ± 221.0), olibanum (749.6 ± 321.7), and musk + olibanum (505.8 ± 523.7), while that of ZO-1 increased by olibaum (443.2 ± 44.9) and decreased by musk + olibanum (213.5 ± 24.9, P < 0.05).

CONCLUSIONIn physiological and pathological conditions, the combination of musk and olibanum acts on the expressions of tight junction proteins in prostate epithelial cells in a selective and dual-targeting manner, promoting their permeability by down-regulating the expression of ZO-1 and maintaining their structural stability by regulating the expressions of Claudin-1, Claudin-3, and Occludin.

Animals ; Claudins ; metabolism ; Down-Regulation ; Epithelial Cells ; drug effects ; Fatty Acids, Monounsaturated ; chemistry ; Frankincense ; chemistry ; Male ; Occludin ; metabolism ; Prostate ; cytology ; Prostatitis ; Rats ; Rats, Sprague-Dawley ; Tight Junction Proteins ; metabolism ; Up-Regulation

Aneurysm, False ; etiology ; surgery ; Blood Loss, Surgical ; Carcinoma ; Carotid Arteries ; Carotid Artery Diseases ; etiology ; surgery ; Humans ; Nasopharyngeal Neoplasms ; radiotherapy ; Rupture

BACKGROUNDInhibition of tumor growth by endostatin has been shown to be an effective strategy in cancer therapy in mice. However, its widespread application has been hampered by difficulties in a large-scale production of the recombinant endostatin protein, rapid loss bioactivity of the protein, and the cumbersome daily administration. These limitations could be resolved by in vivo delivery and expression of the endostatin gene. In this study, we observed the effect and advantage of endostatin gene therapy mediated by a recombinant adenoviral vector (Ad/hEndo) on the growth of hepatocellular carcinoma BEL-7402 xenografted tumors, comparison with recombinant endostatin protein.

METHODSHepatocellular carcinoma BEL-7402 cells were inoculated subcutaneously in the flank of Balb/c nude mice. Nine days after tumor cell inoculation, animals were given a cycle of four courses of intra-tumoral injections of Ad/hEndo of 5 x 10(8) pfu (low-dose group) and 1 x 10(9) pfu (high-dose group) at intervals of six days, respectively. Recombinant human endostatin protein (rhEndo) was administrated daily subcutaneously at a dose of 10 mg.kg(-1).d(-1) at a site nearby the tumor for ten days. The expression of endostatin mRNA in tumor tissue was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) after Ad/hEndo injection. Dynamic changes of concentration of endostatin protein in tumor tissue were quantitated by enzyme-linked immunosorbent assay (ELISA).

RESULTSAfter 4 courses of treatment, the tumor growth rates of high-dose treated group with 1 x 10(9) pfu of Ad/hEndo were inhibited by 42.26% compared with the Ad/LacZ control group (P = 0.001) and by 46.26% compared with the NIH buffer control group (P = 0.003), respectively. However, in this study, Ad/hEndo at low dose of 5 x 10(8) pfu failed to demonstrate significant inhibition of tumor growth, compared with control groups. After daily administration of recombinant human endostatin protein (rhEndo) for 9 days, the ratio of T/C (rhEndo group versus PBS group) was less than 47%. However, two days after rhEndo treatment ceased, the ratio of T/C was more than 50%. The peak of expression of endostatin mRNA in tumor tissue was at 2 or 3 days after administration intratumorally with Ad/hEndo of 1 x 10(9) pfu and gradually dropped undetectable by day 7. Dynamic analysis of endostatin concentration in tumor tissue showed that the highest level of mRNA is up at the third day after injection, and dropped to basal level three weeks later.

CONCLUSIONSEndostatin gene therapy mediated by a recombinant adenoviral vector had significantly inhibited the growth of hepatocellular carcinoma BEL-7402 xenografted tumors at a high dose of 1 x 10(9) pfu compared with other groups. The analysis of dynamic expression of endostatin in vivo indicated that Ad/hEndo had acquired a high-level, relatively long-term expression in vivo and bioactivity capability.

Adenoviridae ; genetics ; Animals ; Cell Line, Tumor ; Endostatins ; analysis ; genetics ; therapeutic use ; Genetic Therapy ; Humans ; Liver Neoplasms, Experimental ; therapy ; Mice ; Mice, Nude ; Neoplasm Transplantation ; RNA, Messenger ; analysis ; Recombinant Proteins ; therapeutic use ; Transplantation, Heterologous

This study was designed to evaluate the role of bcl-2 transcriptional regulation induced by calmodulin I (CaM I) in pressure overload rat hypertrophic hearts. The model of hypertensive Sprague-Dawley rat was established by abdominal aortic constriction. The hearts were collected four weeks after abdominal aortic constriction. Velocity and isopyknic gradient centrifugation was employed to fractionate rat myocardial nuclei. Western blot analysis revealed a marked increase in phosphorylated cAMP response-element binding protein (pCREB) of cardiac hypertrophy group compared with that in control group (P<0.05), while the protein level of cAMP response-element binding protein (CREB) was constant (P>0.05). Immunohistochemistry results showed a significant increase of CaM I protein in cardiac hypertrophy group relative to the control group (P<0.05). Nuclear run off transcription assay displayed a significant increase in bcl-2 mRNA treated with trifluoperazne compared with non-drug treatment (P<0.05). The results obtained suggest that the transcription of bcl-2 is possibly regulated by CaM I hypertrophic rat hearts, and CREB phosphorylation seems to be a minor factor in bcl-2 transcriptional regulation.
Animals ; Aorta, Abdominal ; pathology ; Calmodulin ; physiology ; Cardiomegaly ; metabolism ; physiopathology ; Constriction ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Male ; Phosphorylation ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley