ObjectiveTo investigate the effects of 5-Aza-CdR on the transcriptional regulation through methylation of the DNA promoter protocadherin 8(PCDHg) gene in pancreatic cancer cell line Capan-2.The Capan-2 retardation in growth rate and apoptosis were assessed in when administered 5-Aza-CdR and the chemotherapy agent,gemcitabine.MethodsMTT and flow cytometry were used to analyze the cell growth inhibition and apoptosis when treated with 5-Aza-CdR or in combination with gemcitabine.Methylation-specific PCR,RT-PCR and western blot were performed to detect methylation state,mRNA and protein respectively of PCDH8 gene in 5-Aza-CdR-treated Capan-2cells.Results Capan-2 cells treated with 5-Aza-CdR showed a slower growth rate,and a significant growth inhibition when given both 5-Aza-CdR in combination with gemcitabine.Compared with single drug administration and control,5-Aza-CdR together with gemcitabine can induce a stronger apoptosis signal.Different concentrations 5-Aza-CdR of were able to reverse methylation,restore mRNA and protein levels of PCDH8 in Capan-2.Conclusion5 Aza-CdR may demethylate the PCDH8 gene,which would effectively remove the gene silencing caused by high methylation,and thus induce gene mRNA transcription and protein expression to inhibit cell growth and have collaborative antitumor functions with gemcitabine.

Objective To construct replication deficient recombinant adenovirus of human monocyte chemoattractant protein-1(MCP-1) by homologous recombination.Methods The cDNA of MCP-1 gene was obtained from human liver tissue by using RT-PCR,and was subcloned into a transfer plasmid pAdTrack-CMV.The linearized recombinant transfer plasmid pAdTrack-CMV-MCP-1 was co-transformed with backbone vector pAdEasy-1 into bacteria BJ5183 for recombinant adenoviral plasmid.The recombinant adenoviral plasmid was linearized and then transfected into HEK293 packing cells to produce virus particles.The recombinant adenovirus was detected by using PCR.Results The recombinant adenoviral plasmid was successfully established and confirmed by restriction endonuclease digestion.The expression of green fluorescent protein(GFP) was observed on the 5th day after transfection.The fragment of MCP1 gene was amplified by PCR.Conclusion The achievement of recombinant adenoviral plasmid and recombinant adenovirus of MCP-1 lay a foundation for further investigation of the function and application of MCP-1.

1)were up-regulated in test and 184 genes(RA1.5)and down-regulation(RA

Objective To investigate the antenatal influencing factors associated with birth weight of twins and the risk factors for the occurrence of discordant twins. Methods Totally, 834 twins delivered at the Third Affiliated Hospital from January 2000 to December 2009 and the First Affiliated Hospital of SUN Yat-sen University, from January 2000 to June 2009, were recruited in the study. The diagnosis criteria of discordant twins was intrapair birth weight difference more than 25 %.Antenatal factors on birth weight were retrospectively investigated among twins and discordant twins respectively with Logistic analysis. Results (1) There were several factors associated with birth weight of twins, including maternal age, conceptive style, chorionicity, gestational diabetes mellitus and placental cord insertion ( P = 0. 021, 0. 000, 0. 000, 0. 012 and 0. 017, respectively). While hypertensive disorder in pregnancy, parity and fetal gender differences were not associated with the birth weight of twins (P＞0.05). Hypertensive disorder in pregnancy (OR=2. 600, 95% CI:1. 566-4. 316) and monochorionicity (OR= 1. 833, 95% CI: 1.010-3. 582) were risk factors of discordant twins. All 834 twin pregnancies were divided into 4 groups according to materal age, ＜25 yrs (n=69), 25-30 yrs (n=312), 30-35 yrs (n=325) and ≥35 yrs (n=128). The average birth weight of twins were (2205±483) g, (2347±406) g, (2381±439) g and (2352±455) g. Significant difference was found bewteen every two groups (P＜0. 05) except that between the 30-35 yrs and ≥35 yrs group (P＞0.05). Among twins of different gender (n=270), the average birth weight of boys and girls were (2416±-514) g and (2322±488) g, and that of female twins (n=272) and male twins (n=292) were (2301±418) g and (2381±428) g. Significant difference was shown between every two groups (P＜0.05) except that between the male twins and male fetus of twins of different gender (P＞0.05).Conclusions The fetal birth weight of twins decreases remarkably when maternal age ＜25, or monochorionicity twins, or complicated with marginal or velamentous placental cord insertion or female pairs.However, birth weight of twins would increase if the pregnancy conceived by assisted reproductive technology or complicated with gestational diabetes mellitus. Twin pregnancy complicated with hypertensive disorder in pregnancy or monochorionicity should alert the clinicians of discordance twins.

AIM: The effect of urotensin II (U-II) on proliferation of cultured pulmonary arterial smooth muscle cells (PASMCs) of rabbits and its mechanism are investigated. METHODS: PASMCs were isolated using explant technique. RPASMCs were incubated in serum-free medium with different concentrations of nicardipine, calcimodulin antagonist W 7, PKC inhibitor H 7 or MAPK inhibitor (PD 98059 ), with or without U-II. RPASMC proliferation was examined by MTT [3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay and by the increase in [ 3H]-thymidine incorporation into DNA. RESULTS: U-II (10 -9 mol/L-10 -7 mol/L) increased A value of PASMCs by MTT assay and [ 3H]-thymidine incorporation in PASMCs in a dose-dependent manner. U-II induced a maximal effect at a concentration of 10 -7 mol/L. A value and [ 3H]-thymidine incorporation rose 42 9% and 68 5% ( P

Objective To determine the usefulness of real time three-dimensional transesophageal echocardiography(RT-3D TEE) in the preoperative assessment of mitral valve (MV) pathology by comparing images with surgical findings and to evaluate the function of MV postoperatively. Methods Nineteen consecutive adult patients with established diagnosis of mitral regurgitation(MR) scheduled for surgical correction were enrolled. Intraoperative 2D and 3D transesophageal echocardiography(TEE) were performed. All the 3D images were compared with findings obtained from direct surgical inspection. Postoperative RT-3D TEE was performed immediately to evaluate outcomes of mitral annuloplasty. Results Superb 3D-TEE en face views of the MV were obtained in all patients. Correct diagnoses of MV pathology in agreement with the surgical findings were made in 16 of 19 patients (84. 2%), however in the remaining 3 patients, the diagnoses were incorrect despite good image quality. In one, RT-3D TEE diagnosed prolapse of A2 segment and P2 scallop,which was not confirmed at surgery, but MV degeneration and annular dilatation were observed. In another one, RT-3D TEE revealed prolapse of P3 scallop, while at surgery prolapse involved P2 and P3 with ruptured chordae. In the rest one,surgically detected prolapse of P2 was missed by RT-3D TEE. On the other hand with 2D TEE,the diagnoses correlated poorly with surgical findings,only 10 patients were accurately diagnosed (52.6%). In some patients,2D TEE was able to identify the MVP, but it had difficulty in defining the exact location of the prolapsed segment or scallop. The severity of MR decreased significantly after surgery. MV repair was successful in all patients except one(5.1%), in whom moderate to severe MR was still present and MV replacement was conducted subsequently as an alternative.Conclusions RT-3D TEE provides excellent imaging of MV components, including the anterior and posterior leaflets, as well as annulus and subvalvular structures, which can be especially helpful in planning the most appropriate surgery strategy. RT-3D TEE offers exact anatomic characteristics of mitral annuloplasty rings and bands, providing additional information for the evaluation of surgical outcomes.

OBJECTIVETo study the effection of suppression murine melanoma growth by Intratumor injection of recombinant attenuated salmonella carrying heat shock protein 70 and herpes simplex virus thymidine kinase genes.

METHODSPlasmids PCMV-mtHSP70-IRES-TK were electro-transferred into salmonella typhimurium SL7207 to construct recombinant salmonella typhimurium. In vivo, Recombinant bacteria were injected into the mouse melanoma and the antitumor effection was observed. The survival period was recorded and safety analysis for this vaccine in each group.

RESULTSIn vivo, the mtHSP70/HSV-tk recombinant bacteria can suppress tumor growth significantly and extend survival. After recombinant Salmonella, 10(9) CFU/mL, was administered as an intratumoral injection, No diarrhea were observed. During therapy, body weight did not change markedly.

CONCLUSIONResults of the animal experiment suggests intratumor injection of recombinant attenuated salmonella typhimurium containing mtHSP70 and HSV-tk genes, has targeting ability against B16 tumor cell and could significantly inhibit tumor growth .

Animals ; Bacterial Proteins ; genetics ; immunology ; Cancer Vaccines ; genetics ; immunology ; pharmacology ; Genetic Therapy ; methods ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Melanoma, Experimental ; microbiology ; pathology ; therapy ; Mice ; Mice, Inbred C57BL ; Mycobacterium tuberculosis ; genetics ; Salmonella typhimurium ; genetics ; immunology ; Simplexvirus ; enzymology ; genetics ; Skin Neoplasms ; therapy ; Thymidine Kinase ; genetics ; immunology ; Vaccines, Attenuated ; genetics ; immunology ; pharmacology ; Vaccines, DNA ; genetics ; immunology ; pharmacology