Objective To investigate the expression of protease activated receptors(PARs)in mast cells.Methods Reverse transcription polymerase chain reaction(RT-PCR),flowcytometry and immunofluorescent cell staining were used to detect the expression of PARs in mast cell lines P815 and MC/9 at the levels of protein and mRNA.Results Both the P815 and MC/9 of mast cell lines expressed PAR-1,PAR-2,PAR-3 and PAR-4 at either protein or mRNA level.Conclusion The expression of all the four PARs in mast cells were detectable,which may be of significance for the further study on the function of PARs in mast cells.

Objective To investigate the effect of purified single clonally mesenchymal cells (SCMSCs) on cardiac function in rat models and also try to find out wheter SCMSCs serve as a better source for transplantation than UMSCs, BM-MNCs and PB-MNCs. Methods SCMSCs were isolated, cultured, purified, cloned and expanded. UMSCs were isolated primarily by their tight adherence to the culture dishes. BM-MNCs and PB-MNCs were prepared by Ficoll-Hypaque gradient centrifugation. All cell characters were verified by fluorescence- activated cell sorter (FACS). A total of 5?10~6 PB-MNCs, BM-MNCs, UMSCs, and SCMSCs were transplanted into the ischemic zone immediately after MI. The cardiac function was evaluated by hemodynamic technique 1 month after the transplantation. The vessel density and cell differentiation were determined by histological techniques. Results SCMSCs expressed over 99% of the mesenchymal cell surface protein and none of the hematopoietic stem cell surface protein. Hemodynamics showed that transplantation of snMSC led to the greatest improvement in cardiac function, compared with PB-MNCs, BM-MNCs, and UMSCs transplantation. In consistence with cardiac function recovery, SCMSCs transplantation resulted in the greatest angiogenesis in the ischemic wall, and the greatest number of transplanted SCMSCs expressed these cardiomyocyte proteins, or vascular endothelial cell marker, in comparison with the other heterogeneous cells. Conclusion Transplantation of single clonally purified non-hematopoietic mesenchymal stem cells from human bone marrow showed the greatest imporvement in cardiac function compared to UMSC, BM-MNC, and PB-MNC in this study.

Objective To investigate whether host collateral vessels play a role in the cardiac repair induced by intracoronary injection of bone marrow mononuclear cell (BMMNCs) in a swine myocardial infarction (MI) model. Methods MI was induced in swine models by ligation of left anterior descending artery. Two weeks later, twenty animals with Rentrop score=0 (R0) or Rentrop score=1 (R1) received either intracoronary PBS or BM-MNCs (2?10~8 cells) infusion. Results At 6 weeks after MI, Rentrop score was significantly increased in the R1+BMT group. Ejection fraction and fractional shortening evaluated by echocardiography were increased in both R0+BMT and R1+BMT groups and the greatest improvements were seen in the R1+BMT group. Consistent with the changes in cardiac function and Rentrop score, the greatest degree of angiogenesis and cell count of engrafted BM-MNCs occurred in the R1+BMT group 6 weeks post MI. Conclusion The existence of intrinsic collateral vessels contributes to the beneficial effects for cardiac repair post MI by intracoronary BM-MNCs transplantation.

Objective The 47 myocardial infarction patients (male 41, female 6, average ages 55 7?9 7 years) with silent myocardial ischemia (type Ⅱ SMI) were chosen The relationship was studied between the history of myocardial infarction (MI)and the degree of coronary arterial stenosis and the residual degree of stenosis after percataneous transluminal coronary angioplasty (PTCA) Methods All patients with MI were performed with exercise testing electrocardiogram to investigate the degree of myocardial ischemia The patients were also studied the degree of coronary arterial stenosis before and after PTCA Results The results showed before PTCA, the degree of the patients′ coronary arterial stenosis with a period of no more than three months of MI was higher than that of those patients′ with a period of over three months( P

Objective To evaluate the effect of intravenous Gd-DTPA on 3.0T proton MR spectroscopy (MRS) water suppression and shimming. Methods Prospective study of proton MRS was performed with GE Signa Excite HD 3.0T system and eight-channel phased-array coils with PRESS sequence (head, liver and kidney, respectively). Routine auto prescan program was operated to record full width half maximum (FWHM) and water suppression (WS%). Routine scan was performed after injection of Gd-DTPA, then prescan program was reoperated to record FWHM and WS%. The data of FWHM and WS% in head, liver and kidney were compared between before and after injection of Gd-DTPA with the Wilcoxon matched pairs signed test. Results WS% of spectroscopy of head and liver after administration of Gd-DTPA decreased significantly (T_+=12, T_-=66, P=0.02; T_+=0, T_-=45, P=0.007). The effect of shimming of kidney after administration of Gd-DTPA was poor (T_+=0, T_-=435, P<0.001) and WS% of spectroscopy of kidney after administration of Gd-DTPA decreased significantly (T_+=0, T_-=435, P<0.001). Conclusion WS% of spectroscopy in head, liver and kidney can be impacted negatively by Gd-DTPA. Gd-DTPA has great influence on shimming of spectroscopy of kidney, but has little influence on shimming of spectroscopy of head and liver. It is better to acquire MRS data before administration of contrast medium in kidney.

A novel electrogenerated chemiluminescence (ECL) sensor for the determination of metoclopramide was developed by employing ruthenium complex as an ECL signal producer and an ordered mesoporous carbon (OMC) material as modified material. The ECL sensor was fabricated by adsorption ruthenium complex into a mixture of OMC and Nafion, which showed good electrochemical and ECL behaviors. It was found that the ECL intensity of the sensor fabricated was greatly enhanced in the presence of metoclopramide. Based on this finding, a highly sensitive and reproducible ECL method was developed for the determination of metoclopramide. The result showed that the ECL intensity was linear with the concentration of metoclopramide in the range from 1.0×10-10 to 5.0×10-7M and the detection limit was 3×10-11M. The ECL sensor exhibited a long-term stability and a fine reproducibility with relative standard deviation of 1.0 % for 1.0×10-10M metoclopramide in 18 continuous determinations. The developed method has been applied to the determination of metoclopramide in tablet samples with satisfactory results.

Objective:To investigate the effect of activation of proteinase activated receptor(PAR)2 on mediator release from mast cells.Methods:P815 cells(a mast cell line) were challenged with various concentrations of PAR-2 agonist peptide,trypsin,tryptase or elasetase with or without PAR-2 antagonist peptide.The supernatants were collected and analyzed by enzyme-linked immunosorbent assay(ELISA) to detect the quantity of released IL-4,IL-6 and histamine.Results:PAR-2 agonist peptide,trypsin or tryptase induced a concentration-dependent IL-4,but not histamine release from P815 mast cells.Trypsin and tryptase induced IL-4 release from the mast cells was blocked by addition of PAR-2 antagonist peptide.No IL-4,IL-6 and histamine release was observed when P815 cells were incubated with elasetase.Conclusion:Induction of IL-4 release from mast cells by trypsin and tryptase through activation of PAR-2 added some novel information on the hypothesis of self-amplification mechanism of mast cell activation.

Objective:To investigate the modulatory effect of IL-29 on trypsin-induced protease activated receptors (PARs) ex-pression on P815 mast cell.Methods:After P815 mast cells were challenged with different concentrations of IL-29 alone or combined with trypsin for 2 h, 6 h and 16 h, the challenged cells were collected and analysed by flow cytometry to detect the protein expression of PARs on P815 cells, and analysed by real time RT-PCR to detect the mRNA expression of PARs on P 815 cells.Results:Compared with the corresponding control , IL-29 induced significantly decreased expression of PAR-1 at protein and mRNA level on P815 cells, and upregulated PAR-3, PAR-4 mRNA level on P815 cells, whereas IL-29 did little effect on the expressions of PAR-2,3,4 at protein level on P815 cells accordingly.Preincubation of mast cell with IL-29 did not alter trypsin-induced PAR-1 expression on P815 cells, whereas up-regulated expression of PAR-2, 3, 4 were detected when P815 cell were pre-treated with IL-29 before being challenged with trypsin compared with the corresponding control .Conclusion: IL-29 can upregulate trypsin-induced PARs expression on mast cells through which participated in mast cell related inflammation .

Objective To investigate the potential influence of TNF on the expression of protease activated receptor (PAR)-1,2,3 and 4 by using P815 mast cells. Methods After being challenged with various concentrations of TNF for 2 h, 6 h and 16 h, the P815 mast cells were treated with or without Triton X-100 and the PAR expressions were detected by flow cytometry and immunofluorescent microscopy. Results Compared with the corresponding controls, TNF concentration-dependently upregulated expressions of PAR-2 and PAR-4 both in Triton X-100-treated and the untreated groups, but had no significant effect on the expression of PAR-1 and PAR-3. Moreover, no significantly different expressions of TNF-induced PAR-1, 2, 3 were observed between Triton X-100-treated and the untreated groups, whereas Triton X-100-treated PAR-4 expressions were significantly enhanced by TNF compared with the Triton X-100-untreated ones. Conclusion TNF can up-regulate PAR-2, 4 expression of P815 mast cells but has little effect on the expression of PAR-1, 3 correspondingly. And Triton X-100 treatment had no significant effect on TNF-modulated expression of PARs in P815 mast cells.

Objective To investigate the effects of human IL-18 gene combined with diterpenoid alkaloids in inhibiting the proliferation and inducing the apoptosis of tongue squamous carcinoma cells Tscca.Methods We constructed recombinant plasmid pEGFPN3-IL-18 and tranfected it into tongue squamous carcinoma cells Tscca.The transduction efficiency of the target cells was detected by fluorescent microscopy,cytotoxic effect of IL-18 gene with diterpenoid alkaloids on Tscca was detected by MTT assay,and apoptosis was detected by flow cytometry. Western blot was employed to examine the expression level of cellular signal-regulated kinase Akt/p-Akt.Results The tongue squamous cells Tscca which transfected pEGFPN3-IL-18 had significantly increased apoptosis compared with non-transfected cells (P<0 .05 ).Tongue carcinoma squamous cells cultured with diterpenoid alkaloids at the concentrations of 0 .2 ,0 .4 and 0 .6 mg/mL had significantly increased apoptosis in a dose-dependent manner (P<0.05).Human IL-18 gene combined with diterpenoid alkaloids for 48 hours inhibited significantly Tscca in a concentration-dependent manner compared with diterpenoid alkaloids alone (P<0 .05 ).The two in combination could also decrease the protein level of p-Akt dose-dependently.Conclusion The combination of pEGFPN3-IL-18 and diterpenoid alkaloids has a synergistic effect in inhibiting the growth of tongue squamous carcinoma cells Tscca.