BACKGROUND: Statins can block many intracellular signal transductive pathways and suppress the proliferation of various cells by affecting the synthesis of mevalonic acid and the translation following modification of some membrane-connecting proteins.OBJETCIVE: To investigate the influence of simvastatin on the proliferation of lung fibroblasts, the synthesis of collagen and the secretion of matrix metalproteinase-2 (MMP-2).DESIGN:Completely randomized controlled study.SETTING: At Organ Transplanting Research Institute of Fuwai Cardiovasular Diseases Hospital, Peking Union Medical College.MATERIALS: This study was carried out at the Laboratory of Cardiology of Beijing Union Medical College hospital, Peking Union Medical College from June 2004 to October 2004. Lung fibroblasts derived from neonatal SD rats were co-cultured in vitro with different dosage of simvastatin of 0, 1, 5, 10,50 μmol/L, and 50 μmol/L simvastatin + 200 μmol/L mevalonic acid.METHODS: Lung fibroblasts deriving from neonatal SD rat were co-cultured with different dosage of simvastatine in vitro. Methyl thiazolyl tetrazolium colorinetry was used to detect the cell proliferation, and cell immunohistochemical assay was used to determine the collagen synthesis and meanwhile,MMP-2 content in supernatant was examined with enzyme-linked immunosorbent assay.MAIN OUTCOME MEASURES: The proliferation of fibroblasts, the synthesis of collagen and the secretion of MMP-2 due to different dosage of simvastatin intervention and simvastatin combined with mevalonic acid.presenting the expression of type Ⅰ, Ⅲ of collagen in lung fibroblasts and the level of MMP-2 in 5, 10, 50 μmol/L simvastatin group were obviously lower than those of 0 μmol/L simvastatin group (0. 520 ± 0.010, 0. 334 ± 0.011,0.260±0.012, 0.111±0.011; 0.508±0.011, 0.324±0.014, 0.232±0.015, 0. 083 ±0. 015; 0.445 ±0. 017, 0. 305 ±0. 015, 0.216 ±0. 015,0.068±0.012; 0.561±0.013, 0.361 ±0.012, 0.289±0.012, 0.140value, the mean absorbency( A value) in lung fibroblasts and the level of MMP-2 in 50 μmol/L simvastatin + 200 μmol/L mevalonic acid group were obviously higher than that of the 50 μmol/L simvastatin group(0. 567±0.015, 0.354±0.014, 0.283±0.012, 0.138±0.011, t=4.715-10.950, P ＜ 0.01).CONCLUSION: Simvastatin could suppress the fibroblast proliferation and collagen synthesis, attenuate the secretion of MMP-2 and suppress consequently the adhesion and migration of lung fibroblasts; moreover, it has the capability of anti-cell proliferation by affecting the mevalonic acid pathway.

Objective To investigate the application of delayed sternal closure in the process of open heart surgery.Methods Delayed sternal closure was performed from Mar.2000 to Jun.2005 in 11 patients(4 males and 7 females,aged 16-67 years with an average of 42 years) after cardiac surgery.The indication included: cardiac dilatation(4 cases),intractable arrhythmia(4 cases),continuous bleeding(2 cases) and severe pulmonary edema(1 case).During the open chest period,the wound was covered with 3-layers of latex temporarily,and the delayed sternal closure was performed when bleeding was controlled,heart size reduced,and hemodynamic condition became stable.Results Except two patients died of acute kidney failure during open chest period,delayed sternal closure was successfully carried out in 9 patients 8-43 hours(23.7?11.0) after cardiac surgery.Except one patient died of cardiac arrest 21 days after operation,the 8 others survived and were discharged from ICU 1-8 days(3.9?2.4) after sternal closure,and the wounds healed well.Follow-up for the 8 survivors revealed an improvement of NYHA class(class Ⅰfor 4 cases,class Ⅱfor 2 cases,and class Ⅲ for 2 cases).Conclusion The delayed sternal closure is an effective method in the treatment of postoperative cardiac compression,severe bleeding and arrhythmia,and it does not increase the incidence of complications such as sternal infection.

Objective:To study the antitumor effect and mechanism of cocultured CIK cells modified with IL-24 gene and autologous DCs on HL-60 cells in vitro.Methods:DCs and CIK cells were prepared routinely from human peripheral blood mononuclear cells ( PBMC).IL-24 gene was transferred into CIK cells via electroporation.The cells obtained were named CIK-IL24.RT-PCR and ELISA were used to evaluate expression of IL-24 gene in transfected CIK cells.The phenotypic changes of CIK cells were identified by flowcytometry analysis.The concentration of IFN-γ and TNF-α in supernatant of CIK was determined by ELISA.FCM was used to determine the cytotoxicity of cocultured CIK cells modified with IL-24 gene and autologous DCs against HL-60 cells.Results:Eukaryotic expressing plasmid pcDNA3.0-IL24 was transferred into CIK cells successfully via electroporation.The expressing rate of CD3~+、CD3~+CD56~+ cells had no significant change in CIK cells.However,the rate of CD4~+CD25~+ cells was significantly decreased compared with that of the control group.Expression of adhesion molecules CD54,CXCR4 were significantly increased on CD3+CD56+ cells.CIK-IL24 cells produced markedly higher levels of IFN-γ and TNF-α as compared with the CIK cells.By comparison with non-transfected CIK cells co-cultured with DCs,transfected CIK cells co-cultured with DCs had a significantly higher lytic activity against HL-60 cells.Conclusion:IL-24 gene modification can enhance the anti-tumoral immunity of CIK cells,the mechanism of which might be related to the increased secretion of IFN-γ,TNF-α,up-regulation of adhesion molecule expression,and reduction of the rate of CD4~+CD25~+ cells in CIK cells.

Objective To study the antitumor effect and mechanism of co-cultured cytokine-induced killer(CIK) cells and autologous DC modified with IL-24 gene on A549 cells in vitro. Methods DC and CIK cells were prepared routinely from human peripheral blood mononuclear cells(PBMC). Recombinant adenovirus vector pAdEasy-1-pTrack-CMV-IL-24 was extracted from DH5α, it was lineared with Pac I and transfected into A293 cells, and then the IL-24 recombined adenovirus(Ad-IL-24) was obtained. Ad-IL-24 was used to infect DC. The cells obtained were named DC-IL-24. RT-PCR and ELISA were used to evaluate the expression of IL-24 gene in transfected DC. The phenotypes change of DC were identified by flow cytometry analysis, the concen-tration of IL-12 and TNF-α in supernatant of DC were determined by EIJSA. The ability of CIK producing per-forin was measured by homolysis method. FCM was used to determine the cytotoxicity of cocultured CIK cells and autologous DC modified with IL-24 gene to A549 cells. Results We obtained the high titre of Ad-IL-24.IL-24 gene was transfered into DC successfully via Ad-IL-24. The green fluorescence was observed on DC by fluorescence microscope. The expression rate of CD80, CD83, HI.A-DR, CD40, CXCR4 on DC-IL-24 was sig-nificantly increased compared with that of the control group. DC-IL-24 produced markedly higher levels of IL-12 and TNF-α as compared with DC. DC-IL-24 can enhance the ability of CIK cells producing perforin. On com-parison with non-transfected DC co-cultured with CIK cells, transfected DC co-cultured with CIK cells had a sig-nificantly higher lytic activity against A549 cells. Conclusion IL-24 gene modification can enhance the anti-tu-moral immunity of DC. The mechanism of which might be related to the increased secretion of IL-12 and TNF-α, up-regulation expression of co-stimulatory molecules and MHC Ⅱ class molecules on DC, promoting the acti-vation and maturation of DC, and then enhancing CIK cells to generate specific anti-tumoral immunity.

Objective To summarize the experience of perioperative management of coronary artery bypass graft(CABG) in high-risk patients with coronary heart disease.Methods The clinical data of 164 patients underwent CABG from March 2004 to November 2005 were analyzed.Of 164 patients,151(92.05%) cases had severe coronary heart disease.Results Totally 144 artery-vessels and 353 venous-vessels were transplanted to patients.Combined operative procedures included 11 cases of ventricular aneurysm resect,5 cases of valve replacement,5 cases of valve repair,and 1 case of surgical repair of perforation of ventricular septum.Post-operative complications included 2 cases of low cardiac output,1 case of respiratory failure,1 case of renal failure,and 1 case of cerbral infarction.Two patients died and the operative mortality was 1.22%.Conclusion CABG procedure is safe in the treatment of high-risk patients with coronary heart disease.Proper preoperative and perioperative treatment can improve the curative effects significantly.

Objective: To explore the effect of simvastatin combined with cyclosporin A treatment on the development of obliterative bronchiolitis in a murine heterotopic tracheal transplantation model. Methods: Murine tracheals were heterotopically transplanted from BALB/c donors into C57BL/6 recipients. Transplanted animals received either control chow, chow containing simvastatin, chow containing cyclosporine A, or chow containing simvastatin and cyclosporine A. beginning immediately after transplantation. Epithelial loss and luminal obstruction were analyzed by morphometry. Immunohistochemistry assay was used for quantifying inflammatory cell infiltration and expression of chemokine in tracheal allografts. collagen deposition was studied by picro sirius red staining.Group t test was used to calculate the difference between groups. Results: simvastatin combined with cyclosporin A treatment reduced chemokine(MCP-1, RANTES)release, inhibited CD4⁺ and CD8⁺ T cells and macrophages accumulation in tracheal allografts, resulting in limited bronchial inflammation and diminished epithelial loss. simvastatin plus cyclosporin A treatment also inhibited proliferation of myofibroblast cells, reduced MMP-2 release and decreased the amounts of type I and III collagen deposition, resulting in preserved luminal patency and inhibited development of OB compared with those of controls. Conclusions When simvastatin was used in combination with CsA, the development of OB was significantly inhibited.

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