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1.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 1434-1438, 2009.
Article in Chinese | WPRIM | ID: wpr-405137

ABSTRACT

Objective To investigate the effects of different dialysates on expression of protein kinase C-δ (PKCδ) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKCδ specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKCδ mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group , normal control group and fluid A+powder B+rottlerin group (P<0.05). Compared with normal control group, blank control group and fluid A+powder B+rottlerin group, the expression of PKCδ mRNA and protein of U937 cells in fluid A+powder B group were significantly increased (P<0.05). There was no significant difference in cell apoptosis rates and expression of PKCδ mRNA and protein between fluid A+fluid B group and blank control group, normal control group and fluid A+fluid B+rottlerin group (P>0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKCδ. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.

2.
Chinese Journal of General Surgery ; (12): 496-499, 2008.
Article in Chinese | WPRIM | ID: wpr-396420

ABSTRACT

Objective To investigate the causes of acute renal failure(ARF)after orthtopic liver transplantation(OLT)in patients of acute liver failure(ALF)and the effects of systemic therapy based on continuous renal replacement(CRRT).Methods Clinical data of 412 patients who underwent liver transplantations between January 2001 and June 2006 were analyzed retrospectively (all the cases were followed up to June 2007).According to UNOS grading scale,54 patients were of acute liver failure(UNOS 1 and 2A).Posttransplant ARF developing in 17 cases underwent a systemic therapy based on CRRT as well as anti-rejection,anti-infection and nutrition support.The perioperative courses,complications,causes of death and follow up results were analyzed.Results There were no severe complications during CRRT.Perioperative mortality was 5.4%and 58.8%in patients without ARF and those with ARF respectively.the rate of complications was 35.1%vs 100%.1 year survival rate Was 89.2% vs 41.2%.3 year survival rate was 81.1% vs 41.2%.Condusions The effect of surgery mainly depends on the function of liver and other vital organs.The ALF recipients suffered from a high perioperative mortality,especially those with posttransplant ARL.The systemic therapy based on CRRT benefits patients with postoperative ARF.

3.
Chinese Medical Journal ; (24): 513-517, 2002.
Article in English | WPRIM | ID: wpr-302264

ABSTRACT

<p><b>OBJECTIVE</b>To investigate if tumor necrosis factor-alpha (TNF-alpha) could induce phenotypic transformation or the associated cell function changes of human peritoneal mesothelial cells (HPMCs).</p><p><b>METHODS</b>All tests were performed on the human peritoneal mesothelial cell line (HMrSV5, kindly donated by Prof. Pierre RONCO). Morphological changes of HPMCs were observed by phase contrast microscopy. The expressions of cytokeratin 8, cytokeratin 18 and vimentin were detected by immunocytochemistry. mRNA expressions of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Production of MMP-9 and type I collagen was measured by enzyme-linked immunosorbent assays.</p><p><b>RESULTS</b>TNF-alpha (1 - 10 ng/ml) induced morphological changes in about 50% - 60% of human peritoneal mesothelial cells (HPMCs). The original polygonal cobblestone monolayer was changed into elongated, spindle-shape characteristic of fibroblasts. The original strong positive expression of cytokeratin 8 and cytokeratin 18 was changed in all morphologically changed HPMCs to negative, but the expression of vimentin maintained positive. By 4-hour treatment with TNF-alpha (1 - 10 ng/ml), mRNA expression of MMP9 was significantly up-regulated, and mRNA expression of TIMP-1 and TIMP-2 were down-regulated. We also observed that the production of MMP-9 and type I collagen increased significantly after 24-hour treatment with TNF-alpha.</p><p><b>CONCLUSION</b>24-hour treatment with TNF-alpha, produced a partial transformation of HPMCs into the fibroblast phenotype. TNF-alpha treatment of HPMCs up-regulated the synthesis of MMP-9 and type I collagen, which may facilitate peritoneal extracellular matrix remodeling and fibrogenesis.</p>


Subject(s)
Humans , Cell Line , Cell Size , Collagen Type I , Metabolism , Enzyme-Linked Immunosorbent Assay , Epithelial Cells , Cell Biology , Metabolism , Gene Expression Regulation , Immunohistochemistry , Keratins , Matrix Metalloproteinase 9 , Genetics , Peritoneum , Cell Biology , Metabolism , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Tissue Inhibitor of Metalloproteinases , Genetics , Tumor Necrosis Factor-alpha , Pharmacology , Vimentin
4.
Chinese Journal of Organ Transplantation ; (12): 16-18, 1997.
Article in Chinese | WPRIM | ID: wpr-387584

ABSTRACT

Plasma level of KPTT,factor Ⅷ coagulant activity,factor Ⅷ-related antigen,fib-rinogen,antithrombin Ⅲ activity,antithrombin Ⅲ antigen,tissue-plasminogen activator activity (t-PA:C),plasminogen activator inhibitor activity(PAI:C),PAI:C/t-PA:C,plasmin activity (PLm:C),D-dimer and plasma platelet α granule membrane protein were ditermined in 26 patients treated by long-term peritoneal dialysis(PD group),17 patients with chronic renal failure under-going conservative therapy(CRF group),36 patients on maintenance hemodialysis(HD group) and 20 age & sex-matched normal controls(NC group).The results showed:1.PD group had hypercoagulable state than NC group.The prothrombotic state in PD patients was most severe in contrast with CRF patients and HD patients.2.When PD group was compared with NC group,PLm:C showed no significant differences,but they had secondary fibrinolysis following enhanced coagulation.Moreover,PLm:C in PD group was significantly depressed as compared with in CRF group.3.the activated platelets in the PD patients were significantly elevated as compared with in the normal controls.

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