AIM. To investigate IL-1β＋3 594 in the 5^th intron, IL-10-1 082 and CD14-159 polymorphisms in patients with acute pancreatitis （AP） and septic shock.METHODS： The study induded 215 patients （109 with acute severe...AIM. To investigate IL-1β＋3 594 in the 5^th intron, IL-10-1 082 and CD14-159 polymorphisms in patients with acute pancreatitis （AP） and septic shock.METHODS： The study induded 215 patients （109 with acute severe pancreatitis （SAP）, 106 with acute mild pancreatitis （MAP）） and 116 healthy volunteers. Genomic DNA was prepared from peripheral blood leukocytes. Genotypes and allele frequencies were determined in patients and healthy controls using restriction fragment length polymorphism analysis of PCR products.RESULTS： The frequencies of IL-β＋3 594T, IL-10-1082G and CD14-159T allele were similar in patients with mild or severe pancreatitis and in controls. Within SAP patients, no significant differences were found in the allele distribution examined when etiology was studied again. Patients with septic shock showed a significantly higher prevalence of IL-10-1082G allele than those without shock （X^2 = 5.921,P= 0.015）.CONCLUSION： IL-10-1082G plays an important role in the susceptibility of SAP patients to septic shock. Genetic factors are not important in determination of disease severity or susceptibility to AP.展开更多
Objective: To express human soluble CD14 (sCD14) in eukaryotic cells.Methods: Human sCD14 cDNA was amplified from U937 cells with RT PCR method. The recombinant expression plasmid pEF1/HisC/sCD14 348aa was constructed...Objective: To express human soluble CD14 (sCD14) in eukaryotic cells.Methods: Human sCD14 cDNA was amplified from U937 cells with RT PCR method. The recombinant expression plasmid pEF1/HisC/sCD14 348aa was constructed and the expression in COS 7 cells was carried out using liposome transfection method. The yield was examined with scanning map identification. The expressed product was purified by immuno affinity chromatography. Results: Sequence analysis demonstrated that the amplified gene sequence and those reported by documents were completely identical. sCD14 was expressed with high yield. The expressed product was purified to above 90%. Recombinant sCD14, specifically combinable with endotoxins, had a natural biological activity.Conclusions: Human sCD14 was expressed in COS 7 cells, which laid a foundation for further study.展开更多
基金Supported Dy the Affiliated Hospital of Qindao University Medial College Doctoral Foundation, No. 2003-6
文摘AIM. To investigate IL-1β＋3 594 in the 5^th intron, IL-10-1 082 and CD14-159 polymorphisms in patients with acute pancreatitis （AP） and septic shock.METHODS： The study induded 215 patients （109 with acute severe pancreatitis （SAP）, 106 with acute mild pancreatitis （MAP）） and 116 healthy volunteers. Genomic DNA was prepared from peripheral blood leukocytes. Genotypes and allele frequencies were determined in patients and healthy controls using restriction fragment length polymorphism analysis of PCR products.RESULTS： The frequencies of IL-β＋3 594T, IL-10-1082G and CD14-159T allele were similar in patients with mild or severe pancreatitis and in controls. Within SAP patients, no significant differences were found in the allele distribution examined when etiology was studied again. Patients with septic shock showed a significantly higher prevalence of IL-10-1082G allele than those without shock （X^2 = 5.921,P= 0.015）.CONCLUSION： IL-10-1082G plays an important role in the susceptibility of SAP patients to septic shock. Genetic factors are not important in determination of disease severity or susceptibility to AP.
基金agrantfromtheNationalNatureScienceFoundationofChina (No .394 70 76 1)
文摘Objective: To express human soluble CD14 (sCD14) in eukaryotic cells.Methods: Human sCD14 cDNA was amplified from U937 cells with RT PCR method. The recombinant expression plasmid pEF1/HisC/sCD14 348aa was constructed and the expression in COS 7 cells was carried out using liposome transfection method. The yield was examined with scanning map identification. The expressed product was purified by immuno affinity chromatography. Results: Sequence analysis demonstrated that the amplified gene sequence and those reported by documents were completely identical. sCD14 was expressed with high yield. The expressed product was purified to above 90%. Recombinant sCD14, specifically combinable with endotoxins, had a natural biological activity.Conclusions: Human sCD14 was expressed in COS 7 cells, which laid a foundation for further study.
