The Mutation Of MEFV Gene In Chinese Population And The Role Of Pyrin In The Inflammation.

OBJECTIVE: To study the mutations of MEFV gene in Chinese population and to observe the effects of LPS on the expressions of MEFV mRNA in human peripheral blood leukocytes. METHODS: peripheral venous blood was obtained from 95 Chinese, 49 were from the health volunteers and 46 from burned patients. The genomic DNA was isolated and the 12 common mutations in MEFV gene were detected by reverse-hybridization. The relationship between the mutant type of MEFV gene and the clinical inflammatory presentations in 46 burned patients was also analyzed. The effects of LPS on the expression of MEFV mRNA in human peripheral blood leukocytes were detected with the method of reverse transcription polymerase chain reaction (RT-PCR). The amplification product of RT-PCR was inserted into a plasmid vector pMD18-T, then was cloned into the E. coli competent JM109 cells. RESULTS: 1. Nine mutant types of MEFV gene in Chinese were discovered, they were E148Q homozygous, E148Q heterozygous, P369S heterozygous, M680I heterozygous, E148Q homozygous + P369S heterozygous, E148Q heterozygous + P369S heterozygous, E148Q heterozygous + M680I heterozygous, E148Q homozygous + P369S heterozygous + M680I heterozygous, E148Q heterozygous + M680I heterozygous + M694I heterozygous. E148Q was the most common genotype in all the mutations found, the other genotypes are first reported in Chinese population. 2. No difference of the occurrence of pyemia between the normal group and the MEFV mutant group was observed. Logistic regression analysis showed that the age and the areas of burn influenced mostly the result of burned patient, and the mutation of MEFV gene did not contribute to influence the inflammatory complications. 3. It was found that the unstimulated human peripheral blood leukocytes only expressed a low level of MEFV mRNA. The expression of MEFV mRNA increased when stimulated with LPS at 1μg/mL and 10μg/mL after 2 hours (P<0. 05). However, no difference compared with normal group by LPS at 100μg/mL was observed (P>0. 05). 4. The amplification products of RT-PCR was successfully inserted into the plasmid vector pMD18-T and transformed to E. coli...