AIMTo establish a rapid and simple molecular identification method for six medicinals: Curcuma: C. longa, C. phaeocaulis, C. sichuanensis, C. chuanyujin, C. chuanhuangjiang, and C. chuanezhu in Sichuan Province.

METHODSA molecular approach (trnK nucleotide sequencing) was used in this study.

RESULTSThe sequenced entire chloroplast trnK gene region spanned 2,699-2,705 bp. The matK gene (an intron embodied in trnK gene) sequence and the intron spacer region of the trnK gene have great diversity within these six medicinal Curcuma species. There were six single bases substitutions between trnK coding region and matK region, the 9-bp deletion and 4-bp or 14-bp insertion repeat at some sites of matK region in each taxon.

CONCLUSIONThese relatively variable sequences were potentially informative in the identification for these six Curcuma species at the DNA level.

Base Sequence ; Chloroplasts ; genetics ; Curcuma ; classification ; genetics ; DNA Mutational Analysis ; methods ; DNA, Plant ; genetics ; Genes, Plant ; Introns ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; RNA, Transfer, Lys ; genetics ; Sequence Analysis, DNA ; methods ; Sequence Deletion

PURPOSE: Myoclonic epilepsy with ragged red fiber (MERRF) syndrome is a disease of the mitochondrial encephalomyopathies, characterized by progressive myoclonus (action), epilepsy, cerebellar ataxia, intention tremor, muscle weakness, progressive dementia, sensorineural hearing loss and optic atrophy. Its inheritance is maternally inherited mitochondrial mutation, and its pathologic finding is characterized by ragged red fibers (RRF). Biochemically its defects are diverse. This study was undertaken to investigate the pattern of mitochondrial mutation and characterize the clinical features in Korean patients with MERRF. METHODS: We collected 3 clinically suspected MERRF patients from 2 Korean families who have progressive myoclonus, epilepsy, cerebellar ataxia, intention tremor, muscle weakness, progressive dementia etc. We reviewed their clinical findings, electrophysiologic studies, radiologic findings and pathologic findings, retrospectively. Of the 2 patients (case 1 and case 3) who had undergone muscle biopsy, case 1 showed RRF in modified Gomori trichrome staining, increased mitochondrial number and abnormal inclusion body in EM. To examine the pattern of mitochondrial mutation of these patients, molecular study was carried out in 3 patients, 2 mothers, 2 fathers, and 4 siblings. Their genomic DNAs were isolated from peripheral leukocytes, subsequent PCR-direct nucleotide sequencing and Ban II digestion were followed. RESULTS: All mutations in our cases were A to G point mutations in the tRNALys gene at position 8344. CONCLUSION: We confirmed clinically suspected MERRF patients as MERRF and their mothers and siblings as carriers, on the basis of molecular genetic analysis. This study suggests that the molecular genetic analysis can be utilized to diagnose MERRF patients easily and confirm carriers, especially at the presymptomatic stage before the characteristic pathologic changes appear.
Biopsy ; Cerebellar Ataxia ; Dementia ; Diagnosis* ; Digestion ; DNA ; Epilepsies, Myoclonic* ; Epilepsy ; Fathers ; Hearing Loss, Sensorineural ; Humans ; Inclusion Bodies ; Leukocytes ; MERRF Syndrome ; Mitochondrial Encephalomyopathies ; Molecular Biology* ; Mothers ; Muscle Weakness ; Myoclonus ; Optic Atrophy ; Point Mutation ; Retrospective Studies ; RNA, Transfer, Lys ; Siblings ; Tremor ; Wills

PURPOSE: Myoclonic epilepsy with ragged red fiber (MERRF) syndrome is a disease of the mitochondrial encephalomyopathies, characterized by progressive myoclonus (action), epilepsy, cerebellar ataxia, intention tremor, muscle weakness, progressive dementia, sensorineural hearing loss and optic atrophy. Its inheritance is maternally inherited mitochondrial mutation, and its pathologic finding is characterized by ragged red fibers (RRF). Biochemically its defects are diverse. This study was undertaken to investigate the pattern of mitochondrial mutation and characterize the clinical features in Korean patients with MERRF. METHODS: We collected 3 clinically suspected MERRF patients from 2 Korean families who have progressive myoclonus, epilepsy, cerebellar ataxia, intention tremor, muscle weakness, progressive dementia etc. We reviewed their clinical findings, electrophysiologic studies, radiologic findings and pathologic findings, retrospectively. Of the 2 patients (case 1 and case 3) who had undergone muscle biopsy, case 1 showed RRF in modified Gomori trichrome staining, increased mitochondrial number and abnormal inclusion body in EM. To examine the pattern of mitochondrial mutation of these patients, molecular study was carried out in 3 patients, 2 mothers, 2 fathers, and 4 siblings. Their genomic DNAs were isolated from peripheral leukocytes, subsequent PCR-direct nucleotide sequencing and Ban II digestion were followed. RESULTS: All mutations in our cases were A to G point mutations in the tRNALys gene at position 8344. CONCLUSION: We confirmed clinically suspected MERRF patients as MERRF and their mothers and siblings as carriers, on the basis of molecular genetic analysis. This study suggests that the molecular genetic analysis can be utilized to diagnose MERRF patients easily and confirm carriers, especially at the presymptomatic stage before the characteristic pathologic changes appear.
Biopsy ; Cerebellar Ataxia ; Dementia ; Diagnosis* ; Digestion ; DNA ; Epilepsies, Myoclonic* ; Epilepsy ; Fathers ; Hearing Loss, Sensorineural ; Humans ; Inclusion Bodies ; Leukocytes ; MERRF Syndrome ; Mitochondrial Encephalomyopathies ; Molecular Biology* ; Mothers ; Muscle Weakness ; Myoclonus ; Optic Atrophy ; Point Mutation ; Retrospective Studies ; RNA, Transfer, Lys ; Siblings ; Tremor ; Wills