PURPOSE: We investigated whether the loss of E-cadherin function was related to the peritoneal seeding in colorectal carcinomas. METHODS: Eleven patients who had undergone a palliative resection for a colorectal carcinoma, with peritoneal seeding, were enrolled onto the study. The primary tumors and seeding nodules were analyzed with regarded to mutations in the expressions of the CDH1 and protein of E-cadherin using SSCP, direct sequencing and immunohistochemical staining. RESULTS: In the primary tumors, the E-cadherin was normally expressed in 9 of the 11 cases, with 2 cases showing a reduced expression. In the seeding nodules, the E-cadherin was normally expressed in 6 of the 11 cases, with 5 cases showing a reduced expression. The degree of E-cadherin expression in the seeding nodules was significantly decreased comparing to that in the primary tumors (P<0.001). In the mutational analysis, there were no pathogenic mutations in either the primary tumors or the seeding nodules, with the exception of two silent changes in the ctgggt>ctaggt (intron 2) and GTG>GTA (codon 782). CONCLUSION: The loss of E-cadherin expression might be related to peritoneal seeding. The functional derangement of E-cadherin in peritoneal seeding could possibly be caused by a mechanism, such as promoter methylation, rather than the mutation of the CDH1.
Cadherins* ; Colorectal Neoplasms* ; Humans ; Methylation ; Polymorphism, Single-Stranded Conformational

OBJECTIVETo investigate the sequence polymorphism of mtDNA HV1,HV2 overlapping fragments and coding region encompassing position 8430-8673 in Hebei Han population.

METHODSPolymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combined with sequencing method was used to detect the haplotype distribution of mtDNA in 100 Hebei Han individuals.

RESULTSNinety-one haplotypes were noted in 100 unrelated individuals. The gene diversity is 0.9985 and the random match probability is 0.0115. Compared with the Anderson sequence, 65 sites of different nucleotide sequences were noted, of which 44 sites were previously registered in MITOMAP, 12 sites were not registered and the gene mutations were different from MITOMAP at 9 positions.

CONCLUSIONThe obtained data suggest that these loci are valuable genetic markers for personal identification and thus could be used as basic data for the forensic application of mtDNA in Hebei province.

BACKGROUND: Although abnormalities of p53 gene and their relation to clinicopathologic parameters have been identified in some human malignancies, there is little published data on their prevalence and clinical significance in ampullary adenocarcinoma (AAC). The aim of this study is to determine the prevalence of p53 abnormalities in AAC and to evaluate their relation to clinicopathologic features. METHOD:35 formaline-fixed paraffin-embedded tissues of AAC were examined for detection of p53 abnormalities by both single-strand conformation polymorphism (SSCP) analysis of polymerase chain reaction- amplified DNA fragments corresponding to exons 5-8 and immunohistochemistry (IHC) using monoclonal antibody to p53 protein (Novocastra, DO7), and the association between the p53 abnormalities and clinicopathologic parameters was analyzed. RESULT: In 22.9% of AAC, p53 gene muation was demonstrated by SSCP analysis, mainly at PCR-amplified exon 8 and exon 7. The p53 protein overexpression by IHC was 48.6% of AAC. Six SSCP and IHC-positive (17.2%) cases and 16 normal (45.7%) cases showed concordant results between the methods, although 13 cases (37.1%) showed discordance, including 11 IHC-positive (31.4%) and 2 SSCP-positive (5.7%) cases. Overall, the prevalence of p53 abnormalities was 54.3%. No significant associations between the p53 abnormalities and clinicopathological parameters such as clinical manifestations, histologic differentiation, and tumor stage were observed. CONCLUSION: The p53 abnormalities detected in 55% of AAC are not associated with prognostic factor, suggesting that abnormal p53 gene may play a role in the development of AAC, but not in its invasiveness.
Adenocarcinoma* ; DNA ; Exons ; Genes, p53* ; Humans ; Immunohistochemistry ; Polymorphism, Single-Stranded Conformational ; Prevalence*

PURPOSE: To describe the clinical phenotype of congenital cataract in Korean families and to determine whether the cataract is causally related to other congenital cataract with known gene. METHODS: We examined total 21 patients with congenital cataract and their families. Peripheral blood samples were obtained from congenital cataract patients and their families who visited clinics. The phenotypes were characterized by slit lamp examination and photographs of digital camera. And genetic traits were discriminated by family pedigree analysis. And we performed known mutation specific enzyme digestion, cold - SSCP and sequencing analysis for identification of genetic defects. RESULTS: We found nuclear (42.8%), anterior polar (4.8%), cortical (4.8%), sutural (4.8%) cataract. And the families affected with coexisting cortical and posterior polar opacity, posterior polar and post-subcapsular opacity were two cases (9.4%) respectively. We identified and examined a three-generation family affected with coexisting autosomal dominant zonular cataract and sutural opacity (CCZS, 4.8%: Family Yun). Previously the CCZS had mapped to chromosome 17q11-q12 and reported mutation of the beta A3A1-crystallin (CRYBA1) gene. We had not identified any mutation in CRYAB1 gene. CONCLUSIONS: In conclusion, we reported that nuclear type is very frequent in Korean patients with congenital cataract. And we had not identified any defects in CRYAB1 gene. These results demonstrated another gene caused this congenital cataract with same phenotype. Furthermore it will be necessary to perform the linkage analysis with microsatellite marker.
Cataract* ; Digestion ; Humans ; Microsatellite Repeats ; Pedigree ; Phenotype ; Polymorphism, Single-Stranded Conformational

PURPOSE: Avellino corneal dystrophy (ACD) is the most common form of inherited corneal disorder in Korea. To report 4 cases of ACD concurrent with floppy eyelid syndrome (FES), which had not been previously reported, and to find an additional mutation. METHODS: Five patient in 2 families who were diagnosed as ACD patient were examined whether they had FES. PCR, cold-SSCP and sequencing analysis were performed for identification of genetic defect. RESULTS: Four of 5 ACD patients showed FES which characterized by easily everted eyelid and conjunctival papillary reaction. In one family, succeeding two generations had this feature. We identified R124H mutation in all 5 ACD patients, however, no additional mutation wsa identified in BIGH3 gene. CONCLUSIONS: One case series suggested that there may be some linkage between the genes responsible for ACD and FES.
Eyelids* ; Family Characteristics ; Humans ; Korea ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational

PURPOSE: The purpose of this paper is to identify the forkhead transcription factor gene (FOXL2) mutations in Korean patients with blepharophimosis-ptosis-epicanthus inversus syndrome (BPES). METHODS: We have analyzed the mutations of FOXL2 gene in genomic DNAs extracted from 16 BPES patients and their families by PCR, PCR-SSCP, and sequencing. RESULTS: No deletion in exon 1 to 3 of the FOXL2 gene was observed by PCR. The PCR products were subjected to SSCP analysis and 9 patients showed SSCP shifts. The PCR products showing SSCP shifts were subcloned into plasmid vectors and sequenced to confirm the FOXL2 mutation. In total, 7 mutations (1 nonsense mutation, 1 deletion, and 5 duplications) in exon 2 were identified. CONCLUSIONS: The FOXL2 gene mutations were identified in the Korean BPES patients. Some of the mutations were previously reported and some were new mutations. This study will contribute to the molecular analysis and clinical counseling of BPES patients.
Codon, Nonsense ; Counseling ; DNA ; Exons ; Humans ; Plasmids ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Transcription Factors

BACKGROUND: The 463 codon mutation of katG gene has been reported as an useful marker for the detection of isoniazid(INH) resistant strains of M. tuberculosis. This study aimed to elucidate relationship between 463 mutation in katG gene and INH resistance in M. tuberculosis. METHOD: DNA was extracted from 28 INH susceptible strains(MIC > or = 0.2microg/ml on the Lowenstein Jensen media) and used for amplification of 189bp fragment containing 463 codon by PCR. Amplified fragments were digested by restriction enzyme Msp I, analyzed by single strand conformation polymorphism(SSCP) in the MDE gel and sequenced to prove mutation. RESULT: Only 7(25%) out of 28 were digestible by restriction enzyme Msp I. The SSCP pattern of 21 strains were distinctly different from that of M. tuberculosis H37Rv. Msp I undigestible PCR fragment was substituted at 463 codon from Arg(CGG) to Leu(CTG). CONCLUSION: This finding clearly indicate no relationship between 463 codon mutation of the katG gene and INH resistance.
Codon ; DNA ; Isoniazid* ; Mycobacterium tuberculosis* ; Mycobacterium* ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Tuberculosis

OBJECTIVETo identify the mutations of iduronate-2-sulfatase (IDS) gene in mucopolysaccharidosis type II patients.

METHODSPCR-SSCP analysis was applied to detect the common mutations in the exons 2, 3, 5, 7, 8, 9 in IDS-gene of the patient. DNA sequencing and PCR-RFLP were applied to analyze the mutation detected by PCR-SSCP.

RESULTSA new mutation(1253G-->T) of exon 7 of the IDS gene was found by PCR-SSCP and DNA sequencing in the patient, The PCR-restriction enzyme digestion showed that enzyme digestion location appeared in the patient and his mother, which verified the results of sequencing analysis.

CONCLUSIONThe mutation of patient with MPSII could be detected effectively and quickly by the applications of PCR-SSCP, DNA sequencing and PCR-restriction enzyme digestion analysis, and the new mutation thus detected is necessary for the prenatal diagnosis of the pedigree.

29 isoniazid (INH) resistant isolated strains and INH sensitive reference strain (H37Rv) of Mycobacterium tuberculosis were analysed by polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) and NciI restriction mapping for the detection of mutations in katG gene and inhA gene. The katG gene was divided into 3 parts (Akat, Bkat, Ckat; each part is about 800 bp) and amplified, inhA gene was amplified as a whole. Each of the amplified 800 bp DNA was digested into small fragments of less than 400 bp with restriction enzymes for the direct PCR-SSCP analysis. Firstly, 10 strains were analysed. All the 10 isolates showed clearly distinct SSCP patterns in Bkat from that of the reference strain, but only two isolates showed distinct SSCP patterns in Akat, and no isolated strain showed any distinct SSCP patterns in Ckat. 10 isolates also showed distinct SSCP patterns in inhA. NciI restriction mapping of Bkat showed mutation in codon 463 in 7 strains among 10 isolated strains. With these results an early detection strategy for the INH resistant M. tuberculosis was applied to the rest of 19 isolated INH resistant strains. Firstly, isolates were screened by Ncsl mapping in Bkat, and 13 strains showed mutations in codon 463. Secondly, the rest of 6 INH resistant isolates were analysed by PCR-SSCP with restriction enzyme digestion (PCR-SSCP-RE) in Bkat, and all the strains showed distinct SSCP patterns from that of the INH sensitive reference strain. This proved our strategy as effective and economic and time saving method in early detection of INH resistant M. tuberculosis.
Codon ; Diagnosis* ; Digestion ; DNA ; Isoniazid* ; Korea* ; Mycobacterium tuberculosis* ; Mycobacterium* ; Polymorphism, Single-Stranded Conformational ; Restriction Mapping ; Tuberculosis

β-Amyrin synthase (β-AS) genes of Glycyrrhiza uralensis from 6 different regions were analyzed by PCR-SSCP and sequenced, then the correlationship between β-AS SNP and regions of Glycyrrhiza uralensis were determined. According to the 1 coding single nucleotide polymorphism on the first exon of β-AS gene at 94 bp site, Glycyrrhiza uralensis could be divided into 3 genotypes. In these genotypes, the percentage of 94A type in genuine regions was much higher, and it had significant differences with the percentage in non-genuine regions (P < 0.001). The results of the experiment proved that different β-AS genotypes at 94 bp site from different regions may be one of the important reasons to result in the genuineness of Glycyrrhiza uralensis.
Exons ; Genotype ; Glycyrrhiza uralensis ; classification ; enzymology ; genetics ; Intramolecular Transferases ; genetics ; Plant Proteins ; genetics ; Polymorphism, Single Nucleotide ; Polymorphism, Single-Stranded Conformational