PubMed · 15784673
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William B Hurlbut. 2005-03-17. Altered nuclear transfer.. https://doi.org/10.1056/nejm200503173521121
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BACKGROUND: The genotyping for the study of the SNPs in different complex diseases require a great number of patients. In this sense, the determination of allele frequencies and genotypes requires a rapid and economical procedure. METHODS: The genotype has been carried out by allele-specific PCR in single tube with the discrimination of the products of PCR by its T(m). For this purpose a GC tail was added to 5' extreme of the specific primer. The allele frequencies were also calculated by DNA pooling and QRT-PCR using allele-specific primers. RESULTS: The use of the genotyping in single tube through allele-specific PCR and melting curves has led us to the accurate genotype of three polymorphisms of vdr (cdx-2), osteoprotegerin (A-163G) and ppar-gamma (C-681G) genes in 225 postmenopausal women to be associated to osteoporosis. Only the cdx-2 polymorphism was associated with a reduced bone mineral density (BMD). These data were similar to those obtained when the allele frequencies were calculated using QRT-PCR in DNA pools. CONCLUSIONS: Individual genotyping with allele-specific PCR in single tube and melting curve analysis is a fast, trustworthy and economic method to study any SNP. We propose the following approach to determine the possible association of SNPs with complex and multifactorial diseases like osteoporosis, in which hundreds of individuals should be analyzed: construct control and problem groups, make DNA pools, and calculate pooled allelic frequencies. Genotyping each individual further permits to determine the genotypic distribution when differences in allelic frequencies are observed, thus allowing more complex statistical analyses (including other variables like age, weight, etc.).
BACKGROUND & OBJECTIVE: Intestinal metaplasia (IM) is thought as the precancerous lesion of gastric carcinoma, and CDX2 gene plays important roles in development and differentiation of intestinal epithelium, and maintenance of intestinal phenotype. Recent studies found that CDX2 were expressed aberrantly in IM of chronic atrophic gastritis (CAG) and some gastric carcinomas, which implied that CDX2 may play an important role in IM formation and gastric carcinogenesis. This study was to investigate the roles of CDX2 in the development and progression of IM and gastric carcinogenesis, and determine the correlation of IM to gastric carcinogenesis. METHODS: A tissue microarray containing 46 cases of CAG with IM, 40 cases of gastric carcinoma, and 32 cases of IM foci in paracancerous tissues was constructed. High iron diamine/alcian blue (HID/AB) and HE staining were used to classify IM and gastric carcinoma, and the expression of CDX2 protein and mRNA in different gastric lesions was assessed with immunohistochemistry and in situ hybridization, respectively. RESULTS: The proportion of type III IM was significantly higher in IM foci in paracancerous tissues than in CAG with IM (56.25% vs. 21.74%, P<0.01). The positive rates of CDX2 protein were 69.56% in IM foci in CAG, 53.13% in IM foci in paracancerous tissues, and 42.50% in gastric carcinomas, and the positive rates of CDX2 mRNA were 63.04%, 46.87%, and 35.00%, respectively. The positive rates were significantly lower in gastric cancer than in IM in CAG (P<0.01), but there was no significant difference between gastric cancer and IM foci in paracancerous tissues (P>0.05). The expression of CDX2 protein and mRNA was significantly higher in intestinal-type gastric cancer than in diffuse-type gastric cancer (54.55% vs. 27.78%, 45.45% vs. 22.22%, P<0.05). The expression of CDX2 protein was significantly lower in type III IM than in type I IM (46.42% vs. 79.31%, P<0.05). CONCLUSIONS: CDX2 may play important roles in the development and progression of IM and gastric carcinogenesis.
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