Single-nucleotide polymorphisms and activity analysis of the promoter and enhancer of the pig lactase gene.

Lactose intolerance in northern Europeans is strongly associated with a single-nucleotide polymorphism (SNP) located 14 kb upstream of the human lactase gene: -13,910 C/T. We examined whether SNPs in the 5' flanking region of the pig lactase gene are similar to those in the human gene and whether these polymorphisms play a functional role in regulating pig lactase gene expression. The 5' flanking region of the lactase gene from several different breeds of pigs was cloned and analyzed for gene regulatory activity of a luciferase reporter gene. One SNP was found in the enhancer region (-797 G/A) and two were found in the promoter region (-308G/C and -301 A/G). The promoter C-308,G-301(Pro-CG) strongly promotes the expression of the lactase gene, but the promoter G-308,A-301(Pro-GA) does not. The enhancer A-797(Enh-A) genotype for Pro-GA can significantly enhance promoter activity, but has an inhibitory effect on Pro-CG. The Enhancer G-797(Enh-G) has a significant inhibitory effect on both promoters. In conclusion, the order of effectiveness on the pig lactase gene is Enh-A+Pro-GA>Enh-A/G+Pro-CG>Enh-G+Pro-GA.

Analysis of factors affecting the life quality of the patients with late stomach cancer.

AIMS AND OBJECTIVES: To provide a theoretical basis for the clinical care of patients with late stomach cancer, we investigated the life quality and analysed its related factor in the patients with late stomach cancer. BACKGROUND: Due to the lack of effective screening methods, stomach cancer usually has been in the advanced stage when patients are diagnosed. However, the treatment for late stomach cancer is a tough problem in today's medicine. Chemotherapy or radiotherapy brings patients physiological and psychological distress and heavy financial burden, affecting patients' therapeutic effects, prognosis and life quality. DESIGN: The patients with late stomach cancer were included, and then, questionnaires about the life quality were completed. METHODS: Questionnaires including European Organisation for Research on Treatment of Cancer Quality of Life Questionnaire-Core 30, Social Support Rating Scale, Medical Coping Modes Questionnaire and Self-rating Anxiety Scale were completed by 173 patients with late stomach cancer who received treatment in our hospital between May 2010 and May 2011. Correlation analyses were performed. RESULTS: The overall score of the life quality of the patients with late stomach cancer was only 29·54 ± 12·21. The social support, medical coping modes, anxiety and patients' clinical data (except radiotherapy) markedly affected the overall life quality of the patients with late stomach cancer (p < 0·05). CONCLUSION: The life quality of the patients with late stomach cancer is poor and is associated with many factors. RELEVANCE TO CLINICAL PRACTICE: This study provides a theoretical basis for better nursing the patients with late stomach cancer and improving their life quality.

Expression of differentiation inhibiting factor 1 and vascular endothelial growth factor and the relation between them and microvessel density in gastric cancer tissue.

BACKGROUND/AIMS: To observe the expression of differentiation inhibiting factor 1 (DIF1) and vascular endothelial growth factor (VEGF) and explore the relation between them and microvessel density (MVD) in gastric cancer tissue. METHODOLOGY: The expression of DIF1, VEGF, CD34 and MVD in gastric cancer and peri-cancerous tissues was qualitatively analyzed with immunohistochemistry. The protein expression of DIF1 and VEGF in gastric cancer and peri-cancerous tissue was semi-quantitatively analyzed with Western blot. RESULTS: The expression of DIF1 and VEGF, and MVD were significantly higher in gastric cancer tissue than in peri-cancerous tissue (p<0.05). MVD was significantly higher in DIF-positive gastric cancer tissue than in DIF-negative gastric cancer tissue (p<0.001). CONCLUSIONS: In gastric cancer, DIF1 expression is related to VEGF expression and MVD. DIF1 may promote angiogenesis in gastric cancer.

Promoter methylation of esophageal cancer-related gene 4 in gastric cancer tissue and its clinical significance.

BACKGROUND/AIMS: To observe the promoter methylation of esophageal cancer-related gene 4 (ECRG4) in gastric cancer tissues and explore its clinical significance. METHODOLOGY: ECRG4 promoter methylation was detected with methylation-specific PCR in 49 samples of gastric cancer tissues, 30 samples of peri-cancerous tissues and 15 samples of normal tissues. The relations of ECRG4 promoter methylation to pathology, age, gender and lymph node metastasis were analyzed. RESULTS: The rate of ECRG4 promoter methylation was higher in gastric cancer tissues (69.4% (34/49)) and peri-cancerous tissues (53.3% (16/30)) than in normal tissues (6.7% (1/15)) (p<0.01). The rate of ECRG4 promoter methylation was higher in stage III+IV (80% (24/30)) than in stage I+II gastric cancer tissues (52.6% (10/19)) (p<0.05). The rate of ECRG4 promoter methylation was not related to age, gender and lymph node metastasis (all p>0.05). CONCLUSIONS: Aberrant ECRG4 promoter methylation may be used to monitor early gastric cancer and predict pathological staging. ECRG4 may become a molecular therapeutic target against gastric cancer.

[Relationship between MC4R gene polymorphyism and body weight trait in beagle dogs].

The relationship between MC4R gene polymorphism and body weight in beagle dogs was analyzed. Using gene-specific primers based on canine MC4R exonic sequences a gene fragment was PCR amplified, cloned and sequenced to identify potential polymorphisms. The relationship between a MC4R gene polymorphism detected by PCR-RFLP and canine body weight was analyzed. Three variants were found in beagle dog MC4R DNA sequence, of which two were deletions and one was a transversion which created a PshA I site that could be detected by PCR-RFLP. A statistically significant relationship between this polymorphism and body weight was found. MC4R gene could be a candidate modifier gene for canine body weight.

[The research on the relationship between the polymorphism of T105A locus in MC1R gene and coat color in dogs].

In order to detect the polymorphism of T105A in MC1R gene in dogs and to analyze the relationship between the genetic polymorphisms and phenotypes of dog coat color, the blood samples of 111 cross-breed dogs were taken and their genomic DNAs were extracted. The phenotypes of dog coat color were recorded. The T105A locus of MC1R gene in the canine was detected through the technology of PCR-RFLP. Furthermore, the polymorphic fragments at T105A were sequenced. The relationships between the polymorphism of T105A and coat color trait were analyzed by the statistical methods of bivarate correlation analysis. By the method of PCR-RFLP, the T105A polymorphism was found with two alleles A and B and three genotypes AA, AB and BB. The frequencies of two alleles were 72.97% and 27.03%, respectively. The heterozygosity of T105A locus was 0.39. The frequencies of three genotypes were 55.86%, 34.23% and 9.91%, respectively. According to the results of sequencing, one base change from G to A at the position 105 was found at T105A locus and it altered amino acid at the position 105 from alanine to threonine. According to the statistical analysis, no significant association between the polymorphism of MC1R gene and the coat color was found and the result may be due to the differences of genetic background. Further research on MC1R gene should be done in pure breed dogs.