Professor du Xiaoquan is a famous Chinese medicine in Shaanxi Province,a master's tutor,a professor of Shaanxi University of traditional Chinese Medicine,a director of the Department of traditional Chinese Medicin...Professor du Xiaoquan is a famous Chinese medicine in Shaanxi Province,a master's tutor,a professor of Shaanxi University of traditional Chinese Medicine,a director of the Department of traditional Chinese Medicine,and a professor of Shen Shuwen,a national famous old Chinese medicine.展开更多
Objective To screen coronaryartery disease (CAD) specific expressions and clone their genes. Method Blood samples were collected from CAD and non - CAD patients at the end of coronary angiography. mRNA from samples wa...Objective To screen coronaryartery disease (CAD) specific expressions and clone their genes. Method Blood samples were collected from CAD and non - CAD patients at the end of coronary angiography. mRNA from samples was isolated and converted into cDNA. After ligated with specific linkers, the cDNA was amplified with complementary primers. PCR products from CAD samples were named as tester; the ones from non - CAD samples were named as driver. With different ratio of tester to driver (1 : 100,1: 1, 000, and 1: 10, 000), they were mixed, denatured, and renatured. Single strand cD-NA was eliminated by Mung bean nuclease. Double strand cDNA presented only in tester was amplified, ligated in vector pUC19 and pUC53, and transformed into E. coll DH5a. Strains with inserted cDNA fragments were picked up based on blue and white selection. Insertions were screened by endonuclease digestion and DNA sequencing. Results were compared with DNA sequences of GeneBank. Results: After the selection with representational differential analysis, CAD specific cDNA fragments with different sizes (about 1kb, 0. 75kb, and 0. 6kb) were cloned. Among them, two fragments from unknown genes were identified. One presented a 43. 3 % similarity with part of the rattus norvegicus lipocortin gene. Another presented a 45. 4 % similarity with part of the human polynucleotide kinase 3' - phosphatase gene. Conclusion There are at least two CAD specific - ex- pressions from unknown genes that were partially similar to lipocortin and polynucleotide kinase 3'- phos-phatase genes, respectively. Expression of these genes might affect the formation and progression of plaque within coronary artery.展开更多
It is difficult to make a precise diagnosis of intestinal tuberculosis and to differentiate it from Crohn's disease. For evaluating Polymerase Chain Reaction (PCR) assay in these two aspects, 36 specimens of intes...It is difficult to make a precise diagnosis of intestinal tuberculosis and to differentiate it from Crohn's disease. For evaluating Polymerase Chain Reaction (PCR) assay in these two aspects, 36 specimens of intestinal tuberculosis from surgical resections and endoscopic biopsies and 26 Crohn's disease samples were subjected to PCR assay. 21 specimens of normal colon tissue surrounding cancer were used as the control. Oligonucleotides derived from the IS 6110 sequence, which is repeated in M. tuberculosis chromosome and highly specific for the M. tuberculosis complex, were used as a primer. The amplified PCR products were detected by examination of ethidium-bromide-stained polyacrylamide gels. The specificity of PCR products was confirmed by digestion with Sal 1 restrictive endonuclease and southern blot hybridization using digoxigenin-labeled probe. The results showed that the M. tuberculosis DNA was identified in 27 / 36 intestinal tuberculosis, but none of 26 Crohn's disease. Acid fast bacilli were only found in 16 / 36 intestinal tuberculosis. In conclusion, as a rapid, sensitive, and specific pathogenic method in diagnosis of intestinal tuberculosis, PCR assay has been developed in this study, and is considered valuable in the differentiation between intestinal tuberculosis and Crohn's disease.展开更多
文摘Professor du Xiaoquan is a famous Chinese medicine in Shaanxi Province,a master's tutor,a professor of Shaanxi University of traditional Chinese Medicine,a director of the Department of traditional Chinese Medicine,and a professor of Shen Shuwen,a national famous old Chinese medicine.
文摘Objective To screen coronaryartery disease (CAD) specific expressions and clone their genes. Method Blood samples were collected from CAD and non - CAD patients at the end of coronary angiography. mRNA from samples was isolated and converted into cDNA. After ligated with specific linkers, the cDNA was amplified with complementary primers. PCR products from CAD samples were named as tester; the ones from non - CAD samples were named as driver. With different ratio of tester to driver (1 : 100,1: 1, 000, and 1: 10, 000), they were mixed, denatured, and renatured. Single strand cD-NA was eliminated by Mung bean nuclease. Double strand cDNA presented only in tester was amplified, ligated in vector pUC19 and pUC53, and transformed into E. coll DH5a. Strains with inserted cDNA fragments were picked up based on blue and white selection. Insertions were screened by endonuclease digestion and DNA sequencing. Results were compared with DNA sequences of GeneBank. Results: After the selection with representational differential analysis, CAD specific cDNA fragments with different sizes (about 1kb, 0. 75kb, and 0. 6kb) were cloned. Among them, two fragments from unknown genes were identified. One presented a 43. 3 % similarity with part of the rattus norvegicus lipocortin gene. Another presented a 45. 4 % similarity with part of the human polynucleotide kinase 3' - phosphatase gene. Conclusion There are at least two CAD specific - ex- pressions from unknown genes that were partially similar to lipocortin and polynucleotide kinase 3'- phos-phatase genes, respectively. Expression of these genes might affect the formation and progression of plaque within coronary artery.
文摘It is difficult to make a precise diagnosis of intestinal tuberculosis and to differentiate it from Crohn's disease. For evaluating Polymerase Chain Reaction (PCR) assay in these two aspects, 36 specimens of intestinal tuberculosis from surgical resections and endoscopic biopsies and 26 Crohn's disease samples were subjected to PCR assay. 21 specimens of normal colon tissue surrounding cancer were used as the control. Oligonucleotides derived from the IS 6110 sequence, which is repeated in M. tuberculosis chromosome and highly specific for the M. tuberculosis complex, were used as a primer. The amplified PCR products were detected by examination of ethidium-bromide-stained polyacrylamide gels. The specificity of PCR products was confirmed by digestion with Sal 1 restrictive endonuclease and southern blot hybridization using digoxigenin-labeled probe. The results showed that the M. tuberculosis DNA was identified in 27 / 36 intestinal tuberculosis, but none of 26 Crohn's disease. Acid fast bacilli were only found in 16 / 36 intestinal tuberculosis. In conclusion, as a rapid, sensitive, and specific pathogenic method in diagnosis of intestinal tuberculosis, PCR assay has been developed in this study, and is considered valuable in the differentiation between intestinal tuberculosis and Crohn's disease.
