Clinical Application Study Of Diagnosis And Preoperative Resectability Assessment With Multi-detector Row CT For Hilar Cholangiocarcinoma

Background:The incidence of prostate cancer has been increasing in recent years in Chinese males, In the western countries, the incidence of prostate cancer has been remained in the first position over lung cancer for many years in male malignant carcinomas, and prostate cancer has become the second leading cause of death in male malignant carcinomas. Prostate cancer is a kind of alloplastic disease derived from multiple nidus and developing in random mode. Effective therapeutic methods remain to be found for the advanced prostate cancer with metastasis , especially the androgen-independent and chemical and radio therapy resistant prostate cancer.Microarray technology with high throughput, high specificity and other characteristics has a wide usage in the areas of disease diagnosis. However the major microarray technology currently exists in expensive testing instruments or low sensitivity and other issues, thus limiting the further development of the technologyGold nanoparticles can be combined with silver to form a black particles 106 times bigger than the original, and can be observed with the naked eye, scanned by general scanner to read images.OBJECTIVE:1,Development of nano-gold labeling chip technology2,Establish a reliable marker of prostate cancer gene chip diagnostic gold nanoparticles system.3,Chip with nano-gold labeling to verify the clinical prostate cancer specimens.Methods:So in this study, we applied the gold nanoparticles in microarray technology. To establish more stabilized nanogold-labeled microarray technology,we optimized the spotting buffer,the concentration of nanogold and the time of silver enhancement. In the meantime,we studied the effect of pre-hybridization on hybridization signal,as well as the detection limit of the nanogold-labeled microarray.Application of the labelled Nanogold microarray to detect 11 kinds of genes for prostate cance targets (IGF1, P27, AR, PSMA, KLK3, PCNA, Ki-67, KLK1, KLK2, TMPRSS2, SREBF2) and clinical specimens of prostate cancer, study on each of the gene expressions.RESULTS:The results showed that Labelled Nanogold Microarray detected when the sensitivity of the single-stranded DNA reached to 80 fmol/L ,the hybridization had a good specificity. Among the three kinds of spotting buffer,50%DMSO was the best. Pre-hybridization led to 70% loss of hybridization signal. The time of silver enhancement we chose was 15 minutes,at which the target signal was clear,while the background was low.Under the condition , Application of the labelled Nanogold microarray to detect 11 kinds of genes for prostate cance targets (IGF1, P27, AR, PSMA, KLK3, PCNA, Ki-67, KLK1, KLK2, TMPRSS2, SREBF2) and clinical specimens of prostate cancer, each upregulation gene probe point presented the obvious chromogenic expression,and the different shades.Downregulation gene and the negative control presented no chromogenic expression, This can be figure out by human eyes,to achieve qualitative and quantitative results.CONCLUSION:1,Under the condition , the best kinds of spotting buffer was 50%DMSO, Pre-hybridization led to 70% loss of hybridization signal. The time of silver enhancement we chose was 15 minutes,at which the target signal was clear,while the background was low,the sensitivity of the single-stranded DNA reached to 80 fmol/L.Can be used as a relatively reliable detection of gene chip platform,with high sensitivity and good specificity ,low cost,and it can be used for clinical testing.2,Application of the labelled Nanogold microarray to detect 11 kinds of genes for prostate cance targets (IGF1, P27, AR, PSMA, KLK3, PCNA, Ki-67, KLK1, KLK2, TMPRSS2, SREBF2) and clinical specimens of prostate cancer, each upregulation gene probe point presented the obvious chromogenic expression,and the different shades.Downregulation gene and the negative control presented no chromogenic expression, This can be figure out by human eyes,to achieve qualitative and quantitative results,with high sensitivity and good specificity.So it can serve as an important early diagnosis of prostate cancer reference portfolio and clinical application of prostate cancer.