ABSTRACT
The molecular connectivity index was adopted to explore the characteristics of supramolecular imprinting template of herbs distributed to liver meridian, in order to provide scientific basis for traditional Chinese medicines(TCMs) distributed to liver meridian. In this paper, with "12th five-year plan" national planning textbooks Science of Traditional Chinese Medicine and Chemistry of Traditional Chinese Medicine as the blueprint, literatures and TCMSP sub-databases in TCM pharmacology of northwest science and technology university of agriculture and forestry were retrieved to collect and summarize active constituents of TCM distributed to liver meridian, and calculate the molecular connectivity index. The average molecular connectivity index of ingredients distributed to liver meridian was 9.47, which was close to flavonoid glycosides' (9.17±2.11) and terpenes (9.30±3.62). Therefore, it is inferred that template molecule of liver meridian is similar to physicochemical property of flavonoid glycosides and terpenes, which could be best matched with imprinting template of liver meridian.
ABSTRACT
<p><b>OBJECTIVE</b>To construct a PCR chip with a gene panel for predicting and diagnosing metastatic colorectal cancer.</p><p><b>METHODS</b>The PCR chip was constructed by integrating 29 genes related to colorectal cancer metastasis identified by gene chip analysis and 3 housekeeping genes into a gene panel. The PCR chip was used for detecting the mRNA expressions of the integrated genes in colorectal cell lines, cancerous specimen and adjacent normal mucosa. The primers for amplification were refined and optimized by several rounds of preliminary reactions.</p><p><b>RESULTS</b>The PCR chip containing the 29 candidate genes and 3 housekeeping genes was successfully constructed, which showed specific amplifications of the genes. The results of the PCR chip for detecting the mRNA of the 29 genes related to colorectal cancer metastasis showed a concordance rate of 86% (25 out of 29) with the gene chip data. Application of the PCR chip in the examination of the clinical specimens identified 15 differentially expressed genes between metastatic colorectal cancer and colorectal cancer without metastasis.</p><p><b>CONCLUSION</b>The constructed PCR chip is reliable in the prediction of metastasis of colorectal cancer, and provides a molecular means for evaluating the prognosis of colorectal cancer metastasis.</p>
Subject(s)
Humans , Colorectal Neoplasms , Genetics , Pathology , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Neoplasm Metastasis , Diagnosis , Oligonucleotide Array Sequence Analysis , Polymerase Chain Reaction , Methods , PrognosisABSTRACT
<p><b>OBJECTIVE</b>To identify the enhancers of human lung specific X protein (LUNX) and their regulation at the transcription level in vitro.</p><p><b>METHODS</b>Three enhancer fragments (E1:+3770~+3959bp; E2: +6454~+6555bp; E3: +14553~+14652 bp) predicted by bioinformatics software were isolated from the human genomic DNA by PCR amplification. Luciferase assay was performed to detect the activities of the enhancers in transcriptional regulation.</p><p><b>RESULTS</b>PCR products were confirmed by DNA sequencing. The amplified enhancers digested by Kpn I/Xho I and BamH I/Sal I, to generate the sticky-end fragments were inserted into PGL3-promoter in a reporter vector, and 6 luciferase expression vectors were obtained. All the reporter plasmids and pGL3-promoter were transiently transfected into HEK293 cells with an internal control of pSV-β-Galactosidase reporter vector. The enhancer activity of each construct was evaluated by luciferase assay of the cell extracts after transfection for 48 h. The results showed that the 3 fragments, when located upstream, did not increase transcription of reporter gene, but when at the downstream, E1 and E3 increased the transcription by 2.83 and 1.59 folds of that of pGL3-promoter, respectively.</p><p><b>CONCLUSION</b>LUNX gene sequences from +3770 to +3959 bp and +14553 to +14652 bp possess the capacity to enhance gene transcription.</p>