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Dan Chen,Shi-yang Pan,Erfu Xie,Li Gao,Huaguo Xu,Wenying Xia,Ting Xu,Peijun Huang 대한진단검사의학회 2017 Annals of Laboratory Medicine Vol.37 No.1
Background: Circulating levels of cell-free DNA increase in many pathologic conditions. However, notable discrepancies in the quantitative analysis of cell-free DNA from a large number of laboratories have become a considerable pitfall, hampering its clinical application. Methods: We designed a novel recombinant DNA fragment that could be applied as an internal standard in a newly developed and validated duplex real-time PCR assay for the quantitative analysis of total cell-free plasma DNA, which was tested in 5,442 healthy adults and 200 trauma patients. Results: Compared with two traditional methods, this novel assay showed a lower detection limit of 0.1 ng/mL, lower intra- and inter-assay CVs, and higher accuracy in the recovery test. The median plasma DNA concentration of healthy males (20.3 ng/mL, n=3,092) was significantly higher than that of healthy females (16.1 ng/mL, n=2,350) (Mann-Whitney two-sample rank sum test, P<0.0001). The reference intervals of plasma DNA concentration were 0-45.8 ng/mL and 0-52.5 ng/mL for healthy females and males, respectively. The plasma DNA concentrations of the majority of trauma patients (96%) were higher than the upper normal cutoff values and were closely related to the corresponding injury severity scores (R2=0.916, P<0.0001). Conclusions: This duplex real-time PCR assay with a new internal standard could eliminate variation and allow for more sensitive, repeatable, accurate, and stable quantitative measurements of plasma DNA, showing promising application in clinical diagnosis.
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Hua Guo,Zheng-Wei Xu,Bao-Rong He,Ding-Jun Hao,Wei-Guo Bian 한국조직공학과 재생의학회 2014 조직공학과 재생의학 Vol.11 No.5
A well-defined three-dimensional (3-D) reconstruction of bone-cartilage transitional structures is crucial for the osteochondral restoration. This paper presents an accurate, computationally efficient and semi-automated algorithm for the alignment and segmentation of two-dimensional (2-D) serial to construct the 3-D model of bonecartilage transitional structures. Entire system includes the following five components: (1) image harvest, (2) image registration, (3) image segmentation, (4) 3-D reconstruction and visualization, and (5) evaluation. A computer program was developed in the environment of Matlab for the semi-automatic alignment and automatic segmentation of serial sections. Semi-automatic alignment algorithm based on the position’s cross-correlation of the anatomical characteristic feature points of two sequential sections. A method combining an automatic segmentation and an image threshold processing was applied to capture the regions and structures of interest. SEM micrograph and 3-D model reconstructed directly in digital microscope were used to evaluate the reliability and accuracy of this strategy. The morphology of 3-D model constructed by serial sections is consistent with the results of SEM micrograph and 3-D model of digital microscope.
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