1.Cloning and characterization of the group 5 allergens from Dermatophagoides farinae
Yubao CUI ; Jianglong PENG ; Ying ZHOU ; Ying WANG ; Wei SUN
Chinese Journal of Microbiology and Immunology 2009;29(3):208-212
Objective To obtain the gene coding for the group 5 allergens from Dermatophagoides farinae ( Derf5 ) and predict its molecular characteristics. Methods The total RNA of D. farinae were extracted, and the gene Derf5 was amplified by RT-PCR with the primers designed according to previous sequence published in GenBank. The target gene was linked into pMD19-T Simple plasmid, sequenced and analyzed by bioinformatics software. Results The sequence homology reached to 97.8% between our sequenced result with one complete open reading fragment (ORF) and the reference. The gene encode an extracellular hydrophobic protein with 132 amino acid resides, one signal peptide from 1 to 19 position and one transmembrane domain from 1 to 19 position. The secondary structure was composed of extended strand (1. 52% ), random coil (7.58%) and alpha helix (90.91%). The encoded protein was deduced to have two Casein kinase Ⅱ phosphorylation sites. The similarity of the amino acid sequence of the group 5 allergens were 78% between D. farinae and D. pteronyssinus. Conclusion The gene Derf5 was cloned successfully, and its characteristics was primarily predicted.
2.Plant vector construction and expression of Der f1 allergen of Dermatophagoides pteronyssinu
Jianglong PENG ; Yubao CUI ; Huamin WANG ; Ying ZHOU ; Lina NIU ; Jie WU
Chinese Journal of Immunology 2010;26(3):250-253
Objective:To construct the plant expression vector of Der f1 allergen of Dermatophagoides pteronyssinu and expression in tobacco lamina.Methods:The Der f1 gene was amplified from the glycerin bacterium which contained pET28a(+)-Der f1 plasmid,cloned into the pMD 19-T plasmid,and then sequenced.The Der f1 gene was digested by ClaⅠand SalⅠ,and cloned into potato virus X (PVX) to construct plant expression vector PVX-Der f1,and then was transformed agrobacterium tumefaciens.The positive one was selected to infect tobacco lamina for expressing target protein.The protein was identified and analysed by SDS-PAGEand Western blot.Results:Digestion and sequence analysis confirmed that the plant expression vector was correct,and the SDS-PAGE and Western blot results showed that the molecular weight of the protein was about 34M_r and it could specific binding with positive serum.Conclusion:The plant expression vector of Der f1 is successfully constructed and the recombinant protein is also produced.
3.Antiproliferative and apoptosis effect of atorvastatin on K562 cells
Chunfang KONG ; Jianglong ZHOU ; Weirong DING ; Jianghua DING ; Guoan CHEN ; Hongbo CHENG ; Chenghao JIN
Chongqing Medicine 2018;47(3):299-301,305
Objective To explore the effect of atorvastatin on the proliferation and apoptosis of K562 cells andto investigate its mechanisms.Methods The cells were treated by different concentrations of atorvastatin.The CCK-8 assay was employed to detect the cell proliferation.The cell apoptosis was detected by AnnexinV-FITC/PI dual staining;the flow cytometry was used to detect the cellular cycle;the activities of caspase-3,-8,-9 were detected by the colorimetric method;qRT-PCR was employed to measure the mRNA expression levels of Bcl-2 and PDCD5 in K562 cells.Results Atorvastatin could inhibit the proliferation of K562 cells in a time-and dose-dependent manner(P<0.05);and induced the apoptosis of K562 cells,the percentage of G0/G1 phase cells was increased after atorvastatin treating k562 cells(P<0.01),while the percentage of S phase cells was decreased(P<0.01),moreover which showing the concentration dependence(P<0.01);atorvastatin activated the caspase-3,-8,-9 (P<0.01);down-regulated Bcl-2 mRNA expression and up-regulated PDCD5 mRNA expressionin a concentration dependence(P<0.01).Conclusion Atorvastatin can inhibit the proliferation and induce apoptosis in K562 cells.
4.The scalp localization system of neurosurgery based on augmented reality theory.
Hongli YAN ; Junwen GUAN ; Yonghong LI ; Qiuming ZHANG ; Yicheng ZHOU ; Xuepei LI ; Jianglong XU
Journal of Biomedical Engineering 2019;36(3):428-434
Neurosurgery navigation system, which is expensive and complicated to operate, has a low penetration rate, and is only found in some large medical institutions. In order to meet the needs of other small and medium-sized medical institutions for neurosurgical navigation systems, the scalp localization system of neurosurgery based on augmented reality (AR) theory was developed. AR technology is used to fuse virtual world images with real images. The system integrates computed tomography (CT) or magnetic resonance imaging (MRI) with the patient's head in real life to achieve the scalp positioning. This article focuses on the key points of Digital Imaging and Communications in Medicine (DICOM) standard, three-dimensional (3D) reconstruction, and AR image layer fusion in medical image visualization. This research shows that the system is suitable for a variety of mobile phones, can achieve two-dimensional (2D) image display, 3D rendering and clinical scalp positioning application, which has a certain significance for the auxiliary neurosurgical head surface positioning.
Humans
;
Imaging, Three-Dimensional
;
Magnetic Resonance Imaging
;
Neurosurgery
;
methods
;
Scalp
;
Tomography, X-Ray Computed