2.Cloning of TLR3 Isoform.
Eun Jeong YANG ; Jeon Soo SHIN ; Hyemi KIM ; Hyoung Woo PARK ; Myoung Hee KIM ; Se Jong KIM ; In Hong CHOI
Yonsei Medical Journal 2004;45(2):359-361
Toll-like receptor (TLR) 3 is a member of the TLR family that confers innate immunity by recognizing viral pathogens. Herein, we report that the TLR3 isoform is expressed on human primary cells and cell lines. This isoform has 2, 520 bp cDNAs compared to the 2, 712 bp of full cDNA, is produced by deletion of an intron-like sequence within exon 4 and is co-expressed with wild type TLR3 in primary human astrocytes and glioblastoma cell lines. This finding suggests the TLR3 isoform in astrocytes may have a different immunological role for binding ligands during the immune response in brain.
Astrocytes/*physiology
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Cloning, Molecular
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Human
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Isomerism
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Membrane Glycoproteins/chemistry/*genetics
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Receptors, Cell Surface/chemistry/*genetics
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Support, Non-U.S. Gov't
3.Progress in researches on sperm antigen fertilin beta.
National Journal of Andrology 2004;10(1):52-58
Fertilin beta plays an important role in fertilization by its disintegrin domain as a sperm-specific antigen. This paper reviews its structure, localization and roles in fertilization, and suggests that fertilin beta, as an important target antigen, has a very promising value in the development of human immunocontraceptive vaccine.
ADAM Proteins
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Animals
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Contraception, Immunologic
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Fertilins
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Fertilization
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Humans
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Male
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Membrane Glycoproteins
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chemistry
;
genetics
;
physiology
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Metalloendopeptidases
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chemistry
;
genetics
;
physiology
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Vaccines
;
immunology
4.p47phox, the phagocyte NADPH oxidase/NOX2 organizer: structure, phosphorylation and implication in diseases.
Jamel EL-BENNA ; Pham My Chan DANG ; Marie Anne GOUGEROT-POCIDALO ; Jean Claude MARIE ; Francoise BRAUT-BOUCHER
Experimental & Molecular Medicine 2009;41(4):217-225
Phagocytes such as neutrophils play a vital role in host defense against microbial pathogens. The anti-microbial function of neutrophils is based on the production of superoxide anion (O2(.-)), which generates other microbicidal reactive oxygen species (ROS) and release of antimicrobial peptides and proteins. The enzyme responsible for O2(.-) production is called the NADPH oxidase or respiratory burst oxidase. This multicomponent enzyme system is composed of two transmembrane proteins (p22phox and gp91phox, also called NOX2, which together form the cytochrome b(558)) and four cytosolic proteins (p47phox, p67phox, p40phox and a GTPase Rac1 or Rac2), which assemble at membrane sites upon cell activation. NADPH oxidase activation in phagocytes can be induced by a large number of soluble and particulate agents. This process is dependent on the phosphorylation of the cytosolic protein p47phox. p47phox is a 390 amino acids protein with several functional domains: one phox homology (PX) domain, two src homology 3 (SH3) domains, an auto-inhibitory region (AIR), a proline rich domain (PRR) and has several phosphorylated sites located between Ser303 and Ser379. In this review, we will describe the structure of p47phox, its phosphorylation and discuss how these events regulate NADPH oxidase activation.
*Disease
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Enzyme Activation
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Humans
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Membrane Glycoproteins/chemistry/*metabolism
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NADPH Oxidase/chemistry/genetics/*metabolism
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Phagocytes/cytology/*metabolism
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Phosphorylation
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Protein Conformation
5.Advances in research of SCARB2 functions and related disorders.
Miao HE ; Zhen LIU ; Beisha TANG ; Junling WANG
Chinese Journal of Medical Genetics 2015;32(5):723-727
SCARB2 (scavenger receptor class B, member 2) is a lysosomal membrane glucoprotein, which is encoded by SCARB2 gene. It takes vital parts in the physiological and pathological processes including the transportation of beta-glucocerebrosidase to the lysosome, infection of EV71 and load-induced cardiac myocyte hypertrophy. This article has reviewed the molecular structure and functions of SCARB2 gene and its protein, as well as their relationship with diseases.
Hand, Foot and Mouth Disease
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genetics
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Humans
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Lysosome-Associated Membrane Glycoproteins
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chemistry
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genetics
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physiology
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Myoclonic Epilepsies, Progressive
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genetics
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Parkinson Disease
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genetics
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Receptors, Scavenger
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chemistry
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genetics
;
physiology
6.Polymorphisms of human cytomegalovirus UL139 open reading frame in Hirschsprung's disease.
Zhi-quin MAO ; Ying HUANG ; Qiang RUAN ; Mei SUN ; Ying QI ; Hong GAO
Chinese Journal of Experimental and Clinical Virology 2007;21(4):322-324
OBJECTIVETo explore the UL139 gene polymorphism of human cytomegalovirus (HCMV), and the relationship between polymorphisms of HCMV UL139 ORF and Hirschsprung's disease (HD).
METHODSForty-three specimens of the aganglionic intestinal segment and 6 urine samples of HD infants were amplified by nested polymerase chain reaction (PCR) method. The amplicons were sequenced in both strands directly. The control group consisted of 10 asymptomatic HCMV infected infants, and their urine specimens were also analyzed using the same method.
RESULTSHCMV UL139 genes of 28 clinical strains from HD 1 patients were successfully amplified and sequenced. UL139 was hypervariable and was clustered into 3 major groups and 5 genotypes. The predominant genotype of HCMV in HD infants was UL139 Group 3 (48 percent). Comparison of strains distribution between the two groups did not reach statistical significance using chi square test (chi square=7.378, P=0.194). The results of correlation analysis between UL139 and UL144 genes showed a p value 0.05 by Kendall test. Clinically, strains from the rectosigmoid segment, total colon aganglionosis, and long-segment were distributed sporadically in UL139 genotypes.
CONCLUSIONUL139 gene displayed polymorphisms. No linkage was found between UL139 genotype and clinical phenotype of HD. There was no correlation between HCMV UL144 and UL139.
Cytomegalovirus ; genetics ; Female ; Genotype ; Hirschsprung Disease ; etiology ; virology ; Humans ; Male ; Membrane Glycoproteins ; genetics ; Open Reading Frames ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Viral Proteins ; chemistry ; genetics
7.Rapid Determination of Chimerism Status Using Dihydrorhodamine Assay in a Patient with X-linked Chronic Granulomatous Disease Following Hematopoietic Stem Cell Transplantation.
Hyun Young KIM ; Hee Jin KIM ; Chang Seok KI ; Dae Won KIM ; Keon Hee YOO ; Eun Suk KANG
Annals of Laboratory Medicine 2013;33(4):288-292
Chronic granulomatous disease (CGD) is a rare genetic disease, which is caused by defects in the NADPH oxidase complex (gp91phox, p22phox, p40phox, p47phox, and p67phox) of phagocytes. This defect results in impaired production of superoxide anions and other reactive oxygen species (ROS), which are necessary for killing bacterial and fungal microorganisms and leads to recurrent, life-threatening bacterial and fungal infections and granulomatous inflammation. The dihydrorhodamine (DHR) flow cytometry assay is a useful diagnostic tool for CGD that can detect absent or reduced NADPH oxidase activity in stimulated phagocytes. We report a patient with X-linked CGD carrying a novel mutation of the CYBB gene whose chimerism status following hematopoietic stem cell transplantation (HSCT) has been rapidly determined using the DHR assay. The level of DHR activity correlates well with short tandem repeat PCR analysis. Considering the advantages of this simple, rapid, and cost-effective procedure, serial measurement of DHR assay would facilitate the rapid determination of a patient's engraftment status, as a supplementary monitoring tool of chimerism status following HSCT.
Base Sequence
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*Chimerism
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DNA Mutational Analysis
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Flow Cytometry
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Granulomatous Disease, Chronic/*diagnosis/*enzymology/genetics/surgery
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*Hematopoietic Stem Cell Transplantation
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Homozygote
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Humans
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Infant, Newborn
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Male
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Membrane Glycoproteins/chemistry/*genetics
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Mutation
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NADPH Oxidase/chemistry/*genetics
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Polymerase Chain Reaction
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Rhodamines/chemistry/metabolism
8.Genetic variation of S1 gene hypervariable region I of infectious bronchitis viruses isolated in different periods in Guangxi.
Zheng-Ji WEI ; Ping WEI ; Mei-Lan MO ; Meng LI ; Tian-Chao WEI ; Kang-Ran LI
Chinese Journal of Virology 2008;24(2):126-132
The S1 gene hypervariable region I (HVR I) of 22 infectious bronchitis virus (IBV) strains isolated in Guangxi during the period of 1985-2007 were sequenced and compared to that of the other IBV reference strains and the pigeon coronavirus isolates. A phylogenetic tree based on nucleotide sequences of HVR I of all the IBV showed that they were classified into 5 distinct Clusters. 16 out of 22 IBV isolates were grouped into Cluster I, and had higher homology with pigeon coronavirus isolates but lower homology with the Massachusetts (Mass) type vaccine strains. There were 4 and 3 amino-acid residues inserted at the sites of 33-34 and 34-35 respectively within HVR I in 15 isolates, except in isolate GX-NN6 there had 4 amino-acid residues inserted at the both sites; isolates GX-YL1 and GX-NN2 had close relationship with Mass type vaccine strains, and they shared Cluster II; isolates GX-G and GX-XD of Cluster III had close relationship with the Japanese strain JP Miyazaki 89 which was isolated at the same period; isolates GX-YL6 and GX-NN7 of Cluster V had close relationship with the European strain 4/91. The results showed that there were high phylogenetic diversity among the IBVs prevailed in the field in Guangxi resulting from the commonly occurred mutation or insertion within the S1 gene HVR I of the viruses, and majority of the isolates had lower homology with the commonly used Mass type vaccine strains. There was much higher homology among viruses isolated in the same period of time, but without distinct difference in geographical origins.
Amino Acid Sequence
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Animals
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Chickens
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virology
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Genetic Variation
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Infectious bronchitis virus
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classification
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genetics
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isolation & purification
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Membrane Glycoproteins
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chemistry
;
genetics
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Molecular Sequence Data
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Phylogeny
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Spike Glycoprotein, Coronavirus
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Viral Envelope Proteins
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chemistry
;
genetics
9.The construction of yeast two-hybrid method in the protein-interactions and its significance in hepatic metastasis of colorectal carcinoma.
Fu-Yi ZUO ; Shi-Yong LI ; Ping AN ; Bo YU ; Hui-Yun CAI
Chinese Journal of Surgery 2004;42(11):672-674
OBJECTIVETo construct the yeast two-hybrid system, and screen the proteins which interact with FasL, and investigate the relationship of FasL and hepatic metastasis of colorectal carcinoma.
METHODSWe have cloned the FasL gene into the pGBKT7 vector as the bait, then screened the fetal liver cDNA library, and have got a series of specific proteins that interact with FasL protein. Using the bioinformatics, we analyzed the interacting proteins in the mechanism of hepatic metastasis of colorectal carcinoma.
RESULTSWe have screened several proteins that interaction with FasL protein, including metallothionein 1K, 1G, 2A, cathepsin B, fatty acid synthase, interferon alpha-inducible protein 27, phospholipid scramblase, Ser/Thr-like kinase, anchor attachment protein, fibulin-5.
CONCLUSIONSWe have successfully constructed the yeast two-hybrid system, and preliminary identified that the interaction between FasL, metallothionein, cathepsin and anchor attachment protein is radically related to the hepatic metastasis of colorectal carcinoma.
Cathepsin B ; metabolism ; Cloning, Molecular ; Colorectal Neoplasms ; chemistry ; pathology ; Fas Ligand Protein ; Gene Library ; Humans ; In Vitro Techniques ; Liver Neoplasms ; chemistry ; secondary ; Membrane Glycoproteins ; genetics ; metabolism ; Metallothionein ; metabolism ; Protein Binding ; Tumor Necrosis Factors ; genetics ; metabolism ; Two-Hybrid System Techniques ; Yeasts ; genetics
10.Effect of Herba Epimedii flavone on the osteoblasts metabolism in vitro.
Yi-Heng LIU ; Hai-Ying ZHANG ; Hong-Min ZANG ; Jun-Chang CHENG
China Journal of Chinese Materia Medica 2006;31(6):487-490
OBJECTIVETo explore the effect of Herba Epimedii flavone (HEF) on the osteoblast metabolism in vitro.
METHODOsteoblast were obtained from new born rat calvaria by digestive enzymes. MTF, PNPP and RT-PCR were used to observe the proliferation, activity of ALP and mRNA expression of OPG and RANKL of cultured osteoblasts in vitro.
RESULTIt was found that HEF had the effect on stimulating cell proliferation, activity of ALP and the mRNA expression of OPG of cultured osteoblasts (P < 0.01, P < 0.05).
CONCLUSIONHEF can promote the proliferation, the differentiation and the expression of OPG mRNA of the osteoblasts cultured in vitro.
Alkaline Phosphatase ; metabolism ; Animals ; Animals, Newborn ; Carrier Proteins ; biosynthesis ; genetics ; Cell Proliferation ; drug effects ; Cells, Cultured ; Epimedium ; chemistry ; Flavones ; isolation & purification ; pharmacology ; Glycoproteins ; biosynthesis ; genetics ; Membrane Glycoproteins ; biosynthesis ; genetics ; Osteoblasts ; cytology ; metabolism ; Osteoprotegerin ; Plants, Medicinal ; chemistry ; RANK Ligand ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptors, Cytoplasmic and Nuclear ; biosynthesis ; genetics ; Receptors, Tumor Necrosis Factor ; biosynthesis ; genetics