3.Severe fever with thrombocytopenia syndrome virus nucleoprotein specifically binds to 60kD SSA/Ro protein in host cells.
Bin ZHENG ; Tao WANG ; Shuo ZHANG ; A-Qian LI ; Chuan LI ; Quan-Fu ZHANG ; Mi-Fang LIANG ; De-Xin LI
Chinese Journal of Virology 2014;30(3):233-237
This study aims to investigate whether the nucleoprotein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV) can impact the cellular immunity of host cells. Gene segments that encode the NP and non-structural protein (NSs) of SFTSV were inserted into eukaryotic expression vector VR1012. Host proteins that interact with NP and affect immunity were identified with co-immunoprecipitation (IP), SDS-PAGE, mass spectrometry (MS), and Western blot. Co-localization of NP and the identified host proteins was confirmed by confocal microscopy. A 60kD SSA/Ro, a protein related to immunity, interacted with NP, as found by IP and MS. Confocal microscopy showed that NP and SSA/Ro were co-localized in cytoplasm. These results indicated that SFTSV NP may specifically bind to 60kD SSA/Ro and cause a series of immune responses and clinical symptoms.
Bunyaviridae Infections
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genetics
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metabolism
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virology
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HEK293 Cells
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Humans
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Nucleoproteins
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genetics
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metabolism
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Phlebovirus
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genetics
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metabolism
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Protein Binding
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Ribonucleoproteins
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genetics
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metabolism
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Viral Proteins
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genetics
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metabolism
4.Effects of Taoren-Honghua drug pair on degeneration of cervical disc cartilage endplate in rat model of dynamic and static forces imbalance
Yi-Ru WANG ; De-Zhi TANG ; Qian-Qian LIANG ; Hao XU ; Yong-Jian ZHAO ; Wei-Chao ZHENG
Chinese Pharmacological Bulletin 2018;34(2):279-283
Aim To investigate the effect of TaorenHonghua drug pair on intervertebral disc degeneration (IVDD) in rats.Methods Fifty healthy Wistar rats were randomly divided into control group,model group,sham group,meloxicam group and Taoren-Honghua drug pair group,with 10 rats in each group.We established dynamic and static forces imbalance of cervical disc degeneration model or sham surgery in rats.12 weeks later,rats were intragastrically administered with meloxicam,Taoren-Honghua drug pair or saline for 30 days.C4/5 and C6/7 discs were harvested from rats.ABOG staining was used for observation of intervertebral disc morphology,real time PCR for mRNA expressions of type Ⅱ collagen (Col Ⅱ) and type Ⅹ collagen (Col Ⅹ),and immunohistochemical staining for Col Ⅱ and Col Ⅹ.Results Compared with model group,Col Ⅱ expression increased,while Col X expression decreased in chondrocyte of intervertebral disc in Taoren-Honghua-treated group(P < 0.01).Conclusion Taoren-Honghua drug pair could delay the degeneration of cartilage endplate in rat intervertebral disc.
5.Fetal Membrane Derived Adherent Cells: a Novel Source for Mesenchymal Stem Cells
Zhi-Cheng MA ; Zhi-Gao LONG ; Ling-Qian WU ; Qian PAN ; De-Sheng LIANG ; He-Ping DAI ; Kun XIA ; Jia-Hui XIA ;
China Biotechnology 2006;0(07):-
Objective: To establish a method for isolation and purification of fetal membrane derived adherent cells (FMDACs) , and investigate their biological characteristics. Method: FMDACs were isolated with trypsin inducing and cultured in vitro. FMDACs were induced to differentiate into osteoblasts and adipocytes. FACS and immunocytochemistry technique were used to examine the cell surface antigen. The genetic stability was verified by karyotype analysis. Results: FMDACs were successfully isolated and expanded in vitro. They had strong proliferative ability. FMDACs were positive for CD44 and CD29, but negative for CD34, CD14 and CD45. FMDACs were differentiated into osteoblasts and adipocytes after inducement. The karyotype was stable in the sixth-passaged FMDACs and the tumorigenicity was not found. Conclusion; FMDACs have the possibility of multipotent stem cells, which have strong capacities of self-renewal and multidirectional differentiation. The genetic background of FMDACs is stable. FMDACs may be used as a kind of novel seed cells for tissue engineering.
6.A minidystrophin-EGFP fusion gene expressed in Cos-7 cells mediated by human source vector.
Yu LIANG ; De-sheng LIANG ; Zhi-gang XUE ; Zhi-gao LONG ; Ling-qian WU ; Qian PAN ; Yi-qiao HU ; He-ping DAI ; Kun XIA ; Jia-hui XIA
Chinese Journal of Medical Genetics 2005;22(5):493-496
OBJECTIVETo construct a human source vector containing minidystrophin-EGFP fusion gene and investigate its expression in Cos-7 cells.
METHODSThe recombinant human source vector named pHrnDysG was constructed with PCR-clone methods. Three fragments of dystrophin gene were PCR amplified from normal human dystrophin gene cDNA (GenBank NM04006). These three fragments were ligated to generate a minidystrophin gene. The enhanced green fluorescent protein (EGFP) gene was fused to the C terminal of the minidystrophin gene, and then the pHrnDysG was finally obtained by cloning the fusion gene to pHrneo. Fluorescence microscope and RT-PCR were used to detect the expression of minidystrophin-EGFP fusion gene after the recombinant construct was transfected into Cos-7 cells by lipofectamine.
RESULTSRestrictive enzyme digestion analysis and sequencing confirmed that pHrnDysG vector was constructed successfully. After the recombinant pHrnDysG was transfected to Cos-7 cells, RT-PCR demonstrated that the fusion gene was successfully transcribed, and the green fluorescence was observed at the cell membrane.
CONCLUSIONThe minidystrophin-EGFP fusion gene mediated by pHrneo vector could express in Cos-7 cells and its products' localization in the cell membrane was the same as that of full length dystrophin. These results suggested that the recombinant human source vector pHrnDysG might be potentially used in studies on the gene therapy of Duchenne muscular dystrophy.
Animals ; COS Cells ; Cercopithecus aethiops ; Dystrophin ; genetics ; metabolism ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Models, Genetic ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection
7.Effects of siwu tang on protein expression of bone marrow of blood deficiency mice induced by irradiation.
Ping GUO ; Zeng-chun MA ; Ying-fei LI ; Qian-de LIANG ; Ji-feng WANG ; Sheng-qi WANG
China Journal of Chinese Materia Medica 2004;29(9):893-896
OBJECTIVETo observe the effects of siwu tang on protein expression of bone marrow of blood deficiency mice and provide the theoretical and experimental basis for understanding the molecular mechanism of blood enriching function of siwu tang.
METHODBlood deficiency mice were established by using 3.5 Gy 60Co gamma-ray. With proteomic technologies including two-dimensional electrophoresis, image analysis, in-gel digestion, peptide mass fingerprinting and bioinformatics the proteins of bone marrow of blood deficiency mice were isolated, analyzed, and identified.
RESULTSiwu tang could reverse 10 up-regulated and 4 down-regulated proteins of blood deficiency mice bone marrow. Seven of the proteins were likely to be lymphocyte specific protein 1, proteasome 26S ATPase subunit 4, hematopoietic cell protein-tyrosine phosphatase, glyceraldehyde-3-phosphate dehydrogenase, growth factor receptor binding protein 14 and lgals12 respectively.
CONCLUSIONSiwu tang can regulate the protein expression of bone marrow of blood deficiency mice and thus promote the growth and differentiation of hematopoietic cells and then exert its effects on blood enriching function.
Adaptor Proteins, Signal Transducing ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred C57BL ; Phosphoproteins ; blood ; Proteins ; metabolism ; Proteome ; metabolism ; Proteomics ; methods ; Radiation Injuries, Experimental ; blood ; Whole-Body Irradiation
8.No-flip method versus external method for Shang Ring circumcision: a meta-analysis.
De-Hong CAO ; Liang-Ren LIU ; Lu YANG ; Sheng-Qiang QIAN ; Jun-Hao LEI ; Jiu-Hong YUAN ; Qiang WEI
National Journal of Andrology 2014;20(12):1113-1119
OBJECTIVETo compare the effect and safety of the no-flip method versus the external method in Shang Ring circumcision.
METHODSWe searched relevant randomized controlled trials published in China and abroad comparing the no-flip method and external method of Shang Ring circumcision. Based on the Cochrane Handbook for systematic review, two reviewers independently eval- uated the quality of the included studies and abstracted relevant data, followed by a meta-analysis using the statistical software Review Manager 5.1.0.
RESULTSTotally 7 studies with 1 200 cases were included. Compared with the external method, the no-flip method was associated with a lower total rate of complications (RR = 0.40, 95% CI: 0.18, 0.87, P = 0.02), a lower incidence of postop- erative edema (RR = 0.28, 95% CI: 0.09, 0.81, P = 0.02), and a lower 24 h postoperative pain score (MD = -0.35, 95% CI: -0.55, -0.14, P < 0.001).
CONCLUSIONThe no-flip method of Shang Ring circumcision was superior to the external method for its advantages of fewer complications, lower incidence of postoperative edema, and mild postoperative pain. However, our findings need further support by more high-quality randomized controlled trials.
China ; Circumcision, Male ; adverse effects ; instrumentation ; methods ; Edema ; epidemiology ; Humans ; Male ; Pain Measurement ; Pain, Postoperative ; epidemiology ; Randomized Controlled Trials as Topic
9.Epidemiological investigation on a scrub typhus outbreak in a village from Guangdong province, China.
Jun LIU ; Bang-hua CHEN ; De WU ; Wen-hua LIU ; Li-jun YAO ; Xiao-ting MAO ; Liang-heng XIAO ; Hao-jie ZHONG ; Zhi-qian PENG
Chinese Journal of Epidemiology 2013;34(9):946-947
Aged
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Animals
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China
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epidemiology
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Disease Outbreaks
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Female
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Humans
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Male
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Middle Aged
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Orientia tsutsugamushi
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Scrub Typhus
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epidemiology
10.The effect of Siwu Tang on EPO and G-CSF gene expression in bone marrow of irradiated blood deficiency mice.
Ping GUO ; Qian-De LIANG ; Jian-Jiang HU ; Ji-Feng WANG ; Sheng-Qi WANG
China Journal of Chinese Materia Medica 2005;30(15):1173-1176
OBJECTIVETo observe the blood enriching function of Siwu Tang and its effect on Epo and G-CSF gene expressions in bone marrow of blood deficiency mice, and thus provide the basis for understanding the molecular mechanism of blood enriching function of Siwu Tang.
METHODThe animal model of blood deficiency were established in the mice by using 3.5 Gy60Co gamma-ray. Peripheral blood cells were analyzed and CFU-GM, BFU-E, CFU-E and CFU-mix were counted in bone marrow colony cultured. Both Epo and G-CSF gene expressions in bone marrow were measured with RT-PCR.
RESULTSiwu Tang significantly increased the number of peripheral blood cells and the amount of CFU-GM, BFU-E, CFU-E and CFU-mix in bone marrow and enhanced Epo and G-CSF gene expression in bone marrow in the mice with blood deficiency.
CONCLUSIONThe promotion of Epo and G-CSF gene expressions in bone marrow may be one of the mechanisms underlying the blood enriching function of Siwu Tang decoction.
Animals ; Bone Marrow ; metabolism ; Bone Marrow Cells ; cytology ; Cell Count ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythroid Precursor Cells ; cytology ; Erythropoietin ; biosynthesis ; genetics ; Female ; Granulocyte Colony-Stimulating Factor ; biosynthesis ; genetics ; Leukocyte Count ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred C57BL ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Whole-Body Irradiation