1.Increased activities of Akt in psoriatic epidermis
Xiaoyan ZHANG ; Ping ZHOU ; Liping YOU ; Changan YU ; Lin PAN ; Shengqing MA
Chinese Journal of Dermatology 2009;42(6):413-416
Objective To investigate the significance of Akt in the pathogenesis of psoriasis.Methods Tissue specimens were obtained from involved and uninvolved skin of 30 patients with progressive psoriasis vulgaris and normal skin of 20 human controls.Immunohistochemistry.immunobloting and kinase activity assay were performed to detect the expressions of Akt and phosphorylated Akt as well as Akt activities in these specimens.Immunostaining intensity Was assessed by optical density detection and the results of immunobiot and activity assay by grey scanning.Statistical analyses were performed by variance analysis and student's t test.Results As immunohistochemistry revealed.there was no significant difierence in Akt protein expression among normal epidermis,psoriatic epidermis and uninvolved epidermis(F=0.611,P>0.05):the level of phosphorylated Akt in psoriatic epidermis was significantly higher than that in normal epidermis and psoriatic uninvolved epidermis(F=19.081.P<0.01).while no significant difierence was observed between normal epidermis and psoriatic uninvolved epidermis (t=0.624.P>0.05).Immunoblot showed a significant difierence in phosphorylated Akt(t=237.75.P<0.01)but not in Akt(t=1.378,P>0.05)between psoriatic involved epidermis and normal epidermis.In comparison with normal epidermis,the activity of Akt in psoriatic involved epidermis was increased significantly(t=138.44 1.P<0.0 1).Conclusion The overproliferation of psoriatic keratinocytcs may be associated with increased activation of Akt.
2.Proteomic analysis of chicken peripheral blood mononuclear cells after infection by Newcastle disease virus.
Xiaoyu DENG ; Yanlong CONG ; Renfu YIN ; Guilian YANG ; Chan DING ; Shengqing YU ; Xiufan LIU ; Chunfeng WANG ; Zhuang DING
Journal of Veterinary Science 2014;15(4):511-517
Characteristic clinical manifestations of Newcastle disease include leukopenia and immunosuppression. Peripheral blood mononuclear cells (PBMCs) are the main targets of Newcastle disease virus (NDV) infection. To survey changes in proteomic expression in chicken PBMCs following NDV infection, PBMC proteins from 30 chickens were separated using two-dimensional electrophoresis (2-DE) and subjected to mass spectrometry analysis. Quantitative intensity analysis showed that the expression of 78 proteins increased more than two-fold. Thirty-five proteins exhibited consistent changes in expression and 13 were identified as unique proteins by matrix assisted laser desorption ionization-time of flight mass spectrometer/mass spectrometer including three that were down-regulated and 10 that were up-regulated. These proteins were sorted into five groups based on function: macromolecular biosynthesis, cytoskeleton organization, metabolism, stress responses, and signal transduction. Furthermore, Western blot analysis confirmed the down-regulation of integrin-linked kinase expression and up-regulation of lamin A production. These data provide insight into the in vivo response of target cells to NDV infection at the molecular level. Additionally, results from this study have helped elucidate the molecular pathogenesis of NDV and may facilitate the development of new antiviral therapies as well as innovative diagnostic methods.
Animals
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Avian Proteins/*genetics/metabolism
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*Chickens
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*Gene Expression Regulation
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Leukocytes, Mononuclear/enzymology/virology
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Newcastle Disease/*genetics/virology
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Newcastle disease virus/*physiology
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Poultry Diseases/*genetics/virology
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*Proteome
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Specific Pathogen-Free Organisms
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/veterinary
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Tandem Mass Spectrometry/veterinary
3.Mutation of lambdapL/pR-cI857 system for production of bacterial ghost in Escherichia coli.
Hongliang DONG ; Xian'gan HAN ; Hao BAI ; Liang HE ; Lei LIU ; Rui LIU ; Tongjie CHAI ; Chan DING ; Haiwen LIU ; Shengqing YU
Chinese Journal of Biotechnology 2012;28(12):1423-1430
Bacterial ghost is intact envelope of Gram-negative bacteria, which is produced by the function of the lysis gene E from bacteriophage PhiX174. The expression of the lysis gene E is usually controlled by the thermosensitive lambdapL/pR-cI857 promoter. In this study, we described a mutation (T --> C) at the ninth nucleotide of the OR2 in the lambdapR promoter of the lambdapL/pR-cI857 system by overlap PCR. The bacteriolytic assay showed that the mutation in the lambdapL/pR-cI857 system enhanced the temperature of repressing the expression of gene E up to 37 degrees C. The lysis efficiency of altered lambdapR promoter in Escherichia coli DH5a and avian pathogenic E. coli DE17 was up to 99.9%. The expanded range of temperature will benefit for the production of bacterial ghost.
Bacteriolysis
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physiology
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Bacteriophage lambda
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genetics
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Base Sequence
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Cell Membrane
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physiology
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DNA, Bacterial
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analysis
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Escherichia coli
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genetics
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growth & development
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physiology
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virology
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Gene Expression Regulation
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genetics
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Molecular Sequence Data
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Mutation
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Promoter Regions, Genetic
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genetics
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Viral Proteins
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genetics
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metabolism
4.Research progress in portable diagnosis devices for traumatic intracranial hematoma
Yijie YU ; Zhan CHENG ; Chao CHEN ; Zhihao XU ; Hongkun WANG ; Hongxiang WANG ; Shengqing LYU ; Juxiang CHEN
Chinese Journal of Trauma 2023;39(4):371-377
Traumatic intracranial hematoma progresses rapidly and may cause quick increase of intracranial pressure and even brain hernia, ultimately leading to coma or death. Therefore, traumatic intracranial hematoma needs prompt treatment, but the prerequisite of treatment is early examination and diagnosis. Due to the limited transportation and other reasons, the existing large-scale detection devices such as CT and MRI cannot be deployed on the rescue site or during patient transportation. Instead, the portable diagnosis devices have the characteristics of miniaturization and high flexibility, which is conducive to promoting early detection, assisting diagnosis and further guiding the formulation of treatment plans. At present, more and more attention has been paid to the portable diagnosis devices in the diagnosis of intracranial hematoma. The authors summarized the conventional diagnosis methods and application of portable diagnosis devices for traumatic intracranial hematoma, aiming to provide a reference for the diagnosis of traumatic intracranial hematoma.
5.Chicken RNA-binding protein T-cell internal antigen-1 contributes to stress granule formation in chicken cells and tissues
Yingjie SUN ; Pin ZHANG ; Hang ZHENG ; Luna DONG ; Lei TAN ; Cuiping SONG ; Xusheng QIU ; Ying LIAO ; Chunchun MENG ; Shengqing YU ; Chan DING
Journal of Veterinary Science 2018;19(1):3-12
T-cell internal antigen-1 (TIA-1) has roles in regulating alternative pre-mRNA splicing, mRNA translation, and stress granule (SG) formation in human cells. As an evolutionarily conserved response to environmental stress, SGs have been reported in various species. However, SG formation in chicken cells and the role of chicken TIA-1 (cTIA-1) in SG assembly has not been elucidated. In the present study, we cloned cTIA-1 and showed that it facilitates the assembly of canonical SGs in both human and chicken cells. Overexpression of the chicken prion-related domain (cPRD) of cTIA-1 that bore an N-terminal green fluorescent protein (GFP) tag (pntGFP-cPRD) or Flag tag (pFlag-cPRD) induced the production of typical SGs. However, C-terminal GFP-tagged cPRD induced notably large cytoplasmic granules that were devoid of endogenous G3BP1 and remained stable when exposed to cycloheximide, indicating that these were not typical SGs, and that the pntGFP tag influences cPRD localization. Finally, endogenous cTIA-1 was recruited to SGs in chicken cells and tissues under environmental stress. Taken together, our study provide evidence that cTIA-1 has a role in canonical SG formation in chicken cells and tissues. Our results also indicate that cPRD is necessary for SG aggregation.
Chickens
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Clone Cells
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Cycloheximide
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Cytoplasmic Granules
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Humans
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Protein Biosynthesis
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RNA Precursors
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RNA-Binding Proteins
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T-Lymphocytes