1.Value of interleukin 4 and immunoglobulin E levels monitoring in diagnosis of drug allergy reaction
Zhiyong LI ; Yangling WU ; Shaobin CAI
Chinese Journal of Biochemical Pharmaceutics 2015;(11):30-32
Objective To explore interleukin 4 and immunoglobulin E levels indicators monitoring value in diagnosis of drug allergy reaction . Methods The way of intravenous injection and conventional intraperitoneal injection was used, with the corresponding drugs such as saline, horse serum sensitized, and in different time interleukin -4 and total immunoglobulin E index were recorded in guinea pig serum.Results Injection of saline and Chuankezhi injection, interleukin-4 and total immunoglobulin E protein content , the difference was not statistically significant, but in other drug induced sensitive D8, D14 and D21, interleukin-4 and total immunoglobulin E protein content increased significantly.Conclusion Interleukin 4 and immunoglobulin E levels index detection is the diagnosis of drug allergy is an effective and fast way .
2.Association study between schizophrenia and polymorphism of phosphoserine aminotransferase 1gene
Juan GUO ; Yuantang CHEN ; Changjiang HE ; Li ZHANG ; Yu WU ; Yangling XING ; Lei AO ; Xiang CHEN
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(11):1019-1021
ObjectiveTo detect the association between schizophrenia and polymorphism of phosphoserine aminotransferase 1 ( PSAT1 ) gene.MethodsThe study group included 158 patients with schizophrenia from Xi' an Mental Health Center and the control group included 316 parents.The polymorphism of rs69287125,rs137824326 of phosphoserine aminotransferase 1 gene was detected with PCR methods and SNP typing in all nucleus families by correlation analysis and haplotype relative risk analysis.ResultsThe rs69287125 allele was associated with schizophrenia (P=0.011 ),the G allele was protective factor (Z =-2.31 ) and the A allele was hazarding factor (Z =2.31 ).The rs137824326 allele was associated with schizophrenia (P=0.007 ),the G allele was protective factor ( Z =- 2.54) and the A allele was the hazarding factor( Z =2.54).The haplotypes of A/A and G/G in the rs69287125-rs137824326 were associated with schizophrenia (P =0.021,0.015,Z =2.16,- 1.85).ConclusionThe polymorphism of phosphoserine aminotransferase 1 gene is associated with schizophrenia in Chinese.
3.Association study of schizophrenia and ankyrin repeat and kinase domain containing 1 gene polymorphism
Juan GUO ; Yuantang CHEN ; Changjiang HE ; Li ZHANG ; Yu WU ; Yangling XING ; Lei AO
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(10):912-914
Objective To detect the genetic association between schizophrenia and polymorphism of Ankyrin repeat and kinase domain containing 1 ( ANKK1 ) gene. Methods Observed in a sample of 112 parent/offspring trios where the proband net the American Classification and diagnostic Criteria for Mental Disorders The Forth Revised Edition, criteria for schizophrenia using correlation analysis and haplotype relative risk analysis. The polymorphism of Ankyrin repeat and kinase domain containing 1 gene was detected with PCR methods and SNP typing in all nucleus families. Results The rs2734849 allele was connected with schizophrenia(P= 0. 026). Allele T was protective factor( Z= -2.19) and allele A was the hazard factor( Z=2. 19). The rs4938015,rs7118900 and rs1800497 allele were independence with schizophrenia. Three kinds haplotypes of G/A in the rs7118900 -rs2734849, A/C in the rs2734849 -rs1800497, G/A/C in the rs7118900 -rs2734849 -rs1800497 were associated with schizophrenia ( The P values were 0.032,0. 041,0.046, the genotype frequencies were 0. 36,0.29,0. 17 ).Conclusion It shows an association between schizophrenia and the polymorphism at nucleotide of ankyrin repeat and kinase domain containing 1 gene in Chinese.
4.Immune regulation of hydrogen sulfide in children with acute lymphoblastic leukemia
Shuxu DU ; Yongrui JIA ; Hong TANG ; Yangling SUN ; Wanshui WU ; Liming SUN ; Junbao DU
Chinese Medical Journal 2014;(21):3695-3699
Background Acute lymphoblastic leukemia (ALL) and chemotherapy can cause immune imbalance,and gaseous molecule hydrogen sulfide (H2S) can participate in the process of immune response.This study aimed to investigate the immune regulation of H2S in pediatric ALL.Methods Children (n=78) with ALL admitted during 2010-2013 were included in this study.Two blood samples were collected in period of before chemotherapy,bone marrow remission and two days after chemotherapy,respectively.Serum contents of H2S and cytokines,including interleukin-1β (IL-13),interleukin-2 (IL-2),interferon-γ (IFN-γ),tumor necrosis factor (TNF-α),interleukin-4 (IL-4),interleukin-6 (IL-6),interleukin-10 (IL-10) and macrophage inflammatory protein-1α (MIP-1a),were detected using ELISA method.Stepwise regression was used to analyze the correlation between H2S and cytokines.Furthermore,human Jurkat cells were cultured in vitro,and nucleoprotein of Jurkat cells and peripheral blood mononuclear cells (PBMCs) were collected,contents of cystathionine γ-lyase (CSE) and certain cytokines were measured by Western blotting.Results Serum concentrations of H2S,IL-13,IL-6,IL-10 and MIP-1α in children with ALL were increased significantly (P <0.01),while concentrations of IL-2,TNF-α,IFN-γ and IL-4 decreased obviously (P <0.01).In patients after chemotherapy,concentrations of H2S and IL-10 were decreased significantly (P <0.05),but IL-4 and IFN-γ concentrations increased markedly (P <0.05).At remission stage,H2S,IL-13,IL-4,IL-6,IL-10 and MIP-1α concentrations were further decreased markedly (P <0.05),but concentrations of IL-2,TNF-α and IFN-γ increased again (P <0.05).Protein contents of CSE,IL-10,IL-4 and IL-2 of PBMCs also increased markedly in children with ALL.Moreover,changes of CSE protein contents of PBMCs were consistent with serum H2S contents,and there were significant correlation between H2S and certain cytokines based on stepwise regression analysis.Furthermore,compared with those of PBMCs group,in vitro study indicated that Jurkat cells of H2S group expressed IFN-γ,IL-10,IL-4 and IL-2 protein increased obviously (P <0.05),while IL-4,IL-2 and CSE expression of PPG group decreased markedly (P <0.05).Conclusion Gaseous molecule H2S might participate in the process of immune regulation in pediatric ALL through modulating transcription and expression of cytokines.
5.Changes of sulfur dioxide, nuclear factor-κB, and interleukin-8 levels in pediatric acute lymphoblastic leukemia with bacterial inflammation
Wanshui WU ; Yongrui JIA ; Shuxu DU ; Hong TANG ; Yangling SUN ; Liming SUN
Chinese Medical Journal 2014;(23):4110-4113
Background Bacterial inflammation is a common complication in patients with leukemia,and sulfur dioxide (SO2) is a bioactive molecule in modulating Gram-negative bacilli infection.This study aimed to examine the changes in SO2,nuclear factor-κB (NF-κB),and interleukin-8 (IL-8) levels in pediatric acute lymphoblastic leukemia (ALL) with Gram-negative bacterial inflammation.Methods Fifty-five ALL children were enrolled in this study,including 30 males and 25 females,aged 3-13 years,and the median age was 7.8 years.All these children who accepted chemotherapy for ALL were divided into the control group (before chemotherapy),the infection group (after chemotherapy with infection),and the recovery group (the infection was controlled after 1 week).The serum level of SO2 was detected using high performance liquid chromatography with fluorescence assay,and NF-κB and IL-8 levels were measured by enzyme-linked immunosorbent assay (ELISA).Human THP-1 cells were cultured,induced,and differentiated into macrophages,which were divided into five groups and each group was cultured with different stimulators:lipopolysaccharide (LPS) group,LPS+L-aspartate-β-hydroxamate (HDX) group,LPS+SO2 group,SO2,and control groups.NF-κB level and IL-8 protein contents by ELISA were examined in each group.Results In comparison with those of the control group,levels of serum SO2,NF-κB,and IL-8 of the infection group were significantly increased (P <0.05),while those of the recovery group were significantly decreased (P <0.05).A positive correlation was found between levels of serum SO2 and intracellular NF-κB/IL-8,and the correlation coefficients were 0.671 and 0.798 (P <0.05),respectively.According to the results found in human THP-1 cells,levels of NF-κB and IL-8 in LPS group were significantly increased compared with those of the control group (P <0.05); when compared with those in LPS group,levels of NF-κB in LPS+HDX group further increased significantly (P <0.05); however,the NF-κB levels of LPS+SO2 group decreased significantly (P <0.05).Conclusion SO2 may play an anti-inflammatory role during the process of inflammation by inhibiting the activation and transcription of NF-κB.
6.Process in menstrual blood-derived mesenchymal stem cells for treatment of central nervous system diseases.
Mengmeng LIU ; Xinran CHENG ; Kaikai LI ; Mingrui XU ; Yongji WU ; Mengli WANG ; Qianru ZHANG ; Wenyong YAN ; Chang LUO ; Shanting ZHAO
Chinese Journal of Biotechnology 2018;34(5):644-652
Stem cell research has become a frontier in the field of life sciences, and provides an ideal model for exploring developmental biology problems such as embryogenesis, histiocytosis, and gene expression regulation, as well as opens up new doors for clinical tissue defective and inheritance diseases. Among them, menstrual blood-derived stem cells (MenSCs) are characterized by wide source, multi-directional differentiation potential, low immune rejection characteristics. Thus, MenSCs can achieve individual treatment and have the most advantage of the clinical application. The central nervous system, including brain and spinal cord, is susceptible to injury. And lethality and morbidity of them tops the list of all types of trauma. Compared to peripheral nervous system, recovery of central nervous system after damage remains extremely hard. However, the treatment of stem cells, especially MenSCs, is expected to solve this problem. Therefore, biological characteristics of MenSCs and their treatment in the respect of central nervous system diseases have been reviewed at home and abroad in recent years, so as to provide reference for the treatment of central nervous system diseases.
7.Regulation of SOCS-3, OB, GLUT4 and PPARgamma gene expression by insulin and dexamethasone in porcine primary adipocyte.
Haowei ZHANG ; Jiangwei WU ; Bo WANG ; Zhen LÜ ; Gongshe YANG
Chinese Journal of Biotechnology 2008;24(8):1354-1360
Swine is an ideal model for diabetes studies. Insulin and insulin resistance are closely related with diabetes. To investigate the effect of SOCS-3 in insulin resistance, porcine primary adipocyte was treated with insulin (100 nmol/L) and dexamethasone (300 nmol/L) to induce insulin resistance. The simi-quantitative PCR results suggested that insulin increased GLUT4, PPARgamma and SOCS-3 gene expression in primary culture porcine adipocytes and no change of OB gene expression. Under insulin resistance conditions, SOCS-3 and OB gene expression were up-regulated, whereas GLUT4 and PPARgamma gene expression were down-regulated in primary porcine adipocytes. The overexpression of PPARgamma gene resulted in the increase of GLUT4 expression by insulin. Different expression levels of SOCS-3 determined the inhibitory effects of insulin signaling. Induction of insulin resistance by dexamethasone was not only due to inhibition of glucose transportation, but also repression of insulin signaling. SOCS-3 might be a potential gene to block the insulin resistance.
Adipocytes
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cytology
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metabolism
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Animals
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Cells, Cultured
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Dexamethasone
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pharmacology
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Glucose Transporter Type 4
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biosynthesis
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genetics
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Insulin
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pharmacology
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Insulin Resistance
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Leptin
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biosynthesis
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genetics
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PPAR gamma
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biosynthesis
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genetics
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Suppressor of Cytokine Signaling 3 Protein
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Suppressor of Cytokine Signaling Proteins
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biosynthesis
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genetics
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Swine
8.Construction of SOCS3 recombinant adenovirus and its expression in porcine primary adipocytes.
Shuangjuan YANG ; Chengquan XU ; Jiangwei WU ; Gongshe YANG
Chinese Journal of Biotechnology 2010;26(4):462-469
In order to construct recombinant adenovirus vector expressing Suppressor of cytokine signaling 3 (SOCS3) and obtain infectious adenoviral particles, SOCS3 gene was amplified from plasmid pcDNA3-SOCS3 and subcloned into the adenovirus shuttle plasmid pAdTrack-CMV. After sequence confirmation, the recombinant shuttle plasmid pAdTrack-CMV-SOCS3 was linearized by Pme I, and then transformed into BJ5183 competent cell, the recombinant plasmid pAd-SOCS3 was obtained by homologous recombination between pAdTrack-CMV-SOCS3 and the adenoviral backbone plasmid pAdEasy-1 in BJ5183. The pAd-SOCS3 was linearized by Pac I and transfected into HEK293 cells via liposome. The recombinant adenovirus was packaged and amplified in HEK293 cells. After purifying, virus titer was determined by tissue culture infectious dose 50 (TCID50). Using the recombinant adenoviruses to infect porcine primary adipocytes, the expression of green fluorescent protein (GFP) was observed by fluorescent microscopy, and SOCS3 gene was identified by RT-PCR and Western blotting. Restriction enzyme and PCR analysis demonstrated that the recombinant adenovirus vector was constructed correctly, and the virus titer reached 1.2x10(9) PFU/mL. The result of RT-PCR and Western blotting showed that SOCS3 mRNA and protein expression was remarkably increased in porcine primary adipocytes infected with recombinant adenovirus. In conclusion, this study successfully constructed the recombinant adenovirus containing SOCS3 gene, and can be helpful for further research on the function of SOCS3.
Adenoviridae
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genetics
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metabolism
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Adipocytes
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metabolism
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Animals
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Cloning, Molecular
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Green Fluorescent Proteins
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biosynthesis
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genetics
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HEK293 Cells
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Humans
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RNA, Messenger
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biosynthesis
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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Suppressor of Cytokine Signaling 3 Protein
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Suppressor of Cytokine Signaling Proteins
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biosynthesis
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genetics
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Swine
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Transfection
9.Induction and differentiation of induced pluripotent stem cells into macrophages: a review.
Xinchun YANG ; Xiaolong WU ; Jinlian HUA
Chinese Journal of Biotechnology 2021;37(11):4001-4014
Induced pluripotent stem cells (iPSCs) are a type of cells similar to embryonic stem cells but produced by reprogramed somatic cells. Through in vitro differentiation of iPSCs, we can interrogate the evolution history as well as the various characteristics of macrophages. iPSCs derived macrophages are not only a good model for drug screening, but also an important approach for immunotherapy. This review summarizes the advances, challenges, and future directions in the field of iPSCs-derived macrophages.
Cell Differentiation
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Embryonic Stem Cells
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Induced Pluripotent Stem Cells
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Macrophages
10.Research on secretion expression in Pichia pastoris and function of the HC-pro gene of watermelon mosaic virus.
Jian-Xin ZHANG ; Yun-Feng WU ; Xiu-Min WANG
Chinese Journal of Biotechnology 2007;23(6):1102-1106
HC-pro gene of Watermelon Mosaic virus was obtained by RT-PCR was 1371bp in length. It was cloned into pPI(9K, then the eucaryotic recombinant expression plasmid pPIC9K-WHC was constructed. After being linearized with restriction endonuclease Sal I , the recombinant plasmid was transformed into Pichia pastoris GS115 by electroporation. The high copy transformants with Mut+ /His+ phenotype were selected by RT-PCR and screening on G418, MD and MM medium. Induced by methanol for 5 days, the culture supernatant was analyzed by SDS-PAGE, the results showed that a specific protein with a molecular weight of about 66 kD was expressed. Western blot analysis proved that the expression protein could specifically bind to HC-Pro polyclonal antibody. Far western blot analysis proved that the expression protein could bind to coat protein, given support to "bridge" hypothesis that HC-Pro help aphid transmission of non-persistent viruses.
Citrullus
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virology
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Cysteine Endopeptidases
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biosynthesis
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genetics
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Mosaic Viruses
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genetics
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Pichia
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genetics
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metabolism
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Recombinant Proteins
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biosynthesis
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genetics
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metabolism
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Viral Proteins
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biosynthesis
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genetics