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Interaction of Mina and IL-4 gene and their relevance to asthma children

Chinese Journal of Immunology.2014;(5):684-686. doi:10.3969/j.issn.1000-484X.2014.05.026

Objective:To study the relevance of Mina rs4857304 (G/T) gene locus, IL-4 gene locus rs2243248 (T/G), rs2070874 ( T/C ) single nucleotide polymorphism and allergic asthma pathogenesis of Chinese children , and further analysis of the in-teraction among three loci.Methods:Based on the case-control study, we have analyzed the different distributions , frequencies, inter-action of alleles and genotypes between asthma patients and patients in control group .Results: The Mina rs4857304 sites of patients were compared, and there was statistically significant difference between two groups ( P=0.001, OR =1.773,95%CI =1.256 -2.503);the rs4857304T allelic frequency of patients with asthma was higher than that of the patients in control group , but there was no statistically significant difference about the distribution of different locus genotypes of patients between two groups (P>0.05);MDR interaction analysis have showed that Mina gene loci , rs4857304 and IL-4loci rs2243248, consisting of 2 sites optimal model;Mina loci rs4857304, IL-4loci rs2070874 and rs2243248 that consisting of 3 sites optimal model.There was statistically significant difference be-tween two groups(P<0.05).Conclusion:Allergic asthma would be associated with Mina loci rs4857304, while there might be sig-nificant interactive effects among Mina rs4857304, IL-4loci rs2243248 and rs2070804.

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Expression characteristic of IFNGR in stellate ganglion of female goats

Qiang LI ; Zhihao WANG ; Xiufang JIN ; Yongping XU ; Xiao GUO ; Wendong CHEN ; Wei DONG

Chinese Journal of Immunology.2014;(5):604-608. doi:10.3969/j.issn.1000-484X.2014.05.007

Objective:To detect the existence of IFNGR in the stellate ganglion in female goats .Methods:The stellate ganglia were taken from female goats .The expression of IFNGR was detected by PCR method and immunohistochemical SP staining .Results:The results showed that:IFNGR immunoreactive substances were distributed in neurons , supporting cells and passing fiber , and mainly in the cytomembrane and nuclei of neurons .The relative expression of IFNGR was very significantly higher than that of non-neuronal cells (P<0.01).The sequence of IFNGR1 gene amplified by PCR method contained 376 bp, and IFNGR1 gene of goats exhibited the highest homology with sheep(98%),followed by cattle(97%).Conclusion:The results suggested that the IFNGR in the Stellate Gan-glion of female goats mainly expressed and located in sympathetic postganglionic neurons which were provided with the conditions for the role of IFN-γ, which implied that the Stellate Ganglion may act as the critical point to concert the immune regulation of IFN -γand neu-roregulation of autonomic nerve on cardiovascular system .

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Analyse curative effect and immune indices of tripterygium glycosides on aged glomerulonephritis rats

Shan LIU ; Lunzhi LIU ; Zhihui TAN ; Xiaofen YANG

Chinese Journal of Immunology.2014;(5):627-629. doi:10.3969/j.issn.1000-484X.2014.05.012

Objective:To analyse the curative effect and immune indices of tripterygium glycosides on glomerulonephritis rats . Methods:Aged Wistar rats were randomly divided into 3 groups , Group A:glomerulonephritis model rats received tripterygium glyco-sides treatment;Group B:healthy Wistar rats received no treatment; Group C: glomerulonephritis model rats received no treatment . Results:12 weeks after experiment , Group A and Group C had higher levels of 24-hour urine protein than Group B and these of pre-experiment(P<0.05).Group A had a lower level of 24-hour urine protein than Group C(P<0.05).Group A and Group C had high-er levels of TG, TC than Group B, with lower levels of ALB than Group B (P<0.05).Group A had lower levels of TG, TC than Group C, but a higher level of ALB than Group C (P<0.05).Group A had a higher rate of 0-1 grades in pathology grade than group C(χ2 =6.667,P=0.010).Group C had higher levels of TNF-αand IL-6 than Group B and Group A (P<0.05).Conclusion:Tripterygium glycosides can reduce the urine protein excretion of rats that with mesangial proliferative glomerulonephritis , rectify dyslip-idosis, raise plasma-albumin, decrease the protein expression of TNF-αand IL-6 in glomerulus.

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Syk and VEGF-C in breast cancer's expression with lymph node metastasis

Xifeng ZHANG ; Hongbing ZHAO

Chinese Journal of Immunology.2014;(5):681-683. doi:10.3969/j.issn.1000-484X.2014.05.025

Objective:To explore the relationship of Syk and VEGF-C expression in breast cancer with lymph node metastasis . Methods:The expression of Syk and VEGF-C in 40 cases of breast cancer were determined using immunohistochemistry assay .Then, the positive rates of Syk and VEGF-C were compared with lymph node metastasis .Results:The expression rate of VEGF-C and Syk in breast cancer was respectively 77.5%(31/40), 37.5%(15/40).VEGF-C protein expression was not significant related with the tumor diameter (P>0.05) and histological grade (P>0.05), but was significantly associated with lymph node metastasis (P<0.05).Syk protein expression was not significant related with tumor diameter (P>0.05) and the histological grade(P>0.05), and was associated with lymph node metastasis (P<0.05).The difference of Syk and VEGF-C had statistics significance(P<0.05).Con-clusion:Syk and VEGF-C involved in the lymphatic invasion and metastasis , may be an important indicator of prognosis .

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Construction and identification of lentiviral vector harboring interference RNA targetting murine TNF-αgene

Yingjie ZHAO ; Jibo WANG ; Miaomiao XIN ; Hongda LIANG ; Xiangping LIU ; Kun YANG ; Aihua SUI

Chinese Journal of Immunology.2014;(7):927-932. doi:10.3969/j.issn.1000-484X.2014.07.014

Objective:To construct recombinant lentiviral vectors harboring interference RNA ( RNAi ) targetting murine TNF-αgene,so as to lay the foundation on the RNAi gene therapy.Methods: Three small interfering RNA ( siRNA) sequences targeting murine TNF-αgene ( siRNA1,siRNA2,siRNA3) and negative-control siRNA were designed and synthesized.The inhibition effects of siRNAs on TNF-α,IL-1βand IL-6 secretion of LPS-stimulated RAW264.7 macrophages were observed using real-time PCR and ELISA methods.DNA oligo was designed and synthesized according to the most effective siRNA 2 sequence.The recombinant lentiviral shuttle plasmid expressing short hairpin RNA ( shRNA) was constructed and sequenced.The lentiviral shuttle plasmids with packaging plasmids were transfected into 293T cells to produce lentiviral particles.Results: ①The TNF-αmRNA relative expression levels of siRNA1, siRNA2 and siRNA3 were 0.24±0.01,0.16±0.02,0.19±0.01 respectively,significantly lower than that of negative control (0.95± 0.02) (F=531.3,P<0.001).The inhibition rates at mRNA level were 74.26%,83.09%,79.93%,respectively comparing with negative control.No significance was observed in IL-1βor IL-6 mRNA relative expression change after TNF-αsiRNA transfection ( P>0.05).②The TNF-αprotein expression levels of siRNA1,siRNA2 and siRNA3 were (23.95±1.21),(17.27±1.46),(19.07± 1.57)ng/ml respectively,significantly lower than that of negative control (35.37±2.93)ng/ml (F=18.1,P=0.000 6<0.001).The inhibition rates of protein expression were 32.29%, 51.16%, 46.08%, respectively comparing with negative control.③The PCR product electrophoresis showed that recombinant vectors yielded 343 bp fragments,non-constructed vectors yielded 306 bp fragments.DNA sequencing partially showed insertion sequence.④Lentiviral particles were obtained by transfecting 293T cells with recombinant lentiviral shuttle plasmids and lentiviral packaging plasmids.Cells grew well during virus production with strong fluorescence expression.The titer of concentrated virus was 2×106 TU/μl.Conclusion:The lentiviral vector harboring RNAi targeting murine TNF-αgene has been successfully constructed.

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"Influence of ""Warming yang and Nourishing qi"" acupuncture treatment in serum IL-12 and IL-18 expression levels in experimental autoimmune myasthenia gravis rats and its mechanism"

Hongfeng WANG ; Shi LI ; Li DONG ; Yufeng WANG

Chinese Journal of Immunology.2014;(7):909-912. doi:10.3969/j.issn.1000-484X.2014.07.010

Objective:To observe the influence of the “Warming yang and Nourishing qi” acupuncture treatment in the serum interleukin-12 (IL-12) and interleukin-18 (IL-18) expression levels,in the rats with experimental autoimmune myasthenis gravis (EAMG) and to analyze the mechanism of acupuncture treatment to myasthenis gravis (MG).Methods: AchRα1 129-145 peptide fragments were used to immune the female Lewis rats to establish EAMG rat models.30 successful EAMG model rats were randomly selected and divided into acupuncture treatment group ,drug control group and model control group ( n=10 );the unmodelled 10 rats purchased at the same period were used as blank control group.The rats in acupuncture treatment group were treated by “Warming yang and Nourishing qi” acupuncture treatment,30 min/time,once a day,7 d for a period,1 d rest between two periods,two periods of continuous treatment.The rats in drug control group were treated by the pyridostigmine lavaged treatment , 18.5 mg/( kg · d ) , continuous treatment of 15 d.The rats in the other two groups had no any special treatment.After treatment,the blood samples from the vein of tail root of the rats in 4 groups were collected ,and the serum expression levels of IL-12 and IL-18 were detected with ELISA method.Results:After treatment,compared with model control group ,acupuncture treatment and drug control group the serum IL-12 and IL-18 expression level of the EAMG rats were significantly decreased ( P<0.05 or P<0.01 ).Compared between acupuncture treatment group and drug control group showed that ,the serum IL-12 and IL-18 expression level of the EAMG rats were no significant difference ( P>0.05 ).Conclusion:“Warming yang and Nourishing qi” acupuncture treatment can reduce the serum IL-12 and IL-18 expression levels of EAMG rats to treat MG ,and the effect of treatment was similar to pyridostigmine.

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Influence of leptin on Th1/Th2 balance in obese children with asthma

Fen PENG ; Suying WU ; Zhaomin LIU

Chinese Journal of Immunology.2014;(7):968-971. doi:10.3969/j.issn.1000-484X.2014.07.022

Objective:To investigate the levels of serum leptin and their effect on Th 1/Th2 balance and the association between serum leptin levels and clinical manifestations in obese children with asthma.Methods: All 56 children with physician-diagnosed moderate-to-severe persistent asthma and 56 healthy controls were included.The children with asthma and without asthma were divided into two groups,obese (n=28) and non-obese (n=28) by body mass index,respectively.Asthma severity was assessed by the asthma symptom score.Peripheral blood samples were collected to determine the levels of leptin , IFN-gamma and IL-4.The results were correlated with the parameters studied.Results:Serum leptin levels were significantly higher in the obese asthma group than in the non -obese asthma group ,as well as being significantly higher in the children with asthma than in the controls ( P<0.01 ) , whereas IFN-gamma levels were significantly higher and IL-4 levels were significantly lower in the obese asthma group than in the non-obese asthma group( P<0.01 ).In addition, the obese asthma group showed higher asthma symptom scores and significantly lower FEV 1 (% of predicted) than did the non-obese asthma group(P<0.01).There was a significantly positive correlation between leptin ,IFN-gamma and IL-4 levels only in the obese asthma group ( P<0.01 ).Conclusion:Leptin is involved in the pathogenesis of asthma in obese and non-obese children ,and its effect is more pronounced in the former.In the presence of high leptin levels , only obese children with asthma exhibited Th1 polarization,with higher IFN-gamma levels and greater asthma severity.

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Expression of TSLP and NF-κB in patients with adenomyosis

Ying LIU ; Yunqing REN ; Liping XUE ; Xiaoyan LI ; Binyu SONG

Chinese Journal of Immunology.2014;(7):963-967. doi:10.3969/j.issn.1000-484X.2014.07.021

Objective:To investigate the role of TSLP and NF-κB in the pathogenesis of adenomyosis.Methods:Immunohisto-chemistry was employed to detect the expression of TSLP and NF-κB p65 in the endometrial glandular cell and stromal cell of 35 adeno-myosis patients and 20 hysteromyoma patients as control , and analyze the correlation of the two proteins.The concentration of TSLP was measured by specific ELISA in the serum of healthy subjects besides of the patients with adenomyosis and hysteromyoma .Results:TSLP and NF-κB p65 were both positively expressd in the glandular cells and stromal cells of ectopic and eutopic endometrium of patients with adenomyosis , their expression was significantly higher than that of the control group ( P<0.01 ) , and the expression of TSLP and NF-κB p65 in ectopic endometrium was significantly higher than that in eutopic endometrium ( P<0.01 ).There was positive correlation between TSLP and NF-κB p65 in ectopic endometrium.The concentration of TSLP was higher in the serum of patients with adenomyosis.Conclusion:The TSLP is highly expressed in endometrial glandular and stromal cells of patients with adenomyosis , and there is higher level of TSLP in serum , and there is correlation between the expression of TSLP and NF-κB in ectopic endometrium.

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Effect and clinical significance of immunological state upon telomerase activation of bone marrow mononuclear cells in hemopoietic microenvironment of immune related hematocytopenia (IRHS)

Lifei SUN ; Qiangqiang WU ; Bing HAN ; Hongfeng HAO ; Gang XU ; Yanhui DU ; Hui MING ; Guichen WANG ; Jinbiao ZHANG

Chinese Journal of Immunology.2014;(7):956-962. doi:10.3969/j.issn.1000-484X.2014.07.020

Objective:To study the effect of immunological molecules expressive state upon the telomerase activation ( TA) of bone marrow mononuclear cells ( BMMNC ) in the hemopoietic microenvironment of patients with immune related hematocytopenia ( IRHS) ,and to explore the immunologic mechanism as well as the clinical significance of hematoclasis in marrow of IRHS patients .Methods:①TRAP-PCR-ELISA method was performed to detect the TA of BMMNC in marrow of 366 IRHS patients before and after therapy.②The molecules HLA-DR,anti-hunman IgG,FcγⅡR,mannose receptor ( MR),IL-17A and its receptor ( IL-17AR) were analysed by immunochemistry and immunofluorescence staining .③The flow cytometric ( FCM) was used to analyse the proportion of CD3+CD4+T cells as well as CD3+CD8+T cells ,CD3-CD19+B cells and CD3-CD16/56+NK cells in peripheral blood lymphocyte.60 cases of health examination were selected as the control group , and 30 cases hypoferric anemia patients were selected as disease control.The differences between patient group and control group were analysed with statistic method .Immunochemistry and immunofluo-rescence staining were performed to in situ analyze the activation-characteristics of immunocyte in bone marrow slides of IRHS ,and the dependablity of cellular immunologic injury was also checked.Results: ①The levels of TA was 0.261 7 ±0.021 6 before treatment , higher than the disease control group (0.061 6±0.031 3 ,P<0.01).Among of them HLA-B27+patients were higher than HLA-B27-patients (0.301 3±0.020 6 vs.0.192 3±0.012 9,P<0.05).Serious IRP patients with HLA-B27+IgG+were obviously higher than HLA-B27-IgG+patients (0.401 6±0.017 2 vs.0.221 1±0.011 0,P<0.01).②In marrow of HLA-B27+IgG+patients,both cell immunity and humoral immunity were in disorder in the hemopoietic microenvironment ,and immonocyte in marrow expressed HLA-DR, FcγⅡR,IL-17A,IL-17RA and MR,and Th,Ts,B cell and NK cell in peripheral blood increased in different degree ,inducing the in-flammation of haemocyte and lead to destruction.③Humoral immunity was in the dominant level in morrow;humoral immunity of HLA-B27-IgG+patients,immonocyte expressed FcγⅡR in high level,but IL-17A was seldom expressed,only CD19+B cell was increased slightly ,the antibody dependent cellular cytotoxicity ( ADCC) was the main mode of destruction.After therapy glucocorticoids associated with ciclosporin A ,the TA level of BMMNC decreased to 0 with devitalization.Conclusion: The telomerase activation of bone marrow mononuclear cells in IRHS is related with the immune state of hemopoietic microenvironment and the pathologic lesion degree of hema -topoietic cell in marrow.It is viral infection and immunological activation as well as a variety of inflammatory factors play a part in the immunologic injury that might be an important factor of the enhancement in TA.

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Expression and significance of BAFF/APRIL in childhood acute lymphoblastic leu-kemia

Lihui WU ; Baolan SUN ; Meiyu XU ; Honghua SONG ; Jianhui GU ; Zhiping YANG ; Hongbing NI

Chinese Journal of Immunology.2014;(7):950-955. doi:10.3969/j.issn.1000-484X.2014.07.019

Objective:To investigate the expression and significance of B cell activating factor (BAFF) and a proliferation-inducing ligand ( APRIL) in children with acute lymphoblastic leukemia ( ALL).Methods:The mRNA and protein expressions in ALL.

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