Objective To characterize the immune reaction in SD rats exposed to trichloroethylene （TCE） and to identify the gene expression profiles involved in skin after TCE exposure. Methods Fifteen percent of TCE was inject...Objective To characterize the immune reaction in SD rats exposed to trichloroethylene （TCE） and to identify the gene expression profiles involved in skin after TCE exposure. Methods Fifteen percent of TCE was injected intradermally into the rat back （100 μL/120 g） at intervals of 7 days. Whole blood was collected 24 h after the fifth or seventh intradermic administration of TCE. The percentages of CD4＋ and CD8＋ of T lymphocytes were measured by a flow cytometer. The concentrations of IFN-gamma and 1L-4 in the serum were semi-quantified by ELISA. Total RNAs of skin samples at 3 h or 24 h after the seventh dose of TCE in SD rats were extracted, and gene expression proftles of these tissues were analyszed by rat toxicology U34 array of Affymetrix. Results Obvious decline of CD4＋ in T lymphocytes was observed in the TCE-administer group. No significant concentration differences in IFN-gamma and IL-4 were found between TCE-treated and control rats. Gadd45a and Mel were significantly up regulated in skin tissue 24 h after TCE exposure. The expression regulation of immune response factors was as active as proteins associated with lipid metabolism and synthesis process in these skin samples of SD rats exposed to TCE. Conclusion T-helper type 1 cells mediate immune response can not be elicited in TCE-treated SD rats, but certain immune disorder can be induced.展开更多
It is known to be that lactic acid bacteria induce the IL-12. IL-12 activates NK cells and promotes the production of IFN-<em>γ</em>. IFN-<em>γ</em> activates macrophages, resulting in enhanc...It is known to be that lactic acid bacteria induce the IL-12. IL-12 activates NK cells and promotes the production of IFN-<em>γ</em>. IFN-<em>γ</em> activates macrophages, resulting in enhanced phagocytosis and bactericidal activity. We have been investigating fermented foods that activate the immune function. For that purpose, a specific antibody is required. We tried to express IL-12p35 by the usual method, but IL-12p35 was not expressed at all. In the present study, we constructed, purified human IL-12p35 and obtained a specific antibody against IL-12p35. We also purified human IFN-<em>γ</em> and obtained specific antibody against IFN-<em>γ</em>. We have established a method for expressing poorly expressed proteins. The method we have established can be applied to the purification of poorly expressed proteins and antibody production.展开更多
This work aimed to find quantitative phenotypic traits that can be used to discriminate the levels of resistance/susceptibility to B. bovis and B. bigemina in two groups of cattle presenting the highest (H) or lowest ...This work aimed to find quantitative phenotypic traits that can be used to discriminate the levels of resistance/susceptibility to B. bovis and B. bigemina in two groups of cattle presenting the highest (H) or lowest (L) infection levels and Rhipicephalus microplus ticks count. The animals were selected from a previous study of 50 Canchim (5/8 Charolais/zebu) heifers raised in an endemic area for these parasites. These animals were evaluated regarding their TNFα, IL10, IFN-γ, IL12 and iNOS mRNA levels. No differences were found between these groups regarding TNFα, IFN-γ, IL12β or iNOS transcripts. However, the IL10 transcripts were significantly higher in the H group compared to the L group. Moreover, significant correlation coefficients were observed between B. bovis loads and both IL10 and IFN-γ transcripts, while no correlations were found for B. bigemina loads and all tested immune-related transcripts, suggesting that differential IL10 mRNA profiles were closely associated to B. bovis loads. Our results have contributed to a better understanding of the immune responses against Babesia infection, as we demonstrated that the IL10 cytokine levels might also influence or be influenced by parasitemia levels in persistently infected animals.展开更多
Objective: To demonstrate the utility of DNA vaccines for the tailored methods, the efficacy of enhanced immune responses, and the types of increased im- mune responses. Methods: Four recombinant plasmids constructed ...Objective: To demonstrate the utility of DNA vaccines for the tailored methods, the efficacy of enhanced immune responses, and the types of increased im- mune responses. Methods: Four recombinant plasmids constructed in- cluded the coding regions for the core protein (pC) and for the core, E_1 and E_2 together (pCE_1E_2), IL- 12 p35 and p40. These plasmids were transfected into mammalian cells to test their protein expression and were injected into the quadriceps muscles of BALB/ C mice for measurement of specific antibodies and cytotoxic T-lymphocyte (CTL) responses. Results: All the recombinant plasmids were shown to express specific antigens stably in mammalian cells. Codelivery of pIL-12 expression cassettes with pC and pCE_1E_2 in mice resulted in the enhancement of Ag-dependent CTL responses and the reduction of specific Ab response. The CTL activity was: pC= 18.65%±5.71%, pCE_1E_2=20.07%±11.11%, pC +pIL-12=60.11%±17.37%, pCE_1E_2+pIL-12= 67.48%±15.57%, respectively. The average A val- ues of anti-HCV were pC=0.415±0.127, pCE_1E_2= 0.358±0.096, pC+pIL-12=0.210±0.086, pCE_1E_2 +pIL-12=0.258±0.125. Conclusion: Codelivery of pIL-12 with plasmid DNA can enhance the efficacy of immune responses and shift the type of immune responses.展开更多
Objective:To observe the intervention influence and effect of simvastatin on the expression of interleukin 17(L117),high mobility group protein 1(HMGB1)and TLR4 path in Lupus nephritis(LN)rats.Methods:A total of 28 BS...Objective:To observe the intervention influence and effect of simvastatin on the expression of interleukin 17(L117),high mobility group protein 1(HMGB1)and TLR4 path in Lupus nephritis(LN)rats.Methods:A total of 28 BSXSB male mice with LN(16 weeks)were randomlv divided into observation group and the comparison group,observation group was given 6 mg·kg^(-1)·d^(-1)simvastatin in 0.1%PBS lavage for 4 weeks.the comparison group was not given any trestment.Blood urea nitrogen(BUN)level and urine trace albumin(Scr)level of two groups were determined.The expression of IL17.HMGB1 and TLR4 protein was detected using immune histochemical method,and the kidney histological damage was observed.Results:BNU,LI17.HMGB1,TLR4protein and HMGB1 mRNA in observation group was significantly lower than that in control group(P<0.05):There was no statistical difference of Scr level between two goups(P<0.05).Histological observation showed glomerular lesions integral of observation group was obviously lower than that of control group.Conclusions:Simvastatin can reduce the expression of IL17.HMGB1 and TLR4 protein in LN mice,thereby can inhibit the autoimmune response as a potential treatment function of LN.展开更多
Using restriction endonuclease Hind,. Sal i, the IL-lra cDNA was isolated from PRCra, then by a medium plasmid pUC19, It was cloned into a high level expressive vector PBV220. Therecombinant plasmid pBV-IL--Ira waS tr...Using restriction endonuclease Hind,. Sal i, the IL-lra cDNA was isolated from PRCra, then by a medium plasmid pUC19, It was cloned into a high level expressive vector PBV220. Therecombinant plasmid pBV-IL--Ira waS transrormed Into E. coil DH-So. After induced by temperaIure, a 20KD protein could be detected by SDS-PAGE. Biological assay in vitro showed that the protein could inhibit the activity or IL-l obviously.展开更多
Background: Psoriasis is considered a common skin disease, marked by the production and elevation of inflammatory plaques that regularly shed scales resulting from extensive skin epithelial cell proliferation. T lymph...Background: Psoriasis is considered a common skin disease, marked by the production and elevation of inflammatory plaques that regularly shed scales resulting from extensive skin epithelial cell proliferation. T lymphocytes, neutrophils, and other leukocytes enter the irritated skin, resulting in epidermal keratinocyte hyperplasia, vascular hyperplasia, ectasia, and T lymphocyte, neutrophil other forms of leukocyte infiltration. Aim of the Work: the study aimed to investigate the role of IL-37 and VEGF gene expression in the pathogenesis of psoriasis in Egyptian patients. Methodology: Polymerase chain reaction techniques (PCR) were applied to detect VEGF in the skin homogenates of psoriasis patients. In addition, the ELIZA technique was applied to investigate IL-37 in skin homogenates. One hundred cases have been divided into two groups: 50 healthy volunteers as the group I healthy control;50 psoriasis patients as group II. PCR real-time technology was assessed through extracted DNA samples for VEGF amplification in skin homogenate. Result: Our results revealed that psoriasis patients significantly had a substantial reduction in IL-37 compared to the control group (p Conclusion: There is a significant inverse association between IL-37 and VEGF in psoriasis patients. Study findings revealed that IL-37 gene expression decreases while VEGF gene expression increases in psoriatic individuals. Such a measurement may be beneficial in determining the severity of the condition, as well as taking into consideration of the disease’s diagnosis.展开更多
目的探讨姜黄素对脓毒症肺损伤患者血清NT-pro-BNP、IL-6、IL-10及肺功能的影响。方法收集河北大学附属医院收治的脓毒症肺损伤患者40例,根据用药不同分为对照组和实验组,每组各20例,对照组给予常规治疗,实验组在对照组基础上给予姜黄...目的探讨姜黄素对脓毒症肺损伤患者血清NT-pro-BNP、IL-6、IL-10及肺功能的影响。方法收集河北大学附属医院收治的脓毒症肺损伤患者40例,根据用药不同分为对照组和实验组,每组各20例,对照组给予常规治疗,实验组在对照组基础上给予姜黄素胶囊,治疗连续1周。治疗结束后,对所有患者的血清NT-pro-BNP、IL-6、IL-10及肺功能情况进行检测。结果与对照组治疗后相比,实验组患者血清IL-6、IL-10水平较低(P<0.05),实验组患者的血清NT-pro-BNP水平较低(P<0.05),实验组患者的肺活量(vital capacity,VC)、最大通气量(maximal voluntary ventilation,MVV)、一秒用力呼气量(forced expiratory volume in one second,FEV1)水平较高(P<0.05)。结论姜黄素能够显著降低脓毒症肺损伤患者血清IL-6、IL-10以及NT-pro-BNP水平,改善肺功能。展开更多
The gene encoding human interleukin 10 was amplified with PCR from the pcDNA/3.1 plasmid,and then was cloned into the yeast expression vector pPIC9K containing the AOX1 promoter and the secretion signal sequence codin...The gene encoding human interleukin 10 was amplified with PCR from the pcDNA/3.1 plasmid,and then was cloned into the yeast expression vector pPIC9K containing the AOX1 promoter and the secretion signal sequence coding afactor prepro-leader peptide.Then the yeast expression vector pPIC9K/hIL10 was con-structed and transformed to P.pastoris GS115 strains,screened for multiple inserts,induced with 0.5% methanol in shake flask cultures.Increasing level of rhIL10 was detected in the medium for up to 4 days by SDS-PAGE analysis.Western blot shows that the expressed rhIL10 exhibits high seificity and antigenicity.展开更多
基金This work was supported by "973" Project (No. 2002CB512903) and Shenzhen Bureau of Science and Technology, China (No. 200204121 No. 200304156).
文摘Objective To characterize the immune reaction in SD rats exposed to trichloroethylene （TCE） and to identify the gene expression profiles involved in skin after TCE exposure. Methods Fifteen percent of TCE was injected intradermally into the rat back （100 μL/120 g） at intervals of 7 days. Whole blood was collected 24 h after the fifth or seventh intradermic administration of TCE. The percentages of CD4＋ and CD8＋ of T lymphocytes were measured by a flow cytometer. The concentrations of IFN-gamma and 1L-4 in the serum were semi-quantified by ELISA. Total RNAs of skin samples at 3 h or 24 h after the seventh dose of TCE in SD rats were extracted, and gene expression proftles of these tissues were analyszed by rat toxicology U34 array of Affymetrix. Results Obvious decline of CD4＋ in T lymphocytes was observed in the TCE-administer group. No significant concentration differences in IFN-gamma and IL-4 were found between TCE-treated and control rats. Gadd45a and Mel were significantly up regulated in skin tissue 24 h after TCE exposure. The expression regulation of immune response factors was as active as proteins associated with lipid metabolism and synthesis process in these skin samples of SD rats exposed to TCE. Conclusion T-helper type 1 cells mediate immune response can not be elicited in TCE-treated SD rats, but certain immune disorder can be induced.
文摘It is known to be that lactic acid bacteria induce the IL-12. IL-12 activates NK cells and promotes the production of IFN-<em>γ</em>. IFN-<em>γ</em> activates macrophages, resulting in enhanced phagocytosis and bactericidal activity. We have been investigating fermented foods that activate the immune function. For that purpose, a specific antibody is required. We tried to express IL-12p35 by the usual method, but IL-12p35 was not expressed at all. In the present study, we constructed, purified human IL-12p35 and obtained a specific antibody against IL-12p35. We also purified human IFN-<em>γ</em> and obtained specific antibody against IFN-<em>γ</em>. We have established a method for expressing poorly expressed proteins. The method we have established can be applied to the purification of poorly expressed proteins and antibody production.
文摘This work aimed to find quantitative phenotypic traits that can be used to discriminate the levels of resistance/susceptibility to B. bovis and B. bigemina in two groups of cattle presenting the highest (H) or lowest (L) infection levels and Rhipicephalus microplus ticks count. The animals were selected from a previous study of 50 Canchim (5/8 Charolais/zebu) heifers raised in an endemic area for these parasites. These animals were evaluated regarding their TNFα, IL10, IFN-γ, IL12 and iNOS mRNA levels. No differences were found between these groups regarding TNFα, IFN-γ, IL12β or iNOS transcripts. However, the IL10 transcripts were significantly higher in the H group compared to the L group. Moreover, significant correlation coefficients were observed between B. bovis loads and both IL10 and IFN-γ transcripts, while no correlations were found for B. bigemina loads and all tested immune-related transcripts, suggesting that differential IL10 mRNA profiles were closely associated to B. bovis loads. Our results have contributed to a better understanding of the immune responses against Babesia infection, as we demonstrated that the IL10 cytokine levels might also influence or be influenced by parasitemia levels in persistently infected animals.
文摘Objective: To demonstrate the utility of DNA vaccines for the tailored methods, the efficacy of enhanced immune responses, and the types of increased im- mune responses. Methods: Four recombinant plasmids constructed in- cluded the coding regions for the core protein (pC) and for the core, E_1 and E_2 together (pCE_1E_2), IL- 12 p35 and p40. These plasmids were transfected into mammalian cells to test their protein expression and were injected into the quadriceps muscles of BALB/ C mice for measurement of specific antibodies and cytotoxic T-lymphocyte (CTL) responses. Results: All the recombinant plasmids were shown to express specific antigens stably in mammalian cells. Codelivery of pIL-12 expression cassettes with pC and pCE_1E_2 in mice resulted in the enhancement of Ag-dependent CTL responses and the reduction of specific Ab response. The CTL activity was: pC= 18.65%±5.71%, pCE_1E_2=20.07%±11.11%, pC +pIL-12=60.11%±17.37%, pCE_1E_2+pIL-12= 67.48%±15.57%, respectively. The average A val- ues of anti-HCV were pC=0.415±0.127, pCE_1E_2= 0.358±0.096, pC+pIL-12=0.210±0.086, pCE_1E_2 +pIL-12=0.258±0.125. Conclusion: Codelivery of pIL-12 with plasmid DNA can enhance the efficacy of immune responses and shift the type of immune responses.
基金supported by National Science Foundation of China:81273731
文摘Objective:To observe the intervention influence and effect of simvastatin on the expression of interleukin 17(L117),high mobility group protein 1(HMGB1)and TLR4 path in Lupus nephritis(LN)rats.Methods:A total of 28 BSXSB male mice with LN(16 weeks)were randomlv divided into observation group and the comparison group,observation group was given 6 mg·kg^(-1)·d^(-1)simvastatin in 0.1%PBS lavage for 4 weeks.the comparison group was not given any trestment.Blood urea nitrogen(BUN)level and urine trace albumin(Scr)level of two groups were determined.The expression of IL17.HMGB1 and TLR4 protein was detected using immune histochemical method,and the kidney histological damage was observed.Results:BNU,LI17.HMGB1,TLR4protein and HMGB1 mRNA in observation group was significantly lower than that in control group(P<0.05):There was no statistical difference of Scr level between two goups(P<0.05).Histological observation showed glomerular lesions integral of observation group was obviously lower than that of control group.Conclusions:Simvastatin can reduce the expression of IL17.HMGB1 and TLR4 protein in LN mice,thereby can inhibit the autoimmune response as a potential treatment function of LN.
文摘Using restriction endonuclease Hind,. Sal i, the IL-lra cDNA was isolated from PRCra, then by a medium plasmid pUC19, It was cloned into a high level expressive vector PBV220. Therecombinant plasmid pBV-IL--Ira waS transrormed Into E. coil DH-So. After induced by temperaIure, a 20KD protein could be detected by SDS-PAGE. Biological assay in vitro showed that the protein could inhibit the activity or IL-l obviously.
文摘Background: Psoriasis is considered a common skin disease, marked by the production and elevation of inflammatory plaques that regularly shed scales resulting from extensive skin epithelial cell proliferation. T lymphocytes, neutrophils, and other leukocytes enter the irritated skin, resulting in epidermal keratinocyte hyperplasia, vascular hyperplasia, ectasia, and T lymphocyte, neutrophil other forms of leukocyte infiltration. Aim of the Work: the study aimed to investigate the role of IL-37 and VEGF gene expression in the pathogenesis of psoriasis in Egyptian patients. Methodology: Polymerase chain reaction techniques (PCR) were applied to detect VEGF in the skin homogenates of psoriasis patients. In addition, the ELIZA technique was applied to investigate IL-37 in skin homogenates. One hundred cases have been divided into two groups: 50 healthy volunteers as the group I healthy control;50 psoriasis patients as group II. PCR real-time technology was assessed through extracted DNA samples for VEGF amplification in skin homogenate. Result: Our results revealed that psoriasis patients significantly had a substantial reduction in IL-37 compared to the control group (p Conclusion: There is a significant inverse association between IL-37 and VEGF in psoriasis patients. Study findings revealed that IL-37 gene expression decreases while VEGF gene expression increases in psoriatic individuals. Such a measurement may be beneficial in determining the severity of the condition, as well as taking into consideration of the disease’s diagnosis.
文摘目的探讨姜黄素对脓毒症肺损伤患者血清NT-pro-BNP、IL-6、IL-10及肺功能的影响。方法收集河北大学附属医院收治的脓毒症肺损伤患者40例,根据用药不同分为对照组和实验组,每组各20例,对照组给予常规治疗,实验组在对照组基础上给予姜黄素胶囊,治疗连续1周。治疗结束后,对所有患者的血清NT-pro-BNP、IL-6、IL-10及肺功能情况进行检测。结果与对照组治疗后相比,实验组患者血清IL-6、IL-10水平较低(P<0.05),实验组患者的血清NT-pro-BNP水平较低(P<0.05),实验组患者的肺活量(vital capacity,VC)、最大通气量(maximal voluntary ventilation,MVV)、一秒用力呼气量(forced expiratory volume in one second,FEV1)水平较高(P<0.05)。结论姜黄素能够显著降低脓毒症肺损伤患者血清IL-6、IL-10以及NT-pro-BNP水平,改善肺功能。
文摘The gene encoding human interleukin 10 was amplified with PCR from the pcDNA/3.1 plasmid,and then was cloned into the yeast expression vector pPIC9K containing the AOX1 promoter and the secretion signal sequence coding afactor prepro-leader peptide.Then the yeast expression vector pPIC9K/hIL10 was con-structed and transformed to P.pastoris GS115 strains,screened for multiple inserts,induced with 0.5% methanol in shake flask cultures.Increasing level of rhIL10 was detected in the medium for up to 4 days by SDS-PAGE analysis.Western blot shows that the expressed rhIL10 exhibits high seificity and antigenicity.
