Objective To investigate if immunological factors associated with rheumatoid arthritis(RA) affect the result of human immunodeficiency virus(HIV) screening by electrochemiluminescence immunoassay(ECLIA) and enzyme-lin...Objective To investigate if immunological factors associated with rheumatoid arthritis(RA) affect the result of human immunodeficiency virus(HIV) screening by electrochemiluminescence immunoassay(ECLIA) and enzyme-linked immunosorbent assay(ELISA). Methods 100 RA cases were enrolled from January 2012 to February 2013 into this study. HIV screening was conducted with ECLIA detecting both HIV-1 p24 antigen, HIV-1 and HIV-2 antibodies, with ELISA and colloidal gold method detecting HIV-1 and HIV-2 antibodies. The samples producing positive results were submitted to the Center for Disease Control for confirmation using Western blotting method. The antibody titers of rheumatoid factors(RF) including RF-IgG, RF-IgM, RF-IgA, and CCP-IgG were analyzed by ELISA. Results The HIV positive-rate determined by ECLIA was significantly higher than that by ELISA and colloidal gold method(P<0.01). The false-positive rate of HIV screening was associated with antibody titers of RF-IgG, RF-IgM, RF-IgA, and CCP-IgG in RA(P<0.01). Conclusion Immunological factors, including RF and anti-CCP antibody, may influence the screening of HIV by ECLIA, producing false-positive result.展开更多
Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors(u PAR) in bone marrow fluid and bone marrow tissue from multiple myeloma(MM) patients and assess association of u PAR lev...Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors(u PAR) in bone marrow fluid and bone marrow tissue from multiple myeloma(MM) patients and assess association of u PAR level with prognosis of MM.Methods u PAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow(control) were examined by ELISA.Furthermore,u PAR expression in bone marrow tissue was investigated by RT-PCR and Western blot,respectively.The distribution of u PAR in MM cells was examined using immunofluorescence staining.The pathological changes in different stages of MM patients were studied by HE staining.Results u PAR level in bone marrow fluid of MM patients(1.52±0.32 μg/ml) was found to be higher than that in the control group(0.98±0.15 μg/ml).Interestingly,u PAR protein(0.686±0.075 vs.0.372±0.043,P<0.05) and m RNA(2.51±0.46 vs.4.46±1.15,P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage.The expression of u PAR in MM bone marrow was confirmed by immunofluorescence staining.Moreover,HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma.Conclusion Our study reveals that u PAR expression is positively correlated with the development and progress of MM.展开更多
Objective:To study the cellular immunity status of patientswith recurrent genital herpes. Methods: Serum levels of interlukin-2 and its solublereceptor and interlukin-6 were measured by ELISA in 34patients with recurr...Objective:To study the cellular immunity status of patientswith recurrent genital herpes. Methods: Serum levels of interlukin-2 and its solublereceptor and interlukin-6 were measured by ELISA in 34patients with recurrent genital herpes. Results: Serum levels of IL-2 and IL-6 were significantlydecreased in patients compared to healthy controls (P<0.01),and the level of sIL-2R was significantly increased in patientswith recurrent genital herpes (P<0.01). There were nosignificant differences in all variables amongst patientsregarding relapse stage and remission stage (P>0.05). Conclusion: There was a cellular immune deficiency inpatients with recurrent genital herpes.展开更多
为了测定水中硝基苯胺类化合物,进而开发酶联免疫分析法(EL ISA)快速检测方法,制备了抗硝基苯胺单克隆抗体。以合成的硝基苯胺完全抗原,作为免疫原免疫B a lb/c小鼠,采用B细胞杂交瘤技术,经免疫、融合、筛选和克隆等,得到了抗硝基苯胺...为了测定水中硝基苯胺类化合物,进而开发酶联免疫分析法(EL ISA)快速检测方法,制备了抗硝基苯胺单克隆抗体。以合成的硝基苯胺完全抗原,作为免疫原免疫B a lb/c小鼠,采用B细胞杂交瘤技术,经免疫、融合、筛选和克隆等,得到了抗硝基苯胺单克隆抗体。该抗体亚类为IgG1,制备的单克隆抗体腹水效果评价达10-7,与其他硝基苯胺结构类似物无交叉反应。实验结果表明,用本间接竞争EL ISA方法可以精确地检测不同水样中硝基苯胺残留。展开更多
To yield cholinesterase（ChE） from prokaryotic expression, the ChE gene that belongs to Daphnia magna was amplified by reverse transcription-polymerase chain reaction（RT-PCR） using forward primer 5＇-CCCYGGNGCSAT G...To yield cholinesterase（ChE） from prokaryotic expression, the ChE gene that belongs to Daphnia magna was amplified by reverse transcription-polymerase chain reaction（RT-PCR） using forward primer 5＇-CCCYGGNGCSAT GATGTG-3＇ and reverse primer 5＇-GYAAGTTRGCCCAATATCT-3＇. To express the gene, one sequence of the amplified DNA, which was able to encode a putative protein containing two conserved carboxylesterase domains, was connected to the prokaryotic expression vector PET-29a（＋）. The recombinant vector was transformed into Escherichia coil BL21（DE3）. Protein expression was induced by isopropy-D-thiogalactoside. The expressed ChE was used as an immunogen to immunize BALB/c mice. The obtained antibodies were tested for their specificity towards crude enzymes from species such as Alona milleri, Macrobrachium nipponense, Bombyx mori, Chironomus kiiensis, Apis mellifera, Eisenia foetida, Brachydanio rerio, and Xenopus laevis. Results indicated that the antibodies had specificity suitable for detecting ChE in Daphnia magna. A type of indirect and non-competitive enzyme-linked immunosorbent assay（IN-ELISA） was used to test the immunoreactive content of ChE（ChE-IR） in Daphina magna. The detection limit of the IN-ELISA was found to be 14.5 ng/ml at an antiserum dilution of 1：22 000. Results from tests on Daphnia magna exposed to sublethal concentrations of triazophos indicated a maximal induction of 57.2% in terms of ChE-IR on the second day after the animals were exposed to a concentration of 2.10 μg/L triazophos. Testing on animals acclimatized to a temperature of 16 °C indicated that ChE-IR was induced by 16.9% compared with the ChE-IR content detected at 21 °C, and the rate of induction was 25.6% at 10 °C. The IN-ELISA was also used to test the stability of ChE-IR in collected samples. Repeated freezing and thawing had no influence on the outcome of the test. All these results suggest that the polyclonal antibodies developed against the recombinant ChE are as efficient as those developed against the native ChE in detecting ChE content in Daphnia magna.展开更多
基金Supported by Shanghai Municipal Natural Science Foundation(11ZR1427000)
文摘Objective To investigate if immunological factors associated with rheumatoid arthritis(RA) affect the result of human immunodeficiency virus(HIV) screening by electrochemiluminescence immunoassay(ECLIA) and enzyme-linked immunosorbent assay(ELISA). Methods 100 RA cases were enrolled from January 2012 to February 2013 into this study. HIV screening was conducted with ECLIA detecting both HIV-1 p24 antigen, HIV-1 and HIV-2 antibodies, with ELISA and colloidal gold method detecting HIV-1 and HIV-2 antibodies. The samples producing positive results were submitted to the Center for Disease Control for confirmation using Western blotting method. The antibody titers of rheumatoid factors(RF) including RF-IgG, RF-IgM, RF-IgA, and CCP-IgG were analyzed by ELISA. Results The HIV positive-rate determined by ECLIA was significantly higher than that by ELISA and colloidal gold method(P<0.01). The false-positive rate of HIV screening was associated with antibody titers of RF-IgG, RF-IgM, RF-IgA, and CCP-IgG in RA(P<0.01). Conclusion Immunological factors, including RF and anti-CCP antibody, may influence the screening of HIV by ECLIA, producing false-positive result.
基金Supported by Huzhou Science and Technology Bureau and Huzhou Central Hospital(2014GYB14)
文摘Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors(u PAR) in bone marrow fluid and bone marrow tissue from multiple myeloma(MM) patients and assess association of u PAR level with prognosis of MM.Methods u PAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow(control) were examined by ELISA.Furthermore,u PAR expression in bone marrow tissue was investigated by RT-PCR and Western blot,respectively.The distribution of u PAR in MM cells was examined using immunofluorescence staining.The pathological changes in different stages of MM patients were studied by HE staining.Results u PAR level in bone marrow fluid of MM patients(1.52±0.32 μg/ml) was found to be higher than that in the control group(0.98±0.15 μg/ml).Interestingly,u PAR protein(0.686±0.075 vs.0.372±0.043,P<0.05) and m RNA(2.51±0.46 vs.4.46±1.15,P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage.The expression of u PAR in MM bone marrow was confirmed by immunofluorescence staining.Moreover,HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma.Conclusion Our study reveals that u PAR expression is positively correlated with the development and progress of MM.
文摘Objective:To study the cellular immunity status of patientswith recurrent genital herpes. Methods: Serum levels of interlukin-2 and its solublereceptor and interlukin-6 were measured by ELISA in 34patients with recurrent genital herpes. Results: Serum levels of IL-2 and IL-6 were significantlydecreased in patients compared to healthy controls (P<0.01),and the level of sIL-2R was significantly increased in patientswith recurrent genital herpes (P<0.01). There were nosignificant differences in all variables amongst patientsregarding relapse stage and remission stage (P>0.05). Conclusion: There was a cellular immune deficiency inpatients with recurrent genital herpes.
文摘为了测定水中硝基苯胺类化合物,进而开发酶联免疫分析法(EL ISA)快速检测方法,制备了抗硝基苯胺单克隆抗体。以合成的硝基苯胺完全抗原,作为免疫原免疫B a lb/c小鼠,采用B细胞杂交瘤技术,经免疫、融合、筛选和克隆等,得到了抗硝基苯胺单克隆抗体。该抗体亚类为IgG1,制备的单克隆抗体腹水效果评价达10-7,与其他硝基苯胺结构类似物无交叉反应。实验结果表明,用本间接竞争EL ISA方法可以精确地检测不同水样中硝基苯胺残留。
基金supported by the Zhejiang Provincial Natural Science Foundation(No.LY12B07008),China
文摘To yield cholinesterase（ChE） from prokaryotic expression, the ChE gene that belongs to Daphnia magna was amplified by reverse transcription-polymerase chain reaction（RT-PCR） using forward primer 5＇-CCCYGGNGCSAT GATGTG-3＇ and reverse primer 5＇-GYAAGTTRGCCCAATATCT-3＇. To express the gene, one sequence of the amplified DNA, which was able to encode a putative protein containing two conserved carboxylesterase domains, was connected to the prokaryotic expression vector PET-29a（＋）. The recombinant vector was transformed into Escherichia coil BL21（DE3）. Protein expression was induced by isopropy-D-thiogalactoside. The expressed ChE was used as an immunogen to immunize BALB/c mice. The obtained antibodies were tested for their specificity towards crude enzymes from species such as Alona milleri, Macrobrachium nipponense, Bombyx mori, Chironomus kiiensis, Apis mellifera, Eisenia foetida, Brachydanio rerio, and Xenopus laevis. Results indicated that the antibodies had specificity suitable for detecting ChE in Daphnia magna. A type of indirect and non-competitive enzyme-linked immunosorbent assay（IN-ELISA） was used to test the immunoreactive content of ChE（ChE-IR） in Daphina magna. The detection limit of the IN-ELISA was found to be 14.5 ng/ml at an antiserum dilution of 1：22 000. Results from tests on Daphnia magna exposed to sublethal concentrations of triazophos indicated a maximal induction of 57.2% in terms of ChE-IR on the second day after the animals were exposed to a concentration of 2.10 μg/L triazophos. Testing on animals acclimatized to a temperature of 16 °C indicated that ChE-IR was induced by 16.9% compared with the ChE-IR content detected at 21 °C, and the rate of induction was 25.6% at 10 °C. The IN-ELISA was also used to test the stability of ChE-IR in collected samples. Repeated freezing and thawing had no influence on the outcome of the test. All these results suggest that the polyclonal antibodies developed against the recombinant ChE are as efficient as those developed against the native ChE in detecting ChE content in Daphnia magna.
