AIM： To perform a meta-analysis of the prevalence of anti-ribosomal P （aRP） antibodies in lupus psychosis, and the odds of psychosis in aRP-positive subjects.METHODS： We identifed articles by searching PubMed, Psy...AIM： To perform a meta-analysis of the prevalence of anti-ribosomal P （aRP） antibodies in lupus psychosis, and the odds of psychosis in aRP-positive subjects.METHODS： We identifed articles by searching PubMed, PsychInfo, and ISI, and the reference lists of identifed studies.RESULTS： Twenty-four studies met the inclusion criteria. Positive aRP antibodies were found in 51% （91 of 179 total cases） of cases of lupus psychosis. There was an almost 3.5-fold increased odds of psychosis in aRP-positive patients （OR = 3.46, 95%CI： 1.97-6.09, P 〈 0.001）. The population attributable risk percentage was 36% for aRP antibodies.CONCLUSION： aRP antibodies are common in lupus psychosis, although the potential mechanism（s） underlying this association remain unclear. Given the overlap between the clinical presentation and risk factors for lupus psychosis and schizophrenia, further investigation of aRP antibodies in schizophrenia is warranted.展开更多
P65 protein, the major immunodominant protein of Mycoplasma hyopneu-moniae （Mhp） exhibiting no cross-reaction with other mycoplasmas, is general y used as a target protein for Mhp detection. In this study, BALB/c mi...P65 protein, the major immunodominant protein of Mycoplasma hyopneu-moniae （Mhp） exhibiting no cross-reaction with other mycoplasmas, is general y used as a target protein for Mhp detection. In this study, BALB/c mice were immunized with prokaryotical y expressed P65 recombinant protein to prepare monoclonal anti-body. After screening with Mhp whole-cel protein and P65 protein, a specific hy-bridoma cel line, 3G12, was obtained by ELISA. Identification results indicated that the antibody secreted by 3G12 hybridoma cel s could react with P65 protein and Mhp whole-cel protein. According to indirect ELISA assay, 3G12 cel culture super-natant possessed a titer of 1∶12 800 against P65 protein and 1∶3 200 against Mhp whole-cel protein; 3G12 ascites possessed a titer of above 1∶4 000 000 against P65 protein and above 1∶20 000 against Mhp 168 whole-cel protein. After long-term in vitro culture and continuous freezing-thawing, 3G12 cel line could stably secrete antibodies. A monoclonal antibody against P65 protein and Mhp whole-cel protein was successful y obtained in the present study, which provided basis for further in-vestigating the pathogenic mechanism of Mhp and establishing diagnostic methods of Mycoplasmal pneumonia of swine （MPS）.展开更多
Objective: Conventional immunohistochemistry (IHC) is available to assess P53 mutations, and expensive imported antiP53 monoclonal antibody has been used in China, it is necessary to study a new monoclonal antibody. M...Objective: Conventional immunohistochemistry (IHC) is available to assess P53 mutations, and expensive imported antiP53 monoclonal antibody has been used in China, it is necessary to study a new monoclonal antibody. Methods: The P53 DNA fragment enconding Nterminal 180 amiao acide was obtained by PCR and was cloned into PGEX2T plasmid expressing glutathione Stransferase (GST). The P53GST fusion protein expressed by JM109 was used for immunizing BALB/C mice. We have raised one hybridoma strain secreting McAb to human P53 (named M126). Results: The IHC analysis of 52 paraffinembedded sections from human breast cancer with M126 and PAB1801 (Zymed Co.) has showed that the positive immunoreactions were 25 cases (48%) and 22 cases (42.3%) respectively. The staining of M126 was stronger and preferable to PAB1801. Conclusion: M126 can be instead of PAB1801 for studying immunohistochemical analysis on P53 protein.展开更多
Objective：The aim of this study was to prepare monoclonal antibody against P53, a kind of tumor suppressor protein,and use the antibody initial y in clinical immunoassay. Methods：Monoclonal antibody was prepared and...Objective：The aim of this study was to prepare monoclonal antibody against P53, a kind of tumor suppressor protein,and use the antibody initial y in clinical immunoassay. Methods：Monoclonal antibody was prepared and identified via the classic protocol of monoclonal antibody preparation. Identified monoclonal antibodies were purified by af inity chro-matography. Antibody titer was determined by enzyme linked immunosorbent assay （ELISA）. The specific binding activity of antibody was detected by Western blotting and immunohistochemistry. Results：Three strains of monoclonal antibodies named 1P15, 2P37 and 3P40 were obtained and purified by af inity chromatography. The purity of antibodies was higher than 90%. The titers of antibodies were more than 1：6000. Western blot and immunohistochemistry assay showed that the specific antibody can combine with endogenous P53 protein in the tumor celllines and determine the expression of P53 in tumor tis-sue. Conclusion：Three strains of monoclonal antibodies with high af inity to P53 were successful y established, which can be used for detecting the expression of P53 in tumor cells or tissue.展开更多
The cDNA of amphioxus p23, a highly conserved co-chaperone for Hsp90, was cloned into a bacterial expression vector pGEX - 6P - 1 and the GST-tagged fusion protein was produced in Eschherichia coli cells. The recombin...The cDNA of amphioxus p23, a highly conserved co-chaperone for Hsp90, was cloned into a bacterial expression vector pGEX - 6P - 1 and the GST-tagged fusion protein was produced in Eschherichia coli cells. The recombinant p23 was purified by affinity purification, and its molecular mass was estimated to be approximately 22 kDa by sodium dedecyl sulfate-polyacrylamide gel electrephoresis. The N-terminus of purified p23 was sequenced, and the resulting amino acid sequence matches exactly the predicted residues deduced from the amphioxus p23 gene. Besides, pelyclonal antibodies against the recombinant p23 were generated, and these antibodies not only recognized specifically the fusion protein GST - p23 from induced E. coli cells, purified GST - p23 and p23 protein, but also reacted with the total protein extracted fi＇om the adult amphioxus and formed a single positive band. These results pave the way for identifying its tissue and subcellular localization, and may open the door to clarifying its structure and mechanisms of biological role.展开更多
We are developing methods to quantify antibody interactions that include a quartz crystal microbalance (QCM) system to measure, on a molecular basis, the interaction of p53 and anti-p53 antibodies. Previously, as a mo...We are developing methods to quantify antibody interactions that include a quartz crystal microbalance (QCM) system to measure, on a molecular basis, the interaction of p53 and anti-p53 antibodies. Previously, as a model system, we developed a measurement device consisting of p53 protein (human wild type), characterized by mass spectroscopy and immobilized at various concentrations on a glass slide. The device is designed as a control to be used with immunohistochemical (IHC) assays that incorporate commercially available anti-p53 antibodies and probes. In the current study, p53 protein is covalently immobilized on a silicon dioxide-coated quartz crystal and the resonance frequency shift is followed in-situ. The functionalized sensor is then incubated with the anti-p53 antibody, which provides a direct gravimetric measure of the antibody-antigen binding. This previously described method allows the comparison of the surface immobilized protein concentrations with estimates obtained by fluorescence measurement. The p53 functionalized QCM system offers an independent measure of surface immobilized protein concentration and is essential in development of our IHC calibration prototypes. These results provide a benchmark for comparing antibody systems that may be used in developing other molecular diagnostic assays such as immunocytochemical analysis and the detection of biomarker proteins in blood and urine.展开更多
目的:探讨P选择素、抗心磷脂抗体(anticardiolipin antibody,ACA)及活化蛋白C抵抗(activated protein c resistance,APCR)对下肢创伤骨折术后深静脉血栓形成(deep venous thrombosis,DVT)的预测价值。方法:选择接受手术治疗的下肢创伤...目的:探讨P选择素、抗心磷脂抗体(anticardiolipin antibody,ACA)及活化蛋白C抵抗(activated protein c resistance,APCR)对下肢创伤骨折术后深静脉血栓形成(deep venous thrombosis,DVT)的预测价值。方法:选择接受手术治疗的下肢创伤骨折患者,术后第3天抽取患者静脉血,采用酶联免疫吸附法测定P选择素含量和ACA阳性率,采用Dahlback法测定APCR阳性率。按照《深静脉血栓形成的诊断和治疗指南(第三版)》中DVT的诊断标准判断患者术后2周内是否发生DVT,比较DVT组和无DVT组的血清P选择素含量、血清ACA阳性率及血清APCR阳性率。以是否发生DVT为因变量,以血清P选择素、ACA及APCR为自变量,进行多因素Logistic回归分析。结果:共纳入648例患者,其中31例患者术后2周内发生DVT(DVT组),其余617例均未检出DVT(无DVT组)。2组患者的血清P选择素含量、血清ACA阳性率及血清APCR阳性率比较,差异均有统计学意义(t=26.989,P=0.000;χ^2=0.010,P=0.000;χ^2=12.447,P=0.000)。多因素Logistic回归分析结果显示,血清P选择素、ACA及APCR是下肢创伤骨折术后DVT的危险因素(OR=1.219,P=0.019;OR=1.292,P=0.009;OR=2.430,P=0.012)。结论:P选择素、ACA及APCR是下肢创伤骨折术后DVT的危险因素,可作为预测下肢创伤骨折术后DVT的检测指标。展开更多
基金Supported by The National Institute of Mental Health(1K23MH098014)Georgia Regents Universityhonoraria from Medscape,Insight Consulting,and Decision Resources Group
文摘AIM： To perform a meta-analysis of the prevalence of anti-ribosomal P （aRP） antibodies in lupus psychosis, and the odds of psychosis in aRP-positive subjects.METHODS： We identifed articles by searching PubMed, PsychInfo, and ISI, and the reference lists of identifed studies.RESULTS： Twenty-four studies met the inclusion criteria. Positive aRP antibodies were found in 51% （91 of 179 total cases） of cases of lupus psychosis. There was an almost 3.5-fold increased odds of psychosis in aRP-positive patients （OR = 3.46, 95%CI： 1.97-6.09, P 〈 0.001）. The population attributable risk percentage was 36% for aRP antibodies.CONCLUSION： aRP antibodies are common in lupus psychosis, although the potential mechanism（s） underlying this association remain unclear. Given the overlap between the clinical presentation and risk factors for lupus psychosis and schizophrenia, further investigation of aRP antibodies in schizophrenia is warranted.
基金Supported by National Natural Science Foundation of China(31100136,3111339)Independent Innovation Fund of Agricultural Science and Technology of Jiangsu Province[CX(13)3066]~~
文摘P65 protein, the major immunodominant protein of Mycoplasma hyopneu-moniae （Mhp） exhibiting no cross-reaction with other mycoplasmas, is general y used as a target protein for Mhp detection. In this study, BALB/c mice were immunized with prokaryotical y expressed P65 recombinant protein to prepare monoclonal anti-body. After screening with Mhp whole-cel protein and P65 protein, a specific hy-bridoma cel line, 3G12, was obtained by ELISA. Identification results indicated that the antibody secreted by 3G12 hybridoma cel s could react with P65 protein and Mhp whole-cel protein. According to indirect ELISA assay, 3G12 cel culture super-natant possessed a titer of 1∶12 800 against P65 protein and 1∶3 200 against Mhp whole-cel protein; 3G12 ascites possessed a titer of above 1∶4 000 000 against P65 protein and above 1∶20 000 against Mhp 168 whole-cel protein. After long-term in vitro culture and continuous freezing-thawing, 3G12 cel line could stably secrete antibodies. A monoclonal antibody against P65 protein and Mhp whole-cel protein was successful y obtained in the present study, which provided basis for further in-vestigating the pathogenic mechanism of Mhp and establishing diagnostic methods of Mycoplasmal pneumonia of swine （MPS）.
文摘Objective: Conventional immunohistochemistry (IHC) is available to assess P53 mutations, and expensive imported antiP53 monoclonal antibody has been used in China, it is necessary to study a new monoclonal antibody. Methods: The P53 DNA fragment enconding Nterminal 180 amiao acide was obtained by PCR and was cloned into PGEX2T plasmid expressing glutathione Stransferase (GST). The P53GST fusion protein expressed by JM109 was used for immunizing BALB/C mice. We have raised one hybridoma strain secreting McAb to human P53 (named M126). Results: The IHC analysis of 52 paraffinembedded sections from human breast cancer with M126 and PAB1801 (Zymed Co.) has showed that the positive immunoreactions were 25 cases (48%) and 22 cases (42.3%) respectively. The staining of M126 was stronger and preferable to PAB1801. Conclusion: M126 can be instead of PAB1801 for studying immunohistochemical analysis on P53 protein.
基金Supported by grants from the National Natural Science Foundation of China(No.30973562)National Basic Research Program of China(No.2010CB933904)
文摘Objective：The aim of this study was to prepare monoclonal antibody against P53, a kind of tumor suppressor protein,and use the antibody initial y in clinical immunoassay. Methods：Monoclonal antibody was prepared and identified via the classic protocol of monoclonal antibody preparation. Identified monoclonal antibodies were purified by af inity chro-matography. Antibody titer was determined by enzyme linked immunosorbent assay （ELISA）. The specific binding activity of antibody was detected by Western blotting and immunohistochemistry. Results：Three strains of monoclonal antibodies named 1P15, 2P37 and 3P40 were obtained and purified by af inity chromatography. The purity of antibodies was higher than 90%. The titers of antibodies were more than 1：6000. Western blot and immunohistochemistry assay showed that the specific antibody can combine with endogenous P53 protein in the tumor celllines and determine the expression of P53 in tumor tis-sue. Conclusion：Three strains of monoclonal antibodies with high af inity to P53 were successful y established, which can be used for detecting the expression of P53 in tumor cells or tissue.
文摘The cDNA of amphioxus p23, a highly conserved co-chaperone for Hsp90, was cloned into a bacterial expression vector pGEX - 6P - 1 and the GST-tagged fusion protein was produced in Eschherichia coli cells. The recombinant p23 was purified by affinity purification, and its molecular mass was estimated to be approximately 22 kDa by sodium dedecyl sulfate-polyacrylamide gel electrephoresis. The N-terminus of purified p23 was sequenced, and the resulting amino acid sequence matches exactly the predicted residues deduced from the amphioxus p23 gene. Besides, pelyclonal antibodies against the recombinant p23 were generated, and these antibodies not only recognized specifically the fusion protein GST - p23 from induced E. coli cells, purified GST - p23 and p23 protein, but also reacted with the total protein extracted fi＇om the adult amphioxus and formed a single positive band. These results pave the way for identifying its tissue and subcellular localization, and may open the door to clarifying its structure and mechanisms of biological role.
文摘We are developing methods to quantify antibody interactions that include a quartz crystal microbalance (QCM) system to measure, on a molecular basis, the interaction of p53 and anti-p53 antibodies. Previously, as a model system, we developed a measurement device consisting of p53 protein (human wild type), characterized by mass spectroscopy and immobilized at various concentrations on a glass slide. The device is designed as a control to be used with immunohistochemical (IHC) assays that incorporate commercially available anti-p53 antibodies and probes. In the current study, p53 protein is covalently immobilized on a silicon dioxide-coated quartz crystal and the resonance frequency shift is followed in-situ. The functionalized sensor is then incubated with the anti-p53 antibody, which provides a direct gravimetric measure of the antibody-antigen binding. This previously described method allows the comparison of the surface immobilized protein concentrations with estimates obtained by fluorescence measurement. The p53 functionalized QCM system offers an independent measure of surface immobilized protein concentration and is essential in development of our IHC calibration prototypes. These results provide a benchmark for comparing antibody systems that may be used in developing other molecular diagnostic assays such as immunocytochemical analysis and the detection of biomarker proteins in blood and urine.
文摘目的:探讨P选择素、抗心磷脂抗体(anticardiolipin antibody,ACA)及活化蛋白C抵抗(activated protein c resistance,APCR)对下肢创伤骨折术后深静脉血栓形成(deep venous thrombosis,DVT)的预测价值。方法:选择接受手术治疗的下肢创伤骨折患者,术后第3天抽取患者静脉血,采用酶联免疫吸附法测定P选择素含量和ACA阳性率,采用Dahlback法测定APCR阳性率。按照《深静脉血栓形成的诊断和治疗指南(第三版)》中DVT的诊断标准判断患者术后2周内是否发生DVT,比较DVT组和无DVT组的血清P选择素含量、血清ACA阳性率及血清APCR阳性率。以是否发生DVT为因变量,以血清P选择素、ACA及APCR为自变量,进行多因素Logistic回归分析。结果:共纳入648例患者,其中31例患者术后2周内发生DVT(DVT组),其余617例均未检出DVT(无DVT组)。2组患者的血清P选择素含量、血清ACA阳性率及血清APCR阳性率比较,差异均有统计学意义(t=26.989,P=0.000;χ^2=0.010,P=0.000;χ^2=12.447,P=0.000)。多因素Logistic回归分析结果显示,血清P选择素、ACA及APCR是下肢创伤骨折术后DVT的危险因素(OR=1.219,P=0.019;OR=1.292,P=0.009;OR=2.430,P=0.012)。结论:P选择素、ACA及APCR是下肢创伤骨折术后DVT的危险因素,可作为预测下肢创伤骨折术后DVT的检测指标。
