Objective： In order to evaluate potential application for diagnosis and prognosis of non-small cell-lung cancer （NSCLC）, as well as to determine its role in the pathogenesis of the disease, we prepared anti-human h...Objective： In order to evaluate potential application for diagnosis and prognosis of non-small cell-lung cancer （NSCLC）, as well as to determine its role in the pathogenesis of the disease, we prepared anti-human hnRNPA2/B1 potyclonal antibody. Methods： Prokaryotic expression vector of pET28a （＋）-hnRNP A2/B1 was constructed and bansformed into E.coli BL21. The recombinant protein induced by IPTG was purified and injected to rabbits for antibody preparation. Expression of hnRN P A2/B1 was examined in 45 tissues of NSCLC and 16 inflammatory pseudotumor tissues of lung by immunohistochemistry with the antibody. The commercial hnRNP A2/B1 monoclonal antibody was used as a controI.Results： （1） Polyclonal an-tibody against hnRNP A2/B1 with high title was obtained. （2） The positive staining in NSCLC tissues was 62.22%, which was substantially higher than that in normal tissues （40%, P = 0.035） or inflammatory pseudotumor tissues （31.25%, P=0.033）. （3） Expression of hnRNP A2/B1 positively correlated with age and the history of smoking, whereas it negatively correlated with differentiation staging of tumors. （4） Follow-up study showed that the survival time of patients with positive staining was significantly shorter than that of patients without hnRNP A2/B1 expression （P=0.048）. Conclusion： It is successful to make the recombinant protein and prepare the polyclonal antibody agonist human hnRNP A2/B1. It may be a valuable marker for the diagnosis and prognosis of NSCLC. Our results provide a basis for further study in clinical application.展开更多
AIM: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 ...AIM: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 in liver tissue of patients with hepatic cirrhosis, and the correlation between TIMPs in liver and those in sera so as to discuss whether TIMPs can be used as a diagnosis index of hepatic fibrosis. METHODS: The monoclonal antibodies (McAbs) of TIMP-1 and TIMP-2 were used to sensitize erythrocytes, and solid-phase absorption to sensitized erythrocytes (SPASE) was used to detect TIMP-1 and TIMP-2 in the sera of patients with hepatic cirrhosis. Meanwhile, with the method of in situ hybridization and immunohistochemistry, we studied the mRNA expression and antigen location of TIMP-1 and TIMP-2 in the livers of 40 hepatic cirrhosis patients with pathologic diagnosis. RESULTS: With SPASE, they were 16.4% higher in the acute hepatitis group, 33.3% higher in the chronic hepatitis group, and the positive rates were 73.6% and 61.2% respectively in sera of hepatic cirrhosis patients, which were remarkably higher than those in chronic hepatitis and acute hepatitis group (P【0.001). In 40 samples of hepatic cirrhosis tissues, all of them showed positive expression of TIMP-1 and TIMP-2 mRNA detected with immunohistochemistry or in situ hybridization (positive rate was 100%). Expression of TIMPs in different degrees could be found in liver tissue with cirrhosis. TIMPs were located in cytoplasm of liver cells of patients with hepatic cirrhosis. There was a significant correlation between serum TIMPs level and liver TIMPs level. CONCLUSION: SPASE is a useful method to detect the TIMP-1 and TIMP-2 in sera of patients with hepatic cirrhosis, and TIMP-1 and TIMP-2 can be considered as a useful diagnostic index of hepatic fibrosis, especially TIMP-1.展开更多
Immunomagnetic beads enrichment kit for detection of aflatoxin B1(AFB1) was prepared through reaction of AFB1 and p-phenylenediamine. The catches of AFB1 by the kit were 25 ng/mg. Furthermore, AFB1 was conducted speci...Immunomagnetic beads enrichment kit for detection of aflatoxin B1(AFB1) was prepared through reaction of AFB1 and p-phenylenediamine. The catches of AFB1 by the kit were 25 ng/mg. Furthermore, AFB1 was conducted specific reaction with competitive drugs with similar structure or function to AFB1, including aflatoxin M1, T-2 toxin, ochratoxin A, zearalenone and patulin, and no cross reaction was observed.展开更多
The 2009 H 1N 1 influenza pandemic demonstrated the significance of a global health threat to human beings. Although pandemic H 1N 1 vaccines have been rapidly developed, passive serotherapy may offer superior immedia...The 2009 H 1N 1 influenza pandemic demonstrated the significance of a global health threat to human beings. Although pandemic H 1N 1 vaccines have been rapidly developed, passive serotherapy may offer superior immediate protection against infections in children, the elderly and immune-compromised patients during an influenza pandemic. Here, we applied a novel strategy based on Epstein-Barr virus （EBV）-immortalized peripheral blood memory B cells to screen high viral neutralizing monoclonal antibodies （MAbs） from individuals vaccinated with the 2009 pandemic H1N1 vaccine PANFLU.1. Through a massive screen of 13 090 immortalized memory B-cell clones from three selected vaccinees, seven MAbs were identified with both high viral neutralizing capacities and hemagglutination inhibition （HAl） activities against the 2009 pandemic H 1N 1 viruses. These MAbs may have important clinical implications for passive serotherapy treatments of infected patients with severe respiratory syndrome, especially children, the elderly and immunodeficient individuals. Our successful strategy for generating high-affinity MAbs from EBV-immortalized peripheral blood memory B ceils may also be applicable to other infectious or autoimmune diseases.展开更多
基金a grant from the Provincial Natural Sciences Foundation of Anhui(No.050430703)Major Scientific Research Programs of the Department of Science and Technology of Anhui(No.05023086)
文摘Objective： In order to evaluate potential application for diagnosis and prognosis of non-small cell-lung cancer （NSCLC）, as well as to determine its role in the pathogenesis of the disease, we prepared anti-human hnRNPA2/B1 potyclonal antibody. Methods： Prokaryotic expression vector of pET28a （＋）-hnRNP A2/B1 was constructed and bansformed into E.coli BL21. The recombinant protein induced by IPTG was purified and injected to rabbits for antibody preparation. Expression of hnRN P A2/B1 was examined in 45 tissues of NSCLC and 16 inflammatory pseudotumor tissues of lung by immunohistochemistry with the antibody. The commercial hnRNP A2/B1 monoclonal antibody was used as a controI.Results： （1） Polyclonal an-tibody against hnRNP A2/B1 with high title was obtained. （2） The positive staining in NSCLC tissues was 62.22%, which was substantially higher than that in normal tissues （40%, P = 0.035） or inflammatory pseudotumor tissues （31.25%, P=0.033）. （3） Expression of hnRNP A2/B1 positively correlated with age and the history of smoking, whereas it negatively correlated with differentiation staging of tumors. （4） Follow-up study showed that the survival time of patients with positive staining was significantly shorter than that of patients without hnRNP A2/B1 expression （P=0.048）. Conclusion： It is successful to make the recombinant protein and prepare the polyclonal antibody agonist human hnRNP A2/B1. It may be a valuable marker for the diagnosis and prognosis of NSCLC. Our results provide a basis for further study in clinical application.
基金the Postdoctoral Science Foundation of China,No.1999-10 State Postdoctoral Foundation Commission
文摘AIM: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 in liver tissue of patients with hepatic cirrhosis, and the correlation between TIMPs in liver and those in sera so as to discuss whether TIMPs can be used as a diagnosis index of hepatic fibrosis. METHODS: The monoclonal antibodies (McAbs) of TIMP-1 and TIMP-2 were used to sensitize erythrocytes, and solid-phase absorption to sensitized erythrocytes (SPASE) was used to detect TIMP-1 and TIMP-2 in the sera of patients with hepatic cirrhosis. Meanwhile, with the method of in situ hybridization and immunohistochemistry, we studied the mRNA expression and antigen location of TIMP-1 and TIMP-2 in the livers of 40 hepatic cirrhosis patients with pathologic diagnosis. RESULTS: With SPASE, they were 16.4% higher in the acute hepatitis group, 33.3% higher in the chronic hepatitis group, and the positive rates were 73.6% and 61.2% respectively in sera of hepatic cirrhosis patients, which were remarkably higher than those in chronic hepatitis and acute hepatitis group (P【0.001). In 40 samples of hepatic cirrhosis tissues, all of them showed positive expression of TIMP-1 and TIMP-2 mRNA detected with immunohistochemistry or in situ hybridization (positive rate was 100%). Expression of TIMPs in different degrees could be found in liver tissue with cirrhosis. TIMPs were located in cytoplasm of liver cells of patients with hepatic cirrhosis. There was a significant correlation between serum TIMPs level and liver TIMPs level. CONCLUSION: SPASE is a useful method to detect the TIMP-1 and TIMP-2 in sera of patients with hepatic cirrhosis, and TIMP-1 and TIMP-2 can be considered as a useful diagnostic index of hepatic fibrosis, especially TIMP-1.
基金Supported by Hundred Leading Talents Training Project of Science and Technology Beijing(Z171100001117158)
文摘Immunomagnetic beads enrichment kit for detection of aflatoxin B1(AFB1) was prepared through reaction of AFB1 and p-phenylenediamine. The catches of AFB1 by the kit were 25 ng/mg. Furthermore, AFB1 was conducted specific reaction with competitive drugs with similar structure or function to AFB1, including aflatoxin M1, T-2 toxin, ochratoxin A, zearalenone and patulin, and no cross reaction was observed.
文摘The 2009 H 1N 1 influenza pandemic demonstrated the significance of a global health threat to human beings. Although pandemic H 1N 1 vaccines have been rapidly developed, passive serotherapy may offer superior immediate protection against infections in children, the elderly and immune-compromised patients during an influenza pandemic. Here, we applied a novel strategy based on Epstein-Barr virus （EBV）-immortalized peripheral blood memory B cells to screen high viral neutralizing monoclonal antibodies （MAbs） from individuals vaccinated with the 2009 pandemic H1N1 vaccine PANFLU.1. Through a massive screen of 13 090 immortalized memory B-cell clones from three selected vaccinees, seven MAbs were identified with both high viral neutralizing capacities and hemagglutination inhibition （HAl） activities against the 2009 pandemic H 1N 1 viruses. These MAbs may have important clinical implications for passive serotherapy treatments of infected patients with severe respiratory syndrome, especially children, the elderly and immunodeficient individuals. Our successful strategy for generating high-affinity MAbs from EBV-immortalized peripheral blood memory B ceils may also be applicable to other infectious or autoimmune diseases.
