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Objective:To obtain a high specificity and high affinity anti-human PD-L1 monoclonal antibody which can be used for clinical diagnosis and block PD-L1 and PD-1 binding.Methods:BALB/c mice were immunized with recombinant PD-L1 protein.The positive cell clones stably secreting anti-human PD-L1 monoclonal antibody were obtained by classical hybridoma cell fusion technique.The specificity,affinity,subtype and other characteristics of the antibody were identified by ELISA.Immunofluorescence and indirect immunofluorescence were used to detect the tumor cells.Antibody blocking activity was confirmed by tumor killing test.Results:Two cell strains stably secreting monoclonal antibodies against human PD-LI were screened out.Abl and Ab2 had high titer and affinity.The antibody titers were 1:2.56×106 and 1:3×105,and the affinity was 1.5×109 L/mol and 2.5×10s L/mol respectively.There was no cross reaction between these two antibodies and PD-L2.Immunoblotting,indirect immunofluorescence confirmed that the antibody can be used to the diagnosis.Experiment showed that PD-L1 antibodies can increases tumor-killing activity of CIK cells.Conclusion:Two hybridoma cell lines capable of stably secreting highly specific and high affinity anti-human PD-L1 monoclonal antibody are obtained.They can specifically bind to PD-L1 molecules on tumor cells and can be used to the diagnosis of tumor phenotype and prognosis.Antibody blocking function can be applied to combined CIK cell immunotherapy.
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Objective:To identify enolase,47 kD allergen,from Litopenaeus vannamei by Mass spectrometry. Methods: The proteins were extracted from Litopenaeus vannamei tissue with acetone precipitation method. The protein components were analyzed by SDS-PAGE and Western blot. By using Matrix-Assisted Laser Desorption/Ionization time of flight mass spectrometry ( MALDI-TOF/TOF-MS) ,the 47 kD allergen from Litopenaeus vannamei was identified as enolase. Results:By SDS-PAGE,we proved that the native protein components from Litopenaeus vannamei were completely. According to the Western blot result more than 14 components could react with the positive serum. MALDI-TOF/TOF analysis results showed that the suspected proteins were enolase. A sensitization frequency was 55%. Conclusion:Enolase was identified as a new allergen of Litopenaeus vannamei.
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Objective:To identify the allergens which can react with Chinese allergic patients in shrimp and crab ,and analysis their reaction-rates.This results would provide foundation for further research on allergen-detection and desensitization therapy.Methods:Allergen components in Metapenaeus ensis ,Macrobrachium rosenbergii and Charybdis feriata by using 46 portions of shrimp(crab) allergic patients’ serum IgE in Western blot.Results:The reactions between shrimps and allergic patients ’ serum IgE were stronger than that between crab and serum.32-38 kD Tropomysin (TM),40 kD Arginine kinase (AK),60-80 kD Hemocyanin (Hc) and 21 kD arcoplasmic calcium-binding protein(SCP) were the major allergens in shrimps.TM,AK and Hc were common major allergens among shrimps and crab and TM shared the highest reaction-rate.Compare to the results of some from American researchers , AK,Hc and SCP have higher reaction-rate when react with Chinese patients serum ,and we also found a new allergen in shrimp.Con-clusion:For Chinese patients , shrimps have higher reaction-rate than crabs and the allergens among shrimps and crabs which are roughly same.There are some differences in allergens among different human races.A new allergen with 48 kD was found in this re-search.
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Objective To evaluate the value of serum procalcitonin(PCT)on antibiotics use in treatment of acute exacerbations of chronic obstructive pulmonary disease( AECOPD). Method From May 2004 to December 2006, a total of 235 patients requiring hospitalization for AECOPD were randomly assigned into two groups: standard therapy group(group A, n = 117)and PCT-guided group(group B, n = 118) .PCT levels of all patients were measured after hospital admission by an amplified cryptate emission technology assay. On the base of similarly normal treatment, group A received antibiotics according to the attending physicians,and group B were treated with antibiotics according to serum PCT levels:antibiotic treatment was applied with PCT level ≥0.25 ng/ml and was discouraged with PCT level ＜0.25 ng/ml. Length of hospitalization,clinical efficacy,costs of hospitalization and antibiotics, rate of antibiotics use, hospital mortality,rate of exacerbation and rehospitalization within 1 year were observed. Analyses were performed by t test, Mann-Whitney U test or χ2 test. Results Clinical efficacy, hospital mortality, length of hospitalization, rate of exacerbation and rehospitalization within 1 year were similar in two groups (P =0.635,0.768,0.884,0.747,0.727) ;costs of antibiotics and hospitalization,rate of antibiotics use of PCT-guided group were lower than that of standard therapy group( P = 0.029,0.036,0.014). Conclusions PCT could be used in treatment of AECOPD for antibiotic use after hospital admission,which may reduce antibiotic use and lower costs of antibiotic and hospitalization.
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Objective To study the value of the changes of signal intensity at the jugular bulb in diagnosing sigmoid sinus thrombosis.Methods MR imaging examinations were performed in 27 cases with cerebral venous sinus thrombosis(transverse and sigmoid sinuses).The signal intensity at the jugular bulb was evaluated in comparison with that in 20 healthy persons,and the results were dealed with Wilcoxon rank sum test statistically.Results In sigmoid sinus thrombosis,the signal intensity at ipsilateral jugular bulb was higher than that in healthy group(P