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Objective T9 provide candidate molecules for developing recombinant anti-idiotypic antibody (anti-Id) vaccine of gastric carcinoma by selection of recombinant anti-Id to monoclonal antibody ( McAb) MGb1 directed against the cancer with phage display technique.Methods Balb/c mice were immunized with MGb1 and the mRNA was isolated from the spleens of the immunized mice. The VL and VH cDNAs of the antibody were amplified separately by RT-PCR and assembled into ScFv DNAs with a linker DNA. The ScFv DNAs were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage antibody ScFv library. After four rounds of biopanning to the library with MGb1, the MGb1-positive clones were selected from the enriched phages by ELISA. The types of the anti-Id ScFv displayed on the selected phage clones were preliminarily identified by competition ELISA. Results The VL and VH cDNAs was about 320 bp and 340 bp, respectively. The ScFv DNA were about 750 bp. After four rounds panning to the phage antibody ScFv library with MGb1, 18 MGb1-positive phage clones displayed anti-Id ScFv were selected from 50 pre-selected phage clones, among which 4 clones displayed β or γ type anti-Id ScFv. Conclusion The phagedisplayed anti-Id ScFvs to McAb MGb1 are successfully selected by recombinant phage antibody technique, which might lay a foundation for screening the anti-Id ScFv possessing the characteristics of inducing anti-gastric carcinoma immunity.
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Through introducing and summarizing the application of multimedia technology in the process of biochemistry teaching,the article discusses the contribution of multimedia technology in teaching method,teaching organization,teaching content and so on and points out the importance of multimedia technology in biochemistry teaching.
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Objective To clone the cDNA encoding human HMGB1, express it in E. coli, and identify its biological activity. Methods Human HMGB1 cDNA was amplified by RT-PCR and cloned into vector pUC19. After sequence analysis, the cDNA was ligated into prokaryotic expression vector pQE-80L and induced by IPTG to express HMGB1. The protein was purified with Ni~(2+)-NTA chromatography and polymyxin B affinity column. To identify the function of purified protein, the product was co-cultured with THP1 cells. Results Recombinant expression plasmid pQE-80L/HMGB1 was constructed successfully. After purification, the protein purity reached 96%. The recombinant HMGB1 stimulated THP1 to secrete TNF-? . Conclusion The highly purified HMGB1 was obtained successfully, which showed biological activity. These results lay the foundation for further research on the function of human HMGB1.
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Objective To assess the CT manifestations and diagnostic value in the pancreatic tuberculosis(PTB)with review of the literatures. Methods All cases of PTB proved by surgery or biopsy were examined with plain and enhanced CT scans. Results The CT findings in one case with multiple-nodular type of PTB were diffuse enlargement of the pancreas with multiple, nodular, and low-density lesions; The nodular lesions had peripheral enhancement. 7 cases of local type of PTB encroached on pancreatic head. 4 cases showed local soft tissue masses with multiple flecked calcifications in 2 cases and mild enhancement in one case; Cystic masses was found in 2 cases, with mural calcification in 1 case and multiloculated cystic mass in 1 case, respectively; Massive pancreatic head calcification was demonstrated in one case. In these 8 cases of PTB, the lesion extended out of pancreas in 4 cases, including abdominal tuberculous lymph nodes, tuberculous peritonitis, and hepatosplenic tuberculosis. Conclusion CT findings of PTB were various but had some characteristics. Pancreatic masses with multiple flecked calcification or mild enhancement could suggest the diagnosis. Abdominal tuberculosis accompanied with the pancreatic lesion, especially tuberculous lymph nodes, was highly suggestive of the diagnosis of PTB.
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Objective To assess the imaging and clinical manifestations of gastrointestinal mucosa associated lymphoid tissue(MALT)lymphoma with review of literatures. Methods Six cases of gastrointestinal MALT lymphoma (4 in stomach and 2 in intestine) proved by surgery and biopsy were examined with gastrointestinal barium meal examination (GI), CT or MRI scans. Results Gastrointestinal MALT lymphoma was an indolent disease that patients had a slow natural course, a high response rate to treatment and a long survival. Most cases of gastrointestinal MALT lymphoma had associated with Helicobacter pylori. GI showed thickened folds, multiple mucosal nodularity or polypoid filling defects, multiple point ulcers, and enlarged areae gastricae. CT and MRI revealed diffuse gastrointestinal wall thickening, polypoid lesion, and abdominal lymphadenopathy.Two or more findings and multiplicity of lesions might be the most important imaging features. Conclusions Based on the typical imaging findings combined with clinical characteristics, gastrointestinal MALT lymphoma could be suggested.
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Objective To assess CT, MR manifestations and diagnostic value in hepatic tuberculosis. Methods CT findings in 12 cases and MR findings in 4 cases with hepatic tuberculosis proved by surgery or biopsy were retrospectively analyzed. Results (1) CT findings: one case of serohepatic type of hepatic tuberculosis was multiple nodular lesions in the subcapsule of liver. Parenchymal type were in 10 cases, including multiple,miliary, micronodular and low density lesions with miliary calcifications in 2 cases, singular, low density mass with multiple flecked calcifications in 3 cases, multiple cystic lesions in 1 case, multiple micronodular and low density lesions fusing into multiloculated cystic mass or “cluster” sign in 3 cases, and singular, macronodular and low density lesion with multiple miliary calcifications in 1 case. One case of tuberculous cholangitis showed marked dilated intrahepatic duct with multiple flecked calcifications in the porta hepatis. (2) MR findings in 4 cases were hypointense on both T 1WI and T 2WI in one case, hypointense on T 1WI and hyperintense on T 2WI in 3 cases. Enhanced MR in 3 cases showed slightly peripheral enhancement or with multilocular enhancement. Conclusions Various types of hepatic tuberculosis had their different imaging findings. The typical CT and MR findings could suggest the diagnosis.
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Objective To investigate the correlation of vascular endothelial growth factor (VEGF)-634G/C gene polymorphism to diabetic nephropathy in type 2 diabetes mellitus. Methods VEGF genotype was determined by PCR-RFLP in 98 healthy controls and 216 patients with type 2 diabetes including 104 cases without nephropathy and 112 cases with nephropathy. VEGF genotype and allele frequencies were compared among these groups.Results (1)Individuals with CC genotype had higher serum VEGF level than those with CG and GG groups. (2)The genotype CC distribution and C allele frequency in diabetic nephropathy were significantly higher than those in cases without nephropathy group and in controls. (3)Compared with genotype GG and CG class, the morbidity of diabetic nephropathy in genotype CC class was significantly increased. (4) Logistic regression analysis revealed that serum VEGF level and its CC genotype, SBP, HbA1C, LDL-C and BMI were important risk factors of type 2 diabetic nephropathy. Conclusions VEGF -634G/C polymorphism is associated with the development of type 2 diabetic nephropathy. C allele is probably a susceptible gene of type 2 diabetic nephropathy.
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This article is to report an ultratnicro-quantitative method to determine the tissue content of NAD+. The cycling mixture of nitrotetrazolium blue chloride, phenagene methosulfate, lactate and lactic dehydrogenase is used. The procedure of this assay is very simple and easy to perform. It is sensitive enough to detect as little an amount as 1?10-11 mole per ml of NAD+.