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To prepare and purify the rabbit anti-TgMIC16 polyclonal antibody, so as to apply it in subcellular localization.New Zealand white rabbits were immunized with purified recombinant TgMIC16 mixing with the same volume of Freund’s adjuvant for three times, respectively. The rabbit serum was collected on the 14th day after the last immunization. The polyclonal antibody in rabbit serum was purified with Protein A affinity purification column. ELISA and Western blotting were used to detect the antibody titer and specificity of polyclonal antibody. The polyclonal antibody was used to the localization of TgMIC16 by the immunofluorescence method.Indirect ELISA showed that the antibody titer was 1∶512 000. Western blotting showed that the recombinant TgMIC16 protein was recognized by the specific polyclonal antibody. IFA showed that TgMIC16 was located in the microneme of Toxoplasma gondii.The rabbit anti-TgMIC16 is prepared and purified, and successfully applied to immunofluorescence localization of TgMIC16 in T. gondii.
ABSTRACT
OBJECTIVE@#To investigate the effects and mechanisms of anthocyanin from Ligustrum vicaryi on chronic inflammatory pain induced by complete Freund's adjuvant.@*METHODS@#Thirty male SD rats were randomly divided into three groups (=10):normal saline control group (NS), chronic inflammatory pain model group(Mod, injected with complete Freund's adjuvant(CFA) 100 μl to the left hind leg), anthocyanin treatment group(Ant, dosed with anthocyanins (90 m), mechanical pain threshold (MPT), and left toe volume in each group were measured before modeling and 1,3,5,7,9,11,13 days after operation. Antioxidant indexes in serum were mensurated by spectrophotometer, and the total capsaicin receptor (TRPV1) and phosphorylated capsaicin receptor (p-TRPV1) in hippocampus were detected by Western blot.@*RESULTS@#In comparison with controls, HPT and MPT were improved (<0.05),toe swelling was reduced(<0.05), the serum level of SOD was increased (<0.01), while the levels of MDA and NO were decreased (<0.05), the ratio of P-TRPV1/TRPV1 protein was depressed in Mod rat hippocampal region treated with anthocyanin.@*CONCLUSIONS@#The results show that anthocyanins has an analgesic effect on chronic inflammatory pain induced by CFA, and its mechanism may be related to the improvement of antioxidant capacity and the reduction of TRPV1 phosphorylation.
Subject(s)
Animals , Male , Rats , Anthocyanins , Freund's Adjuvant , Inflammation , Pain , Rats, Sprague-Dawley , TRPV Cation ChannelsABSTRACT
The bioinformatics software was used to predict the B cell and T cell epitopes of Toxoplasma gondii microneme 16 (TgMIC16) followed by epitopes display on the yeast of Saccharomyces cerevisiae.B and T cell epitopes of TgMIC16 were predicted by DNAStar and IEDB,and software of SYFPEITHI and BIMAS,respectively.According to the predicted results,the antigenic epitope region was expressed with pCTCON2/EBY100 display system.The expression protein was detected by indirect immunofluorescence (IFA) and flow cytometry.Results showed that there were potential B/T cell epitopes in TgMIC16 and concentrated in the aa343-625 region.The epitope region was successfully displayed on the surface of yeast cells,and the optimal induction time was 24 hours.The study would lay the foundation for the development and efficacy evaluation of the yeast carrier vaccine in the further research.
ABSTRACT
The bioinformatics software was used to predict the B cell and T cell epitopes of Toxoplasma gondii microneme 16 (TgMIC16) followed by epitopes display on the yeast of Saccharomyces cerevisiae.B and T cell epitopes of TgMIC16 were predicted by DNAStar and IEDB,and software of SYFPEITHI and BIMAS,respectively.According to the predicted results,the antigenic epitope region was expressed with pCTCON2/EBY100 display system.The expression protein was detected by indirect immunofluorescence (IFA) and flow cytometry.Results showed that there were potential B/T cell epitopes in TgMIC16 and concentrated in the aa343-625 region.The epitope region was successfully displayed on the surface of yeast cells,and the optimal induction time was 24 hours.The study would lay the foundation for the development and efficacy evaluation of the yeast carrier vaccine in the further research.
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Objective: To evaluate the clinical efficacy and safety of neoadjuvant chemoradiotherapy (nCRT) versus neoadjuvant radiotherapy (nRT) for patients with stages II-III rectal cancer. Methods: A computer-based online search of PubMed, EMBase, Cochrane Library, China Journal Full-text Database, Wanfang Database and Chinese sci-tech periodical full-text database was performed to include randomized controlled trials (RCTs) about nCRT vs nRT (including neoadjuvant short-course radiotherapy, nSRT) in patients with stages II-III rectal cancer in accordance with the inclusion and exclusion criteria. The relevant literatures and conference proceedings were also manually searched to include eligible RCTs. After evaluating the quality of included trials and extracting data, the data analysis was conducted using RevMan 5.2 software. Results: A total of six randomized controlled trials (RCTs) involving 2 602 patients with stages II-III rectal cancer were included. The results of Meta-analysis showed that the nCRT group had higher radical resection rate (odds ratio: 1.62, 95% confidence interval: 1.14-2.30; P = 0.008), pathological complete response rate (odds ratio: 3.17, 95% confidence interval: 2.28-4.40; P < 0.000 01), and rate of severe side effects (grades 3-4) (odds ratio: 2.66, 95% confidence interval: 1.29-5.46; P = 0.008), as compared with nRT group. The five-year local recurrence (LR) rate in the nCRT group was obviously lower than that in the nRT group (odds ratio: 0.51, 95% confidence interval: 0.39-0.67, P < 0.000 01). Regarding nCRT vs nSRT, the radical resection rate (odds ratio: 2.40, 95% confidence interval: 1.25-4.63; P = 0.009) and the pathological complete response rate (odds ratio: 4.28, 95% confidence interval: 1.72-10.68, P = 0.002) were also higher in the nCRT group than those in the nSRT group. Furthermore, there were no differences in two-, three- and five-year survival rates between nCRT and nRT groups (P < 0.05). Conclusion: As compared with nRT, the nCRT can obviously improve the radical resection rate and complete pathologic response rate, and reduce the local recurrence rate in patients with stages II-III rectal cancer, but it can cause more severe toxicities.
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BACKGROUND:Simulative dynamics provides advantages of repeatable and non-invasive to a model. Additional y, structural model of individualism improves the reliability of Finite Elemental Analysis. It is an optimal attempt to analyze mechanics of knee joint after virtual replacement surgery. OBJECTIVE:To achieve dynamic information of contact stress upon knee joint surface by finite element analysis surgery for total knee arthroplasty postoperatively, and to provide objective data for further“surgery plan”. METHODS:After scanning affected knee joint by CT/MR and scanning knee prosthesis by laser instrument, a model composed of prosthesis, knee joint as wel as its ligament was rebuilt computational y;dynamic lines were measured. After prosthesis instal ation&osteotomy performed by facility of Simulation in Mimics according to knee joint replacement standard, this model was imported into ANSYS so as for Meshing, Material assignment, load applied. Stress distribution was analyzed by Finite Element Method. RESULTS AND CONCLUSION:The best finite element model of postoperative TKA 3D knee joint was obtained;dynamic data were tested to be approximately agreeable to those previous studies of direct measurement upon prosthesis. The experiment of analyzing structural deformation, stress distribution&internal energy change benefits to search the best position for prosthesis instal ation, osteotomy and surgical result prediction. Thus, these are indispensable data in surgery plan.