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The aim of this study was to identify antigens for a vaccine or drug target to control rabbit coccidiosis. A combination of 2-dimensional electrophoresis, immunoblotting, and mass spectrometric analysis were used to identify novel antigens from the sporozoites of Eimeria stiedae. Protein spots were recognized by the sera of New Zealand rabbits infected artificially with E. stiedae. The proteins were characterized by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS) analysis in combination with bioinformatics. Approximately 868 protein spots were detected by silver-staining, and a total of 41 immunoreactive protein spots were recognized by anti-E. stiedae sera. Finally, 23 protein spots were successfully identified. The proteins such as heat shock protein 70 and aspartyl protease may have potential as immunodiagnostic or vaccine antigens. The immunoreactive proteins were found to possess a wide range of biological functions. This study is the first to report the proteins recognized by sera of infected rabbits with E. stiedae, which might be helpful in identifying potential targets for vaccine development to control rabbit coccidiosis.
Subject(s)
Rabbits , Coccidiosis , Computational Biology , Eimeria , Electrophoresis , HSP70 Heat-Shock Proteins , Immunoblotting , Mass Spectrometry , SporozoitesABSTRACT
Objective To study the effect of microRNA-204 (miR-204) on the biological characteristics of breast cancer cells.Methods Real-time PCR was used to detect the expression of miR-204 in human breast cancer cell MDA-MB-231 after transfection of miR-204 mimics and inhibitor for 48 h.Flow cytometry was used to analyse the effect of miR-204 on the proliferation and apoptosis of MDA-MB-231 cells.The effect of miR-204 on the migration of MDA-MB-231 cells was detected by Transwell migration assay.Results Real-time PCR analysis showed that miR-204 mimics and inhibitors had significant effect compared with normal control group(P<0.01).Flow cytometry analysis showed that compared with normal control group,the number of G1 phase cells of miR-204 mimics group was significantly decreased(P<0.01),while the number of G2/M cells of miR-204 mimics group was significantly increased(P<0.01).In contrast,the number of G1 phase cells of miR-204 inhibitor group was significantly increased(P<0.01),while the number of G2/M cells of miR-204 inhibitor group was significantly decreased(P<0.01).miR-204 mimics group significantly promoted apoptosis,while the inhibitor group significantly inhibited apoptosis(P<0.01).Transwell migration analysis showed that the number of cells of miR-204 mimics group were significantly reduced,while the number of cells was significantly increased in the inhibitor group(P<0.01).Conclusion We find miR-204,which can promote cell apoptosis and inhibit cell proliferation and migration,is a negative factor in the breast cancer cell line MDA-MB-231.
ABSTRACT
Advanced gastric cancer without distant metastasis remains a potentially curable disease, but the prognosis is poor in this condition because of the high unresectability rate at presentation and the high recurrence rate after radical surgery. Administration of neoadjuvant chemotherapy has several potential benefits for advanced gastric cancer. This treatment can decrease tumor stage and improve R0 resection rate. Neoadjuvant chemotherapy has higher patient tolerability and a higher rate of chemotherapy completion than adjuvant chemotherapy. In vivo drug sensitivity tests can also be conducted to avoid unnecessary surgeries. Although high-intensi-ty chemotherapy results in a high overall response rate, a few advanced gastric patients can achieve a pathologically complete re-sponse. However, no standardized treatment has been achieved. This article introduces five cases of advanced gastric cancer treated with neoadjuvant chemotherapy in the Affiliated Tumor Hospital of Guangxi Medical University. The five cases achieved a pathological complete response. This article also aims to explore the clinicopathological characteristics of these patients, proper cooperative treat-ment practices, and prognostic factors for the benefit of future patients.
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Background Mutant C57BL/6 mouse with corneal opacity (B6-Co) appears eye open at birth (EOB) phenotype,which is a good animal model in the study of developmental mechanism of eyelid.Investigating the relationship between serum response factor (SRF) and EOB phenotype can provide theoretical support for the research on the mechanism of innate defects in eyelid development in humans.Objective This study was to assess the dynamic expressions of SRF in eyelid of embryonic B6-Co mouse.Methods Total RNA was extracted from B6 and B6-Co mice eyelid tissue at embryonic day 16.5 (E16.5 d),E17.5 d and E18.5 d.The relative expression levels of SRF mRNA and protein in the eyelid tissue of B6 and B6-Co embryonic mice were assayed by real-time quantitative PCR and Western blot,respectively.In situ expressions of SRF protein in eyelid of B6-Co mice and B6 mice were detected using immunofluorescence technique.The use and care of the animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals of Nantong University.Results The relative expression levels ofSRF mRNA in the eyelids were 0.41±0.06 and 0.24±0.17 in E16.5 d and E17.5 d of B6-Co mice,showing a significant decline in comparison with 1.03 ±0.17 and 1.01 ±0.09 in the B6 mice (P =0.025,0.017).The expression levels of SRF protein in the eyelids of E16.5 d and E17.5 d B6-Co mice were 0.08±0.01 and 0.08± 0.01,which were significantly lower than 0.12 ±0.03 and 0.13 ± 0.02 of B6 mice (P =0.036,0.024).However,there were no significant differences in the expression levels of SRF mRNA and protein in E18.5 d between the B6-Co mice and B6 mice (P =0.387,0.774).Immunofluorescence assay displayed that SRF was expressed in the keratinocytes of eyelids in both mice,but the fluorescence intensity was weaker in the B6-Co mice.Conclusions SRF probably interrupts the developing process of eyelid in early embryo of B6-Co mice.
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Objective To study the clone sequencing and expression of fibroblast growth factor 10(Fgf10) gene in corneal opaci-ty (B6-Co) mouse .Methods Normal mice mate with B6-Co mice ,the skin tissue separation from B6 and B6-Co mice at embryo 16 . 5 d ,total RNA extraction and reverse transcription ,the target gene was fragment amplification by RT-PCR ,connection with T vec-tor ,transformed to competent cells ,selection positive clone ,sequencing analysis .The gene expression in B6-Co mice was detected by real-time PCR .Results The base A inserted between 1 914 and 1 915 in Fgf10 gene by sequencing .The expression of Fgf10 was significant down regulation in B6-Co mice by real-time PCR(P< 0 .05) .Conclusion Fgf10 is relevant with phenotype of B6-Co mouse ,and the regulation mechanism was expected further study .