Rabbit hemorrhagic disease (RHD) is contagious and highly lethal. Commercial vaccines against RHD are produced from the livers of experimentally infected rabbits. Although several groups have reported that recombinant subunit vaccines against rabbit hemorrhagic disease virus (RHDV) are promising, application of the vaccines has been restricted due to high production costs or low yield. In the present study, we performed codon optimization of the capsid gene to increase the number of preference codons and eliminate rare codons in Spodoptera frugiperda 9 (Sf9) cells. The capsid gene was then subcloned into the pFastBac plasmid, and the recombinant baculoviruses were identified with a plaque assay. As expected, expression of the optimized capsid protein was markedly increased in the Sf9 cells, and the recombinant capsid proteins self-assembled into virus-like particles (VLPs) that were released into the cell supernatant. Rabbits inoculated with the supernatant and the purified VLPs were protected against RHDV challenge. A rapid, specific antibody response against RHDV was detected by an ELISA in all of the experimental groups. In conclusion, this strategy of producing a recombinant subunit vaccine antigen can be used to develop a low-cost, insect cell-derived recombinant subunit vaccine against RHDV.
Animals ; Antigens, Viral/genetics/metabolism ; Caliciviridae Infections/prevention & control/*veterinary/virology ; Capsid Proteins/*genetics/metabolism ; Cell Culture Techniques/*methods ; Codon/genetics/metabolism ; Enzyme-Linked Immunosorbent Assay/veterinary ; *Gene Expression Regulation, Viral ; Hemorrhagic Disease Virus, Rabbit/*genetics/immunology ; *Rabbits ; Recombinant Proteins/genetics/metabolism ; Sf9 Cells ; Spodoptera ; Viral Structural Proteins/*genetics/metabolism ; Viral Vaccines/genetics/immunology

OBJECTIVE：To study the effects of Plantago asiatica polysaccharide on the proliferation ，migration and invasion of breast cancer cells ，and to investigate its mechanism preliminarily. METHODS ：Using human breast cancer cell MDA-MB- 231 as subjects ，MTT method was adopted to detect the effects of different concentrations of P. asiatica polysaccharide（8，16，32，64 mg/L）on the cell proliferation ability ，and survival rate of the cells was calculated. Scratch test and Transwell invasion test were used to detect the effects of different concentrations of P. asiatica polysaccharide（8，16 mg/L）on cell migration ability and invasion ability. Western blot assay was used to detect the expression of epithelial-mesenchymal transition （EMT）-related proteins [matrix metalloproteinase- 2（MMP-2），MMP-9，E-cadherin，N-cadherin，vimentin]. RESULTS ：Results of MTT assay showed that survival rate of the cells in 32，64 mg/L P. asiatica polysaccharide groups were significantly lower than control group （P＜0.05 or P＜0.01），so that 8，16 mg/L，which did not affect the cell survival rate ，were used as the follow-up drug concentrations. Compared with control group ，relative mobility （12，24 h），relative invasion rate and relative expression of MMP- 2，MMP-9， N-cadherin and vimentin protein were decreased significantly in 8，16 mg/L P. asiatica polysaccharide groups （P＜0.05 or P＜ 0.01），while relative expression of E-cadherin protein was increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：P. asiatica polysaccharide can inhibit the proliferation of breast cancer cells MDA-MB- 231，and inhibit the migration and invasion of the cells by regulating the expression of metastasis and EMT-related proteins.

Non-small-cell lung cancer (NSCLC), the most common type of lung cancer accounting for 85% of the cases, is often diagnosed at advanced stages owing to the lack of efficient early diagnostic tools. 5-Hydroxymethylcytosine (5hmC) signatures in circulating cell-free DNA (cfDNA) that carries the cancer-specific epigenetic patterns may represent the valuable biomarkers for discriminating tumor and healthy individuals, and thus could be potentially useful for NSCLC diagnosis. Here, we employed a sensitive and reliable method to map genome-wide 5hmC in the cfDNA of Chinese NSCLC patients and detected a significant 5hmC gain in both the gene bodies and promoter regions in the blood samples from tumor patients compared with healthy controls. Specifically, we identified six potential biomarkers from 66 patients and 67 healthy controls (mean decrease accuracy >3.2, P < 3.68E-19) using machine-learning-based tumor classifiers with high accuracy. Thus, the unique signature of 5hmC in tumor patient's cfDNA identified in our study may provide valuable information in facilitating the development of new diagnostic and therapeutic modalities for NSCLC.
5-Methylcytosine ; analogs & derivatives ; blood ; Biomarkers, Tumor ; blood ; genetics ; Carcinoma, Non-Small-Cell Lung ; blood ; diagnosis ; genetics ; Case-Control Studies ; Circulating Tumor DNA ; blood ; DNA Methylation ; Epigenomics ; Female ; Humans ; Lung Neoplasms ; blood ; diagnosis ; genetics ; Male ; Middle Aged