The Role of H2-Calponin Antigen in Cancer Metastasis: Presence of Autoantibodies in Liver Cancer Patients

To investigate the potential of H2-calponin (CNN2) as a serum biomarker for hepatocellular carcinoma (HCC), this study employed the serological analysis of recombinantly expressed cDNA clone (SEREX) technique to identify the presence of CNN2 antibody in the serum of patients with HCC and other tumors. The CNN2 protein was produced through genetic engineering and used as an antigen to determine the positive rate of serum CNN2 autoantibodies via indirect enzyme-linked immunosorbent assay (ELISA). In addition, the mRNA and protein expressions of CNN2 in cells and tissues were evaluated using RT-PCR, in situ RT-PCR, and immunohistochemistry methods. The HCC group exhibited a significantly higher positive rate of anti-CNN2 antibody (54.8%) compared to gastric cancer (6.5%), lung cancer (3.2%), rectal cancer (9.7%), hepatitis (3.2%), liver cirrhosis (3.2%), and normal tissues (3.1%). The positive rates of CNN2 mRNA in HCC with metastasis, non-metastatic HCC, lung cancer, gastric cancer, nasopharyngeal cancer, liver cirrhosis, and hepatitis were 56.67%, 41.67%, 17.5%, 10.0%, 20.0%, 53.13%, and 41.67%, respectively. Meanwhile, the positive rates of CNN2 protein were 63.33%, 37.5%, 17.5%, 27.5%, 45%, 31.25%, and 20.83%, respectively. The down-regulation of CNN2 could inhibit the migration and invasion of liver cancer cells. CNN2 is a newly identified HCC-associated antigen that is implicated in the migration and invasion of liver cancer cells, making it a promising target for liver cancer therapy

RPTOR blockade suppresses brain metastases of NSCLC by interfering the ceramide metabolism via hijacking YY1 binding

Abstract Background Ceramide metabolism is crucial in the progress of brain metastasis (BM). However, it remains unexplored whether targeting ceramide metabolism may arrest BM. Methods RNA sequencing was applied to screen different genes in primary and metastatic foci and whole-exome sequencing (WES) to seek crucial abnormal pathway in BM + and BM-patients. Cellular arrays were applied to analyze the permeability of blood–brain barrier (BBB) and the activation or inhibition of pathway. Database and Co-Immunoprecipitation (Co-IP) assay were adopted to verify the protein–protein interaction. Xenograft and zebrafish model were further employed to verify the cellular results. Results RNA sequencing and WES reported the involvement of RPTOR and ceramide metabolism in BM progress. RPTOR was significantly upregulated in BM foci and increased the permeability of BBB, while RPTOR deficiency attenuated the cell invasiveness and protected extracellular matrix. Exogenous RPTOR boosted the SPHK2/S1P/STAT3 cascades by binding YY1, in which YY1 bound to the regions of SPHK2 promoter (at -353 ~ -365 nt), further promoting the expression of SPHK2. The latter was rescued by YY1 RNAi. Xenograft and zebrafish model showed that RPTOR blockade suppressed BM of non-small cell lung cancer (NSCLC) and impaired the SPHK2/S1P/STAT3 pathway. Conclusion RPTOR is a key driver gene in the brain metastasis of lung cancer, which signifies that RPTOR blockade may serve as a promising therapeutic candidate for clinical application

Epidemiology of pseudorabies in intensive pig farms in Shanghai, China: herd-level prevalence and risk factors

Pseudorabies (PR), also known as Aujeszky's disease, is a highly contagious disease affecting pigs and a wide range of animals. Pseudorabies is enzootic in many countries. In China, it is a priority animal disease for control and eradication, however the data on disease frequency in intensive pig farms and the information on associated risk factors is inadequate. A cross-sectional study of intensive pig farms (≥350 sows) in Shanghai was conducted to determine herd-level prevalence of PRV and associated risk factors. Following a two-stage random sampling design, a total of 1349 sow serum samples were tested by gpI-ELISA from a total of 91 intensive pig farms in Shanghai. A herd was classified as positive if at least one PRV test-positive sow was present. Information on putative risk/protective factors was collected using questionnaires to pig farm owners or veterinarians. A logistic regression model was built to identify risk/protective factors for herd positivity. The results indicated that the herd-level true prevalence was 67.6% (95% CI：57.0–77.0). In the multivariable logistic regression model using backward stepwise procedure, two risk factors were found to be significantly associated with herd positivity: ‘Breeding with introduced sows in the last 12 months’ (OR = 3.5, 95%CI:1.2, 10.3) and ‘Presence of stray dogs or cats’ (OR = 4.0, 95%CI: 1.2, 12.6). The multivariable logistic model fitted the data well. Hosmer-Lemeshow goodness of fit test showed χ = 10.86 (df = 8, p = 0.21 > 0.05) and the predictability (area under the ROC curve) was 0.86. This study suggested that PR was highly endemic in intensive pig farms in Shanghai. The risk and protective factors identified in this study could be useful to improve the prevention policy of PR in Shanghai and other areas of China