Whole-Genome Analysis of Termite-Derived Bacillus velezensis BV-10 and Its Application in King Grass Silage.

Bacillus velezensis (B. velezensis) is a cellulose-degrading strain that has the potential as an additive in fermented feed. B. velezensis BV-10 was isolated and screened from the termite gut. We sequenced the whole genome of this new source of B. velezensis to reveal its potential for use in cellulose degradation. Whole-genome sequencing of B. velezensis BV-10 showed that it has a circular chromosome of 3929792 bp containing 3873 coding genes with a GC content of 45.51% and many genes related to cellulose, hemicellulose, and lignin degradation. King grass silage was inoculated with B. velezensis BV-10 and mixed with other feed additives to assess the effect of B. velezensis BV-10 on the fermentation quality of silage. Six treatment groups were established: the control, B. velezensis BV-10, molasses, cellulase, B. velezensis BV-10 plus molasses, and B. velezensis BV-10 plus cellulase groups. After 30 days of silage-fermentation testing, B. velezensis BV-10 was found to rapidly reduce the silage pH value and significantly reduce the acid-detergent fiber (ADF) content (p < 0.05). The addition of B. velezensis BV-10 plus molasses and cellulase in fermented feed significantly reduced the silage neutral-detergent fiber and ADF content and promoted organic-acid accumulation (p < 0.05). The above results demonstrate that B. velezensis BV-10 promotes the fermentation quality of silage and that this effect is greater when other silage-fermentation additives are included. In conclusion, genes involved in cellulose degradation in B. velezensis BV-10 were identified by whole-genome sequencing and further experiments explored the effects of B. velezensis BV-10 and different feed additives on the fermentation quality of king grass silage, revealing the potential of Bacillus velezensis as a new silage additive.

The Cancers-Specific Survival of Metastatic Pulmonary Carcinoids and Sites of Distant Metastasis: A Population-Based Study.

BACKGROUND: Lung cancer is the leading cause of cancer-related deaths and pulmonary carcinoids (PCs) account for almost 2% of all pulmonary malignancies. However, few published articles have reported prognosis and related factors of pulmonary carcinoid patients. MATERIAL AND METHOD: The Surveillance, Epidemiology, and End Results (SEER) database was used to collect data of patients diagnosed with metastatic PCs from 2010 to 2016. The prognosis and survival of these patients were compared by employing Cox proportional hazards and the Kaplan-Meier survival analysis. RESULTS: A total of 1763 patients were analyzed. The liver (668, 25.6%) was shown to be the most common metastatic site in the isolated organ metastasis cohort, followed by the lung (636, 24.4%), bone (562, 21.6%), and brain (460, 17.6%). Among the patients, the tumor metastasized to a single distant site included the liver, bone, lung, and brain. Cancer-specific survival (CSS) in metastatic PCs is determined by the site of metastasis and the total number of such sites. Pulmonary carcinoid patients with isolated liver metastasis manifested more favorable survival rates in comparison to patients having isolated metastasis in the lung, brain, or bone. The median CSS was 45, 7, 6, 5 months (P = 0.011). The number of distant metastatic sites and the location of distant metastasis were found to be independent risk factors for CSS. For patients with distant isolated metastasis, liver metastasis (P < 0.0001) had better CSS in comparison to those with bone metastasis. When compared to patients whose carcinoids had metastasized to the bones, patients with a brain (P = 0.273) or lung (P = 0.483) metastasis had the same CSS. CONCLUSION: Cancer-specific survival in metastatic PCs depends on the site of metastasis and the total number of such locations. PC patients with isolated liver metastasis manifested more favorable survival in comparison to patients with isolated metastasis in the lung, brain, or bone.

LIMD2 targeted by miR­34a promotes the proliferation and invasion of non­small cell lung cancer cells.

A previous study indicated that LIM domain containing 2 (LIMD2) is an oncogene in a variety of human cancers, including breast, bladder and thyroid cancers, and melanoma; however, the role of LIMD2 in non­small cell lung cancer (NSCLC) remains largely unknown. In the present study, by reverse transcription­quantitative polymerase chain reaction (RT­qPCR) analysis, it was demonstrated that LIMD2 was significantly upregulated in NSCLC tissues compared with adjacent normal tissues. Consistently, LIMD2 was also upregulated in NSCLC cell lines. Furthermore, the present study reported that knockdown of LIMD2 significantly inhibited the proliferation and invasion of H1299 and A549 cells by Cell Counting Kit­8 and Transwell assays. In addition, the expression of LIMD2 was determined to be regulated by microRNA (miR)­34a in the present study. RT­qPCR and western blot analysis indicated that overexpression of miR­34a notably reduced the mRNA and protein expression levels of LIMD2 in H1299 and H549 cells. Additionally, the present study reported an inverse correlation between the expression of LIMD2 and miR­34a in NSCLC tissues. A luciferase reporter assay also demonstrated that miR­34a directly targeted the mRNA expression of LIMD2 in NSCLC cells. Finally, miR­34a was revealed to possess a tumor suppressive role in NSCLC cells. Collectively, the results of the present study revealed that LIMD2 promoted NSCLC progression and was regulated by miR­34a.

Effect of miR-483-5p on apoptosis of lung cancer cells through targeting of RBM5.

RBM5 has been reported to be a candidate tumor suppressor gene which plays an important role in the induction of apoptosis. In this study, we investigated the effect of miR-483-5p on apoptosis of lung cancer cells and the underlying mechanism. We found that the expression of miR-483-5p mRNA was significantly up-regulated in lung cancer compared with adjacent para-cancerous tissues by using real-time PCR. Silencing miR-483-5p promoted A549 cell apoptosis and enhanced caspase-3 activity by flow cytometry with annexin V-FITC/PI staining and caspase-3 activity report kit. Western blotting demonstrated that miR-483-5p mimicked down-regulated RBM5 protein expression and miR-483-5p inhibitor up-regulated RBM5 protein expression. With additional bioinformatics analysis, we confirmed that RBM5 is a target gene of miR-483-5p and is favored for treating NSCLC. The immunohistochemical pattern of RBM5 could be used to predictoutcome for NSCLC. In conclusion, our results support that RBM5 expression can be regulated by miR-483-5p which is a prognostic marker for NSCLC patients. miR-483-5p inhibitor plays a role in lung cancer through targeting RBM5 to induce apoptosis.

Five miRNAs Considered as Molecular Targets for Predicting Esophageal Cancer.

BACKGROUND: Esophageal cancer (EC) is one of the most aggressive malignant gastrointestinal tumors; however the traditional therapies for EC are not effective enough. Great improvements are needed to explore new and valid treatments for EC. We aimed to screen the differentially expressed miRNAs (DEMs) in esophageal cancer and explore the pathogenesis of esophageal cancer along with functions and pathways of the target genes. MATERIAL AND METHODS: miRNA high-throughput sequencing data were downloaded from The Cancer Genome Atlas (TCGA), then the DEMs underwent principal component analysis (PCA) based on their expression value. Following that, TargetScan software was used to predict the target genes, and enrichment analysis and pathway annotation of these target genes were done by DAVID and KEGG, respectively. Finally, survival analysis between the DEMs and patient survival time was done, and the miRNAs with prediction potential were identified. RESULTS: A total of 140 DEMs were obtained, 113 miRNAs were up-regulated including hsa-mir-153-2, hsa-mir-92a-1 and hsa-mir-182; while 27 miRNAs were down-regulated including hsa-mir comprising 29a, hsa-mir-100 and hsa-mir-139 and so on. Five miRNAs (hsa-mir-103-1, hsa-mir-18a, hsa-mir-324, hsa-mir-369 and hsa-mir-320b-2) with diagnostic and preventive potential were significantly correlated with survival time. CONCLUSIONS: The crucial molecular targets such as p53 may provide great clinical value in treatment, as well to provide new ideas for esophageal cancer therapy. The target genes of miRNA were found to play key roles in protein phosphorylation, and the functions of the target genes during protein phosphorylation should be further studied to explore novel treatment of EC.

Nonhematotoxic naphthalene diimide modified by polyamine: synthesis and biological evaluation.

With the aim of up-regulating antitumor efficacy and down-regulating adverse effects, three types of aromatic imide and diimides were designed to couple with different polyamines. The in vitro assays revealed that two naphthalene diimide-polyamine conjugates could inhibit the growth of multiple cancer cell lines more potently than amonafide. 9f, the most potent compound, was verified to efficiently induce apoptosis via a ROS mediated mitochondrial pathway in a preliminary mechanistic study. The comprehensive in vivo trials on three H22 tumor transplant models demonstrated that 9f improved the indexes in terms of inhibitive effect and lifespan extension and reduced the hematotoxicity which is one of main drawbacks of amonafide. More importantly, the obviously elevated ability in preventing lung cancer metastasis was observed, which increased the value of 9f as a promising lead compound. This work supported that the versatile function of polyamines may endow some intriguing biological features to the parent drugs.