ABSTRACT
Lymantria dispar (Linnaeus, 1758), which is commonly known as spongy moth, with two subspecies, is found in Asia: Lymantria dispar asiatica and Lymantria dispar japonica, collectively referred to as the Asian spongy moth (ASM). The subspecies Lymantria dispar dispar occurs in Europe and is commonly known as the European spongy moth (ESM). The ASM is on the quarantine list of many countries because it induces greater economic losses than the ESM. Accurate identification is essential to prevent the invasion of ASM into new areas. Although several techniques for identifying ASMs have been developed, the recent discovery of complex patterns of genetic variation among ASMs in China as well as new subspecies in some areas has necessitated the development of new, improved identification techniques, as previously developed techniques are unable to accurately identify ASMs from all regions in China. Here, we demonstrate the efficacy of an improved technique for the identification of the ASM using ASM-specific primers, which were designed based on cytochrome oxidase I sequences from samples obtained from all sites where ASMs have been documented to occur in China. We show that these primers are effective for identifying a single ASM at all life stages and from all ASM populations in China, and the minimum detectable concentration of genomic DNA was 30 pg. The inclusion of other Lymantria samples in our analysis confirmed the high specificity of the primers. Our improved technique allows the spread of ASMs to be monitored in real time and will help mitigate the spread of ASMs to other areas.
ABSTRACT
Cigarette smoke is a major cause of chronic obstructive pulmonary disease (COPD). Circular RNAs (circRNAs) are involved in regulating various biological processes. This study aimed to explore the role and molecular basis of hsa_circ_0006872 in cigarette smoke extract (CSE)-induced cell injury. HPMECs and BEAS-2B cells were treated with CSE to mimic COPD in vitro. The levels of hsa_circ_0006872 and miR-145-5p were measured by quantitative real-time polymerase chain reaction. Cell proliferation was assessed via Cell Counting Kit-8 (CCK-8) and colony formation assays. Flow cytometry was used to evaluate apoptosis and cell cycle. The levels of inflammatory factors were assayed via enzyme-linked immunosorbent assay (ELISA). The levels of oxidative stress markers were determined via commercial kits. The interaction between hsa_circ_0006872 and miR-145-5p was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation assay. Protein expression was measured using Western blot assay. Hsa_circ_0006872 level was elevated in COPD patients and was negatively correlated with miR-145-5p level. CSE exposure promoted apoptosis, inflammation and oxidative stress of HPMECs and BEAS-2B cells, while hsa_circ_0006872 down-regulation undermined the effects. In addition, hsa_circ_0006872 silencing inhibited CSE-induced cell injury via regulating miR-145-5p. Moreover, CSE contributed to the activation of NF-κB pathway through hsa_circ_0006872/miR-145-5p axis. Hsa_circ_0006872 facilitated CSE-triggered apoptosis, inflammation and oxidative stress in HPMECs and BEAS-2B cells by regulating miR-145-5p/NF-κB pathway.
