NAT10-mediated mRNA N4-acetylcytidine modification promotes bladder cancer progression.

BACKGROUND: Dysregulation of the epitranscriptome causes abnormal expression of oncogenes in the tumorigenic process. Previous studies have shown that NAT10 can regulate mRNA translation efficiency through RNA acetylation. However, the role of NAT10-mediated acetylation modification in bladder cancer remains elusive. METHODS: The clinical value of NAT10 was estimated according to NAT10 expression pattern based on TCGA data set and the tumor tissue array. Acetylated RNA immunoprecipitation sequencing was utilized to explore the role of NAT10 in mRNA ac4C modification. Translation efficiency and mRNA stability assay were applied to study the effect of NAT10-deletion on target genes. The nude mouse model and genetically engineered mice were conducted to further verify the characteristics of NAT10 in promoting BLCA progression and regulating downstream targets. RESULTS: NAT10 was essential for the proliferation, migration, invasion, survival and the stem-cell-like properties of bladder cancer cell lines. NAT10 was responsible for mRNA ac4C modification in BLCA cells, including BCL9L, SOX4 and AKT1. Deficient NAT10 in both xenograft and transgenic mouse models of bladder cancer reduced the tumor burden. Furthermore, acetylated RNA immunoprecipitation sequencing data and RNA immunoprecipitation qPCR results revealed that NAT10 is responsible for a set of ac4C mRNA modifications in bladder cancer cells. Inhibition of NAT10 led to a loss of ac4C peaks in these transcripts and represses the mRNA's stability and protein expression. Mechanistically, the ac4C reduction modification in specific regions of mRNAs resulting from NAT10 downregulation impaired the translation efficiency of BCL9L, SOX4 and AKT1 as well as the stability of BCL9L, SOX4. CONCLUSIONS: In summary, these findings provide new insights into the dynamic characteristics of mRNA's post-transcriptional modification via NAT10-dependent acetylation and predict a role for NAT10 as a therapeutic target in bladder cancer. HIGHLIGHTS: NAT10 is highly expressed in BLCA patients and its abnormal level predicts bladder cancer progression and low overall survival rate. NAT10 is necessary and sufficient for BLCA tumourigenic properties. NAT10 is responsible for ac4C modification of target transcripts, including BCL9L, SOX4 and AKT1. NAT10 may serve as an effective and novel therapeutic target for BLCA.

[Start-up and Performance Optimization of a CANON Pilot Reactor].

A completely autotrophic nitrogen removal over nitrite (CANON) reactor was established by seeding ordinary activated sludge with reject water as the influent at (30±3)℃ from a sewage treatment plant in Qingdao. To solve bacterial loss and optimize reactor performance, a suspension of biological carriers was added to the CANON reactor. The result showed that the reactor was successfully started 130 days later. The total nitrogen removal load was up to 0.03 kg·(m3·d)-1, and the average variation ratio of nitrate and ammonia (RNaA) was 0.09, which was close to theoretical value 0.11. The CANON active sludge reactor ran for 300 days. During the stable operation period, the total nitrogen removal rate was stable at 0.20 kg·(m3·d)-1. Red granular sludge was mixed with the effluent of the system, and the particle size of granular sludge was between 1 and 3 mm. The suspension carriers were added to the CANON reactor with a filling rate of 30%. The fillers added to the moving bed biofilm reactor (MBBR) were mature fillers in the nitrifying reactor of the laboratory. The accumulation rate of nitrification was greater than 95%, and the ammonia-oxidized surface load reached 2.0 g·(m2·d)-1. After 30 days of operation and culturing, the system was successfully converted to a pure membrane system, and the biofilm on the surface of the carrier turned pale red. The total nitrogen removal load was up to 0.17 kg·(m3·d)-1. The average RNaA was 0.14, which was slightly higher than the theoretical value of 0.11. This suggested that the CANON sludge adapted to the environment in the MBBR and began to enter a stable stage. The CANON-MBBR ran for 200 days. During the stable operation period, the total nitrogen removal rate was stable at 1.15 kg·(m3·d)-1. The biofilm was bright brick red with a thickness of 150-250 µm. MLSS and MLVSS on the carriers were approximately 10200 mg·m-2 and 9000 mg·m-2, respectively, and the total biomass in the system was approximately 1.5 kg. Through high-throughput sequencing, AOB and AnAOB were found to be the dominant bacteria species on the suspension carrier, with a relative abundance of 26.24% and 30.08%, respectively, and nitrate oxidizing bacteria were successfully suppressed. The above results showed that CANON-MBBR with high-density polyethylene filler as the suspension carrier had good nitrogen-removal efficiency and was conducive to the stable operation of the autotrophic nitrogen removal process.

Mitochondrial genome of the garfish Hyporhamphus quoyi (Beloniformes: Hemiramphidae) and phylogenetic relationships within Beloniformes based on whole mitogenomes.

Mitochondrial DNA (mtDNA) can provide genome-level information (e.g. mitochondrial genome structure, phylogenetic relationships and codon usage) for analyzing molecular phylogeny and evolution of teleostean species. The species in the order Beloniformes have commercial importance in recreational fisheries. In order to further clarify the phylogenetic relationship of these important species, we determined the complete mitochondrial genome (mitogenome) of garfish Hyporhamphus quoyi of Hemiramphidae within Beloniformes. The mitogenome was 16,524 bp long and was typical of other teleosts mitogenomes in size and content. Thirteen PCGs started with the typical ATG codon (with exception of the cytochrome coxidase subunit 1 (cox1) gene with GTG). All tRNA sequences could be folded into expected cloverleaf secondary structures except for tRNASer (AGN) which lost a dihydrouracil (DHU) stem. The control region was 866 bp in length, which contained some conserved sequence blocks (CSBs) common to Beloniformes. The phylogenetic relationship between 26 fish Beloniformes species was analyzed based on the complete nucleotide and amino acid sequences of 13 PCGs by two different inference methods (Maximum Likelihood and Bayesian Inference). Phylogenetic analyses revealed Hemiramphidae as the sister group to Exocoetidae and it is a paraphyletic grouping. Our results may provide useful information on mitogenome evolution of teleostean species.

WITHDRAWN: Correlation of SPINK1 gene polymorphisms in canine pancreatitis.

Ahead of Print article withdrawn by publisher.

Early exposure of rotating magnetic fields promotes central nervous regeneration in planarian Girardia sinensis.

Magnetic field exposure is an accepted safe and effective modality for nerve injury. However, it is clinically used only as a supplement or salvage therapy at the later stage of treatment. Here, we used a planarian Girardia sinensis decapitated model to investigate beneficial effects of early rotary non-uniform magnetic fields (RMFs) exposure on central nervous regeneration. Our results clearly indicated that magnetic stimulation induced from early RMFs exposure significantly promoted neural regeneration of planarians. This stimulating effect is frequency and intensity dependent. Optimum effects were obtained when decapitated planarians were cultured at 20 °C, starved for 3 days before head-cutting, and treated with 6 Hz 0.02 T RMFs. At early regeneration stage, RMFs exposure eliminated edema around the wound and facilitated subsequent formation of blastema. It also accelerated cell proliferation and recovery of neuron functionality. Early RMFs exposure up-regulated expression of neural regeneration related proteins, EGR4 and Netrin 2, and mature nerve cell marker proteins, NSE and NPY. These results suggest that RMFs therapy produced early and significant benefit in central nervous regeneration, and should be clinically used at the early stage of neural regeneration, with appropriate optimal frequency and intensity.