Toxicity and risk priority ranking of polybrominated diphenyl ethers (PBDEs): A relative receptor-bound concentration approach.

Toxicity and risk priority ranking of chemicals are crucial to management and decision-making. In this work, we develop a new mechanistic ranking approach of toxicity and risk priority ranking for polybrominated diphenyl ethers (PBDEs) based on receptor-bound concentration (RBC). Based on the binding affinity constant predicted using molecular docking, internal concentration converted from human biomonitoring data via PBPK model, and the receptor concentration derived from the national center for biotechnology information (NCBI) database, the RBC of 49 PBDEs binding to 24 nuclear receptors were calculated. 1176 RBC results were successfully obtained and analyzed. High brominated PBDEs, including BDE-201, BDE-205, BDE-203, BDE-196, BDE-183, BDE-206, BDE-207, BDE-153, BDE-208, BDE-204, BDE-197, and BDE-209, exerted more potent than low brominated congeners (BDE-028, BDE-047, BDE-099, and BDE-100) at the same daily intake dose in terms of toxicity ranking. For risk ranking, with human biomonitoring serum data, the relative RBC of BDE-209 was significantly greater than that of any others. For receptor prioritization, constitutive androstane receptor (CAR), retinoid X receptor alpha (RXRA), and liver X receptor alpha (LXRA) may be the sensitive targets for PBDEs to trigger effects in the liver. In summary, high brominated PBDEs are more potent than low brominated congeners, thus, besides BDE-047 and BDE-099, BDE-209 should be priority controlled. In conclusion, this study provides a new approach for toxicity and risk ranking of groups of chemicals, which can readily be used for others.

MiR-3017b contributes to metamorphosis by targeting sarco/endoplasmic reticulum Ca2+ ATPase in Tribolium castaneum.

In recent years, increasing numbers of microRNAs (miRNAs) have been reported to regulate insect metamorphosis. One thousand, one hundred fifty-four miRNAs have been previously identified from Tribolium castaneum by high-throughput sequencing; however, little is known about which miRNAs can participate in metamorphosis, leaving the role of miRNAs in regulating the underlying mechanism elusive. Here, we report the participation of miR-3017b in the metamorphosis of T. castaneum. Temporal profiles revealed that miR-3017b was highly expressed at the late larval stage, but significantly decreased at the early pupal stage. Overexpression of miR-3017b caused larval to pupal to adult metamorphosis arrested. Dual-luciferase reporter assay and miRNA-mRNA interaction assay illustrated that miR-3017b interacts with the coding sequence of sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) and suppresses its expression. Knockdown of SERCA caused metamorphosis arrested, similar to that observed in miR-3017b overexpression beetles. Further functional mechanism analyses revealed that 20-hydroxyecdysone application downregulates miR-3017b and up-regulates SERCA expression. The expression level of downstream genes in the 20E pathway was disrupted after overexpressing miR-3017 and the knockdown of SERCA. These results provided evidence miR-3017b-SERCA contributes to metamorphosis by regulating the 20E pathway in T. castaneum. It could advance our understanding of the coordination of 20E and miRNA regulation in insect metamorphosis.

Effect of High Suspension and Low Incision Surgery Based on Traditional Ligation of Chinese Medicine in Treatment of Mixed Haemorrhoids: A Multi-centre, Randomized, Single-Blind, Non-inferiority Clinical Trial.

OBJECTIVE: To observe the clinical effect of high suspension and low incision (HSLI) surgery on mixed haemorrhoids, compared with Milligan-Morgan haemorrhoidectomy. METHODS: A multi-centre, randomized, single-blind, non-inferiority clinical trial was performed. Participants with mixed haemorrhoids from Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing Rectum Hospital, Air Force Medical Center of People's Liberation Army of China, and Puyang Hospital of Traditional Chinese Medicine were enrolled from September 2016 to March 2018. By using a blocked randomization scheme, participants were assigned to two groups. The experimental group was treated with HSLI, while the control group was treated with Milligan-Morgan haemorrhoidectomy. The primary outcome was the clinical effect evaluated at 12 weeks after operation. The secondary outcomes included the number of haemorrhoids treated during the operation, pain scores, use of analgesics, postoperative oedema, wound healing, incidence of anal stenosis, anorectal manometry after operation, as well as surgical duration, length of stay and total hospitalization expenses. A safety evaluation was also conducted. RESULTS: In total, 246 eligible participants were enrolled, with 123 cases in each group. There was no significant difference in the clinical effect between the two groups (100.00% vs. 99.19%, P>0.05). Compared with the control group, the number of external haemorrhoids treated during the operation and the pain scores after operation were significantly reduced in the experimental group (P<0.05 or P<0.01); the patient number with wound healing at 2 weeks after operation and the functional length of anal canal at 12 weeks after operation were significantly increased in the experimental group (P<0.05). There was no significant difference in the incidence of anal stenosis, the numbers of patients using analgesics and patients with postoperative oedema between the two groups after operation (P>0.05). The surgical duration and length of stay in the experimental group were significantly longer than those in the control group, and the total hospitalization expense was significantly higher than that in the control group (all P<0.05). No adverse events were reported in either group during the whole trial or follow-up period. CONCLUSION: HSLI had the advantages of preserving the skin of anal canal completely, alleviating postsurgical pain and promoting rapid recovery after operation. (Registration No. ChiCTR1900022883).

The complete mitochondrial genome of Anas penelope (Anatidae: Anas).

The complete mitochondrial DNA genome of the Eurasian Wigeon, Anas penelope, was mapped by the next-generation sequencing and Mega 7.0. The circular mitogenome (16,596 bp in length) contains 13 protein-coding genes, 2 rRNA genes (12S ribosomal RNA and 16S ribosomal RNA), 22 tRNA genes and a control region. The content of four base pairs of the complete mitochondrial DNA is 28.9% of A, 22.3%of T, 32.7%of C and 16.1% of G. To validate the phylogenetic relationship, 25 published complete mitochondrial genomes of Anseriformesalong with the genome of Terek sandpiper were used to construct the phylogenetic tree.

Characterization and functional analysis of hsp18.3 gene in the red flour beetle, Tribolium castaneum.

Small heat shock proteins (sHSPs) are diverse and mainly function as molecular chaperones to protect organisms and cells from various stresses. In this study, hsp18.3, one Tribolium castaneum species-specific shsp, has been identified. Quantitative real-time polymerase chain reaction illustrated that Tchsp18.3 is expressed in all developmental stages, and is highly expressed at early pupal and late adult stages, while it is highly expressed in ovary and fat body at the adult period. Moreover, it was up-regulated 4532 ± 396-fold in response to enhanced heat stress but not to cold stress; meanwhile the lifespan of adults in ds-Tchsp18.3 group reduced by 15.8% from control group under starvation. Laval RNA interference (RNAi) of Tchsp18.3 caused 86.1% ± 4.5% arrested pupal eclosion and revealed that Tchsp18.3 played an important role in insect development. In addition, parental RNAi of Tchsp18.3 reduced the oviposition amount by 94.7%. These results suggest that Tchsp18.3 is not only essential for the resistance to heat and starvation stress, but also is critical for normal development and reproduction in T. castaneum.

Preparation and properties of cryogel based on poly(hydroxypropyl methacrylate).

A novel supermacroporous poly(hydroxypropyl methacrylate) (p(HPMA)) cryogel was synthesized by cryogelation method at -16 °C. In this synthesis process, HPMA was used as a monomer, and N,N'-methylenebisacrylamide (MBAAm) was used as cross-linker; the reaction was carried out in the presence of redox initiator pair N,N,N',N'-tetramethylene diamine (TEMED) and ammonium persulfate (APS). The effect of monomer concentration, cross-linker content, cooling rate, and dioxane co-solvent were determined with respect to the pore structure, mechanical behavior, swelling degree, and porosity of cryogel. The ESEM images indicate that the pore wall structure of cryogels was rough; moreover, small holes were present in the pore walls of cryogels. The result of compression test indicates that cryogels can be compressed by at least 80% without any breakdown. The result of swelling kinetics indicates that cryogels attain swelling equilibrium in 10 s. Furthermore, p(HPMA)-Cu2+ cryogel was prepared by loading Cu2+ ions on functionalized poly(hydroxypropyl methacrylate)-iminodiacetic acid (p(HPMA)-IDA) cryogel. We investigated the adsorption of bovine serum albumin (BSA) on cryogels. The results indicate that compared to Freundlich isotherm, Langmuir isotherm could more suitably describe the adsorption process of BSA on cryogels. Meanwhile, the adsorption capacity of p(HPMA)-Cu2+ cryogel was significantly greater than that of p(HPMA) cryogel. The maximum adsorption capacity of BSA on p(HPMA)-Cu2+ cryogel, which was treated with 1 M Cu2+ ions, was as high as 196.87 mg/g cryogel (equivalent to 20.48 mg/mL cryogel) at 25 °C and pH = 7.8; therefore, the maximum adsorption capacity of BSA on p(HPMA)-Cu2+ cryogel was 4.35 times higher than that of p(HPMA) cryogel. Thus, the adsorption capacity of cryogels was strongly influenced by Cu2+ concentration, moreover, temperature changes clearly affected the adsorption capacity of p(HPMA)-Cu2+cryogel. The adsorption capacity at 25 °C was twice as that at 15 °C. By calculating Gibbs free energy change (∆G) of adsorption, we found that the adsorption process was spontaneous; moreover, adsorption process occurred better at higher temperature.

Comparative RNA-sequencing analysis of ER-based HSP90 functions and signal pathways in Tribolium castaneum.

Tribolium castaneum, the red flour beetle, is a major agriculture pest that damages stored grains and cereal products. Heat-shock protein 90 of T. castaneum (Tchsp90) has been reported to play pivotal roles in heat stress response, development, reproduction, and life span. However, the signaling pathway of Tchsp90 remains unclear. Thus, the global transcriptome profiles between RNA interference (RNAi)-treated insects (ds-Tchsp90) and control insects of T. castaneum were investigated and compared by RNA sequencing. In all, we obtained 14,145,451 sequence reads, which assembled into 13,243 genes. Among these genes, 461 differentially expressed genes (DEGs) were identified between the ds-Tchsp90 and control samples. These DEGs were classified into 44 gene ontology (GO) functional groups, including the cellular process, the response to stimulus, the immune system process, the development process, and reproduction. Interestingly, knocking down the expression of Tchsp90 suppressed both the DNA replication and cell division signaling pathways, which most likely modulated the effects of Tchsp90 on development, reproduction, and life span. Moreover, the DEGs encoding AnnexinB9, frizzled-4, sno, Fem1B, TSL, and CSW might be related to the regulation of the development and reproduction of ds-Tchsp90 insects. The DEGs including TLR6, PGRP2, defensin1, and defensin2 were involved in heat stress and immune response simultaneously, which suggested that cross talk might exist between immunity and stress response. Additionally, RNAi of Tchsp90 altered large-scale serine protease (sp) gene expression patterns and amplified the SP signaling pathway to regulate the development and reproduction as well as the stress response and innate immunity in T. castaneum. All these results shed new light onto the regulatory mechanism of Tchsp90 involved in insect physiology and could further facilitate research into appropriate and sustainable pest control management.

MicroRNA-dependent regulation of metamorphosis and identification of microRNAs in the red flour beetle, Tribolium castaneum.

To date, although some microRNAs (miRNAs) have been discovered in the holometabolism insect Tribolium castaneum, large numbers of miRNAs still require investigation. Knocking down Dicer-1 (Dcr-1) and Argonaute-1 (Ago-1) in late larvae impaired miRNA synthesis, affected the juvenile hormone pathway by up-regulating Methoprene-tolerant (Met) and Krüppel-homolog1 (Kr-h1) transcript levels, and resulted in a series of defects in T. castaneum development and metamorphosis. Thus, high-throughput Illumina/Solexa sequencing was performed with a mixed sample of eight key developmental stages of T. castaneum. In total, 1154 unique miRNAs were discovered containing 274 conserved miRNAs belong to 68 miRNA families, 108 known candidate miRNAs and 772 novel miRNAs. Genome locus analysis showed that miRNA clusters are more abundant in T. castaneum than other species. The results indicated that RNAi of Dcr-1 and Ago-1 in T. castaneum resulted in miRNA-induced metamorphosis defects. Furthermore, large numbers of novel miRNAs were discovered in T. castaneum and localized to T. castaneum genome loci.

Genome-wide DNA methylomes from discrete developmental stages reveal the predominance of non-CpG methylation in Tribolium castaneum.

Cytosine DNA methylation is a vital epigenetic regulator of eukaryotic development. Whether this epigenetic modification occurs in Tribolium castaneum has been controversial, its distribution pattern and functions have not been established. Here, using bisulphite sequencing (BS-Seq), we confirmed the existence of DNA methylation and described the methylation profiles of the four life stages of T. castaneum. In the T. castaneum genome, both symmetrical CpG and non-CpG methylcytosines were observed. Symmetrical CpG methylation, which was catalysed by DNMT1 and occupied a small part in T. castaneum methylome, was primarily enriched in gene bodies and was positively correlated with gene expression levels. Asymmetrical non-CpG methylation, which was predominant in the methylome, was strongly concentrated in intergenic regions and introns but absent from exons. Gene body methylation was negatively correlated with gene expression levels. The distribution pattern and functions of this type of methylation were similar only to the methylome of Drosophila melanogaster, which further supports the existence of a novel methyltransferase in the two species responsible for this type of methylation. This first life-cycle methylome of T. castaneum reveals a novel and unique methylation pattern, which will contribute to the further understanding of the variety and functions of DNA methylation in eukaryotes.