Mosquito E-20-Monooxygenase Gene Knockout Increases Dengue Virus Replication in Aedes aegypti Cells.

E-20-monooxygenase (E20MO) is an enzymatic product of the shade (shd) locus (cytochrome p450, E20MO). Initially discovered in Drosophila, E20MO facilitates the conversion of ecdysone (E) into 20-hydroxyecdysone (20E) and is crucial for oogenesis. Prior research has implicated 20E in growth, development, and insecticide resistance. However, little attention has been given to the association between the E20MO gene and DENV2 infection. The transcriptome of Ae. aegypti cells (Aag2 cells) infected with DENV2 revealed the presence of the E20MO gene. The subsequent quantification of E20MO gene expression levels in Aag2 cells post-DENV infection was carried out. A CRISPR/Cas9 system was utilized to create an E20MO gene knockout cell line (KO), which was then subjected to DENV infection. Analyses of DENV2 copies in KO and wild-type (WT) cells were conducted at different days post-infection (dpi). Plasmids containing E20MO were constructed and transfected into KO cells, with pre- and post-transfection viral copy comparisons. Gene expression levels of E20MO increased after DENV infection. Subsequently, a successful generation of an E20MO gene knockout cell line and the verification of code-shifting mutations at both DNA and RNA levels were achieved. Furthermore, significantly elevated DENV2 RNA copies were observed in the mid-infection phase for the KO cell line. Viral RNA copies were lower in cells transfected with plasmids containing E20MO, compared to KO cells. Through knockout and plasmid complementation experiments in Aag2 cells, the role of E20MO in controlling DENV2 replication was demonstrated. These findings contribute to our understanding of the intricate biological interactions between mosquitoes and arboviruses.

Aedes aegypti Beta-1,3-Glucan-Binding Protein Inhibits Dengue and ZIKA Virus Replication.

GNBPB6, a beta-1,3-glucan-binding protein, was identified in the transcriptome of Aedes aegypti (A. aegypti) with dengue (DENV), Zika (ZIKV), and chikungunya viruses (CHIKV). In this study, we not only clarified that DENV2 and ZIKV regulate the changes in GNBPB6 expression but also identified the relationship of this gene with viral infections. The changes in GNBPB6 expression were quantified and showed a decrease in A. aegypti cells (Aag2 cells) at 2 dpi and 3 dpi and an increase at 4 dpi and 5 dpi (p < 0.05). A significant increase was observed only at 5 dpi after DENV2 infection. Subsequently, a GNBPB6 knockout (KO) cell line was constructed using the CRISPR/Cas9 system, and the DENV2 and ZIKV RNA copies, along with cell densities, were quantified and compared between the KO and wild type (WT) cells at different dpi. The result showed that DENV2 and ZIKV RNA copies were significantly increased in the KO cell line with no significant change in cell growth. Finally, DENV2 copies decreased after GNBPB6 was complemented in the KO. In conclusion, GNBPB6 knockout and complementation in Aag2 cells revealed that GNBPB6 can inhibit the replication of both DENV2 and ZIKV. These results contribute to subsequent research on mosquito-virus interactions.

Microplastics affect mosquito from aquatic to terrestrial lifestyles and are transferred to mammals through mosquito bites.

Microplastics (MPs) transfer from the environment to living organisms is a nonignorable global problem. As a complete metamorphosis insect, the larvae and adult Culex quinquefasciatus mosquito live in aquatic and terrestrial environments, respectively, where they easily access MPs. However, little is known about mosquitoes' potential role in MPs accumulation throughout ecosystems. Therefore, we conducted a study with different MPs particle sizes (0.1/1/10 µm) and concentrations (0.5/5/50 µg/mL) on Cx. quinquefasciatus to address this issue. Once exposed at the young larval stage, MPs could accompany the mosquitoes their entire life. The fluorescence signals of MPs in the larvae were mainly located in the intestines. Its intensity increased (from 3.72 × 106 AU to 5.45 × 107 AU) as the concentrations of MPs increases. The fluorescence signals of MPs were also detected in the blood and skin tissues of mice bitten by adult mosquitoes with MPs containing in their bodies. Mosquitos exposed to MPs showed longer larval pupation and eclosion time as well as lower adult body weight. In addition, MPs significantly reduced the lethal effect of pyrethroid insecticides (97.77 % vs. 48.88 %, p < 0.05) with 15.1 % removal of the deltamethrin concentration. After MPs exposure, the relative abundance of the Cx. quinquefasciatus gut microbiome, such as Wolbachia spp., Elizabethkingia spp., and Asaia spp., changed as the MPs size and concentration changes. Mosquitoes provide a new pathway for MPs accumulation and transfer to higher-level living organisms. Moreover, MPs significantly reduce the control effect of deltamethrin, providing new guidelines for mosquito insecticide application in MPs contamination circumstances.

The status and risk factors for anxiety/depression in patients with atrophic chronic gastritis: a cross-sectional study.

BACKGROUND: Atrophic chronic gastritis (ACG) is a preneoplastic condition of gastric carcinoma. Numerous studies have shown anxiety and depression can affect gastrointestinal function, which may promote gastrointestinal disorders development and progression. Thus, we hypothesized that anxiety and depression may enhance the development and progression of ACG. In this study, we aimed to analyse risk factors for anxiety and depression in ACG patients and integrate these risk factors to construct an effective clinical prediction model. METHODS: In total, 118 ACG patients were included from July 2021 to May 2022. Anxiety and depression were assessed utilizing the Generalized Anxiety Disorder-7 (GAD-7) and Patient Health Questionnaire-9 (PHQ-9). Data were collected on demographic characteristics, lifestyle, and dietary habits. Risk factors for anxiety and depression were explored with univariate analysis and multivariate stepwise logistic regression, and risk prediction models were built. RESULTS: Among 118 ACG patients, 36.4% had anxiety, 25.4% had depression, and 21.2% had both anxiety and depression. Poor sleep quality [odd ratio (OR) 4.32, 95% confidence interval (CI): 1.60-11.65, P=0.004] was positively associated with risk of anxiety, while smoking (OR 0.15, 95% CI: 0.03-0.68, P=0.014) and weekly exercise time (OR 0.89, 95% CI: 0.79-0.99, P=0.037) were negatively associated with risk of anxiety. The area under the receiver operating characteristic (ROC) curve was 80.3%, 95% CI: [0.722-0.885]. The sensitivity was 72.1%, and the specificity was 78.7%. Poor sleep quality (P<0.001, OR 23.89, 95% CI: 4.05-141.05), high salt diet (P=0.004, OR 6.94, 95% CI: 1.86-25.96), family history of tumours (P=0.020, OR 6.10, 95% CI: 1.33-27.93), and abdominal pain (P=0.018, OR 4.44, 95% CI: 1.29-15.23) were positively associated with the risk of depression, with an area under the ROC curve of 77.3%, 95% CI: 0.687-0.860. The sensitivity was 83.3%, and the specificity was 62.5%. CONCLUSIONS: Potential anxiety and depression in ACG patients can be identified early by referring to risk factors and protective factors. The prediction model could be used to detect anxiety and depression in ACG patients at their earliest stage and provide meaningful suggestions for ACG patients.

Association between trichlorophenols and neurodegenerative diseases: A cross-sectional study from NHANES 2003-2010.

PURPOSE OF THE RESEARCH: To evaluate the association of the exposure of trichlorophenols (TCPs) on the morbidity and mortality of patients with Parkinson's disease (PD) and Alzheimer's disease (AD) using the data from the National Health and Nutrition Examination Survey (NHANES) 2003-2010. Multivariable logistic regression models and COX regression were used to evaluate the association between TCP exposure and the AD and PD risk. Least Absolute Shrinkage and Selection Operator (LASSO) methods were used to screen latent covariates. PRINCIPAL RESULTS: A total of 6333 participants over the age of 18 years were included in the analysis. After the adjustments for major confounders, participants with higher concentrations of urinary 2,4,6-TCP had higher risk of AD (odds ratios (ORs), 3.19; 95% CI: 1.07, 9.45) than the group below the limit of detection (LOD). Compared to group of below the LOD, higher urinary concentrations of 2,4,5-TCP was associated with higher risk of all-cause mortality in PD patients (log-rank P = 0.022) and all participants (log-rank P < 0.001) without adjustments for confounders. In addition, a higher risk of all-cause mortality in all participants with high urinary concentrations of 2,4,6-TCP (log-rank P = 0.001) was found without adjustments for confounders. With the adjustments for major confounders, participants with higher concentrations of urinary 2,4,5-TCP had a higher risk of death in patients with PD (hazard ratios (HRs), 53.19; 95% CI: 2.82, 1004.13) than in the group below the LOD. MAJOR CONCLUSIONS: Exposure to high concentration of 2,4,6-TCP may increase the risk of AD, and the level of 2,4,5-TCP may be associated with the risk of death in patients with PD. Our findings reveal the potential toxicity of TCPs, highlight the potential impact of TCPs on neurodegenerative diseases, and express concerns regarding the use of organochlorine pesticides.

Long Non-Coding RNAs as Biomarkers and Therapeutic Targets in Sepsis.

Sepsis, an infection-induced systemic inflammatory disorder, is often accompanied by multiple organ dysfunction syndromes with high incidence and mortality rates, and those who survive are often left with long-term sequelae, bringing great burden to social economy. Therefore, novel approaches to solve this puzzle are urgently needed. Previous studies revealed that long non-coding RNAs (lncRNAs) have exerted significant influences on the process of sepsis. The aim of this review is to summarize our understanding of lncRNAs as potential sepsis-related diagnostic markers and therapeutic targets, and provide new insights into the diagnosis and treatment for sepsis. In this study, we also introduced the current diagnostic markers of sepsis and discussed their limitations, while review the research advances in lncRNAs as promising biomarkers for diagnosis and prognosis of sepsis. Furthermore, the roles of lncRNAs in sepsis-induced organ dysfunction were illustrated in terms of different organ systems. Nevertheless, further studies should be carried out to elucidate underlying molecular mechanisms and pathological process of sepsis.

Expression Profiling of Long Noncoding RNA and Messenger RNA in a Cecal Ligation and Puncture-Induced Colon Injury Mouse Model.

BACKGROUND: Emerging evidence reveals that long noncoding RNAs (lncRNAs) play important roles in the pathogenesis of sepsis. However, the detailed regulatory mechanisms of lncRNAs or whether certain lncRNA could serve as a biomarker in the septic colon remains unclear. The aim of this study was to investigate the profiles of lncRNAs and mRNAs in the septic colon through whole-transcriptome RNA sequencing and to reveal the associated regulatory mechanism. METHOD AND RESULT: We established a mouse model of sepsis by cecal ligation and puncture (CLP). Colon samples were collected upon CLP or sham surgery after 24 h. Whole-transcriptome RNA sequencing was performed to profile the relative expressions of lncRNAs and mRNAs. 808 lncRNAs and 1509 mRNAs were differentially found in the septic group compared with the sham group. Bioinformatics analysis including Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis (KEGG) was performed to predict the potential functions of these RNAs. GO analysis showed that the altered lncRNAs were enriched and involved in multiple immune responses, which may be a response to sepsis stress. KEGG analysis indicated that upregulated lncRNAs were significantly enriched in the p53 signaling pathway, NF-κB signaling pathway, and HIF-1 signaling pathway. Downregulated lncRNAs were mostly found to be involved in tight junction, leukocyte transendothelial migration, and HIF-1 signaling pathway. CONCLUSION: Our results indicate that these altered lncRNAs and mRNAs may have crucial roles in the pathogenesis of sepsis. This study could contribute to extending the understanding of the function of lncRNAs in sepsis, which may help in searching for new diagnostic biomarkers and therapeutic targets to treat sepsis.

Two zinc(II) coordination complexes based on an asymmetric multidentate ligand: syntheses, structures, selective fluorescence sensing of iron(III) ions and thermal analyses.

The rational selection of ligands is vitally important in the construction of coordination complexes. Two novel ZnII complexes, namely bis(acetato-κO)bis[1-(1H-benzotriazol-1-ylmethyl)-2-propyl-1H-imidazole-κN3]zinc(II) monohydrate, [Zn(C13H15N5)2(C2H3O2)2]·H2O, (1), and bis(azido-κN1)bis[1-(1H-benzotriazol-1-ylmethyl)-2-propyl-1H-imidazole-κN3]zinc(II), [Zn(C13H15N5)2(N3)2], (2), constructed from the asymmetric multidentate imidazole ligand, have been synthesized under mild conditions and characterized by elemental analyses, IR spectroscopy and single-crystal X-ray diffraction analysis. Both complexes exhibit a three-dimensional supramolecular network directed by different intermolecular interactions between discrete mononuclear units. The complexes were also investigated by fluorescence and thermal analyses. The experimental results show that (1) is a promising fluorescence sensor for detecting Fe3+ ions and (2) is effective as an accelerator of the thermal decomposition of ammonium perchlorate.