We combine the labeling of newly transcribed RNAs with 5-ethynyluridine with the characterization of bound proteins. This approach, named capture of the newly transcribed RNA interactome using click chemistry (RICK), systematically captures proteins bound to a wide range of RNAs, including nascent RNAs and traditionally neglected nonpolyadenylated RNAs. RICK has identified mitotic regulators amongst other novel RNA-binding proteins with preferential affinity for nonpolyadenylated RNAs, revealed a link between metabolic enzymes/factors and nascent RNAs, and expanded the known RNA-bound proteome of mouse embryonic stem cells. RICK will facilitate an in-depth interrogation of the total RNA-bound proteome in different cells and systems.
Background and Purpose The coronavirus disease 2019 (COVID-19) pandemic presents an unprecedented health crisis to the entire world. As reported, the body mass index (BMI) may play an important role in COVID-19; however, this still remains unclear. The aim of this study was to explore the association between BMI and COVID-19 severity and mortality. Methods The Medline, PubMed, Embase and Web of science were systematically searched until August 2020. Random-effects models and dose-response meta-analysis were used to synthesize the results. Combined odds ratios (ORs) with their 95% confidence intervals (CIs) were calculated, and the effect of covariates were analyzed using subgroup analysis and meta-regression analyses. Results A total of 16 observational studies involving 109,881 patients with COVID-19 were included in the meta-analysis. The pooled results showed that patients with a BMI≥30kg/m 2 had a 2.35-fold risk(OR = 2.35, 95%CI = 1.64 − 3.38, P < 0.001) for critical COVID-19 and a 2.68-fold risk for COVID-19 mortality (OR = 2.68, 95%CI = 1.65 −4.37, P < 0.001 ) compared with patients with a BMI <30kg/m 2 . Subgroup analysis results showed that patients with obesity and age > 60 years was associated with a significantly increased risk of critical COVID-19(OR = 3.11, 95%CI = 1.73 − 5.61, P < 0.001) and COVID-19 mortality(OR = 3.93, 95%CI = 2.18 − 7.09, P < 0.001). Meta-regression analysis results also showed that age had a significant influence on the association between BMI and COVID-19 mortality(Coef.=0.036, p=0.048). Random-effects dose-response meta-analysis showed a linear association between BMI and both critical COVID-19(P non-linearity = 0.242) and mortality (P non-linearity = 0.116). The risk of critical COVID-19 and mortality increased by 9%(OR = 1.09, 95%CI = 1.04 −1.14, P < 0.001) and 6%(OR = 1.06, 95%CI = 1.02 −1.10, P = 0.002) for each 1 kg/m 2 increase in BMI, respectively. Conclusions Evidence from this meta-analysis suggested that a linear dose-response association between BMI and both COVID-19 severity and mortality. Further, obesity(BMI≥30kg/m 2 ) was associated with a significantly increased risk of critical COVID-19 and in-hospital mortality of COVID-19.
Aims: As reported, hypertension may play an important role in adverse outcomes of coronavirus disease-2019 , but it still had many confounding factors. The aim of this study was to explore whether hypertension is an independent risk factor for critical COVID-19 and mortality. Data synthesis: The Medline, PubMed, Embase, and Web of Science databases were systematically searched until November 2020. Combined odds ratios (ORs) with their 95% confidence interval (CIs) were calculated by using random-effect models, and the effect of covariates was analyzed using the subgroup analysis and meta-regression analysis. A total of 24 observational studies with 99,918 COVID-19 patients were included in the meta-analysis. The proportions of hypertension in critical COVID-19 were 37% (95% CI: 0.27 À0.47) when compared with 18% (95% CI: 0.14 À0.23) of noncritical COVID-19 patients, in those who died were 46% (95%CI: 0.37 À0.55) when compared with 22% (95% CI: 0.16 À0.28) of survivors. Pooled results based on the adjusted OR showed that patients with hypertension had a 1.82-fold higher risk for critical COVID-19 (aOR: 1.82; 95% CI: 1.19 À 2.77; P Z 0.005) and a 2.17-fold higher risk for COVID-19 mortality (aOR: 2.17; 95% CI: 1.67 À 2.82; P < 0.001). Subgroup analysis results showed that male patients had a higher risk of developing to the critical condition than female patients (OR: 3.04; 95%CI: 2.06 À 4.49; P < 0.001) and age >60 years was associated with a significantly increased risk of COVID-19 mortality (OR: 3.12; 95% CI: 1.93 À 5.05; P < 0.001). Meta-regression analysis results also showed that age (Coef. Z 2.3Â10 À2 , P Z 0.048) had a significant influence on the association between hypertension and COVID-19 mortality. Conclusions: Evidence from this meta-analysis suggested that hypertension was independently associated with a significantly increased risk of critical COVID-19 and inhospital mortality of COVID-19.
Pyrethroids are the major class of insecticides used for mosquito control. Excessive and improper use of insecticides, however, has resulted in pyrethroid resistance, which has become a major obstacle for mosquito control. The development of pyrethroid resistance is a complex process involving many genes, and information on post-transcription regulation of pyrethroid resistance is lacking. In this study, we extracted RNA from mosquitoes in various life stages (fourth-instar larvae, pupae, male and female adult mosquitoes) from deltamethrin-sensitive (DS) and resistant (DR) strains. Using Illumina sequencing, we obtained 13760296 and 12355472 reads for DS-strains and DR-strains, respectively. We identified 100 conserved miRNAs and 42 novel miRNAs derived from 21 miRNA precursors in Culex pipiens. After normalization, we identified 28 differentially expressed miRNAs between the two strains. Additionally, we found that cpp-miR-71 was significant down regulated in female adults from the DR-strain. Based on microinjection and CDC Bottle Bioassay data, we found that cpp-miR-71 may play a contributing role in deltamethrin resistance. The present study provides the firstly large-scale characterization of miRNAs in Culex pipiens and provides evidence of post-transcription regulation. The differentially expressed miRNAs between the two strains are expected to contribute to the development of pyrethroid resistance.
Background MicroRNAs have been implicated in many biological pathways involved in tumourigenesis and can serve as prognostic biomarkers in many cancer types. The present study aims at evaluating the prognostic significance of miR-425-5p in cervical cancer. Methods Real-time polymerase chain reaction was performed to assess the expression levels of miR-425-5p in 35 pairs of cervical cancer tissues and their matched normal tissues as well as serum samples from 40 cervical cancer patients, 13 benign cervical disease patients and 32 healthy controls. The association between miR-425-5p expression levels in tissue and serum, and clinicopathological factors was examined. The correlation between serum miR-425-5p expression levels and overall survival of cervical cancer patients was assessed by Kaplan-Meier analysis and Cox proportional hazards model. Results MiR-425-5p expression levels were significantly increased in cervical cancer tissues compared with matched non-cancerous tissues. Higher expression of miR-425-5p was positively associated with high tumour stage ( P = 0.0003) and positive lymph node metastasis ( P = 0.0107). Serum concentrations of miR-425-5p in cervical cancer patients were significantly higher compared with benign cervical disease and healthy controls. Moreover, the up-regulation of serum miR-425-5p occurred more frequently in cervical cancer patients with high TNM stage ( P = 0.0003) and positive lymph node metastasis ( P = 0.0037). Kaplan-Meier analysis showed that high serum miR-425-5p expression levels predicted poor survival ( P = 0.0571). Cox proportional hazards risk analysis demonstrated that miR-425-5p was an independent prognostic factor for cervical cancer. Conclusion Our study suggests that miR-425-5p is up-regulated in cervical cancer and serum miR-425-5p may serve as a potential prognostic biomarker for cervical cancer.
Insecticide resistance has been a major public health challenge. It is impendent to study the mechanism on insecticide resistance. In our previous study, 14 differentially accumulated insect cuticle proteins (ICPs) based on insecticide resistance proteomes and transcriptomes were found in the deltamethrin-resistant (DR) and -susceptible (DS) strains of Culex pipiens pallens. To investigate if these ICPs are associated with deltamethrin resistance, different transcriptional levels of the 14 ICPs were detected in the DS and DR strains from laboratory and field populations by using quantitative real-time polymerase chain reaction (qRT-PCR). The expression levels of the 14 ICPs were also measured after short-term exposure of the DS strain to deltamethrin. The full-length complementary DNA (cDNA) of CpCPLCG5 gene, which encodes one of the 14 ICPs, was cloned from Cx. pipiens pallens. Homology analysis and phylogenetic analysis were carried out with some other insects. Furthermore, small interfering RNA (siRNA) was used to knockdown the expression level of CpCPLCG5 gene for characterizing its contribution to deltamethrin resistance. The results showed that the expression level of CpCPLCG5 gene was higher in DR strain than in DS strain both in laboratory and field populations while the other 13 ICPs were downregulated. The full-length cDNA of CpCPLCG5 gene was 732 bp, with the ORF of 390 bp and deduced 129 amino acids (GenBank/KF723314,2013). Knockdown of CpCPLCG5 gene increased the susceptibility of the DR strain while the expression level of the other 13 ICPs elevated. Our findings indicate that the cuticle proteins are associated with deltamethrin resistance in Cx. pipiens pallens.
BackgroundAnopheles sinensis is an important mosquito vector of Plasmodium vivax, which is the most frequent and widely distributed cause of recurring malaria throughout Asia, and particularly in China, Korea, and Japan.ResultsWe performed 454 next-generation sequencing and obtained a draft sequence of A. sinensis assembled into scaffolds spanning 220.8 million base pairs. Analysis of this genome sequence, we observed expansion and contraction of several immune-related gene families in anopheline relative to culicine mosquito species. These differences suggest that species-specific immune responses to Plasmodium invasion underpin the biological differences in susceptibility to Plasmodium infection that characterize these two mosquito subfamilies.ConclusionsThe A. sinensis genome produced in this study, provides an important resource for analyzing the genetic basis of susceptibility and resistance of mosquitoes to Plasmodium parasites research which will ultimately facilitate the design of urgently needed interventions against this debilitating mosquito-borne disease.
BackgroundMosquito control based on chemical insecticides is considered as an important element in the current global strategies for the control of mosquito-borne diseases. Unfortunately, the development of pyrethroid resistance in important vector mosquito species jeopardizes the effectiveness of insecticide-based mosquito control. To date, the mechanisms of pyrethroid resistance are still unclear. Recent advances in proteomic techniques can facilitate to identify pyrethroid resistance-associated proteins at a large-scale for improving our understanding of resistance mechanisms, and more importantly, for seeking some genetic markers used for monitoring and predicting the development of resistance.MethodsWe performed a quantitative proteomic analysis between a deltamethrin-susceptible strain and a deltamethrin-resistant strain of laboratory population of Culex pipiens pallens using isobaric tags for relative and absolute quantitation (iTRAQ) analysis. Gene Ontology (GO) analysis was used to find the relative processes that these differentially expressed proteins were involved in. One differentially expressed protein was chosen to confirm by Western blot in the laboratory and field populations of Cx. pipiens pallens.ResultsWe identified 30 differentially expressed proteins assigned into 10 different categories, including oxidoreductase activity, transporter activity, catalytic activity, structural constituent of cuticle and hypothetical proteins. GO analysis revealed that 25 proteins were sub-categorized into 35 hierarchically-structured GO classifications. Western blot results showed that CYP6AA9 as one of the up-regulated proteins was confirmed to be overexpressed in the deltamethrin-resistant strains compared with the deltamethrin-susceptible strains both in the laboratory and field populations.ConclusionsThis is the first study to use modern proteomic tools for identifying pyrethroid resistance-related proteins in Cx. pipiens. The present study brought to light many proteins that were not previously thought to be associated with pyrethroid resistance, which further expands our understanding of pyrethroid resistance mechanisms. CYP6AA9 was overexpressed in the deltamethrin-resistant strains, indicating that CYP6AA9 may be involved in pyrethroid resistance and may be used as a potential genetic marker to monitor and predict the pyrethroid resistance level of field populations.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-015-0709-5) contains supplementary material, which is available to authorized users.
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