Application Value of Next-Generation Sequencing of Bronchial Alveolar Lavage Fluid in Emergency Patients with Infection.

This study was performed to explore the application value of next-generation sequencing (NGS) of bronchial alveolar lavage fluid (BALF) in emergency patients with infection. In this regard, a total of 52 patients with infection who were diagnosed and treated in the emergency department of our hospital from September 2019 to September 2021 were selected as the research objects. The BALF of the patients was analyzed by NGS, and the results were compared with the pathogen detection results of traditional microbial culture of the patients to analyze the diagnostic value of NGS in patients with infection. The results showed that among the 52 patients, 47 were positive by NGS and 13 were positive by traditional microbial culture. The pathogen detection rate of NGS was higher than that of traditional microbial culture[90.4% (47/52) VS 25% (13/52), χ2=45.539, P<0.001], and was able to detect viruses, fungi and other special pathogens that were difficult to be detected by traditional microbial culture, such as Chlamydia psittaci. A total of 129 pathogens were detected in the NGS test results of 47 of these patients, including bacteria, fungi, viruses and mycoplasma/chlamydia. 14 pathogens were detected in the conventional microbiological cultures of 13 patients, including bacteria and fungi. Overall, compared with traditional microbial culture methods, NGS detection has higher accuracy and can effectively detect pathogens that cannot be detected by traditional microbial culture. It has a high application value in the diagnosis of pathogens and can provide clinical guidance for the diagnosis of pathogens in patients with emergency infections.

Identification of key biomarkers in ischemic stroke: single-cell sequencing and weighted co-expression network analysis.

PURPOSE: At present, there is a lack of accurate early diagnostic markers for ischemic stroke. METHODS: By using dimensionality reduction cluster analysis, differential expression analysis, weighted co-expression network analysis, protein-protein interaction network analysis, cell heterogeneity and key pathogenic genes were identified in ischemic stroke. Immunomicroenvironment analysis was used to explore the immune landscape and immune associations of key genes in ischemic stroke. The analysis platform we use is R software (version 4.0.5). PCR experiments were used to verify the expression of key genes. RESULTS: Single cell sequencing data in ischemic stroke can be annotated as fibroblast cells, pre-B cell CD34, neutrophils cells, bone marrow (BM), keratinocytes, macrophage, neurons and mesenchymal stem cells (MSC). By the intersection of differential expression analysis and WGCNA analysis, 385 genes were obtained. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that these genes were highly correlated with multiple functions and pathways. Protein-protein interaction network analysis revealed that MRPS11 and MRPS12 were key genes, both of which were down-regulated in ischemic stroke. The Pseudo-time series analysis found that the expression of MRPS12 decreased gradually with the differentiation of pre-B cell CD34 cells in ischemic stroke, suggesting that the downregulation of MRPS12 expression may play an important role in ischemic stroke. At last, PCR showed that MRPS11 and MRPS12 were significantly down-regulated in peripheral blood of patients with ischemic stroke. CONCLUSIONS: Our study provides a reference for the study of pathogenesis and key targets of ischemic stroke.

Phytosphingosine alleviates Staphylococcus aureus-induced mastitis by inhibiting inflammatory responses and improving the blood-milk barrier in mice.

Mastitis is one of the common diseases in dairy cows which threatens the health of cows and impacts on economic benefits seriously. Recent studies have been showed that Subacute Ruminal Acidosis (SARA) increased the susceptibility of cow mastitis. SARA leads the disturbance of the rumen microbiota, and the rumen bacterial disordered community is an important endogenous factor of cow mastitis. That is to say, cows which suffer from SARA have a disordered rumen microbiota, a prolonged decline in ruminal PH and a high level of lipopolysaccharide (LPS) in the rumen, blood. Therefore, ruminal metabolism is closely related to the rumen microbiota. However, the specific mechanism of SARA and mastitis still not clear. We found an intestinal metabolite according to the metabonomics, which is correlated to inflammation. Phytophingosine (PS), a product from rumen fluid and milk of the cows which suffer from SARA and mastitis. It has the effect of killing bacteria and anti-inflammatory. Emerging evidences indicate that PS can alleviate inflammatory diseases. However, how PS affects mastitis is largely unknown. In this study, we explored the concrete role of PS on Staphylococcus aureus (S. aureus) -induced mastitis in mice. We found that PS obviously decreased the level of the proinflammatory cytokines. Meanwhile, PS also significantly relieved the mammary gland inflammation caused by S. aureus and restored the function of the blood-milk barrier. Here, we showed that PS increased the expression of the classic Tight-junctions (TJs) proteins including ZO-1, Occludin and Claudin-3. Moreover, PS improves S. aureus-induced mastitis by inhibiting the activation of the NF-κB and NLRP3 signaling pathways. These data indicated that PS relieved S. aureus-induced mastitis effectively. This also provides a reference for exploring the correlation between the intestinal metabolism and inflammation.

Transcriptome analysis revealed the mechanism of Luciobarbus capito (L. capito) adapting high salinity: Antioxidant capacity, heat shock proteins, immunity.

Salinity has a significant influence on the physiology of freshwater aquatic organisms. However, there are few studies on the hematology and immunology of freshwater fish under high salinity. In the current study, we aimed to analyze the adaptive effect of salt stress on L. capito spleen immune function and hematology using transcriptomic analysis. We replicated a L. capito acute salinity stress model, and collected blood and spleens from freshwater and saltwater fish. It was found that salinity affected significantly the numbers of leukocytes, lymphocytes, neutrophils, and red blood cells, as well as the content of haemoglobin. Salt treatment resulted in a significant increase in the expression of HSP70, HSP90, CAT, SOD, and GPX1 genes in L. capito spleens. Transcriptomic analysis revealed a total of 546 differentially expressed genes (DEGs) in spleens, including 224 up-regulated DEGs and 322 down-regulated DEGs. In addition, GO enrichment analysis revealed immune system process, multicellular organismal process, and biological regulation of genes with the most differences in biological processes. KEGG enrichment analysis showed that the regulation of lipolysis in adipocyte, FoxO signaling pathway, Hematopoietic cell lineage signaling pathway, and HIF-1 signaling pathway were significantly enriched. L. capito adapted oxidative to high salinity through FoxO signaling pathway and immune to high salinity through Hematopoietic cell lineage signaling pathway. At the same time, we selected 10 DEGs for qRT-PCR detection, and the results showed that the qRT-PCR results were consistent with our RNA-Seq results, indicating that transcriptome sequencing was accurate and reliable. In conclusion, our results demonstrated that the improvement of antioxidant capacity, heat shock protein and immunity are involved in the molecular mechanism of L. capito adapting to high salinity. Our findings provided a rationale for further study on high salinity adaptation and related enrichment pathways.

Characterization of the tenofovir resistance-associated mutations in the hepatitis B virus isolates across genotypes A to D.

Tenofovir is wildly used to treat chronic hepatitis B (CHB) infection due to good potency and high genetic barrier to drug resistance. To date, it remains controversial whether hepatitis B virus (HBV) could be resistant to tenofovir. This study used multiple HBV isolates across genotypes A to D to characterize the reported tenofovir resistance-associated mutations identified from a few clinical case reports. Our data demonstrated that all the rtS78T, rtA194T, and CYEI (S106C, H126Y, D134E, and L269I) mutants exhibited no resistance to tenofovir in vitro. In contrast, the quadruple mutation MLVV (rtL180M, rtT184L, rtA200V, and rtM204V) decreased tenofovir susceptibility, with increased half maximal efficient concentration ranging from 3.28-fold to 5.34-fold, but it severely impaired viral fitness by reducing both replication capacity and infectivity of the mutants. Interestingly, basal core promoter (BCP) mutations A1762T/G1764A and precore (PC) mutation G1896A restored the replication capacity of the MLVV mutant to a level even higher than the wild-type virus without BCP and PC mutations. In conclusion, our data support the role of tenofovir as a first-line agent to treat CHB infection but also indicate tenofovir-resistant mutants could emerge due to multiple mutations accumulated during long-term therapy, particularly in hepatitis B e antigen-negative patients.

Effects of exposure to cadmium (Cd) and selenium-enriched Lactobacillus plantarum in Luciobarbus capito: Bioaccumulation, antioxidant responses and intestinal microflora.

Cadmium (Cd) is a dangerous pollutant with multiple toxic effects on aquatic animals, and it exists widely in the environment. Selenium (Se) is a biologically essential trace element. Interactions between heavy metals and selenium can significantly affect their biological toxicity, although little is known about the mechanism of this antagonism. Lactobacillus is one of the dominant probiotics, given that a certain dose promotes host health. In this study, we evaluated the protective effect of a dietary probiotic supplementation, Se-enriched Lactobacillus plantarum (L. plantarum), on the bioaccumulation, oxidative stress and gut microflora of Luciobarbus capito exposed to waterborne Cd. Fish were exposed for 28 days to waterborne Cd at 0.05 mg/L and/or dietary Se-enriched L. plantarum. Exposure to Cd in water leads to Cd accumulation in tissues, oxidative stress and significant changes in gut microflora composition. Adding Se-enriched L. plantarum to the diet can reduce the accumulation of Cd in tissues, enhance the activity of antioxidant enzymes, and reverse changes in intestinal microbial composition after Cd exposure. The results obtained indicate that Se-enriched L. plantarum provides significant protection against the toxicity of Cd by inhibiting bioaccumulation. Selenium reduced oxidative stress by increasing the activity of glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and malondialdehyde (MDA). Se-enriched L. plantarum can reduce the increase in the number of pathogenic Aeromonas caviae bacteria in the intestine caused by Cd stress and increase the number of Gemmobacter to regulate the microbial population. The results of this study show that Se-enriched L. plantarum dietary supplements can effectively protect Luciobarbus capito against Cd toxicity at subchronic levels.

Selenium-enriched Bacillus subtilis reduces the effects of mercury-induced on inflammation and intestinal microbes in carp (Cyprinus carpio var. specularis).

Mercury (Hg) is a global pollutant that affects the health of humans and ecosystems. Selenium (Se) is an essential trace element for many organisms including humans. Bacillus subtilis is one of the main probiotics used in aquaculture, and has a certain adsorption effect on heavy metals. The interaction between Hg and Se was rigorously studied, especially due to the observation of the protective effect of Se on Hg toxicity. The objective of this study was to research the effects of Hg, Se, and B. subtilis on inflammation and intestinal microbes in common carp. The common carp was exposed to Hg (0.03 mg/L), and 105 cfu/g Se-rich B. subtilis was added to the feed. After 30 days of feeding, samples were taken to evaluate the growth performance, serological response, inflammatory response, and intestinal microbial changes. In this study, when fish were exposed to Hg, the growth performance of the Se-rich B. subtilis plus 0.03 mg/L Hg fish group was lower than that of the control group and higher than 0.03 mg/L Hg; The levels of serum immunoglobulin M (IgM) and lysozyme (LZM) decreased, but after supplementation with Se-rich B. subtilis, the levels of LZM and IgM increased; Hg treatment significantly upregulated the mRNA expression of interleukin-1ß (IL-1ß), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and nuclear factor-kB (NF-κB P65), but downregulated the mRNA expression of interleukin-10 (IL-10), transforming growth factor-ß (TGF-ß) and NF-kappa-B inhibitor alpha (IkBα). However, compared with the Hg group, the Se-rich B. subtilis plus Hg group can significantly increase the mRNA expression levels of IL-1ß, IL-8, TNF-α, and NF-κB P65, but reduce the regulation of IL-10, TGF-ß, and IkBα expression. Through the analysis of the microbiological, we found that the Hg group was mainly composed of Aeromonas sobria and Aeromonas hydrophila. However, in the Se-rich B. subtilis treatment group, we found that Aeromonas sobria was significantly less than the Hg group. Se-rich B. subtilis improves Hg-induced intestinal microbial changes, alleviates the abundance of Aeromonas, and alleviates the inflammation of the fish. The results of this study show that Se-rich B. subtilis dietary supplements can effectively protect common carp against Hg toxicity.

Dynamic distribution of nasal microbial community in yaks (Bos grunniens) at different ages.

The significance of microbial community structure has been extensively recognized due to its key roles in metabolism, immunity, and health maintenance. Importantly, increasing evidence indicated that the dynamic distribution of microbial community structure can be used for evaluating the health condition of host. Yaks (Bos grunniens), mainly inhabiting in high-altitude hypoxic environment, are characterized by excellent adaptability and strong resistance. Currently, it has been determined that yaks possessed the complicated gastrointestinal microbial ecosystem, whereas not much is known about the nasal microbial community structure of yaks. Therefore, this study was performed to compare and analyze the differences in nasal microbiota of yaks with different ages by high-throughput sequencing. In this study, a total of 487,168 and 486,498 high-quality sequences were achieved from YYG (1-month-old yaks) and AYG (1-year-old yaks), respectively. Additionally, 5,340 operational taxonomic units (OTUs) were identified and 657 OTUs were in common among all samples. Proteobacteria and Firmicutes were the two most predominant phyla in all samples. Moreover, Actinobacteria and Bacteroidetes were the tertiary dominant phyla in YYG and AYG, respectively. At the level of genus, Moraxella, Faucicola, and Mannheimia were the most preponderant bacterial genera in the young and adult yaks. As compared to the AYG, the proportions of Actinobacillus, Parabacteroides, and Haemophilus in the YYG were significantly increased, whereas the Rhizobacter was decreased. In conclusion, this study firstly compared and investigated the distribution of nasal microbiota in yaks with different ages. Results demonstrated that age was an important factor affecting the nasal microbiota. Moreover, the current study will provide a theoretical basis for the further study on the microbial community structure of yaks.

Derivation and validation of the CANP scoring model for predicting the neurological outcome in post-cardiac arrest patients.

OBJECTIVES: To establish and validate a prognostic scoring model in a Chinese population to predict the neurological outcome among comatose survivors of cardiac arrest (CA). METHODS: 159 CA patients between January 2016 and November 2020 were recruited in this retrospective study. In the derivation cohort, prognostic factors available from arrest circumstances and early in-hospital indicators were measured. The Cardiac Arrest Neurological Prognosis (CANP) score was then constructed to predict unfavorable outcomes at 30 days after CA. The assessment of predictive effectiveness of this scoring model was conducted in both derivation and validation cohorts. RESULTS: Witnessed status, bystander cardiopulmonary resuscitation, initial rhythm, duration of resuscitation, Glasgow Coma Scale motor score, pupillary/corneal reflex, gray-white matter ratio and neuron-specific enolase exhibit strong correlations with the neurological outcomes in the derivation cohort (all p<0.05), and a risk scoring model for the prediction of an unfavorable outcome was created using these factors. In the validation cohort, significantly higher CANP scores were noted in the unfavorable outcome group. A CANP score ≥5 was associated with unfavorable neurological outcomes (sensitivity 68.8%, specificity 100%). CONCLUSION: The CANP score was established and validated for predicting the possible neurological prognosis in comatose post-CA survivors.

TUG1 knockdown suppresses cardiac fibrosis after myocardial infarction.

Cardiac fibrosis is involved in myocardial remodeling following acute myocardial infarction (AMI), which can result in heart failure, arrhythmias and even sudden cardiac death. Investigating the molecular mechanisms of cardiac fibrosis in acute myocardial infarction (AMI) is essential for better understanding this pathology. The current study aims to investigate the effect of TUG1 on cardiac fibrosis after AMI and elucidated the underlying molecular mechanism of AMI. Rats were randomly divided into four groups (sham-operation group, myocardial infarction group (AMI group), si-NC treated group and si-TUG1 treated group). The biological behavior of cardiac fibroblasts treated with TGF-ß1after being transfected by si-TUG1 or miR-590 mimic or miR-590 inhibitor or FGF1 mimic or a combination was evaluated using the cell counting kit-8 (CCK8) and Transwell assays. SatarBase v2.0 was used to predict the target microRNAs binding site candidates with TUG1 and FGF1. Western blot and recovery experiments were used to explore the potential mechanism. TUG1 expression was up-regulated and knockdown of TUG1 improved cardiac function in AMI rats. Knockdown of TUG1 suppressed cell viability and migration and improved collagen production of TGF-ß1 treated cardiac fibroblasts. SatarBase v2.0 showed TUG1 served as a sponge for miR-590 and FGF1 is a direct target of miR-590. TUG1 expression was increased in AMI tissue and cardiac fibroblasts treated with TGF-ß1. TUG1 knockdown suppressed the biological process of cardiac fibroblasts treated with TGF-ß1 by sponging miR-590.

Long non-coding RNA OIP5-AS1 aggravates acute lung injury by promoting inflammation and cell apoptosis via regulating the miR-26a-5p/TLR4 axis.

BACKGROUND: Acute lung injury (ALI) is a pulmonary disorder that leads to acute respiration failure and thereby results in a high mortality worldwide. Increasing studies have indicated that toll-like receptor 4 (TLR4) is a promoter in ALI, and we aimed to explore the underlying upstream mechanism of TLR4 in ALI. METHODS: We used lipopolysaccharide (LPS) to induce an acute inflammatory response in vitro model and a murine mouse model. A wide range of experiments including reverse transcription quantitative polymerase chain reaction, western blot, enzyme linked immunosorbent assay, flow cytometry, hematoxylin-eosin staining, RNA immunoprecipitation, luciferase activity and caspase-3 activity detection assays were conducted to figure out the expression status, specific role and potential upstream mechanism of TLR4 in ALI. RESULT: TLR4 expression was upregulated in ALI mice and LPS-treated primary bronchial/tracheal epithelial cells. Moreover, miR-26a-5p was confirmed to target TLR4 according to results of luciferase reporter assay. In addition, miR-26a-5p overexpression decreased the contents of proinflammatory factors and inhibited cell apoptosis, while upregulation of TLR4 reversed these effects of miR-26a-5p mimics, implying that miR-26a-5p alleviated ALI by regulating TLR4. Afterwards, OPA interacting protein 5 antisense RNA 1 (OIP5-AS1) was identified to bind with miR-26a-5p. Functionally, OIP5-AS1 upregulation promoted the inflammation and miR-26a-5p overexpression counteracted the influence of OIP5-AS1 upregulation on cell inflammatory response and apoptosis. CONCLUSION: OIP5-AS1 promotes ALI by regulating the miR-26a-5p/TLR4 axis in ALI mice and LPS-treated cells, which indicates a promising insight into diagnostics and therapeutics in ALI.

The dynamics of select cellular responses and cytokine expression profiles in mice infected with juvenile Clonorchis sinensis.

Clonorchiasis is a zoonotic disease that can result in chronic infection in humans. The causative agent, Clonorchis sinensis (C. sinensis), is believed to primarily induce a Th2 immune response in infected mice. However, few studies have profiled host immune responses to C. sinensis infection during the juvenile phase. In the present study, the dynamics of select cellular responses and cytokine expression profiles during juvenile C. sinensis infection were investigated. The flow cytometry results showed that the CD4+ T cells percentage was significantly decreased between 12 days post-infection (dpi) and 24 dpi in the peripheral blood, and the CD8+ T cells percentage was significantly elevated after 3 dpi. The ratio of CD4+/CD8+ T cells was also significantly decreased after 3 dpi. Furthermore, we observed that the proportion of CD14+ monocyte-macrophages in the peripheral blood was significantly increased between 1 dpi and 12 dpi and peaked at 6 dpi. The percentage of classically activated macrophages (M1) and alternatively activated macrophages (M2) in the liver was significantly increased between 18 dpi and 30 dpi. qRT-PCR results showed that the expression levels of iNOS in the liver were significantly elevated after 3 dpi, and Arg-1 expression was significantly increased beginning at 12 dpi. ELISA results showed that the serum levels of the Th1 cytokines IFN-γ and IL-2 peaked at 6 dpi and decreased thereafter. Furthermore, the Th2 cytokines IL-4 and IL-13 began to be expressed and peaked at 24 dpi and 30 dpi, respectively. In addition, the levels of the Treg cytokines IL-10 and TGF-ß1 were significantly increased beginning at 6 dpi until 30 dpi. In the liver homogenate, the expression of IFN-γ, IL-2, and IL-4 mainly occurred before 6 dpi. IL-13 expression was significantly increased at 30 dpi. IL-10 and TGF-ß1 levels were significantly increased at 12 dpi and 24 dpi, and expression peaked at 24 dpi and 30 dpi, respectively. This study provides a fundamental characterization for the future analysis of host-parasite interactions and immune responses in hosts infected with juvenile C. sinensis.

First case report of Metorchis orientalis from Black Swan.

Metorchis orientalis belongs to the genus Metorchis of Opisthorchiidae, which mainly parasitizes in liver and bile ducts of waterfowl, causing liver dysfunction of the host. It has been reported that M. orientalis also infects humans. As a natural species in Australia and a popular ornamental animal, Black Swan (Cygnus atratus) has been imported into many countries. At present there has been no report of M. orientalis infection in Black Swan. In the present study M. orientalis infection in Black Swan was identified by a combination of different techniques, including morphological observation and molecular analysis. M. orientalis adults were found in the gallbladder and bile duct of a three-year-old female Black Swan, which was further confirmed by internal transcribed spacer (ITS) sequence analysis. In addition, the intermediate and definitive hosts of M. orientalis from the 'Qing' lake (a man-made lake in Changchun, China) that Black Swan lived were investigated and the infection route was preliminarily determined. Parafossarulus striatulus functioned as the first intermediate host which contained M. orientalis DNA, and fishes such as Pseudorasbora parva and Rhodeinae served as the second intermediate hosts with M. orientalis metacercariae in the fish flesh. M. orientalis eggs were found in the feces of three other Swans and six ducks that lived in the 'Qing' lake. This was the first reported case about M. orientalis infection of Black Swan. Our study described the course of the infection and provided new information about potential carriers and disseminators of M. orientalis.

The immune protection induced by a serine protease from the Trichinella spiralis adult administered as DNA and protein vaccine.

Trichinellosis is caused by Trichinella spiralis (T. spiralis), which is an important public health problem. In this study, a gene encoding a serine protease from adult worms of T. spiralis (Ts-Adsp) was screened from a cDNA library of adult worms and was cloned and expressed in a prokaryotic expression system. The gene Ts-Adsp was subcloned into the eukaryotic expression vector pcDNA3.1(+), which was named pcDNA3.1(+)-Adsp. Previous studies have found that recombinant Ts-Adsp protein (rTs-Adsp) can elicit partial protection against T. spiralis infection in mice. Our aim was to explore the protective effect of combining a DNA vaccine with the rTs-Adsp protein against T. spiralis. One week after the last vaccination, the serum and spleen were obtained. The levels of IgG, IgG1 and IgG2a and cytokine production in serum and spleen cells were analyzed. The results showed that the levels of humoral and cell-mediated immune responses increased in the pcDNA3.1(+)-Adsp/rTs-Adsp group mice and demonstrated that a Th1/Th2 mixed immune response was induced by pcDNA3.1(+)-Adsp/rTs-Adsp after vaccination. Moreover, mice vaccinated with pcDNA3.1(+)-Adsp/rTs-Adsp displayed a 69.50% reduction in muscle larvae burden. This study suggested that mixed immunity could improve the muscle larvae reduction rate.

Overexpression of tripartite motif containing 26 inhibits non-small cell lung cancer cell growth by suppressing PI3K/AKT signaling.

It has been reported that tripartite motif containing 26 (TRIM26) is involved in the tumorigenesis of some cancers, but its function in non-small cell lung cancer (NSCLC) is still unclear. In this study, we found that TRIM26 was markedly down-regulated in both of NSCLC tumor tissues and cell lines. Additionally, high expression of TRIM26 in NSCLC patients predicted a positive index for patients' overall survival. What is more, overexpression of TRIM26 significantly suppressed NSCLC cell growth. Our further studies indicated that overexpression of TRIM26 inhibited the phosphorylation of PI3K p85 and AKT. And overexpressed TRIM26 regulated cell cycle-related genes' expression, including downregulating CDK4, Cyclin A, Cyclin D1, Cyclin D3, and Cyclin E, and upregulating p27 expression. Finally, we found that TRIM26 up-regulated PTEN expression by stabilizing PTEN protein in NSCLC cells. Collectively, our present study indicated that TRIM26 was decreased in NSCLC and overexpression of TRIM26 inhibited NSCLC cell growth by suppressing PI3K/AKT pathway, which suggested that TRIM26 could be as a potential target for the treatment of NSCLC in the future.

Long noncoding RNA NEAT1 sponges miR-495-3p to enhance myocardial ischemia-reperfusion injury via MAPK6 activation.

The biological function of long noncoding RNA NEAT1 has been revealed in a lot of diseases. Nevertheless, it is still not yet clear whether NEAT1 can modulate the process of myocardial ischemia-reperfusion injury (M-I/R). Here, we reported that NEAT1 was able to sponge miR-495-3p to contribute to M-I/R injury through activating mitogen-activated protein kinase 6 (MAPK6). First, elevated expression of NEAT1 was revealed in M-I/R injury mice, meanwhile, lactate dehydrogenase (LDH) and creatine kinase-muscle/brain (CK-MB) were also upregulated in the serum. Meanwhile, as previously reported, miR-495 serves as a tumor suppressor or an oncogenic miRNA in different types of cancer. Currently, we found miR-495-3p was remarkably reduced in M-I/R mice. Additionally, NEAT1 was significantly induced whereas miR-495-3p was greatly reduced by H2 O2 treatment in H9C2 cells. Moreover, loss of NEAT1 in H9C2 cells could repress the viability and proliferation of cells. For another, overexpression of NEAT1 exhibited an opposite phenomenon. Furthermore, LDH release and caspase-3 activity were obviously triggered by upregulation of NEAT1 while suppressed by NEAT1 knockdown. miR-495-3p was indicated and validated as a target of NEAT1 using the analysis of bioinformatics. Interestingly, we observed that miR-495-3p mimics repressed tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and IL-18 protein expression while their levels were enhanced by the inhibition of miR-495-3p in H9c2 cells. Subsequently, it was manifested that MAPK6 was a target of miR-495-3p, which could exert a lot in the NEAT1/miR-495-3p-mediated M-I/R injury. Overall, our results implied that NEAT1 contributed to M-I/R injury via the modulation of miR-495-3p and MAPK6.

Ginsenoside Rg1 attenuates cardiomyocyte apoptosis and inflammation via the TLR4/NF-kB/NLRP3 pathway.

Sepsis-induced myocardial dysfunction (SIMD) causes high mortality in seriously ill patients. Ginsenoside Rg1 has been proven to have effective anti-inflammatory and antiapoptotic properties. However, the specific role of Rg1 in SIMD and the molecular mechanism remain unclear. Hence, we aimed to investigate the latent effects of ginsenoside Rg1 against SIMD and explore its underlying mechanisms. Male C57BL/6J mice and neonatal rat cardiomyocytes (NRCMs) were used as in vivo and in vitro models, respectively. Western blot analysis was used to detect the level of protein expression, and reverse transcription polymerase chain reaction was conducted to determine the messenger RNA expression of inflammatory factors. The terminal deoxynucleotidyl transferase-mediated nick end labeling assay and flow cytometry were used to determine the apoptosis rate. Echocardiography was performed to assess cardiac function. The results showed that Rg1 improved cardiac function and attenuated lipopolysaccharide (LPS)-induced apoptosis and inflammation in mice. In addition, in NRCMs, Rg1 downregulated the expression of LPS-induced inflammatory cytokines and reversed the increased expression of Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), and NOD-like receptor 3 (NLRP3). In addition, treatment with TLR4 small interfering RNA (siRNA), a p-NF-κB inhibitor, or NLRP3 siRNA suppressed LPS-induced apoptosis and inflammation. In conclusion, Rg1 can attenuate LPS-induced inflammation and apoptosis both in NRCMs and septic mice and restore impaired cardiac function. Moreover, Rg1 may exert its effect via blocking the TLR4/NF-κB/NLRP3 pathway.

The MOV10 helicase restricts hepatitis B virus replication by inhibiting viral reverse transcription.

Interferons inhibit viruses by inducing antiviral protein expression. One of the interferon-induced antiviral proteins, human Moloney leukemia virus 10 (MOV10), a superfamily 1 RNA helicase, has been shown to inhibit retroviruses and several RNA viruses. However, it remains undetermined whether MOV10 also inhibits DNA viruses, including hepatitis B virus (HBV). Here, we report that MOV10 dramatically reduces the levels of intracellular HBV DNA, resulting in significant inhibition of both the HBV experimental strain and the clinical isolates. Mechanistic experiments revealed that MOV10 interacts with HBV RNA and blocks the early step of viral reverse transcription, thereby impairing viral DNA synthesis, without affecting viral gene expression and pregenomic RNA encapsidation. Moreover, mutation of the helicase domain of MOV10 caused loss of binding to HBV RNA and of the anti-HBV activity. Together, our results indicate that MOV10 restricts HBV replication, insights that may open new avenues to the development of anti-HBV therapeutics.

The Effects of Dimethyl Fumarate on Atherosclerosis in the Apolipoprotein E-Deficient Mouse Model with Streptozotocin-Induced Hyperglycemia Mediated By the Nuclear Factor Erythroid 2-Related Factor 2/Antioxidant Response Element (Nrf2/ARE) Signaling Pathway.

BACKGROUND This study aimed to investigate the effects of dimethyl fumarate (DMF) on thoracic aortic atherosclerosis in the apolipoprotein E (apo-E)-deficient mouse model with streptozotocin (STZ)-induced hyperglycemia, and the signaling pathways involved. MATERIAL AND METHODS Eight-week-old ApoE-/- male mice (n=30) were randomly divided into three groups: the Control group (ApoE-/-) (n=10); the diabetic model (STZ) group (n=10); and the DMF-treated (25 mg/kg) diabetic model (DMF+STZ) group (n=10). The area of the thoracic aortic atherosclerosis was determined by histology. Reactive oxygen species (ROS) levels in mouse serum and homogenates of the thoracic aorta were determined by colorimetry. Levels of nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase subunit gp91phox were detected by immunological hybridization, and levels of heme oxygenase-1 (HO-1) were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS Compared with the Control group, in the STZ group, the area of aortic atherosclerosis was significantly increased, the levels of serum and aortic ROS, HO-1, nuclear factor-kappaB (NF-kappaB), intercellular adhesion molecule 1 (ICAM-1), and gp91phox were increased, and nuclear factor erythroid 2-related factor 2 (Nrf2), endothelial nitric oxide synthase (eNOS), and phosphorylated eNOS (p-eNOS) were significantly reduced. Compared with the STZ group, in the DMF+STZ group, the area of aortic atherosclerosis was significantly reduced, the levels of serum and aortic ROS, HO-1, NF-kappaB, ICAM-1, and gp91phox were significantly reduced, and Nrf2, eNOS, and p-eNOS were significantly increased. CONCLUSIONS In the apo-E-deficient mouse model with STZ-induced hyperglycemia, DMF reduced the development of atherosclerosis of the thoracic aorta through the nuclear factor erythroid 2-related factor 2/antioxidant response element (Nrf2/ARE) signaling pathway.

Satellite-based assessment of electricity restoration efforts in Puerto Rico after Hurricane Maria.

A real-time understanding of the distribution and duration of power outages after a major disaster is a precursor to minimizing their harmful consequences. Here, we develop an approach for using daily satellite nighttime lights data to create spatially disaggregated power outage estimates, tracking electricity restoration efforts after disasters strike. In contrast to existing utility data, these estimates are independent, open, and publicly-available, consistently measured across regions that may be serviced by several different power companies, and inclusive of distributed power supply (off-grid systems). We apply the methodology in Puerto Rico following Hurricane Maria, which caused the longest blackout in US history. Within all of the island's settlements, we track outages and recovery times, and link these measures to census-based demographic characteristics of residents. Our results show an 80% decrease in lights, in total, immediately after Hurricane Maria. During the recovery, a disproportionate share of long-duration power failures (> 120 days) occurred in rural municipalities (41% of rural municipalities vs. 29% of urban municipalities), and in the northern and eastern districts. Unexpectedly, we also identify large disparities in electricity recovery between neighborhoods within the same urban area, based primarily on the density of housing. For many urban areas, poor residents, the most vulnerable to increased mortality and morbidity risks from power losses, shouldered the longest outages because they lived in less dense, detached housing where electricity restoration lagged. The approach developed in this study demonstrates the potential of satellite-based estimates of power recovery to improve the real-time monitoring of disaster impacts, globally, at a spatial resolution that is actionable for the disaster response community.