Integrated analysis of miRNA-mRNA expression of newly emerging swine H3N2 influenza virus cross-species infection with tree shrews.

BACKGROUND: Cross-species transmission of zoonotic IAVs to humans is potentially widespread and lethal, posing a great threat to human health, and their cross-species transmission mechanism has attracted much attention. miRNAs have been shown to be involved in the regulation of IAVs infection and immunity, however, few studies have focused on the molecular mechanisms underlying miRNAs and mRNAs expression after IAVs cross-species infection. METHODS: We used tree shrews, a close relative of primates, as a model and used RNA-Seq and bioinformatics tools to analyze the expression profiles of DEMs and DEGs in the nasal turbinate tissue at different time points after the newly emerged swine influenza A virus SW2783 cross-species infection with tree shrews, and miRNA-mRNA interaction maps were constructed and verified by RT-qPCR, miRNA transfection and luciferase reporter assay. RESULTS: 14 DEMs were screened based on functional analysis and interaction map, miR-760-3p, miR-449b-2, miR-30e-3p, and miR-429 were involved in the signal transduction process of replication and proliferation after infection, miR-324-3p, miR-1301-1, miR-103-1, miR-134-5p, miR-29a, miR-31, miR-16b, miR-34a, and miR-125b participate in negative feedback regulation of genes related to the immune function of the body to activate the antiviral immune response, and miR-106b-3p may be related to the cross-species infection potential of SW2783, and the expression level of these miRNAs varies in different days after infection. CONCLUSIONS: The miRNA regulatory networks were constructed and 14 DEMs were identified, some of them can affect the replication and proliferation of viruses by regulating signal transduction, while others can play an antiviral role by regulating the immune response. It indicates that abnormal expression of miRNAs plays a crucial role in the regulation of cross-species IAVs infection, which lays a solid foundation for further exploration of the molecular regulatory mechanism of miRNAs in IAVs cross-species infection and anti-influenza virus targets.

SOV sensitizes gastric cancer cells to radiation by suppressing radiation-induced autophagy in vitro and in vivo.

Vanadium is a transition metal that naturally occurs in the environment and has a variety of biological and physiological impacts on humans. Sodium orthovanadate (SOV), a well-known chemical compound of vanadium, has shown notable anti-cancer activity in various types of human malignancies. However, the effect of SOV on stomach cancer is yet undetermined. Furthermore, only a few studies have investigated the association of SOV and radiosensitivity with stomach cancer. Our study has investigated the ability of SOV to increase the sensitivity of gastric cancer cells to radiation. To detect autophagy triggered by ionizing radiation and the influence of SOV on cell radiosensitivity, the Cell Counting Kit-8 (CCK8) test, EDU staining experiment, colony formation assay, and immunofluorescence were performed. The possible synergistic effects of SOV and irradiation were examined in vivo using a xenograft mouse model of stomach cancer cells. Both in vitro and in vivo studies showed that SOV markedly reduced the growth of stomach cancer cells and improved their radiosensitivity. Our results showed that SOV increased gastric cancer cells' radiosensitivity, thereby blocking the radiation-induced autophagy-related protein, ATG10. Thus, SOV can be considered a potential agent for radiosensitizing gastric cancer.

Pathogenicity and anti-infection immunity of animal H3N2 and H6N6 subtype influenza virus cross-species infection with tree shrews.

Animal influenza viruses can spread across species and pose a fatal threat to human health due to the high pathogenicity and mortality. Animal models are crucial for studying cross-species infection and the pathogenesis of influenza viruses. Tupaia belangeri (tree shrew) has been emerging as an animal model for multiple human virus infections recently because of the close genetic relationship and phylogeny with humans. So far, tree shrew has been reported to be susceptible to human influenza virus subtype H1N1, avian influenza viruses subtype H9N2, subtype H5N1, and subtype H7N9. However, the pathogenicity, infection, and immunity of swine and land avian influenza viruses with low pathogenicity and the potential to jump to humans remain largely unexplored in the tree shrew model. Previously, our team has successfully isolated the newly emerging swine influenza virus subtype H3N2 (A/Swine/GX/NS2783/2010, SW2783) and avian influenza virus subtype H6N6 (A/CK/ZZ/346/2014, ZZ346). In this study, we observed the pathogenicity, immune characteristics, and cross-species infection potential ability of SW2783 and ZZ346 strains in tree shrew model with 50% tissue culture infective dose (TCID50), hematoxylin and eosin (HE) staining, immunohistochemistry (IHC), real-time quantitative PCR (qRT-PCR) and other experimental methods. Both animal-borne influenza viruses had a strong ability on tissue infection in the turbinate and the trachea of tree shrews in vitro, in which SW2783 showed stronger replication ability than in ZZ346. SW2783 and ZZ346 both showed pathogenic ability with infected tree shrews model in vivo without prior adaptive culture, which mainly happened in the upper respiratory tract. However, the infection ability was weak, the clinical symptoms were mild, and the histopathological changes in the respiratory tract were relatively light. Furthermore, innate immune responses and adaptive immunity were observed in the tree shrew model after the infection of SW2783 and ZZ346 strains. We observed that the unadapted SW2783 and ZZ346 virus could transmit among tree shrews by direct contact. We also observed that SW2783 virus could transmit from tree shrews to guinea pigs. These results indicated that both animal-borne influenza viruses could induce similar pathogenicity and immune response to those caused by human-common influenza viruses. Tree shrews may be an excellent animal model for studying the interaction between the influenza virus and the host and the cross-species infection mechanism of the animal influenza virus.

Influenza A (H6N6) viruses isolated from chickens replicate in mice and human lungs without prior adaptation.

The H6H6 subtype avian influenza virus (AIV) is currently prevalent in wild birds and poultry. Its host range has gradually expanded to mammals, such as swines. Some strains have even acquired the ability to bind to human-like SAα-2,6 Gal receptors, thus increasing the risk of animal to human transmission. To investigate whether the H6N6 AIV can overcome interspecies barriers from poultry to mammals and even to humans, we have assessed the molecular characteristics, receptor-binding preference, replication in mice and human lungs of three chicken-originated H6N6 strains. Among these, the A/CK/Zhangzhou/346/2014 (ZZ346) virus with the P186T, H156R, and S263G mutations of the hemagglutinin molecule showed the ability to bind to avian-like SAα-2,3 Gal and human-like SAα-2,6 Gal receptors. Moreover, H6N6 viruses, especially the ZZ346 strain, could replicate and infect mice and human lungs. Our study showed the H6N6 virus binding to both avian-like and human-like receptors, confirming its ability to cross the species barrier to infect mice and human lungs without prior adaptation. This study emphasizes the importance of continuous and intense monitoring of the H6N6 evolution in terrestrial birds.

M6 A demethylase fat mass and obesity-associated protein regulates cisplatin resistance of gastric cancer by modulating autophagy activation through ULK1.

Drug resistance is an important factor for treatment failure of gastric cancer. N6 -methyladenosine (m6 A) is the predominant mRNA internal modification in eukaryotes. The roles of m6 A modification in drug resistance of gastric cancer remains unclear. In the present study, the m6 A methylated RNA level was significantly decreased while the expression of m6 A demethylase fat mass and obesity-associated protein (FTO) was obviously elevated in cisplatin-resistant (SGC-7901/DDP) gastric cancer cells. Knockdown of FTO reversed cisplatin resistance of SGC-7901/DDP cells both in vitro and in vivo, which was attributed to the inhibition of Unc-51-like kinase 1 (ULK1)-mediated autophagy. Mechanistically, ULK1 expression was regulated in an FTO-m6 A-dependent and YTHDF2-mediated manner. Collectively, our findings indicate that the FTO/ULK1 axis exerts crucial roles in cisplatin resistance of gastric cancer.

How Wealth Inequality Affects Happiness: The Perspective of Social Comparison.

Since Easterlin pointed out that economic growth in nations does not guarantee increasing happiness for the average citizen, the underlying reason has remained controversial. The present study focuses on income inequality to explain the "Easterlin Paradox," ignoring the permanent inequality that long-term wealth accumulation brings. Based on social comparison theory, the literature aims to determine how wealth inequality, which accompanies economic growth, diminishes one's happiness (inequality aversion). Specifically, we conduct this study in which we split the wealth inequality into the upward wealth inequality and the downward wealth inequality as measures of upward comparison and downward comparison, respectively. The upward wealth inequality measures the average gap between one and the better-off in wealth while the downward wealth inequality measures the average gap between one and the worse-off in wealth. Furthermore, the heterogeneity of the area of respondent is analyzed and the family life cycle is tested as a moderator. The main findings of the paper are as follows: (1) The empirical test results of hypothesis 1 indicate that the upward wealth inequality aversion (jealousy effect: people envy who is richer than themselves) is stronger than the downward wealth inequality inclination (proud effect: people enjoy having a superior position in the wealth hierarchy). It is due to the psychological preference: loss aversion. As an increase in upward distance implies a loss in relative status and an increase in downward distance implies a gain in relative status, people focus more on loss rather than gain. (2) The empirical test results of hypothesis 2 indicate that residents who live in rural areas do not have a proud effect as much as those who live in urban areas. There is a huge urban-rural wealth gap in China. With the expansion of the social network, people living in rural areas realize that even he is almost the rich in rural areas but still the lower classes in the whole society. It is hard for rural residents to have a proud effect. (3) The empirical test results of hypothesis 3 indicate that family members have the strongest upward inequality aversion in the middle-stage phase of the life cycle (when the family head is approximately 50). During the family life cycle, inequality aversion will be different in different life stages due to the changes in economic status expectations. At the beginning of the family life cycle, family members assume their life has limitless possibilities, and they have high expectations for the future. Logically, they can be easily satisfied by achieving a little more than their peers. In later periods, with increasing age, the members will pay more attention to health instead of wealth. The results shed light on how macroeconomics influence changes in individual psychology.

MicroRNA-204 plays a role as a tumor suppressor in Newcastle disease virus-induced oncolysis in lung cancer A549 cells.

Tumor development and progression are closely associated with various microRNAs (miRNAs/miRs). We have previously shown that Newcastle disease virus (NDV) strain 7793 induces oncolysis in lung cancer. However, how NDV exerts its oncolytic effect on lung cancer remains to be investigated. The present study assessed the role of miR-204 in the NDV-induced oncolysis of lung cancer A549 cells by oncolysis induction in vitro. miR-204 was significantly upregulated in NDV-treated A549 cells. Overexpression or inhibition of miR-204 was significantly associated with NDV-induced oncolysis in A549 cells. Caspase-3 and Bax, major regulators of the apoptosis pathway, were regulated by miR-204, and the association between caspase-3-related apoptosis and miR-204 was identified in NDV-mediated oncolysis. These data demonstrated that miR-204 as a tumor suppressor played a role in NDV-induced oncolysis in lung cancer cells. The present study demonstrates the potential of strategies using miRs to improve oncolytic NDV potency, and highlights miR-204 as a tumor suppressor in NDV-induced oncolysis of lung cancer cells.

A Half-Century Studies on Epidemiological Features of Ancylostomiasis in China: A Review Article.

BACKGROUND: Ancylostomiasis is a prevalent and global parasitic disease, including China. A systematic review is significant to understand the epidemiological features of hookworm and provide guidance for prevention and treatment. METHODS: We systematically searched academic databases and assessed 944 papers published from 1955-2015 to establish the comprehensive analysis of prevalence of hookworm disease in China. We searched Chinese databases, including CNKI, Wanfang and VIP, for literature with the subject word "Ancylostomiasis and hookworm". The data were analyzed with SPSS 19.0 software using Spearman correlation analysis. Results were statistically significant for a P-value of <0.01. RESULTS: The search yielded 532,151 cases from epidemiological investigation and 7294 cases based on hospital diagnosis. Hookworm infection was highest (15.83%) in Fujian province, with high rates also found in East China, Southwest China, Central China and Southern China and lower rates in Northwest China, North China and Northeast China. In terms of occupation, farmers had the highest proportion of infections (72.54%). There was no correlation between epidemiological investigations and hospital-diagnosed cases. However, there was significant positive correlation between hospital-diagnosed cases and misdiagnosed cases. The proportion of hospital-misdiagnosed cases was 32.80%. CONCLUSION: Ancylostomiasis is a serious public health problem that negatively influences health and hinders socioeconomic development. Positive measures are required by both health services and individuals to prevent and control hookworm disease.

Diagnostic Value of Serum MiR-1 in Patients with Acute Kawasaki Disease.

BACKGROUND: The current study aims to investigate the expression of miR-1 in serum of patients with Kawasaki disease (KD) and its clinical significance. METHODS: The serum samples of 33 patients with KD and 15 healthy people were collected from January 2017 to June 2017 at the Affiliated Hospital of North China University of Science and Technology. The expression of serum miR-1 was detected by real-time quantitative PCR (RT-qPCR). The diagnostic value of miR-1 as a marker of KD disease was evaluated by receiver operating characteristic (ROC) curve. RESULTS: The level of miR-1 in serum of children with acute KD was significantly higher than that of healthy children, but it decreased to normal level in convalescence. ROC curves showed that the area under the curve (AUC) of miR-1 for KD diagnosis was 0.754 (95% CI: 0.541 - 0.952). When the critical value (diagnostic threshold) was 1.08, the diagnostic sensitivity and specificity were 0.867 and 0.735, respectively. In addition, we further divided the KD patients according to clinical characteristics. Here, we showed that the expression of miR-1 was higher in the serum of KD patients with higher platelet level. CONCLUSIONS: In summary, serum miR-1 in patients with acute KD was significantly increased, which may be one of the potential serum markers for the diagnosis of KD.

Integration analysis of a miRNA-mRNA expression in A549 cells infected with a novel H3N2 swine influenza virus and the 2009 H1N1 pandemic influenza virus.

Swine are reservoirs for anthropogenic/zoonotic influenza viruses, and the prevalence and repeated introduction of the 2009 H1N1 pandemic influenza virus (pdm/09) into pigs raises the possibility of generating novel swine influenza viruses with the potential to infect humans. However, studies aiming to identify miRNAs involved in the transfer of novel swine influenza virus infection to human cells are rare. In this investigation, from the view of small RNA, microarrays and high-throughput sequencing were used to detect differentially expressed miRNAs and mRNAs after human lung epithelial cells were infected with the following three stains of influenza viruses: a novel H3N2 swine influenza virus reassorted with pdm/09 fragments, pdm/09 and classical swine influenza virus. A miRNA-mRNA interaction map was generated to show the correlation between miRNAs related to infection by the viruses with human infective potential/capability. The expression of 4 miRNAs (hsa-miR-96-5p, hsa-miR-140-5p, hsa-miR-30a-3p and hsa-miR-582-5p) and 5 relevant mRNAs (RCC1, ERVFRD-1, RANBP1, SCARB2 and RPS29) was determined. The integration analysis indicated that these candidates have rarely been reported to be associated with influenza virus. Focusing on miRNA expression changes could reveal novel reassortant viruses with human infective potential that may provide insight into future pandemics.

Integrated analysis of microRNA-mRNA expression in A549 cells infected with influenza A viruses (IAVs) from different host species.

Although several miRNAs have been demonstrated to be involved in the influenza virus replication cycle, the identification of miRNAs and mRNAs that are expressed in A549 cells infected with influenza A viruses (IAVs) from different host species has remained poorly studied. To investigate the molecular mechanisms associated with the differential expression of miRNAs during influenza A virus infection, we performed global miRNA and mRNA expression profiling in A549 cells infected with human-origin seasonal influenza A virus H3N2 (Human_Br07), swine-origin influenza A virus H1N1 (SW_3861) or avian-origin influenza A virus H3N2 (AVI_9990). The miRNA and mRNA expression profiles were obtained by microarray and high-throughput sequencing analyses, respectively. The integrated analysis of differentially expressed miRNAs (DEMs) and differentially expressed genes (DEGs) was performed using bioinformatics tools, and the expression of miRNAs and mRNAs was validated by real-time quantitative polymerase chain reaction (RT-qPCR). We identified 20 miRNAs (6 upregulated and 14 downregulated) and 1286 mRNAs (935 upregulated and 351 downregulated) exhibiting the same differential expression trends in three infected groups of cells compared with an uninfected control. An integrated analysis of these expression profiles identified 79 miRNA-mRNA pairs associated with the influenza A reference pathway, and 107 miRNA-mRNA interactions were correlated with the defense of the virus. Additionally, the obtained results were supported by an RT-qPCR analysis of 8 differentially expressed miRNAs (hsa-miR-210-3p, hsa-miR-296-5p, hsa-miR-371a-5p, hsa-miR-762, hsa-miR-937-5p, hsa-miR-1915-3p, hsa-miR-3665, and hsa-miR-1290) and 13 differentially expressed mRNAs (IFNL1, CXCL10, RSAD2, MX1, OAS2, IFIT2, IFI44 L, MX2, XAF1, NDRG1, FGA, EGLN3, and TFRC). Our findings indicate that dysregulated miRNA expression plays a crucial role in infection caused by IAVs originating from different species and provide a foundation for further investigations of the molecular regulatory mechanisms of miRNAs involved in influenza A virus infection.

The hemagglutinin-neuramidinase protein of Newcastle disease virus upregulates expression of the TRAIL gene in murine natural killer cells through the activation of Syk and NF-κB.

Newcastle disease virus (NDV) is responsible for tumoricidal activity in vitro and in vivo. However, the mechanisms that lead to this activity are unclear. Natural killer cells are able to induce apoptosis of tumor cells through multiple pathways, including the tumor necrosis factor-related apoptosis-inducing ligand-death receptor pathway. We previously showed that exposure of NK and T cells to NDV resulted in enhanced tumoricidal activity that was mediated by upregulated expression of the TRAIL gene, via an interferon gamma -dependent pathway. Other pathways involved in the upregulated expression of TRAIL are yet to be identified. In the current study, we used mice in which the IFN-γ receptor one gene was inactivated functionally. We identified an IFN-γ-independent TRAIL pathway in the NDV-stimulated NK cells. Hemagglutinin-neuramidinase induced expression of the TRAIL gene in IFN-R1-/- NK cells by binding to the NKp46 receptor. This upregulation was inhibited by pretreatment of NDV with a neutralizing monoclonal antibody against HN, or desialylation of NK cells. Phosphorylation of spleen tryosine kinases and IκBα was increased in HN-induced IFN-R1-/- NK cells. Treatment with the HN neutralizing monoclonal antibody, pharmacological disialylation, or a Syk inhibitor decreased Syk and IκBα phosphorylation levels. We concluded that killer activation receptors pathway is involved in the IFN-γ-independent TRAIL expression of NDV-stimulated NK cells, and these are activated by Syk and NF-κB.

[Newcastle disease virus enhances tumoricidal activity of mouse NK cells against mouse Novikoff hepatoma cells via up-regulating expression of TRAIL on the NK cells].

OBJECTIVE: To observe the effect of intraperitoneal injection of Newcastle disease virus (NDV) on tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) expression in mouse spleen NK cells and NK cells-mediated tumoricidal activity against mouse Novikoff hepatoma cell line, and explore the role of interferon (IFN)-γ in NDV-induced TRAIL expression and tumoricidal activity. METHODS: NDV was injected intraperitoneally to BALB/c mice and IFN-γ receptor-deficient (IFN-γR-/-) B6.129S7 mice. Twelve hours after injection, the concentration of IFN-γ in peripheral blood from BALB/c mice was determined by ELISA. Mouse spleen NK cells were separated. The mRNA and protein expression of TRAIL in NK cells were detected through reverse transcription PCR (RT-PCR) and Western blotting. Lactate dehydrogenase (LDH) release assay was used to determine the cytotoxic activity of NK cells against mouse hepatoma cells. RESULTS: NDV injection increased the IFN-γ concentration in peripheral blood of BALB/c mice, induced up-regulation of TRAIL at the mRNA and protein levels in mouse spleen NK cells, and enhanced the killing ability of mouse spleen NK cells towards Novikoff hepatoma cells. Blocking TRAIL by neutralizing antibody suppressed the cytotoxic activity of NK cells against Novikoff hepatoma cells. Furthermore, NDV injection in IFN-γR-/- B6.129S7 mice did not make significant difference from control group in TRAIL expression in spleen NK cells, and the tumoricidal activity of IFN-γR-/- B6.129S7 mouse spleen NK cells against Novikoff hepatoma cells was significantly lower than that of BALB/c mouse NK cells. CONCLUSION: Intraperitoneal injection with NDV could enhance tumoricidal activity of mouse spleen NK cells in vitro, and one of the mechanisms might be that NDV injection up-regulates TRAIL expression in NK cells through the IFN-γ receptor pathway.

TRAIL is involved in the tumoricidal activity of mouse natural killer cells stimulated by Newcastle disease virus in vitro.

Newcastle disease virus (NDV) is a potential antitumor agent, and its antitumor effect has been evaluated in preclinical tests. However, the mechanisms of NDV-based antitumor therapy are still not completely clear. In the present study we found that NDV-stimulation enhanced the killing ability of mouse spleen natural killer (NK) cells towards mouse hepatoma cell lines, and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) plays an important role in this tumoricidal activity. NDV stimulation induced up-regulation of TRAIL both at the mRNA and protein levels in NK cells. Blocking TRAIL by antibody (Ab) almost completely eliminated the killing effect of NK cells on hepatoma cell lines. Furthermore, neutralizing interferon (IFN)-γ by Ab could inhibit TRAIL expression and tumoricidal activity of NDV-stimulated NK cells. These results indicated a substantial role of TRAIL as an effector molecule in NDV-induced NK cells mediated tumoricidal activity. The NDV stimulation triggered TRAIL expression in mouse spleen NK cells could be mediated by IFN-γ induction.

Clinical study of isosorbide mononitrate treatment for angina pectoris in coronary heart disease.

The aim of this study was to observe and investigate the clinical efficacy of an intravenous drip of isosorbide mononitrate (ISMN) for the treatment of angina pectoris in coronary heart disease. A total of 102 patients with angina pectoris in coronary heart disease were divided into two groups. For the treatment group (n=51), 20 mg ISMN was added to 250 ml 0.9% normal saline and administered by intravenous drip for 14 consecutive days, twice daily. For the control group (n=51), 10 mg glyceryl trinitrate was added to 250 ml 0.9% normal saline and administered by intravenous drip for 14 consecutive days, twice daily. The clinical efficacy and adverse reactions were compared between the two groups. The disease symptoms of the two groups were improved. Compared with the control group, the clinical efficacy and electrocardiogram examination results of the treatment group were significantly improved (P<0.05). The intravenous formulation of ISMN is an effective treatment for angina pectoris in coronary heart disease and it has the advantages of fewer adverse reactions and higher safety.