Lnc-PLCB1 is stabilized by METTL14 induced m6A modification and inhibits Helicobacter pylori mediated gastric cancer by destabilizing DDX21.

Helicobacter pylori (H. pylori) infection is considered to be an important factor in gastric cancer (GC). Long noncoding RNA (lncRNA) and m6A modification are involved in the occurrence and development of GC, but the role of lncRNA m6A modification in the development of GC mediated by H. pylori is still unclear. Here, we found that H. pylori infection downregulated the expression of lnc-PLCB1 through METTL14-mediated m6A modification and IRF2-mediated transcriptional regulation. Overexpression of lnc-PLCB1 inhibited the proliferation and migration of GC cells, while downregulation of lnc-PLCB1 promoted the proliferation and migration ability of GC cells. In addition, clinical analysis showed that lnc-PLCB1 is lower in GC tissues than in normal tissues. Further study found that lnc-PLCB1 reduced the protein stability of its binding protein DEAD-box helicase 21 (DDX21) and then downregulated the expression of CCND1 and Slug, thereby playing tumour suppressing role in the occurrence and development of GC. In conclusion, the METTL14/lnc-PLCB1/DDX21 axis plays an important role in H. pylori-mediated GC, and lnc-PLCB1 can be used as a new target for GC treatment.

Advanced Lung Cancer Inflammation Index for Predicting Prognostic Risk for Patients with Acute Coronary Syndrome Undergoing Percutaneous Coronary Intervention.

Purpose: The decreased advanced lung cancer inflammation index (ALI), defined as body mass index (BMI) * albumin (Alb)/neutrophil-to-lymphocyte ratio (NLR), is an independent prognostic risk factor for overall survival in gastric, lung, and colorectal cancers. This study aimed to investigate the value of ALI in predicting the risk of major adverse cardiovascular events (MACEs) in patients with acute coronary syndrome (ACS). Patients and Methods: A total of 1624 patients with ACS undergoing percutaneous coronary intervention (PCI) were consecutively enrolled between January 2016 and December 2018. Follow-up data were collected at 1, 3, 6, and 12 months and annually thereafter. The primary endpoints were MACEs. All endpoints were defined as all-cause mortality, recurrent angina pectoris, restenosis/intra stent thrombosis, stroke, heart failure, and all-cause bleeding. Results: The MACEs group and non-MACEs group showed significant differences in patients with age >65 years (28 [50.0%] vs 319 [23.7%]), history of heart failure (16 [28.6%] vs 127 [9.4%]), history of ischemic stroke (14 [25.0%] vs 186 [13.8%]), history of cardiogenic shock (6 [10.71%] vs 16 [1.19%]), left ventricular ejection fraction <40% (8 [14.29%] vs 33 [2.46%]), and ALI <343.96 (44 [78.65%] vs 680 [50.60%]) (all p<0.001). The optimal cut-off value for ALI was 334.96. The area under the curve (AUC) of the 1-, 2-, 3-, and 5-year was 0.560, 0.577, 0.665, and 0.749, respectively. The survival rate was significantly lower in the low ALI group than in the high ALI group (log-rank p<0.001). Low ALI was an independent risk factor for the long-term prognosis of patients with ACS after PCI, univariate HR: 3.671, 95% CI: 1.938-6.953, p<0.001; multivariate HR: 3.009, 95% CI: 1.57-5.769, p=0.001. Conclusion: ALI score less than 334.96 is an independent prognostic risk factor for patients with ACS undergoing PCI and may be a novel marker for clinical practice.

The Prognostic Value of Hematologic Inflammatory Markers in Patients With Acute Coronary Syndrome Undergoing Percutaneous Coronary Intervention.

The aggregate index of systemic inflammation (AISI), systemic inflammation response index (SIRI), and neutrophil-to-lymphocyte*platelet ratio (NLRP) are novel indices that simultaneously reflect the inflammatory and immune status. However, the role of these indices in acute coronary syndrome (ACS) patients undergoing percutaneous coronary intervention (PCI) remains unclear. We aimed to elucidate the predictive value of AISI, SIRI, and NLRP in patients with ACS undergoing PCI. A total of 1558 patients with ACS undergoing PCI were consecutively enrolled from January 2016 to December 2018. The AISI, SIRI, NLRP, systemic immune-inflammatory index, derived neutrophil-to-lymphocyte ratio, neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, and monocyte-to-lymphocyte ratio cutoff values for predicting major adverse cardiovascular events (MACE) were calculated using receiver-operating characteristic curves, and Spearman's test was used to analyze correlations between these indices. Kaplan-Meier curves and Cox regression models were used for survival analyses, and the endpoint was a MACE, which included all-cause mortality and rehospitalization for severe heart failure during the follow-up period. The Kaplan-Meier curves showed that higher AISI, SIRI, and NLRP values were associated with a higher risk of MACE (all P < .001). The association between AISI, SIRI, and NLRP and ACS prognosis was stable in various subgroups according to sex, age, smoking, dyslipidemia, hypertension, diabetes mellitus, history of stroke, and heart failure (P for interaction > .05). Increasing tertiles of AISI, SIRI, and NLRP significantly increased the MACE risk (P for trend < .05). AISI, SIRI, and NLRP may be suitable laboratory markers for identifying high-risk patients with ACS after PCI.

Tumour-suppressing functions of the lncRNA MBNL1-AS1/miR-889-3p/KLF9 axis in human breast cancer cells.

This study aimed to explore the role and potential mechanism of the long non-coding (lncRNA) MBNL1-AS1 in human breast cancer. We included 80 patients with breast cancer in this study. Breast cancer cell lines, including MCF7, SKBR3, MDA-MB-231 and MDA-MB-415, and the normal human breast cell line MCF10A were used in this study. MBNL1-AS1, miR-889-3p mimics, si-Krüppel-like factor 9 (KLF9) or their controls were transfected in the cells. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), Western blotting and immunohistochemistry assay were performed to detect the expression of MBNL1-AS1, miR-889-3p and KLF9. Cell proliferation, invasion and migration were detected. Luciferase reporter gene and pull-down assay were performed to verify the target relationship among MBNL1-AS1, miR-889-3p and KLF9. Glycolysis was also detected after transfection. The expression of the lncRNA MBNL1-AS1 was low in the breast cancer tissues and cells. Lower expression levels of the lncRNA MBNL1-AS1 were associated with poor prognosis of breast cancer. Overexpression of the lncRNA MBNL1-AS1 decreased proliferation, invasion, migration and glycolysis of breast cancer cells. The lncRNA MBNL1-AS1 could interact with miR-889-3p, and KLF9 was the downstream target of miR-889-3p. Moreover, miR-889-3p was negatively correlated with KLF9 and lncRNA MBNL1-AS1. Both miR-889-3p and si-KLF9 could reverse the overexpression of lncRNA MBNL1-AS1 in breast cancer development. The lncRNA MBNL1-AS1 decreased proliferation, invasion, migration and glycolysis of breast cancer via the miR-889-3p/KLF9 axis, which might be a potential biomarker for the diagnosis of breast cancer.

Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2.

Pseudoviruses are useful virological tools because of their safety and versatility, especially for emerging and re-emerging viruses. Due to its high pathogenicity and infectivity and the lack of effective vaccines and therapeutics, live SARS-CoV-2 has to be handled under biosafety level 3 conditions, which has hindered the development of vaccines and therapeutics. Based on a VSV pseudovirus production system, a pseudovirus-based neutralization assay has been developed for evaluating neutralizing antibodies against SARS-CoV-2 in biosafety level 2 facilities. The key parameters for this assay were optimized, including cell types, cell numbers, virus inoculum. When tested against the SARS-CoV-2 pseudovirus, SARS-CoV-2 convalescent patient sera showed high neutralizing potency, which underscore its potential as therapeutics. The limit of detection for this assay was determined as 22.1 and 43.2 for human and mouse serum samples respectively using a panel of 120 negative samples. The cutoff values were set as 30 and 50 for human and mouse serum samples, respectively. This assay showed relatively low coefficient of variations with 15.9% and 16.2% for the intra- and inter-assay analyses respectively. Taken together, we established a robust pseudovirus-based neutralization assay for SARS-CoV-2 and are glad to share pseudoviruses and related protocols with the developers of vaccines or therapeutics to fight against this lethal virus.

Evaluation of miR-331-3p and miR-23b-3p as serum biomarkers for hepatitis c virus-related hepatocellular carcinoma at early stage.

BACKGROUND AND AIMS: This study aimed to investigate the diagnostic values of serum miR-331-3p and miR-23b-3p as tumor markers for the diagnosis of hepatocellular carcinoma (HCC) at early stage. METHODS: A total of 191 subjects were enrolled and consisted of 45 healthy controls (HC), 106 hepatitis c virus (HCV)-related chronic liver disease (CLD) patients, and 40 early-stage HCC patients. CLD patients were subdivided according to Metavir fibrosis-scoring. Serum miR-331-3p and miR-23b-3p were measured. The area under curves (AUC) was calculated for each microRNA and compared with that for alpha-fetoprotein (AFP) in the detection of HCC at early stage. RESULTS: Serum miR-331-3p was significantly higher in early-stage HCC than that in CLD and HC respectively, and it decreased significantly after surgery in early-stage HCC. Contrarily, serum miR-23b-3p was significantly lower in early-stage HCC and increased significantly after surgery. Further, receiver operating characteristic analysis demonstrated AUC was 0.806 (95%CI: 0.728-0.883; sensitivity: 85.85%, specificity: 65.00%) for serum miR-23b-3p in discriminating early-stage HCC from CLD patients, higher than that for AFP (AUC:0.660, 95%CI: 0.556-0.764; sensitivity: 70.00%, specificity: 56.60%). In discrimination early-stage HCC from severe fibrosis/cirrhosis (F3 + F4) patients, both miR-23b-3p (AUC: 0.796, 95%CI: 0.703-0.889; sensitivity: 85.11%, specificity: 65.00%) and miR-331-3p (AUC:0.832, 95%CI: 0.812-0.953; sensitivity: 75.00%, specificity: 85.11%) had better diagnostic performances than AFP (AUC:0.632, 95%CI: 0.512-0.753; sensitivity: 50.00%, specificity: 55.32%). Serum miR-331-3p levels also showed a significant correlation with BCLC stages of HCC. CONCLUSION: Serum miR-331-3p and miR-23b-3p could be used as novel invasive biomarkers in the early detection of HCC in high-risk patients.

LncRNA PVT1 regulates growth, migration, and invasion of bladder cancer by miR-31/ CDK1.

PURPOSE: The aim of our study was to validate the sway of long noncoding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) on the metabolism and growth of bladder cancer cells by microRNA-31 (miR-31)/cyclin-dependent kinase 1 ( CDK1). METHODS: The Gene Expression Omnibus database was used for analyzing the differentially expressed lncRNA and messenger RNA (mRNA) in bladder cancer tissues, with the highly expressed lncRNA PVT1 and mRNA CDK1 screened out. The expression level of PVT1 was detected by quantitative reverse-transcription polymerase chain reaction, cell viability by Cell Counting Kit-8 assay, cell proliferation and scratch by 5-bromo-2'-deoxyuridine assay, cell migration and invasion by transwell assays, the expression level of CDK1 by immunohistochemistry and western blot analysis, transcription factor targeting by dual-luciferase assay, and the effect of PVT1 on bladder cancer growth by nude mice tumor formation experiment. RESULTS: LncRNA PVT1 and mRNA CDK1 had a higher expression in bladder cancer cells than that in neighboring tissues. Activity, proliferation, colony formation, migration, and invasion of bladder cancer cell were noticeably reduced by the PVT1 inhibitor than that of control group. PVT1 and CDK1 have binding sites with miR-31. When miR-31 decreased, CDK1 mRNA and protein levels increased in vivo experiments in nude mice. When PVT1 was downregulated, the tumor size was significantly reduced and tumor proliferation was curbed. Immunohistochemistry showed that the positive rate of CDK1 and Ki-67 decreased and the expression of miR-31 increased after PVT1 was inhibited. CONCLUSIONS: LncRNA PVT1 was overexpressed in bladder cancer cells, and it was downregulated miR-31 to enhance CDK1 expression and facilitate bladder cancer cells proliferation, migration, and invasion.

[Clinical value of angiogenin in predicting the prognosis of patients with idiopathic pulmonary fibrosis].

OBJECTIVE: To explore the relationship between angiogenin-1/2 (Ang-1/2) and clinical parameters of idiopathic pulmonary fibrosis (IPF), and to assess the value of Ang-1/2 in predicting the prognosis of patients with IPF. METHODS: A retrospective analysis was conducted. Ninety-one patients diagnosed as IPF by high resolution CT (HRCT) and lung biopsy admitted to Daqing Oil Field General Hospital from March 2014 to January 2015 were enrolled. The general data, serum parameters and pulmonary function parameters of all patients were collected. After treatment, all of the 91 patients were followed-up to 2 years. The patients were divided into favorable prognosis group and unfavorable prognosis group according to follow-up results. The differences in all parameters between the two groups were compared. The relationship between Ang-1, Ang-2 and lung function parameters was analyzed by Pearson correlation analysis. Cox proportional hazard regression model was used to evaluate the effect of clinical parameters on the prognosis of patients with IPF. The effect of Ang-2 in predicting prognosis of patients with IPF was analyzed by receiver operating characteristic (ROC) curve. RESULTS: During the 2-year follow-up period, 30 of 91 patients showed a favorable prognosis, and 55 showed an unfavorable prognosis with a poor prognosis rate of 64.71%, and 6 patients withdrew from the study due to loss of follow-up and death. Compared with the favorable prognosis group, Ang-2 level in the unfavorable prognosis group was significantly increased (µg/L: 2.88±1.63 vs. 1.89±1.22, t = 2.909, P = 0.005), but Ang-1 only showed a slight increase (µg/L: 28.70±14.26 vs. 25.62±11.95, t = 1.005, P = 0.318). The results of Pearson correlation analysis showed that Ang-2 level was negatively correlated with forced expiratory volume in 1 second (FVC1) and the percentage of carbon monoxide diffusing capacity accounting for the expected value (DLCO%: r value was -0.227 and -0.206, and P value was 0.147 and 0.253, respectively), but no significant correlation between the level of Ang-1 and FVC1 as well as DLCO% was found (r value was -0.153 and -0.121, and P value was 0.147 and 0.253, respectively). Cox proportional hazard regression model analysis showed that the prognosis of patients with IPF was significantly affected by smoking time and Ang-2 (both P < 0.05), and the influence of Ang-2 was greater [relative risk (RR): 1.236 vs. 1.006, P = 0.037]. Age, gender, smoking and the levels of FVC1, DLCO% and Ang-1 had no significant effect on the prognosis of IPF patients (all P > 0.05). Prognostic analysis showed that the area under ROC curve (AUC) of Ang-2 for predicting prognosis of patients with IPF was 0.692, and the best diagnostic point was 0.35 µg/L, the sensitivity was 61.8%, the specificity was 73.3%, the positive predictive value was 69.8%, and the negative predictive value was 65.7% which indicated that Ang-2 could predict the prognosis of patients with IPF. CONCLUSIONS: Ang-2 could assess the prognosis of patients with IPF, which is expected to be used as an indicator of predicting the prognosis of patients with IPF.

Host Responses and Regulation by NFκB Signaling in the Liver and Liver Epithelial Cells Infected with A Novel Tick-borne Bunyavirus.

Infection in humans by severe fever with thrombocytopenia syndrome virus (SFTSV), a novel bunyavirus transmitted by ticks, is often associated with pronounced liver damage, especially in fatal cases. Little has been known, however, about how liver cells respond to SFTSV and how the response is regulated. In this study we report that proinflammatory cytokines were induced in liver tissues of C57/BL6 mice infected with SFTSV, which may cause tissue necrosis in mice. Human liver epithelial cells were susceptible to SFTSV and antiviral interferon (IFN) and IFN-inducible proteins were induced upon infection. We observed that infection of liver epithelial cells led to significant increases in proinflammatory cytokines and chemokines, including IL-6, RANTES, IP-10, and MIP-3a, which were regulated by NFκB signaling, and the activation of NFκB signaling during infection promoted viral replication in liver epithelial cells. Viral nonstructural protein NSs was inhibitory to the induction of IFN-ß, but interestingly, NFκB activation was enhanced in the presence of NSs. Therefore, NSs plays dual roles in the suppression of antiviral IFN-ß induction as well as the promotion of proinflammatory responses. Our findings provide the first evidence for elucidating host responses and regulation in liver epithelial cells infected by an emerging bunyavirus.

Prognostic value of serum leptin in advanced lung adenocarcinoma patients with cisplatin/pemetrexed chemotherapy.

Cisplatin/pemetrexed chemotherapy has been established as a standard treatment in lung adenocarcinoma. However, the response to the cisplatin/pemetrexed combination varies considerably among patients due to individual variations. Thus, novel biomarkers are required to aid the prediction of the response to the cisplatin/pemetrexed combination. We hypothesized that leptin expression may be a determinant for prognosis in lung adenocarcinoma patients with cisplatin/pemetrexed chemotherapy. Serum from consenting patients with lung adenocarcinoma were obtained for the measurement of leptin and associated tumor biomarkers. Leptin expression was measured by radioimmunoassay. Carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), CA15-3, CA125, CA72-4, cytokeratin 19 fragment (CYFRA21-1) and neuron-specific enolase (NSE) expression were determined by electrochemiluminescence immunoassays. Serum squamous cell carcinoma antigen levels were measured using a microparticle enzyme immunoassay. The associations between serum leptin and tumor biomarker expression were evaluated by Spearman's correlation analysis. Serum CEA, CA19-9, CA15-3, CA125, CA72-4, CYFRA21-1 and NSE levels showed no obvious difference among patients. However, a trend towards an improved prognosis was observed in patients with lower serum leptin at diagnosis and an increase during cisplatin/pemetrexed chemotherapy. The results indicated that the serum leptin level has prognostic indications in patients with advanced lung adenocarcinoma during cisplatin/pemetrexed chemotherapy, which indicates that it may be a useful marker for the prognosis of cancer patients undergoing chemotherapy treatment.

Robust antiviral responses to enterovirus 71 infection in human intestinal epithelial cells.

Enterovirus 71 (EV71) is a single-stranded RNA virus that belongs to Picornaviridae family. It causes the hand-foot-and-mouth disease and fatal neurological diseases in young children and infants. The mechanism of EV71 pathogenesis remains obscure. The intestinal tract is the initial site of EV71 replication, but no or only mild gastrointestinal symptoms are observed clinically, suggesting that host immune responses of the intestinal epithelium to EV71 may be unique, which, however, remains rarely investigated. In this study, we showed that human intestinal epithelial cells HT-29 were susceptible to EV71, and the infected cells exhibited cytopathic effects (CPEs) and were prone to apoptosis. TLR-7 and TLR-8 were induced significantly post infection and may be pivotal in the induction of IFN-ß and host innate immune responses against EV71. Among proinflammatory responses in EV71-infected intestinal epithelial cells, IL-6, CCL5, and IP10 were up-regulated and may play a key role in intestinal pathogenicity. We examined extrinsic and intrinsic apoptotic pathways and found that both were activated in EV71 infection. The mitochondria-mediated intrinsic pathway may also be activated through Bid cleaved by active caspase-8. Robust induction of IFN-ß in human intestinal epithelial cells contradicts the finding that IFN induction was suppressed in other types of the cells, suggesting that mild gastrointestinal symptoms may be the result of sufficient local antiviral inductions. Our study has demonstrated a unique way of antiviral responses in human gut different from other tissue cells in response to EV71, which may account for mild symptoms in intestinal tract. This finding will broaden our understanding of host defense mechanism and the pathogenesis of EV71 infection.

SH2-B beta expression in alveolar macrophages in BAL fluid of asthmatic guinea pigs and its role in NGF-TrkA-mediated asthma.

BACKGROUND AND OBJECTIVE: Nerve growth factor (NGF)/tyrosine kinase receptor A (TrkA) signalling may play an important role in the pathogenesis of asthma, and SH2-B beta, a TrkA-binding protein, modulates the NGF signalling pathway. In this study, SH2-B beta expression in alveolar macrophages (AM) in guinea pig BAL fluid and its role in asthma pathogenesis through the NGF-TrkA signalling pathway were investigated. METHODS: Guinea pigs were randomized into five groups: control, a model of asthma, anti-SH2-B beta antibody treatment, anti-NGF antibody treatment and anti-TrkA antibody treatment. The asthmatic model was established in guinea pigs by inhalation of ovalbumin. Specific anti-SH2-B beta, anti-NGF and anti-TrkA antibodies were administered and AM were isolated from BAL fluid to assess SH2-B beta expression using an immunofluorescence assay. SH2-B beta and TrkA protein expression were determined by western blotting, IL-1 beta and IL-4 levels in the BAL fluid supernatants were determined by ELISA, and pathological changes in the bronchi and lung tissues were examined by HE staining. RESULTS: Lymphocyte, eosinophil and total inflammatory cell numbers in BAL fluid were significantly higher in the asthma model group than in the other groups (P < 0.01). NGF expression in the asthma model group was significantly higher than that in the PBS control group (P < 0.01). SH2-B beta was expressed in AM of control animals and expression was significantly higher in the asthma model than in the other groups (P < 0.01). TrkA protein expression was significantly higher in the asthma model group than in the PBS group (P < 0.01), and treatment with anti-NGF antibody resulted in significant reduction of TrkA expression (P < 0.01). CONCLUSIONS: SH2-B beta is expressed in AM of normal guinea pigs, and SH2-B beta may participate in asthma pathogenesis through the NGF-TrkA signalling pathway.