The N-Terminal Region of Middle East Respiratory Syndrome Coronavirus Accessory Protein 8b Is Essential for Enhanced Virulence of an Attenuated Murine Coronavirus.

Middle East respiratory syndrome coronavirus (MERS-CoV) is a beta coronavirus that emerged in 2012, causing severe pneumonia and renal failure. MERS-CoV encodes five accessory proteins. Some of them have been shown to interfere with host antiviral immune response. However, the roles of protein 8b in innate immunity and viral virulence was rarely studied. Here, we introduced individual MERS-CoV accessory protein genes into the genome of an attenuated murine coronavirus (Mouse hepatitis virus, MHV), respectively, and found accessory protein 8b could enhance viral replication in vivo and in vitro and increase the lethality of infected mice. RNA-seq analysis revealed that protein 8b could significantly inhibit type I interferon production (IFN-I) and innate immune response in mice infected with MHV expressing protein 8b. We also found that MERS-CoV protein 8b could initiate from multiple internal methionine sites and at least three protein variants were identified. Residues 1-23 of protein 8b was demonstrated to be responsible for increased virulence in vivo. In addition, the inhibitory effect on IFN-I of protein 8b might not contribute to its virulence enhancement as aa1-23 deletion did not affect IFN-I production in vitro and in vivo. Next, we also found that protein 8b was localized to the endoplasmic reticulum (ER)/Golgi membrane in infected cells, which was disrupted by C-terminal region aa 88-112 deletion. This study will provide new insight into the pathogenesis of MERS-CoV infection. IMPORTANCE Multiple coronaviruses (CoV) cause severe respiratory infections and become global public health threats such as SARS-CoV, MERS-CoV, and SARS-CoV-2. Each coronavirus contains different numbers of accessory proteins which show high variability among different CoVs. Accessory proteins are demonstrated to play essential roles in pathogenesis of CoVs. MERS-CoV contains 5 accessory proteins (protein 3, 4a, 4b, 5, 8b), and deletion of all four accessory proteins (protein 3, 4a, 4b, 5), significantly affects MERS-CoV replication and pathogenesis. However, whether ORF8b also regulates MERS-CoV infection is unknown. Here, we constructed mouse hepatitis virus (MHV) recombinant virus expressing MERS-CoV protein 8b and demonstrated protein 8b could significantly enhance the virulence of MHV, which is mediated by N-terminal domain of protein 8b. This study will shed light on the understanding of pathogenesis of MERS-CoV infection.

Prognosis and management of new-onset atrial fibrillation in critically ill patients.

INTRODUCTION: The prognosis of new-onset atrial fibrillation (AF) compared with that of preexisting and non-AF remains controversial. The purpose of this study was to evaluate the effect of new-onset AF compared with preexisting and non-AF on hospital and 90-day mortality. METHODS: A retrospective cohort study was performed using data obtained from the Medical Information Mart for Intensive Care III database. The primary outcome was 90-day mortality. Secondary outcomes included hospital mortality, hospital and intensive care unit (ICU) length of stay, and acute kidney injury. Logistic and Cox regression analyses were performed to evaluate the relationship between new-onset AF and study outcomes. RESULTS: A total of 38,159 adult patients were included in the study. The incidence of new-onset AF was 9.4%. Ninety-day mortality, hospital mortality, and hospital and ICU length of stay in patients with new-onset and preexisting AF were significantly increased compared with those in patients with non-AF patients (all p < 0.001). After adjusting for patient characteristics, new-onset AF remained associated with increased 90-day mortality compared with non-AF (adjusted hazard ratio (HR) 1.37, 95% confidence interval (CI) 1.26 to 1.50; p < 0.01) and preexisting AF (adjusted HR 1.12; 95%-CI 1.02 to 1.23; p < 0.01). Patients in the surgical intensive care unit (SICU) had significantly higher 90-day mortality than patients in the coronary care unit (adjusted HR 1.30; 95% CI 1.31 to 1.51; p < 0.001). CONCLUSIONS: Critically ill patients with new-onset AF have significantly increased hospital and 90-day mortality compared with patients with preexisting and non-AF. Patients with new-onset AF in the ICU, especially those in the SICU, require robust management measures.

Clinical and Ultrasonographic Features of Papillary Thyroid Carcinoma Located in the Isthmus.

ABSTRACT: The aim of this research was to investigate the clinical and ultrasonographic features of papillary thyroid carcinoma (PTC) in the isthmus. A total of 823 patients with 823 PTCs including 133 in the isthmus and 690 in the lateral lobe were included in our study. All patients were confirmed by postoperative pathology. The clinical and ultrasonographic characteristics were retrospectively analyzed and compared. Univariate analysis and multivariate logistic regression analysis were performed. Multifactor analyses showed that PTC in the isthmus was significantly different from PTC originating from the lateral lobe in aspect ratio, microcalcification, extrathyroid extension, lymph node metastases, and lymph node density ( P < 0.05, for all). There were no significant differences in age, sex, tumor size, margin, halo, echogenicity, and homogeneity ( P > 0.05, for all). The results indicated that the sonographic appearances of PTC in the isthmus were relatively atypical; however, it had a higher incidence of extrathyroidal extension, central lymph node metastasis, and a tendency of higher lymph node density. Therefore, more careful ultrasound evaluation should be performed for these nodules.

Diagnostic value of qualitative and quantitative parameters of contrast-enhanced ultrasound for differentiating differentiated thyroid carcinomas from benign nodules.

Objective: To explore the diagnostic value of contrast-enhanced ultrasound (CEUS) of qualitative and quantitative parameters for differentiating differentiated thyroid cancers from benign nodules. Method: A total of 290 thyroid nodules that were pathologically confirmed were enrolled in this study. The univariate analysis was performed for the clinical characteristics and CEUS qualitative and quantitative parameters of the inside and peripheral zone of nodules, including age, gender, nodule size, intensity of enhancement, homogeneity, wash-in and wash-out patterns, margin after CEUS, ring enhancement, peak intensity, sharpness, time to peak(TP), and area under the curve(AUC), and the meaningful indicators in the single-factor analysis were further included in multivariate logistic regression analysis. Results: Multivariate analysis showed that there were significant differences in age (p=0.031), nodule size (p<0.001), heterogeneous enhancement (p<0.001), hypo-enhancement (p=0.001), unclear margin after CEUS(p=0.007), inside peak (p<0.001), and outside sharpness(p<0.001) between benign and malignant nodules. However, there were no significant differences in gender, ring enhancement, wash-in, wash-out, outside TP, outside AUC between benign and malignant thyroid nodules (P>0.05, for all). Conclusion: CEUS might be useful in the differential diagnosis of differentiated thyroid cancers and benign nodules, which could provide a certain basis for clinical treatment.

Characterization of aging cancer-associated fibroblasts draws implications in prognosis and immunotherapy response in low-grade gliomas.

Background: Due to the highly variable prognosis of low-grade gliomas (LGGs), it is important to find robust biomarkers for predicting clinical outcomes. Aging cancer-associated fibroblasts (CAFs) within the senescent stroma of a tumor microenvironment (TME) have been recently reported to play a key role in tumor development. However, there are few studies focusing on this topic in gliomas. Methods and Results: Based on the transcriptome data from TCGA and CGGA databases, we identified aging CAF-related genes (ACAFRGs) in LGGs by the weighted gene co-expression network analysis (WGCNA) method, followed by which LGG samples were classified into two aging CAF-related gene clusters with distinct prognosis and characteristics of the TME. Machine learning algorithms were used to screen out eight featured ACAFRGs to characterize two aging CAF-related gene clusters, and a nomogram model was constructed to predict the probability of gene cluster A for each LGG sample. Then, a powerful aging CAF scoring system was developed to predict the prognosis and response to immune checkpoint blockage therapy. Finally, the ACAFRGs were verified in two glioma-related external datasets. The performance of the aging CAF score in predicting the immunotherapy response was further validated in two independent cohorts. We also confirmed the expression of ACAFRGs at the protein level in glioma tissues through the Human Protein Atlas website and Western blotting analysis. Conclusion: We developed a robust aging CAF scoring system to predict the prognosis and immunotherapy response in LGGs. Our findings may provide new targets for therapeutics and contribute to the exploration focusing on aging CAFs.

Spontaneous coronary dissection should not be ignored in patients with chest pain in autosomal dominant polycystic kidney disease: A case report.

BACKGROUND: When autosomal dominant polycystic kidney disease (ADPKD) presents with acute coronary syndrome (ACS), the possibility of spontaneous coronary artery dissection (SCAD) should be highly considered. In some cases, SCAD is considered an extrarenal manifestation of ADPKD depending on the pathological characteristics of the unstable arterial wall in ADPKD. CASE SUMMARY: Here, we report a 46-year-old female patient with ADPKD who presented with ACS. Coronary angiography revealed no definite signs of dissection, while intravascular ultrasound revealed a proximal to distal dissection of the left circumflex. After a careful conservative medication treatment, the patient exhibited favorable prognosis. CONCLUSION: In cases of ADPKD co-existing with ACS, differential diagnosis of SCAD should be considered. Moreover, when no clear dissection is found on coronary angiography, IVUS should be performed to prevent missed diagnosis.

A predictive nomogram of bleeding risk in patients with atrial fibrillation after drug-eluting stent implantation.

BACKGROUND: The use of anticoagulants and antiplatelet therapies is associated with a higher risk of bleeding in atrial fibrillation (AF) patients after percutaneous coronary intervention, especially after stent implantation. However, no accurate bleeding risk prediction tool has been developed for these patients. The aim of this study was thus to establish a bleeding risk prediction model (predictive nomogram) for patients with AF after stent implantation. METHODS: Construction of the predictive nomogram was based on a retrospective study, which enrolled 943 AF patients who underwent drug-eluting stent implantation between May 2012 and September 2016. A range of factors, including demographics, comorbidities, medication strategies, arterial access, and laboratory tests, were collected as baseline data. The least absolute shrinkage and selection operator (LASSO) and multivariate logistic regression analysis were used to identify the key clinical features for construction of the predictive nomogram. The concordance index (C-index) and internal validation were used to evaluate the efficacy of the nomogram. RESULTS: Of the 943 AF patients that underwent stent implantation, the occurrence of bleeding events was 8.2% (77 out of 943). Key predictors included the number of antiplatelet drugs, peptic ulcer, cerebral infarction, type 2 diabetes, thrombocytopenia, anemia, prior myocardial infarction, sex (male), use of anticoagulant drugs, liver dysfunction, hypertension, and acute myocardial infarction. These predictors were used to construct the nomogram. The C-index for the prediction of bleeding risk by the nomogram was 0.841 (95% CI: 0.79-0.89), which indicated good discrimination and calibration. The C-index of internal validation was 0.795, which demonstrated good efficacy of the model. CONCLUSIONS: This study suggests that our novel nomogram can accurately predict bleeding risk in AF patients after stent implantation during hospitalization, thereby helping to avoid complications. The nomogram may also be helpful for the creation of individualized post-discharge medication strategies.

Multi-platform omics analysis reveals molecular signature for COVID-19 pathogenesis, prognosis and drug target discovery.

Disease progression prediction and therapeutic drug target discovery for Coronavirus disease 2019 (COVID-19) are particularly important, as there is still no effective strategy for severe COVID-19 patient treatment. Herein, we performed multi-platform omics analysis of serial plasma and urine samples collected from patients during the course of COVID-19. Integrative analyses of these omics data revealed several potential therapeutic targets, such as ANXA1 and CLEC3B. Molecular changes in plasma indicated dysregulation of macrophage and suppression of T cell functions in severe patients compared to those in non-severe patients. Further, we chose 25 important molecular signatures as potential biomarkers for the prediction of disease severity. The prediction power was validated using corresponding urine samples and plasma samples from new COVID-19 patient cohort, with AUC reached to 0.904 and 0.988, respectively. In conclusion, our omics data proposed not only potential therapeutic targets, but also biomarkers for understanding the pathogenesis of severe COVID-19.

Maximum blood glucose levels during hospitalisation to predict mortality in patients with acute coronary syndrome: a retrospective cohort study.

OBJECTIVE: The aim in this study was to stratify maximum blood glucose levels to identify the the best cut-off value of glucose levels to predict mortality in acute coronary syndrome (ACS) patients, regardless of whether they had diabetes. DESIGN: A retrospective cohort study. SETTING: All clinical data were obtained from the 'Medical Information Mart for Intensive Care III' database. PARTICIPANTS: A total of 3078 patients with ACS were included in the study. We divided the patients into four levels based on their maximum blood glucose levels (glucosemax), then analysed the relationship between each group with mortality. RESULTS: Among enrolled patients, 2780 and 298 were survivors and non-survivors, respectively. Blood glucose levels and mortality showed a 'tick' type relationship, with levels 3 and 4 found to be closely associated with increased hospital mortality (p<0.05), relative to level 1 (<6.1 mmol/L), used as the reference group. No significant association was observed in mortality between level 2 and level 1 (p=0.095). In addition, we found a gradual increase in OR for level 2 (OR: 2.42, 95% CI 0.86 to 6.80, p=0.095), level 3 (OR: 4.33, 95% CI 1.55 to 12.13, p=0.005) and level 4 (OR: 7.27, 95% CI 2.56 to 20.62, p<0.001), relative to level 1. Based on receiver operating characteristic curves, the optimal cut-off value for predicting mortality were 11.5 (area under curve (AUC)=0.724), 11.2 (AUC=0.729), 13.4 (AUC=0.638), 15.8 (AUC=0.717) and 11.3 mmol/L (AUC=0.764) in all ACS, acute myocardial infarction, unstable angina, diabetes and non-diabetes patients, respectively. The results of subgroup analysis suggested that in patients with significantly elevated blood glucose, the mortality of non-diabetes was higher than patients with diabetes (OR: 0.42, 95% CI 0.31 to 0.57, p<0.001). CONCLUSION: Overall, glucosemax ≥11.5 mmol/L had a significant association with increased mortality in patients with ACS. Non-diabetes ACS patients need a more robust blood glucose management strategy compared with diabetes counterparts.

Kinetics of viral load and antibody response in relation to COVID-19 severity.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent for coronavirus 2019 (COVID-19) pneumonia. Little is known about the kinetics, tissue distribution, cross-reactivity, and neutralization antibody response in patients with COVID-19. Two groups of patients with RT-PCR-confirmed COVID-19 were enrolled in this study: 12 severely ill patients in intensive care units who needed mechanical ventilation and 11 mildly ill patients in isolation wards. Serial clinical samples were collected for laboratory detection. Results showed that most of the severely ill patients had viral shedding in a variety of tissues for 20-40 days after onset of disease (8/12, 66.7%), while the majority of mildly ill patients had viral shedding restricted to the respiratory tract and had no detectable virus RNA 10 days after onset (9/11, 81.8%). Mildly ill patients showed significantly lower IgM response compared with that of the severe group. IgG responses were detected in most patients in both the severe and mild groups at 9 days after onset, and remained at a high level throughout the study. Antibodies cross-reactive to SARS-CoV and SARS-CoV-2 were detected in patients with COVID-19 but not in patients with MERS. High levels of neutralizing antibodies were induced after about 10 days after onset in both severely and mildly ill patients which were higher in the severe group. SARS-CoV-2 pseudotype neutralization test and focus reduction neutralization test with authentic virus showed consistent results. Sera from patients with COVID-19 inhibited SARS-CoV-2 entry. Sera from convalescent patients with SARS or Middle East respiratory syndrome (MERS) did not. Anti-SARS-CoV-2 S and N IgG levels exhibited a moderate correlation with neutralization titers in patients' plasma. This study improves our understanding of immune response in humans after SARS-CoV-2 infection.

Self-Assembled Injectable Nanocomposite Hydrogels Coordinated by in Situ Generated CaP Nanoparticles for Bone Regeneration.

Due to the great similarity to the natural extracellular matrix and minimally invasive surgeries, injectable hydrogels are appealing biomaterials in cartilage and bone tissue engineering. Nevertheless, undesirable mechanical properties and bioactivity greatly hamper their availability in clinic applications. Here, we developed an injectable nanocomposite hydrogel by in situ growth of CaP nanoparticles (ICPNs) during the free-radical polymerization of dimethylaminoethyl methacrylate (DMAEMA) and 2-hydroxyethyl methacrylate (HEMA) matrix (PDH) for bone regeneration. The ICPNs are self-assembled by incorporation of poly-l-glutamic acid (PGA) with abundant carboxyl functional groups during the formation of carboxyl-Ca2+ coordination and further CaP precipitation. Furthermore, the carboxyl groups of PGA could interact with the tertiary amines of DMAEMA fragments and thus improve the mechanical strength of hydrogels. Upon mixing solutions of DMAEMA and HEMA bearing PGA, Ca2+, and PO43-, this effective and dynamic coordination led to the rapid self-assembly of CaP NPs and PDH nanocomposite hydrogels (PDH/mICPN). The obtained optimal nanocomposite hydrogels exhibited suitable injectable time, an enhanced tensile strength of 321.1 kPa, and a fracture energy of 29.0 kJ/m2 and dramatically facilitated cell adhesion and upregulated osteodifferentiation compared to hydrogels prepared by blending ex situ prefabricated CaP NPs. In vivo experiments confirmed the promoted osteogenesis, which shows a striking contrast to pure PDH hydrogels. Additionally, the methacrylate groups on the monomers could easily be functionalized with aptamers and thereby facilitate recognition and capturing of bone marrow stromal cells both in vitro and in vivo and strengthen the bone regeneration. We believe that our conducted research about in situ self-assembled CaP nanoparticle-coordinated hydrogels will open a new avenue for bone regeneration in the future endeavors.