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Abstract Background Syndrome coronavirus-2 (SARS-CoV-2) has developed various strategies to evade the antiviral impact of type I IFN. Non-structural proteins and auxiliary proteins have been extensively researched on their role in immune escape. Nevertheless, the detailed mechanisms of structural protein-induced immune evasion have not been well elucidated. Methods Human alveolar basal epithelial carcinoma cell line (A549) was stimulated with polyinosinic-polycytidylic acid (PIC) and independently transfected with four structural proteins expression plasmids, including nucleocapsid (N), spike (S), membrane (M) and envelope (E) proteins. By RT-qPCR and ELISA, the structural protein with the most pronounced inhibitory effects on IFN-β induction was screened. RNA-sequencing (RNA-Seq) and two differential analysis strategies were used to obtain differentially expressed genes associated with N protein inhibition of IFN-β induction. Based on DIANA-LncBase and StarBase databases, the interactive competitive endogenous RNA (ceRNA) network for N protein-associated genes was constructed. By combining single-cell sequencing data (GSE158055), lncRNA-miRNA-mRNA axis was further determined. Finally, RT-qPCR was utilized to illustrate the regulatory functions among components of the ceRNA axis. Results SARS-CoV-2 N protein inhibited IFN-β induction in human alveolar epithelial cells most significantly compared with other structural proteins. RNA-Seq data analysis revealed genes related to N protein inhibiting IFNs induction. The obtained 858 differentially expressed genes formed the reliable ceRNA network. The function of LINC01002-miR-4324-FRMD8 axis in the IFN-dominated immune evasion was further demonstrated through integrating single-cell sequencing data. Moreover, we validated that N protein could reverse the effect of PIC on LINC01002, FRMD8 and miR-4324 expression, and subsequently on IFN-β expression level. And LINC01002 could regulate the production of FRMD8 by inhibiting miR-4324. Conclusion SARS-CoV-2 N protein suppressed the induction of IFN-β by regulating LINC01002 which was as a ceRNA, sponging miR-4324 and participating in the regulation of FRMD8 mRNA. Our discovery provides new insights into early intervention therapy and drug development on SARS-CoV-2 infection.

Abstract Background Beyond the observed alterations in cellular structure and mitochondria, the mechanisms linking rare genetic mutations to the development of heart failure in patients affected by desmin mutations remain unclear due in part, to the lack of relevant human cardiomyocyte models. Methods To shed light on the role of mitochondria in these mechanisms, we investigated cardiomyocytes derived from human induced pluripotent stem cells carrying the heterozygous DES E439K mutation that were either isolated from a patient or generated by gene editing. To increase physiological relevance, cardiomyocytes were either cultured on an anisotropic micropatterned surface to obtain elongated and aligned cardiomyocytes, or as a cardiac spheroid to create a micro-tissue. Moreover, when applicable, results from cardiomyocytes were confirmed with heart biopsies of suddenly died patient of the same family harboring DES E439K mutation, and post-mortem heart samples from five control healthy donors. Results The heterozygous DES E439K mutation leads to dramatic changes in the overall cytoarchitecture of cardiomyocytes, including cell size and morphology. Most importantly, mutant cardiomyocytes display altered mitochondrial architecture, mitochondrial respiratory capacity and metabolic activity reminiscent of defects observed in patient’s heart tissue. Finally, to challenge the pathological mechanism, we transferred normal mitochondria inside the mutant cardiomyocytes and demonstrated that this treatment was able to restore mitochondrial and contractile functions of cardiomyocytes. Conclusions This work highlights the deleterious effects of DES E439K mutation, demonstrates the crucial role of mitochondrial abnormalities in the pathophysiology of desmin-related cardiomyopathy, and opens up new potential therapeutic perspectives for this disease.

Abstract Background As a histone methyltransferase, suppressor of variegation 3–9 homolog 1 (SUV39H1) plays an important role in the occurrence and development of cancer. To explore the mechanism and biological function of SUV39H1 in hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC) can gain an insight into the pathogenesis of HBV-HCC. Methods The effect of HBV infection on SUV39H1 in hepatoma cells was detected. CCK-8, colony growth assay and wound healing assay were used to assess the proliferation and migration of HBV-positive hepatoma cells. RNA sequencing (RNA-seq) was applied to find differential genes and enriched pathways. The serum SUV39H1 level in HBV-HCC patients was detected and its correlation with clinical indicators was analyzed. Results SUV39H1 was increased by HBV infection and promoted the proliferation and migration of hepatoma cells. SUV39H1 could upregulate the expression of mitochondrial oxidative phosphorylation (OXPHOS) pathway-related genes. OXPHOS pathway inhibitors could reduce the capacity of proliferation and migration of hepatoma cells after overexpressing SUV39H1. Serum SUV39H1 levels were higher in chronic hepatitis B (CHB) patients than in healthy controls and higher in HBV-HCC patients than in CHB patients. In the diagnosis of HCC, the predictive value of SUV39H1 combined with alpha-fetoprotein (AFP) was better than that of AFP alone. Conclusion SUV39H1 is regulated by HBV infection and promotes the proliferation and migration of hepatoma cells by targeting OXPHOS pathway. It indicates that SUV39H1 may be a new biomarker of the diagnosis of HCC.

Abstract Background Large-scale medical equipment, which is extensively implemented in medical services, is of vital importance for diagnosis but vulnerable to various anomalies and failures. Most hospitals that conduct regular maintenance have been suffering from medical equipment-related incidents for years. Currently, the Internet of Medical Things (IoMT) has emerged as a crucial tool in monitoring the real-time status of the medical equipment. In this paper, we develop an IoMT system of Computed Tomography (CT) equipment in the West China Hospital, Sichuan University and collected the system status time-series data. Novel multivariate time-series classification models and frameworks are proposed to predict the anomalies of CT equipment. The important features that are closely related to the equipment anomalies are identified with the model. Methods We extracted the real-time CT equipment status time-series data of 11 equipment between May 19, 2020 and May 19, 2021 from the IoMT, which includes the equipment oil temperature, anode voltage, etc. The arcs are identified as labels of anomalies due to their relationship with decreased imaging quality and CT equipment failures. To improve prediction accuracy, the statistics and transformations of the raw historical time-series data segment in the sliding time window are used to construct new features. Due to the particularity of time-series data, two frameworks are proposed for splitting the training and test sets. Then the Decision Tree, Support Vector Machine, Logistic Regression, Naive Bayesian, and K-Nearest Neighbor classification models are used to classify the system status. We also compare our model to state-of-the-art models. Results The results show that the anomaly prediction accuracy and recall of our method are 79% and 77%, respectively. The oil temperature and anode voltage are identified as the decisive features that may lead to anomalies. The proposed model outperforms the others when predicting the anomalies of the CT equipment based on our dataset. Conclusions The proposed method could predict the state of CT equipment and be used as a reference for practical maintenance, where unexpected anomalies of medical equipment could be reduced. It also brings new insights into how to handle non-uniform and imbalanced time series data in practical cases.

Abstract Objective This study aimed to assess the feasibility of software-aided selection of monoenergetic level for acute necrotising pancreatitis (ANP) depiction compared to other automatic image series generated using dual-energy computed tomography (CT). Methods The contrast-enhanced dual-source dual-energy CT images in the portal venous phase of 48 patients with ANP were retrospectively analysed. Contrast-to-noise ratio (CNR) of pancreatic parenchyma-to-necrosis, signal-to-noise ratio (SNR) of the pancreas, image noise, and score of subjective diagnosis were measured, calculated, and compared among the CT images of 100 kV, Sn140 kV, weighted-average 120 kV, and optimal single-energy level for CNR. Results CNR of pancreatic parenchyma-to-necrosis in the images of 100 kV, Sn140 kV, weighted-average 120 kV, and the optimal single-energy level for CNR was 5.18 ± 2.39, 3.13 ± 1.35, 5.69 ± 2.35, and 9.99 ± 5.86, respectively; SNR of the pancreas in each group was 6.31 ± 2.77, 4.27 ± 1.56, 7.21 ± 2.69, and 11.83 ± 6.30, respectively; image noise in each group was 18.78 ± 5.20, 17.79 ± 4.63, 13.28 ± 3.13, and 9.31 ± 2.96, respectively; and score of subjective diagnosis in each group was 3.56 ± 0.50, 3.00 ± 0.55, 3.48 ± 0.55, and 3.88 ± 0.33, respectively. The four measurements of the optimal single-energy level for CNR images were significantly different from those of images in the other three groups (P

Abstract Background High-resolution computed tomography (HRCT) is recommended diagnosing and monitoring connective tissue disease-associated interstitial lung disease (CTD-ILD). Quantitative computed tomography has the potential to precisely assess the radiological severity of CTD-ILD, but has still been under study. Objective To investigate whether dual-energy computed tomography (DECT), a novel quantitative technique, can be used for quantitative severity assessment in CTD-ILD. Methods This cross sectional study recruited adult CTD-ILD patients who underwent DECT scans from the ICE study between October 2019 and November 2021. DECT parameters, including effective atomic number (Zeff), lung (lobe) volume, and monochromatic CT number (MCTN) of each lung lobe, were evaluated. CTD-ILD was classified into extensive CTD-ILD and limited CTD-ILD by staging algorithm using combined forced vital capacity (FVC)%predicted and total extent of ILD (TEI) on CT. Dyspnea, cough, and life quality were scored by Borg dyspnea score, Leicester cough questionnaire (LCQ), and short-form 36 health survey questionnaire (SF-36), respectively. Results There was a total of 147 patients with DECT scans enrolled. Higher Zeff value (3.104 vs 2.256, p

Abstract Background Silent information regulator 6 (SIRT6) is a mammalian homolog of the nicotinamide adenine dinucleotide (NAD)-dependent deacetylase sirtuin family. Prior evidences suggested that the anti-inflammatory function of SIRT6 after spinal cord and brain injury, and it plays a crucial role in macrophages polarization of adipose tissue and skin. However, the role of SIRT6 in macrophages involved peripheral nerve injury is still unknown. Given the prominent role of macrophages in peripheral nerve recovery, we aim to investigate the role of SIRT6 in the regulation of phenotypes shift and functions in macrophages after peripheral nerve injury. Results In the present study, we first identified a significant increase of SIRT6 expression during nerve degeneration and macrophages phagocytosis. Next, we found nerve recovery was delayed after SIRT6 silencing by injected shRNA lentivirus into the crushed sciatic nerve, which exhibited a reduced expression of myelin-related proteins (e.g., MAG and MBP), severer myoatrophy of target muscles, and inferior nerve conduction compared to the shRNA control injected mice. In vitro, we found that SIRT6 inhibition by being treated with a selective inhibitor OSS_128167 or lentivirus transfection impairs migration and phagocytosis capacity of bone marrow-derived macrophages (BMDM). In addition, SIRT6 expression was discovered to be reduced after M1 polarization, but SIRT6 was enhanced after M2 polarization in the monocyte-macrophage cell line RAW264.7 and BMDM. Moreover, SIRT6 inhibition increased M1 macrophage polarization with a concomitant decrease in M2 polarization both in RAW264.7 and BMDM via activating NF-κB and TNF-α expression, and SIRT6 activation by UBCS039 treatment could shift the macrophages from M1 to M2 phenotype. Conclusion Our findings indicate that SIRT6 inhibition impairs peripheral nerve repair through suppressing the migration, phagocytosis, and M2 polarization of macrophages. Therefore, SIRT6 may become a favorable therapeutic target for peripheral nerve injury.

It has been demonstrated that arterial stiffness is linked to arterial fibrosis manifested by increased collagen and other extracellular matrix synthesis in smooth muscle cells (SMC). Transmembrane receptors integrins mediating cell-cell and cell-matrix signalling pathways are involved in tissue fibrosis. We study the role of one integrin subunit av in angiotensin II (AngII)-induced SMC proliferation and arterial fibrosis and stiffness using SMC specific knock-out av mouse model (avSMKO) induced in adult mice by injection of tamoxifen. There is no difference in vascular fibrosis in basal conditions between control and mutant mice. However, decreased arterial fibrosis is observed in avSMKO mutant mice after 28-day perfusion of AngII. Analysis of RNA from aorta of control and mutant mice by Affymetrix microarrays indicated an alteration of the TGF-β pathway in AngII-treated mutant mice. In order to examine the mechanism associated to the decreased fibrosis in vascular SMC from avSMKO mice, isolated VSMC from the aorta of control and avSMKO mice were treated with TGF-β1 or AngII. The results indicated that these two treatments increased the expression of collagen, fibronectin and integrin αν at the protein and RNA levels as well as the phosphorylation of ERK and smad2/3 in the control cells. Inactivation of integrin αν partly inhibited all above effects induced by TGF-β1 and AngII. Our study indicates a role of av and TGF-β1 in the arterial fibrogenesis. Therefore, av signaling could be a therapeutic target against arterial fibrosis and stiffness in pathological conditions.

Integrin αv functions as a receptor for adhesion proteins and is expressed at high density in vascular smooth muscle cells (VSMC)1,2,3,4,5 whose phenotypic modulation plays a crucial role in arterial ageing and atherosclerosis.6,7. Our aim was to define the arterial phenotype in mice conditionally inactivated for the integrin αv subunit in VSMC8,9,10(αv SMKO) and its role in angiotensin II (AngII)-induced arterial fibrosis. Transgenic mice αv SMKO and their control littermates (WT) were treated with two doses of AngII, low (0.3 mg/kg/day) and high (1.5 mg/kg/day), for 4 weeks. At baseline, blood pressure was lower in αvSMKO compared to WT mice. Carotid distensibility was increased in αv SMKO mice (13.3±0.7 vs 10.3±0.6 mmHg−1.10−3). With low dose AngII isobaric distensibility remained higher in αvSMKOmice (12.4±1.2 vs 10.7±1.0 mmHg−1.10−3).With high dose AngII the increase in collagen content in carotid media was lower in αvSMKOthan in WT (19 vs 35%) for a similar increase in blood pressure (30 mmHg) and arterial wall hypertrophy. Collagen immunostaining and fluorescence measurements (multiphoton microscopy second harmonic generation) confirmed that high dose AngII induced lower increases in collagen content in αvSMKO mice versus WT (8.9±1.7 vs 14.2±1.4 greyscale mean/pixel). The combination of similar arterial wall hypertrophy with less fibrosis in mutant mice explains an increased distensibility in response to AngII. The αv subunit regulates AngII-induced arterial fibrosis as determined by collagen staining, immunostaining and fluorescence. Pharmacological targeting of vascular αv integrin may have clinical applications in the treatment of patients with fibrosis associated with hypertension and atherosclerosis.

Integrin avb3 is expressed at high density in vascular smooth muscle cells (VSMCs). It functions as a receptor for adhesion proteins in VSMCs which phenotypic modulation plays a pivotal role in arteriosclerosis and atherosclerosis. The aim was to study the role of integrin avb3 in angiotensin II (Ang II)-induced arterial fibrosis in mice and in human samples of atherosclerotic arteries in situ. Transgenic mice conditionally inactivated for integrin av subunit in VSMCs (avSMKO) were treated with Ang II (1,5 mg/kg/day) for 4 weeks. Immunostained slices of atherosclerotic plaques at different stages of development and primary cultures of human aortic VSMCs were used. At baseline, blood pressure was lower in avSMKO compared to control (WT) mice. Isobaric carotid distensibility was increased and remained higher in avSMKO in response to Ang II. The increase in collagen content in response to Ang II was lower in avSMKO than in WT (15 vs 36%) for similar increase in blood pressure (20 mmHg) and arterial wall hypertrophy. The immunohistochemistry of aortic slices showed stronger staining for integrin avb3 in atherosclerotic plaques compared to healthy aortas. In VSMC cultures, the mRNA of av was decreased. In conclusion, these results show that avb3 is strongly expressed in neointimal proliferation and in fibrous plaques. The av integrin subunit seems to regulate arterial fibrosis in response to hypertension and plaque growth. Low RNA quantities of av subunit of VSMCs contrasted with strong protein staining in plaques suggesting the participation of inflammatory cells in the synthesis of this integrin.

Abstract Purpose Products enriched with probiotics have always been fashionable. Weizmannia coagulans has become a hot research topic in the academic community due to their multiple functional properties and high resistance to stress, which can retain their activity in a variety of harsh environments. This review aims to evaluate the probiotic effects of different strains of Weizmannia coagulans in animals and humans and to inspire better exploitation of the value of this strain. Methods This review summarizes the latest research progress of Weizmannia coagulans from two major applications in animal breeding and human health. Results The functional properties of Weizmannia coagulans are extensively recognized. In animals, the strain can promote nutrient absorption, reduce mortality, and enhance the slaughter rate in livestock and poultry. In humans, the strain can be used to treat gastrointestinal disorders, immunomodulation, depressive symptoms, and non-alcoholic fatty liver. Weizmannia coagulans is projected as an ideal substitute for antibiotics and other chemical drugs. Conclusion Despite the outstanding functional properties of Weizmannia coagulans, there are numerous strains of Weizmannia coagulans and significant differences between strains in functional and physiological properties. Currently, there are few literature reports on the probiotic mechanism and functional gene identification of Weizmannia coagulans, which is crucial for the commercialization of Weizmannia coagulans and the benefit of human society.

Abstract Background We undertook a comparative biomechanical study of type B1 fractures around femoral prostheses following cemented hip arthroplasty using the Ortho-Bridge System (OBS) and a locking compression plate/locking attachment plate structure (LCP + LAP). We aimed to investigate the biomechanical characteristics and advantages of the OBS compared with LCP + LAP when treating this fracture type. Methods An OBS fixation model was designed based on OBS and LCP + LAP fixation characteristics. The LCP + LAP combination (Group A) and three different OBS combinations (Groups B, C, and D) were used to fix a B1 fracture model with a femoral periprosthetic fracture. Axial compression and torsion experiments were then performed using simple and comminuted fracture models. The axial compression failure experiment was carried out, and the model stiffness during axial compression, torsion angle in torsion test, and vertical load in the final failure test were collected. Results When simulating simple oblique fractures, no significant difference was found among the four groups in terms of stiffness in the axial compression experiment (P = 0.257). The torsion angle of the LCP + LAP system was significantly higher compared with the OBS system (P