Pathogenicity and transcriptomic profiling reveals immunology molecular hallmarks after CA10 virus infection.

BACKGROUND: Hand, foot and mouth disease (HFMD) is a common infectious disease caused by viral infection by a variety of enteroviruses, with coxsackievirus A 10 (CA10) having become more prevalent in recent years. METHODS: In this study, models of CA10 infection were established in 7-day-old Institute of Cancer Research (ICR) mice by intraperitoneal injection to analyze the pathogenicity of the virus. RNA sequencing analysis was used to screen the differentially expressed genes (DEGs) after CA10 infection. Coxsackievirus A 16 (CA16) and enterovirus 71 (EV71) infections were also compared with CA10. RESULTS: After CA10 virus infection, the mice showed paralysis of the hind limbs at 3 days post infection and weight loss at 5 days post infection. We observed viral replication in various tissues and severe inflammatory cell infiltration in skeletal muscle. The RNA-sequencing analysis showed that the DEGs in blood, muscle, thymus and spleen showed heterogeneity after CA10 infection and the most up-regulated DEGs in muscle were enriched in immune-related pathways. Compared with CA16 and EV71 infection, CA10 may have an inhibitory effect on T helper (Th) cell differentiation and cell growth. Additionally, the common DEGs in the three viruses were most enriched in the immune system response, including the Toll-like receptor pathway and the nucleotide-binding and oligomerization domain (NOD)-like pathway. CONCLUSIONS: Our findings revealed a group of genes that coordinate in response to CA10 infection, which increases our understanding of the pathological mechanism of HFMD.

Cepharanthine analogs mining and genomes of Stephania accelerate anti-coronavirus drug discovery.

Cepharanthine is a secondary metabolite isolated from Stephania. It has been reported that it has anti-conronaviruses activities including severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Here, we assemble three Stephania genomes (S. japonica, S. yunnanensis, and S. cepharantha), propose the cepharanthine biosynthetic pathway, and assess the antiviral potential of compounds involved in the pathway. Among the three genomes, S. japonica has a near telomere-to-telomere assembly with one remaining gap, and S. cepharantha and S. yunnanensis have chromosome-level assemblies. Following by biosynthetic gene mining and metabolomics analysis, we identify seven cepharanthine analogs that have broad-spectrum anti-coronavirus activities, including SARS-CoV-2, Guangxi pangolin-CoV (GX_P2V), swine acute diarrhoea syndrome coronavirus (SADS-CoV), and porcine epidemic diarrhea virus (PEDV). We also show that two other genera, Nelumbo and Thalictrum, can produce cepharanthine analogs, and thus have the potential for antiviral compound discovery. Results generated from this study could accelerate broad-spectrum anti-coronavirus drug discovery.

Pathogenicity and landscape of differential gene expression in mice orally infected with clinical coxsackievirus A6 (CA6).

Hand, foot, and mouth disease (HFMD) is caused by more than 20 pathogenic enteroviruses belonging to the Picornaviridae family and Enterovirus genus. Since the introduction of the enterovirus-71 (EV71) vaccine in 2016, the number of HFMD cases caused by EV71 has decreased. However, cases of infections caused by other enteroviruses, such as coxsackievirus A6 (CA6) and coxsackievirus A10, have been increasing accordingly. In this study, we used a clinical isolate of CA6 to establish an intragastric infection mouse model using 7-day-old mice to mimic the natural transmission route, by which we investigated the differential gene expression profiles associated with virus infection and pathogenicity. After intragastric infection, mice exhibited hind limb paralysis symptoms and weight loss, similar to those reported for EV71 infection in mice. The skeletal muscle was identified as the main site of virus replication, with a peak viral load reaching 2.31 × 107 copies/mg at 5 dpi and increased infiltration of inflammatory cells. RNA sequencing analysis identified differentially expressed genes (DEGs) after CA6 infection. DEGs in the blood, muscle, brain, spleen, and thymus were predominantly enriched in immune system responses, including pathways such as Toll-like receptor signaling and PI3K-Akt signaling. Our study has unveiled the genes involved in the host immune response during CA6 infection, thereby enhancing our comprehension of the pathological mechanism of HFMD.IMPORTANCEThis study holds great significance for the field of hand, foot, and mouth disease (HFMD). It not only delves into the disease's etiology, transmission pathways, and severe complications but also establishes a novel mouse model that mimics the natural coxsackievirus A6 infection process, providing a pivotal platform to delve deeper into virus replication and pathogenic mechanisms. Additionally, utilizing RNA-seq technology, it unveils the dynamic gene expression changes during infection, offering valuable leads for identifying novel therapeutic drug targets. This research has the potential to enhance our understanding of HFMD, offering fresh perspectives for disease prevention and treatment and positively impacting children's health worldwide.

Global analysis of lysine acetylation in the brain cortex of K18-hACE2 mice infected with SARS-CoV-2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected hundreds of millions of people all over the world and thus threatens human life. Clinical evidence shows that SARS-CoV-2 infection can cause several neurological consequences, but the existing antiviral drugs and vaccines have failed to stop its spread. Therefore, an understanding of the response to SARS-CoV-2 infection of hosts is vital to find a resultful therapy. Here, we employed a K18-hACE2 mouse infection model and LC-MS/MS to systematically evaluate the acetylomes of brain cortexes in the presence and absence of SARS-CoV-2 infection. Using a label-free strategy, 3829 lysine acetylation (Kac) sites in 1735 histone and nonhistone proteins were identified. Bioinformatics analyses indicated that SARS-CoV-2 infection might lead to neurological consequences via acetylation or deacetylation of important proteins. According to a previous study, we found 26 SARS-CoV-2 proteins interacted with 61 differentially expressed acetylated proteins with high confidence and identified one acetylated SARS-CoV-2 protein nucleocapsid phosphoprotein. We greatly expanded the known set of acetylated proteins and provide the first report of the brain cortex acetylome in this model and thus a theoretical basis for future research on the pathological mechanisms and therapies of neurological consequences after SARS-CoV-2 infection.

Proteomic and phosphoproteomic characteristics of the cortex, hippocampus, thalamus, lung, and kidney in COVID-19-infected female K18-hACE2 mice.

BACKGROUND: Neurological damage caused by coronavirus disease 2019 (COVID-19) has attracted increasing attention. Recently, through autopsies of patients with COVID-19, the direct identification of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in their central nervous system (CNS) has been reported, indicating that SARS-CoV-2 might directly attack the CNS. The need to prevent COVID-19-induced severe injuries and potential sequelae is urgent, requiring the elucidation of large-scale molecular mechanisms in vivo. METHODS: In this study, we performed liquid chromatography-mass spectrometry-based proteomic and phosphoproteomic analyses of the cortex, hippocampus, thalamus, lungs, and kidneys of SARS-CoV-2-infected K18-hACE2 female mice. We then performed comprehensive bioinformatic analyses, including differential analyses, functional enrichment, and kinase prediction, to identify key molecules involved in COVID-19. FINDINGS: We found that the cortex had higher viral loads than did the lungs, and the kidneys did not have SARS-COV-2. After SARS-CoV-2 infection, RIG-I-associated virus recognition, antigen processing and presentation, and complement and coagulation cascades were activated to different degrees in all five organs, especially the lungs. The infected cortex exhibited disorders of multiple organelles and biological processes, including dysregulated spliceosome, ribosome, peroxisome, proteasome, endosome, and mitochondrial oxidative respiratory chain. The hippocampus and thalamus had fewer disorders than did the cortex; however, hyperphosphorylation of Mapt/Tau, which may contribute to neurodegenerative diseases, such as Alzheimer's disease, was found in all three brain regions. Moreover, SARS-CoV-2-induced elevation of human angiotensin-converting enzyme 2 (hACE2) was observed in the lungs and kidneys, but not in the three brain regions. Although the virus was not detected, the kidneys expressed high levels of hACE2 and exhibited obvious functional dysregulation after infection. This indicates that SARS-CoV-2 can cause tissue infections or damage via complicated routes. Thus, the treatment of COVID-19 requires a multipronged approach. INTERPRETATION: This study provides observations and in vivo datasets for COVID-19-associated proteomic and phosphoproteomic alterations in multiple organs, especially cerebral tissues, of K18-hACE2 mice. In mature drug databases, the differentially expressed proteins and predicted kinases in this study can be used as baits to identify candidate therapeutic drugs for COVID-19. This study can serve as a solid resource for the scientific community. The data in this manuscript will serve as a starting point for future research on COVID-19-associated encephalopathy. FUNDING: This study was supported by grants from the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences, the National Natural Science Foundation of China, and the Natural Science Foundation of Beijing.

Ultrahigh performance heat spreader based on gas-liquid exfoliation boron nitride nanosheets.

Due to their high thermal conductivity and insulation performance, boron nitride nanosheets (BNNS) have great promise to fabricate thermal management equipment for application in power electronics. The liquid-phase exfoliation route has been regarded as the most commonly used approach to produce single and few-layered BNNS for many research fields. However, this process takes a long time, and the production yield is extremely low. In this work, an efficient technique to obtain few-layered (mostly < 5 layers), high-yield (∼33%), and plane-defect-free BNNS by the combination of liquid N2 (L-N2) gasification and liquid exfoliation was developed. The as-obtained BNNS suspensions could be vacuum filtered to make a thermal conductive film named a BNNS heat spreader which possessed a superior thermal conductivity of 61.2 W m-1 K-1 at room temperature. In addition, we also proved that the thermal conductivity of the BNNS heat spreader increased with the increase of density, creating an approach for fine tuning the thermal property of this heat spreader.

Hydroxylated graphene-based flexible carbon film with ultrahigh electrical and thermal conductivity.

Graphene-based films are widely used in the electronics industry. Here, surface hydroxylated graphene sheets (HGS) have been synthesized from natural graphite (NG) by a rapid and efficient molten hydroxide-assisted exfoliation technique. This method enables preparation of aqueous dispersible graphene sheets with a high dispersed concentration (∼10.0 mg ml-1) and an extraordinary production yield (∼100%). The HGS dispersion was processed into graphene flexible film (HGCF) through fast filtration, annealing treatment and mechanical compression. The HGS endows graphene flexible film with a high electrical conductivity of 11.5 × 104 S m-1 and a superior thermal conductivity of 1842 W m-1 K-1. Simultaneously, the superflexible HGCF could endure 3000 repeated cycles of bending or folding. As a result, this graphene flexible film is expected to be integrated into electronic packaging and high-power electronics applications.

Coxsackievirus A16 infection stimulates imbalances of T cells in children.

Immune reaction plays a crucial role in the regulation of the progression of Coxsackievirus A16 (CA16)-infected hand, foot and mouth disease (HFMD). However, no details of T-cell subset frequency or imbalance during the CA16 infection process have been revealed. In the present study, whether CA16-induced HFMD changes the frequency of different T-cell subsets and associated immune mediators was determined in children. The results indicate that the percentages of Th1 and Tc1 cells were significantly increased in children with HFMD compared with those in healthy children. In addition, the Th1/Th2 ratio and interferon (IFN)-γ levels were significant higher in children with HFMD. Furthermore, the percentage of Th17 cells and the Th17/Treg ratio as well as interleukin (IL)-17A levels were higher in HFMD cases. In conclusion, the present study demonstrated the dysregulation of T-cell subsets following CA16 infection. The Th1/Th2 and Th17/Treg ratios were imbalanced following infection. Also, the imbalance Th1/Th2 and Th17/Treg ratios contributed to the increased levels of IFN-γ and IL-17A. Based on this information, the present study provides new insights for the future study of CA16-induced HFMD and offers new data of diagnostic and therapeutic value for CA16 infection.

[Cancer of the nasal cavity].

OBJECTIVE: To analyze the factors affecting prognosis of patients with nasal carcinoma. METHODS: 163 patients treated from 1985 to 1998 were analyzed. The survival analysis was performed by Kaplan-Meier estimate and the comparison between groups by Log-rank test. Multivariate analysis was carried out by Cox proportional hazard model. RESULTS: The overall 5-year survival rate was 58.2%. The 5-year survival rate was 55.8% in squamous-cell carcinoma, 44.0% in adenocarcinoma, 59.7% in undifferentiated carcinoma, 76.3% in adenoid cystic carcinoma, 71.4% in mucoepidermoid carcinoma, 25.0% in rhabdomyosarcoma, 26.7% in malignant melanoma, 50.0% in neuroblastoma (P > 0.05). Patients with cervical metastasis gave a 5-year survival of 53.5% while those without gave 58.9% (P > 0.05). Patients with involvement of sphenoidal sinus or maxillary sinus gave the worse survival. The 5-year survival rate was 73.8% in patients whose cancer completely disappeared after treatment. It was 41.6% in patients whose cancer incompletely disappeared, and 34.3% in patients whose cancer remained refractory (P < 0.01). The 5-year survival was 78.3% in stage I disease, 56.4% in stage II disease, 54.2% in stage III and 35.9% in stage IV (P < 0.05). The 5-year survival rate of patients who were treated with radiotherapy only was 56.9%. That of patients who were treated with surgery only was 56.6%. That with chemotherapy only was 25.0% whereas that of patients treated with combination treatment was 61.8% (P > 0.05). So far, 85 patients have died up to writing this report, 57.6% (49 patients) of recurrence or uncontrolled. CONCLUSION: Clinical stage, immediate therapeutic response and involvement of sphenoidal or maxillary sinus; but not the pathologic type, the presence of cervical metastasis nor the method of treatment, are the factors affecting the prognosis of patients with nasal carcinoma. Recurrence and uncontrolled disease are the cause of death.

[Prognosis and treatment for 98 patients with carcinoma of nasal cavity].

BACKGROUND & OBJECTIVE: Carcinoma of nasal cavity is a rare disease without standard treatment. This study was designed to evaluate treatment outcome, and prognostic factors of patients with carcinoma of nasal cavity. METHODS: Records of 98 patients with carcinoma of nasal cavity were reviewed,43 patients received radiotherapy alone, 55 patients received surgery plus radiotherapy. Survival analysis was performed by Kaplan-Meier method,differences between groups were tested by log-rank test,multivariate analysis was carried out by Cox proportional hazard model. RESULTS: The overall 5-, and 10-year survival rates were 65.3%, and 37.1%. The 5-, and 10-year survival rates of patients with tumor of stage I, II were 87.3%, and 51.1%, those of patients with tumor of stage III, IV were 56.1%, and 32.2% (P=0.02). The patients with squamous cell carcinoma had lower survival rates than those with adenocarcinoma (P< 0.01). There was significant difference in survival rates between patients received radiotherapy alone and patients received radiotherapy plus surgery in advanced lesions (P=0.04) and in squamous cell carcinoma (P< 0.01), but not in early lesions (P=0.41) and in adenocarcinoma (P=0.73). Patients who were initially diagnosed cervical lymph node metastasis had a reduced survival rate compared with node-negative patients (P=0.01). In Cox's regression, clinical stage and node-positive were independent prognostic factors. CONCLUSIONS: The criteria should be taken into account when choosing treatment method for patients with carcinoma of nasal cavity: radiotherapy plus surgery is preferred for advanced lesions and for squamous cell carcinoma, radiotherapy alone is better for early lesions and for adenocarcinoma. Clinical stage, and node-positive may be independent prognostic factors of patients with carcinoma of nasal cavity.