GMI, a fungal immunomodulatory protein, ameliorates SARS-CoV-2 envelope protein-induced inflammation in macrophages via inhibition of MAPK pathway

Clinically, COVID-19 is often accompanied by a severe immune response (cytokine storm) which produces a large number of cytokines, such as TNF-α, IL-6 and IL-12, and consequently causes acute respiratory distress syndrome (ARDS). GMI is a type of fungal immunomodulatory protein that is cloned from Ganoderma microsporum and acts as modulating immunocyte for various inflammatory diseases. This study identifies GMI as a potential anti-inflammatory agent and determines the effects of GMI on the inhibition of SARS-CoV-2-induced cytokine secretion. Functional studies showed that SARS-CoV-2 envelop (E) protein induces inflammatory process in murine macrophages RAW264.7 and MH-S cells and in phorbol 12-myristate 13-acetate (PMA)-stimulated human THP-1 cells. GMI exhibits a strong inhibitory effect for SARS-CoV-2-E-induced pro-inflammatory mediators, including NO, TNF-α, IL-6, and IL-12 in macrophages. GMI reduces SARS-CoV-2-E-induced intracellular inflammatory molecules, such as iNOS and COX-2, and inhibits SARS-CoV-2-E-stimulated phosphorylation of ERK1/2 and P38. GMI also downregulates pro-inflammatory cytokine levels in lung tissue and serum after the mice inhale SARS-CoV-2-E protein. In conclusion, this study shows that GMI acts as an agent to alleviate SARS-CoV-2-E-induced inflammation.

SO2 compliance monitoring and emission characteristics analysis of navigating ships: A case study of Shanghai waters in emission control areas, China

For the purpose of monitoring the emission compliance of navigating ships in key waters of Shanghai, the present study adopts a sniffing method involving an unmanned aerial vehicle (UAV) platform-based ship exhaust mini-sniffing system measuring the concentrations of SO2 and CO2 within the plumes of navigating ships, then estimating the fuel sulfur content (FSC) of ship. The proposed method aims to provide a low-cost, non-contact method to assess the compliance level of FSC of navigating ships. Experimental tests were performed on ships in the Yangtze River Estuary and the core section of the Huangpu River, the key waters of Shanghai. 679 ships were monitored in the experiment, of which 13 ships were monitored by UAV with FSC greater than 0.5% (m/m). Due to the prevention and control of the COVID-19 pandemic, 8 monitored ships were intercepted by maritime law enforcement(MLE), and fuel oil samples were collected and analyzed in laboratory. The results show that the FSC of the 8 ships was greater than the limit of 0.5% (m/m). Finally, the monitoring results are comprehensively analyzed according to different ship lengths, types, nationalities and water areas. From the experimental results, it can be concluded that the FSC of Chinese cargo ships with length in the range of 80–160 m is more likely to exceed the standard 0.5%(m/m). The probability of exceeding the standard 0.5%(m/m) of navigating ships in the Yangtze River is higher than that of the Huangpu River. Overall, the presented study provides a reference for ship supervision in emission control areas (ECAs), which can improve the efficiency of MLE.

GMI, a protein from Ganoderma microsporum, induces ACE2 degradation to alleviate infection of SARS-CoV-2 Spike-pseudotyped virus.

BACKGROUND: Severe Acute Respiratory Syndrome Coronavirus Type 2 (SARS-CoV-2) induces a global serious pandemic and is responsible for over 4 million human deaths. Currently, although various vaccines have been developed, humans can still get SARS-CoV-2 infection after being vaccinated. Therefore, the blocking of SARS-CoV-2 infection may be potential therapeutic strategies. Ganoderma microsporum immunomodulatory protein (GMI), a small fungal protein, is cloned from Ganoderma microsporum. It exhibits anti-cancer and immunomodulatory functions. Currently, it is still unclear whether GMI involves in interfering with viral infection. PURPOSE: This study aimed to examine the potential functions and mechanisms of GMI on inhibiting SARS-CoV-2 pseudovirus infection. METHODS: The effects of GMI were examined in vitro on ACE2 overexpressing HEK293T (HEK293T/ACE2) cells exposed to SARS-CoV-2 Spike lentiviral pseudovirus encoding a green fluorescent protein (GFP) gene. The infection efficacy was determined using fluorescence microscopy and flow cytometry. The protein level of ACE2 was verified by Western blot. The effects of GMI on cell viability of HEK293T/ACE2 and lung epithelial WI38-2RA cells were determined by MTT assay. Mice received GMI via nebulizer. RESULTS: GMI did not affect the cell viability of HEK293T/ACE2, WI38-2RA and macrophages. Functional studies showed that GMI inhibited GFP expressing SARS-CoV-2 pseudovirus from infecting HEK293T/ACE2 cells. GMI slightly interfered the interaction between ACE2 and Spike protein. GMI interacted with S2 domain of Spike protein. Specifically, GMI dramatically reduced ACE2 expression in HEK293T/ACE2 and WI38-2RA cells. Mechanistically, GMI induced ACE2 degradation via activating protein degradation system, including proteasome and lysosome. Abolishing proteasome and lysosome by MG132 and bafilomycin A1, respectively, rescued GMI-reduced ACE2 levels. In addition, GMI triggered dynamin and lipid raft-mediated ACE2 endocytosis. ACE2 levels were downregulated in the lung tissue after the mice inhaling GMI. CONCLUSIONS: GMI prevents SARS-CoV-2 pseudovirus infection via induction of ACE2 degradation in host cells. Our findings suggest that GMI will be a potential prevention agent to alleviate SARS-CoV-2 infection.

The Traditional Chinese Medicine Formula Jing Guan Fang for Preventing SARS-CoV-2 Infection: From Clinical Observation to Basic Research.

COVID-19 is a global epidemic. Developing adjuvant therapies which could prevent the virus from binding to cells may impair viral infection. This study produces a traditional Chinese medicine formula, Jing Guan Fang (JGF), based on ancient medical texts, and examines the efficacy and the mechanism by which JGF prevents viral infections. JGF reduces COVID-19 like symptoms. Functional studies show that JGF inhibits the formation of syncytium and reduces the formation of viral plaque. JGF is not toxic in vitro and in vivo. Mechanistically, JGF induces lysosomal-dependent ACE2 degradation and suppresses mRNA and the protein levels of TMPRSS2 in human lung WI-38 and MRC-5 cells. Mice that inhale JGF exhibit reduced ACE2 and TMPRSS2 protein levels in lung tissues. Together, these findings suggest that JGF may improve the COVID-19 like symptoms and inhibit viral infection. Moreover, JGF may be applicable as an adjuvant preventive strategy against SARS-CoV-2 infection in addition to the use of vaccines.

Impacts and strategies for the construction of the disease control and prevention system in the COVID-19 outbreak / 上海预防医学

The outbreak of COVID-19 has exposed many shortcomings in disease control and prevention system (DCPS) of China. Resolving the problems and strengthening the DCPS became the top priority in China’s public policy agenda. This paper revealed the problems of the system regarding policy-making, regulations, operation mechanism and staff, and proposed several strategies from three aspects of legal construction, management system, and operation mechanism, including: 'Prevention first' should be incorporated into the national legal system, all the departments should be engaged in 'Healthy China 2030' initiative, laws and regulations should be amended, new disease prevention and control management institutions should be set up, a high-quality professional team should be retained, the regional health information exchange channels should be strengthened, the coordinated mechanism for disease prevention and control should be normalized, the long-term investment mechanism should be established, and the equipment renewal and reserve system should be improved.