Phillyrin (KD-1) exerts anti-viral and anti-inflammatory activities against novel coronavirus (SARS-CoV-2) and human coronavirus 229E (HCoV-229E) by suppressing the nuclear factor kappa B (NF-κB) signaling pathway.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has extensively and rapidly spread in the world, causing an outbreak of acute infectious pneumonia. However, no specific antiviral drugs or vaccines can be used. Phillyrin (KD-1), a representative ingredient of Forsythia suspensa, possesses anti-inflammatory, anti-oxidant, and antiviral activities. However, little is known about the antiviral abilities and mechanism of KD-1 against SARS-CoV-2 and human coronavirus 229E (HCoV-229E). PURPOSE: The study was designed to investigate the antiviral and anti-inflammatory activities of KD-1 against the novel SARS-CoV-2 and HCoV-229E and its potential effect in regulating host immune response in vitro. METHODS: The antiviral activities of KD-1 against SARS-CoV-2 and HCoV-229E were assessed in Vero E6 cells using cytopathic effect and plaque-reduction assay. Proinflammatory cytokine expression levels upon infection with SARS-CoV-2 and HCoV-229E infection in Huh-7 cells were measured by real-time quantitative PCR assays. Western blot assay was used to determine the protein expression of nuclear factor kappa B (NF-κB) p65, p-NF-κB p65, IκBα, and p-IκBα in Huh-7 cells, which are the key targets of the NF-κB pathway. RESULTS: KD-1 could significantly inhibit SARS-CoV-2 and HCoV-229E replication in vitro. KD-1 could also markedly reduce the production of proinflammatory cytokines (TNF-α, IL-6, IL-1ß, MCP-1, and IP-10) at the mRNA levels. Moreover, KD-1 could significantly reduce the protein expression of p-NF-κB p65, NF-κB p65, and p-IκBα, while increasing the expression of IκBα in Huh-7 cells. CONCLUSIONS: KD-1 could significantly inhibit virus proliferation in vitro, the up-regulated expression of proinflammatory cytokines induced by SARS-CoV-2 and HCoV-229E by regulating the activity of the NF-кB signaling pathway. Our findings indicated that KD-1 protected against virus attack and can thus be used as a novel strategy for controlling the coronavirus disease 2019.

Generation of a Broadly Useful Model for COVID-19 Pathogenesis, Vaccination, and Treatment.

COVID-19, caused by SARS-CoV-2, is a virulent pneumonia, with >4,000,000 confirmed cases worldwide and >290,000 deaths as of May 15, 2020. It is critical that vaccines and therapeutics be developed very rapidly. Mice, the ideal animal for assessing such interventions, are resistant to SARS-CoV-2. Here, we overcome this difficulty by exogenous delivery of human ACE2 with a replication-deficient adenovirus (Ad5-hACE2). Ad5-hACE2-sensitized mice developed pneumonia characterized by weight loss, severe pulmonary pathology, and high-titer virus replication in lungs. Type I interferon, T cells, and, most importantly, signal transducer and activator of transcription 1 (STAT1) are critical for virus clearance and disease resolution in these mice. Ad5-hACE2-transduced mice enabled rapid assessments of a vaccine candidate, of human convalescent plasma, and of two antiviral therapies (poly I:C and remdesivir). In summary, we describe a murine model of broad and immediate utility to investigate COVID-19 pathogenesis and to evaluate new therapies and vaccines.

Liu Shen capsule shows antiviral and anti-inflammatory abilities against novel coronavirus SARS-CoV-2 via suppression of NF-κB signaling pathway.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has spread worldwide through person-to-person contact, causing a public health emergency of international concern. At present, there is no specific antiviral treatment recommended for SARS-CoV-2 infection. Liu Shen capsule (LS), a traditional Chinese medicine, has been proven to have a wide spectrum of pharmacological properties, such as anti-inflammatory, antiviral and immunomodulatory activities. However, little is known about the antiviral effect of LS against SARS-CoV-2. Herein, the study was designed to investigate the antiviral activity of SARS-CoV-2 and its potential effect in regulating the host's immune response. The inhibitory effect of LS against SARS-CoV-2 replication in Vero E6 cells was evaluated by using the cytopathic effect (CPE) and plaque reduction assay. The number of virions of SARS-CoV-2 was observed under transmission electron microscope after treatment with LS. Proinflammatory cytokine expression levels upon SARS-CoV-2 infection in Huh-7 cells were measured by real-time quantitative PCR assays. The results showed that LS could significantly inhibit SARS-CoV-2 replication in Vero E6 cells, and reduce the number of virus particles and it could markedly reduce pro-inflammatory cytokines (TNF-α, IL-6, IL-1ß, IL-8, CCL-2/MCP-1 and CXCL-10/IP-10) production at the mRNA levels. Moreover, the expression of the key proteins in the NF-κB/MAPK signaling pathway was detected by western blot and it was found that LS could inhibit the expression of p-NF-κB p65, p-IκBα and p-p38 MAPK, while increasing the expression of IκBα. These findings indicate that LS could inhibit SARS-CoV-2 virus infection via downregulating the expression of inflammatory cytokines induced virus and regulating the activity of NF-κB/MAPK signaling pathway in vitro, making its promising candidate treatment for controlling COVID-19 disease.

Liquefaction of Biomass and Upgrading of Bio-Oil: A Review.

The liquefaction of biomass is an important technology to converse the biomass into valuable biofuel. The common technologies for liquefaction of biomass are indirect liquefaction and direct liquefaction. The indirect liquefaction refers to the Fischer-Tropsch (F-T) process using the syngas of biomass as the raw material to produce the liquid fuel, including methyl alcohol, ethyl alcohol, and dimethyl ether. The direct liquefaction of biomass refers to the conversion biomass into bio-oil, and the main technologies are hydrolysis fermentation and thermodynamic liquefaction. For thermodynamic liquefaction, it could be divided into fast pyrolysis and hydrothermal liquefaction. In addition, this review provides an overview of the physicochemical properties and common upgrading methods of bio-oil.

Depression can be prevented by astaxanthin through inhibition of hippocampal inflammation in diabetic mice.

The critical factor considered in a depression induced by diabetes is the inflammation eliciting hippocampal, amygdala and thalamic neuronal injury. Therefore, inhibiting inflammatory reactions in the brain and reducing neuronal injury can alleviate depression in rodents suffering from diabetes mellitus. The oral administration of astaxanthin has been employed in emotional disorders and diabetic complications due to its anti-depressant, anti-inflammatory and anti-apoptotic functions. However, it has not been reported whether astaxanthin can improve diabetes-related depression-like behavior, and its potential mechanisms have not been elucidated. The objective of the present study is to elucidate the effect of astaxanthin on depression in diabetic mice and to understand the underlying molecular mechanisms. In this study, experimental diabetic mice were given a single intraperitoneal injection of streptozotocin (STZ, 150mg/kg, dissolved in citrate buffer) after fasting for 12h. The diabetic model was assessed 72h after STZ injection, and mice with a fasting blood glucose level more than or equal to 16.7mmol/L were used in this study, and oral astaxanthin (25mg/kg) was provided uninterrupted for ten weeks. Depression-like behavior was evaluated by the tail suspension test (TST) and forced swimming test (FST). The glial fibrillary acidic protein (GFAP) and cleaved caspase-3-positive cells were measured by immunohistochemistry, and the western blotting was used to test the protein levels of interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and cyclooxygenase (COX-2). The results showed that astaxanthin had an anti-depressant effect on diabetic mice. Furthermore, we observed that astaxanthin significantly reduced the number of GFAP-positive cells in the hippocampus and hypothalamus, and also the expression of cleaved caspase-3 in the hippocampus, amygdala and hypothalamus was decreased as well. Moreover, astaxanthin could down-regulate the expression of IL-6, IL-1ß and COX-2 in the hippocampus. These findings suggest that the mechanism of astaxanthin in preventing depression in diabetic mice involves the inhibition of inflammation/inflammation inhibition, thereby protecting neurons in hippocampus, amygdala and hypothalamus against hyperglycemic damage.

[Analysis of volatile compounds of fresh tea leaves from yaoluoping nature preserve by SDE-GC-MS].

OBJECTIVE: To study the volatile compounds of fresh tea leaves from Yaoluoping Nature Preserve, and to provide scientific basis for the quality and medicinal value of tea from high mountainous area. METHODS: The volatile compounds were extracted from fresh tea leaves by simultaneous distillation and extraction, and analyzed by gas chromatography-mass spectrum (GC-MS). RESULTS: 41 volatile compounds were identified from two tea cultivars, Shifoxiang and Shifocui. Shifoxiang had 32 kinds of volatile compounds and Shifocui had 38 kinds of volatile compounds. The main volatile components of Shifoxiang and Shifocui were green leaf volatiles. The kind and relative content of terpenes of Shifocui were more than that of Shifoxiang. Also the kind and relative content of ketones of Shifocui were more than that of Shifoxiang. There was one kind of heterocyclic compound of Shifocui. CONCLUSION: The main volatile components of Shifoxiang and Shifocui had an important role for the formation of unique aroma of high mountain ous tea. The composition and proportion of volatile components between Shifoxiang and Shifocui had certain differences. Some trace components may be associated with defense in the process of tea plant growth. The functional components from the volatile components had good medicinal value, worthy of further research.

[Nitrogen uptake and denitrification study on the joint treatment of aquatic vegetation and immobilized nitrogen cycling bacteria in Taihu Lake].

Undisturbed sediment cores were collected from Meiliang Bay, Taihu Lake. Immobilized nitrogen cycling bacteria (INCB), Elodea nuttallii were added to four groups of restoration incubation chambers respectively to explore the nitrogen removal mechanism in different restoration treatments. uN tracer and isotope pairing technique were used to determine the rates of plant uptake and denitrification in different treatments. The results showed that denitrification rates were significantly different among the treatments, while cores with addition of both INCB and Elodea nuttallii achieved the highest denitrification rate of 99.35 µmol · (m2 · h)( -1) and plant uptake rate of 36.55 µg · (m2 · h) -1. Elodea nuttallii in the cores could assimilate nitrate itself and enhance coupled nitrification- denitrification. Compared with plant uptake, denitrification was the main pathway of nitrogen removal. The results also showed that the combination of Elodea nuttallii and INCB could promote benthic nitrogen removal and purification of water body.

[Effect of warmth-promotion needling on cerebral SOD, MDA and AChE in vascular dementia rats].

OBJECTIVE: To observe the effect of warmth-promotion needling on cerebral malondialdehyde (MDA) content, superoxide dismutase (SOD) and acetylcholinesterase (AChE) activity so as to explore its underlying mechanism in improving vascular dementia (VD). METHODS: Forty Wistar rats were randomized into control, model, medication, needle-twirling and warmth-promotion groups with 8 cases in each. VD model was established by occlusion of the bilateral common carotid arteries and reperfusion in combination with intraperitoneal injection of sodium nitroprusside (2.5 mg/kg). Warmth-promotion needling was applied to "Dazhui" (GV 14), "Baihui" (GV 20) and "Shuigou" (GV 26). Rats of medication group were intragastric perfusion of Nimodipine suspension (0.0108 g/kg). The treatment was given once daily continuously for 15 days. SOD and AChE activity and MDA content of the brain (right side) tissue were detected with purine oxidase method, hydroxylamine chromatometry and thio-malonylurea method separately. RESULTS: In comparison with control group. SOD activity of model group decreased significantly (P < 0.01), AChE activity and MDA content increased considerably (P < 0.01); while compared with model group, SOD activity of warmth-promotion, needle-twirling and medication groups raised evidently (P < 0.01), AChE activity and MDA content of the later 3 groups lowered significantly (P < 0.05, 0.01). The effects of warmth-promotion group were significantly superior to that of medication group in lowering MDA content and to those of needle-twirling group in raising SOD activity and lowering AChE activity and MDA content (P < 0.05). CONCLUSION: Warmth-promotion needling can function well in resisting lipid peroxidation injury and lowering AChE activity in VD rats, which may contribute to its effect in improving learning-memory ability.

[Experimental study on the prevention and treatment effects of warm-reinforcing needling for hyperlipoidemia in the rabbit].

OBJECTIVE: To investigate the effect of warm-reinforcing needling (WRN) on serum lipid metabolism in hyperlipoidemia (HL) rabbits. METHODS: A total of 32 rabbits were randomly and evenly divided into control group, WRN group, model group and uniform reinforcing-reducing needling (URRN) group. HL model was established by feeding the rabbits with hypercholesterol and high-fat forage continuously for 7 weeks. For rabbits of WRN and URRN groups, "Zusanli" (ST 36), "Sanyinjiao" (SP 6), "Xuehai" (SP 10) were punctured and stimulated manually with WRN and URRN techniques respectively, once daily, continuously for 7 weeks. At the end of experiments, blood samples were collected from the heart, and centrifuged for detecting serum total cholesterol (TC) triglyceride (TG) and high density lipoprotein (HDL) with enzyme method, and platelet aggregation rate (PAR) with an intelligent blood agglutometer. Low density lipoprotein (LDL) = TC- (HDL + TG/2.2). RESULTS: Compared with normal control group, serum TC, TG and LDL contents in model group, WRN and URRN groups increased significantly (P < 0.01), and HDL content of model group decreased considerably (P < 0.01); while in comparison with model group, serum TC, LDL and TG contents of WRN and URRN groups decreased considerably (P < 0.01), and serum HDL levels of WRN and URRN groups increased significantly (P < 0.01). Serum TC content of WRN group was markedly lower than that of URRN group (P < 0.05), and HDL of WRN group was remarkably higher than that of URRN group (P < 0.01). In comparison with normal control group, 1 min, 5 min and maximum PARs in model, WRN and URRN groups increased significantly (P < 0.01), while in comparison with model group, 1 min, 5 min and maximum PARs of WRN and URRN groups decreased considerably (P < 0.01). In addition, 1 min and maximum PARs in WRN group were markedly lower than those in URRN group (P < 0.05). Light microscopic observation showed that the apoptotic foam cells in the lipid plaque of aorta wall in model group appeared and were obviously more than those of WRN and URRN groups. CONCLUSION: Acupuncture can effectively lower serum TC, LDL and TG levels, raise serum HDL content, and suppress platelet aggregation, and the therapeutic effect of WRN is significantly superior to that of URRN, which may contribute to the action of acupuncture in regulating blood lipid and inhibiting the formation of atheromatous plaque.

[Effects of "warmth-producing needling for removing obstruction" on hippocampal morphological changes and learning-memory abilities in vascular dementia rats].

OBJECTIVE: To observe the effects of "Warmth-producing Needling for Removing Obstruction" (WPNRO) on the pathomorphological changes of brain and learning-memory abilities in vascular dementia (VD) rats. METHODS: A total of 50 Wistar rats were randomly divided into normal control, model, medication, needle-twirling, and WPNRO groups, with 10 cases in each group. VD model was established by repeated cerebral ischemia-reperfusion via repeated occlusion of the bilateral common carotid arteries under anesthesia (3% embutal 40 mg/kg, i.p.). "Dazhui" (GV 14), "Baihui"(GV 20) and "Shuigou"(GV 26) were punctured with filiform needles and stimulated with twirling technique and WPNRO technique respectively and continuously for 1 min. Animals of medication group were fed with Nimodipine (0.0108 g/kg). The treatment was conducted once daily continuously for 15 days. The rats' learning and memory results were detected respectively with step-down avoidance test in the first 6 days' training and 24 h later. At the end of experiments and after decapitation, the right brain was taken, cut into sections (5 microm) and stained with H & E method for observing structural changes of hippocampus. RESULTS: 1) Behavior reactions: compared with control group, the latency for finding the safe platform and the times of error in model group increased significantly, and compared with model group, both latencies and times of error in WPNRO, needle-twirling and medication groups decreased considerably (P < 0.05, 0.01). In comparison with needle-twirling group, both latencies and times of error in WPNRO group were significantly fewer (P < 0.05, 0.01), and no significant differences were found between WPNRO and medication groups in these two indexes (P > 0.05). It indicated that acupuncture particularly WPNRO could improve both learning and memory abilities in VD rats. 2) Structural changes of neurons in CA1 region of hippocampus: in normal group, the neurons arranged in order and closely, and were normal in the structure; in model group, the neurons arranged obviously in disorder, had karyopyknosis and hyperplasia in glial cells and decreased in the number. In WPNRO group, no evident cellular karyopyknosis or hyperplasia of glial cells was found, and the structure and number of cells were close to those of normal group. In acupuncture and medication groups, fewer neurons, cellular karyopyknosis and hyperplasia of more glial cells were found. CONCLUSION: "Warming-producing Needling for Removing Obstruction" can significantly relieve cerebral ischemia induced disturbance of learning and memory and lessen structural injury of hippocampal neurons in rats.