Olgotrelvir as a Single-Agent Treatment of Nonhospitalized Patients with Covid-19.

BACKGROUND: Olgotrelvir is an oral antiviral with dual mechanisms of action targeting severe acute respiratory syndrome coronavirus 2 main protease (i.e., Mpro) and human cathepsin L. It has potential to serve as a single-agent treatment of coronavirus disease 2019 (Covid-19). METHODS: We conducted a phase 3, double-blind, randomized, placebo-controlled trial to evaluate the efficacy and safety of olgotrelvir in 1212 nonhospitalized adult participants with mild to moderate Covid-19, irrespective of risk factors, who were randomly assigned to receive orally either 600 mg of olgotrelvir or placebo twice daily for 5 days. The primary and key secondary end points were time to sustained recovery of a panel of 11 Covid-19-related symptoms and the viral ribonucleic acid (RNA) load. The safety end point was incidence of treatment-emergent adverse events. RESULTS: The baseline characteristics of 1212 participants were similar in the two groups. In the modified intention-to-treat population (567 patients in the placebo group and 558 in the olgotrelvir group), the median time to symptom recovery was 205 hours in the olgotrelvir group versus 264 hours in the placebo group (hazard ratio, 1.29; 95% confidence interval [CI], 1.13 to 1.46; P<0.001). The least squares mean (95% CI) changes of viral RNA load from baseline were -2.20 (-2.59 to -1.81) log10 copies/ml in olgotrelvir-treated participants and -1.40 (-1.79 to -1.01) in participants receiving placebo at day 4. Skin rash (3.3%) and nausea (1.5%) were more frequent in the olgotrelvir group than in the placebo group; there were no treatment-related serious adverse events, and no deaths were reported. CONCLUSIONS: Olgotrelvir as a single-agent treatment significantly improved symptom recovery. Adverse effects were not dose limiting. (Funded by Sorrento Therapeutics, a parent company of ACEA Therapeutics; ClinicalTrials.gov number, NCT05716425.).

Olgotrelvir, a dual inhibitor of SARS-CoV-2 Mpro and cathepsin L, as a standalone antiviral oral intervention candidate for COVID-19.

BACKGROUND: Oral antiviral drugs with improved antiviral potency and safety are needed to address current challenges in clinical practice for treatment of COVID-19, including the risks of rebound, drug-drug interactions, and emerging resistance. METHODS: Olgotrelvir (STI-1558) is designed as a next-generation antiviral targeting the SARS-CoV-2 main protease (Mpro), an essential enzyme for SARS-CoV-2 replication, and human cathepsin L (CTSL), a key enzyme for SARS-CoV-2 entry into host cells. FINDINGS: Olgotrelvir is a highly bioavailable oral prodrug that is converted in plasma to its active form, AC1115. The dual mechanism of action of olgotrelvir and AC1115 was confirmed by enzyme activity inhibition assays and co-crystal structures of AC1115 with SARS-CoV-2 Mpro and human CTSL. AC1115 displayed antiviral activity by inhibiting replication of all tested SARS-CoV-2 variants in cell culture systems. Olgotrelvir also inhibited viral entry into cells using SARS-CoV-2 Spike-mediated pseudotypes by inhibition of host CTSL. In the K18-hACE2 transgenic mouse model of SARS-CoV-2-mediated disease, olgotrelvir significantly reduced the virus load in the lungs, prevented body weight loss, and reduced cytokine release and lung pathologies. Olgotrelvir demonstrated potent activity against the nirmatrelvir-resistant Mpro E166 mutants. Olgotrelvir showed enhanced oral bioavailability in animal models and in humans with significant plasma exposure without ritonavir. In phase I studies (ClinicalTrials.gov: NCT05364840 and NCT05523739), olgotrelvir demonstrated a favorable safety profile and antiviral activity. CONCLUSIONS: Olgotrelvir is an oral inhibitor targeting Mpro and CTSL with high antiviral activity and plasma exposure and is a standalone treatment candidate for COVID-19. FUNDING: Funded by Sorrento Therapeutics.

Carboxymethyl chitosan/sodium alginate hydrogel films with good biocompatibility and reproducibility by in situ ultra-fast crosslinking for efficient preservation of strawberry.

Strawberry is a seasonal and regional fruit. Thus, strawberry waste caused by spoilage and decay is an urgent problem that must be solved. Developing hydrogel films (HGF) for multifunctional food packaging can effectively slow down strawberry. Based on the carboxymethyl chitosan/sodium alginate/citric acid with excellent biocompatibility, preservation effect, and ultrafast (10 s) coating on the strawberry surface, HGF specimens were designed and prepared through the electrostatic interaction of opposite charges between polysaccharides. The prepared HGF specimen exhibited excellent low moisture permeability and antibacterial properties. Its lethality rates against both Escherichia coli and Staphylococcus aureus were more than >99 %. The HGF could keep strawberries fresh for up to 8, 19, and 48 days at 25.0, 5.0, and 0 °C, respectively, by delaying the fruits' ripening, dehydration, microbial invasion, and respiration rate. The HGF dissolved and regenerated five times still exhibited good performance. The water vapor transmission rate of the regenerative HGF could reach 98 % of that of the original HGF. The regenerative HGF could maintain the freshness of strawberries for up to 8 days at 25.0 °C. This study provides new insight into an alternative film design for convenient, green, and renewable alternative films to delay perishable fruit spoilage.

Enhanced antibacterial effect and biodegradation of coating via dual-in-situ growth based on carboxymethyl cellulose.

The lack of antimicrobial effect of commercial paper coating for food packaging makes it difficult to prevent food spoilage and harms the environment by non-biodegradation. Herein, carboxymethyl cellulose (CMC) provides negatively charged sites for anchoring Ag+ and Zn2+ to grow AgNPs and ZIF-8 in situ on its molecular chains. The ZIF-8/AgNPs@CMC paper coating has excellent synergistic antibacterial activity to prolong the shelf-life of food. It not only has good thermal stability but binds closely to the paper and its adhesion force reaches 628.9 nN. Besides, the ZIF-8/AgNPs@CMC coated paper has better mechanical properties, water vapor barrier, and resists water solubility. Interestingly, due to the confinement effect of ZIF-8, the cumulative release of AgNPs after 168 h is only 2.66 % to avoid possible food safety risks. Especially, the coating can be almost biodegraded in the soil after 30 days, which provides the possibility to replace the non-biodegradable coatings in food packaging.

Hybridization of carboxymethyl chitosan with bimetallic MOFs to construct renewable metal ion "warehouses" with rapid sterilization and long-term antibacterial effects.

Pathogens transmitted through the water environment pose a great threat to human health. Hence developing more reliable and efficient antibacterial materials to eliminate bacterium in water environments is urgent. Herein, we posed a novel strategy of interweaving carboxymethyl chitosan (CMCS) and Ag/Cu-MOFs to construct renewable Ag/Cu-BTC@CMCS composite beads with rapid sterilization, long-term antibacterial effects and high biosafety. Characterizations revealed that CMCS and bimetallic MOFs act as the "warehouses" of metal ions and played key roles in anchoring, storage, delivery, and controlled release of metal ions. The synergistic antibacterial effect achieved by the combination of Ag+ and Cu2+ provides the composite beads with high antibacterial efficiency, resulting in low minimum inhibitory concentrations (0.32 mg/mL against E. coli and 0.16 mg/mL against S. aureus) and over 99.9 % bacteria killing rate. Benefiting from the rapid release of metal ions from polymer chains and the long-term release from MOFs, the composite beads can effectively sterilize the simulated swimming pool water in 2 h and persistently inhibit bacterial reproduction over 48 h, and show a safe level of residual heavy metals because of the chelation of CMCS. This work provides new insights and promises a strategy for the design and commercial application of novel water fungicides.

XueFu ZhuYu Decoction Alleviates Cardiopulmonary Bypass-Induced NLRP3 Inflammasome-Dependent Pyroptosis by Inhibiting IkB-α/NF-κB Pathway in Acute Lung Injury Rats.

XueFu ZhuYu Decoction (XFZYD) is an effective prescription that is widely used to improve blood circulation by removing blood stasis. This study aimed to investigate the effects and the underlying molecular mechanisms of XFZYD on lung pyroptosis in cardiopulmonary bypass- (CPB-) induced acute lung injury (ALI) rats. A rat model of ALI was induced by CPB treatment after XFZYD, Ac-YVAD-CMK, and Bay-11-7082 administration. The respiratory index (RI) and oxygenation index (OI) were determined at each time point. The levels of interleukin (IL)-1ß, IL-6, IL-18, and TNF-α in serum and lung were measured by enzyme-linked immunosorbent assays (ELISA). Moreover, the protein levels, neutrophil counts, and total cell of bronchoalveolar lavage fluid (BALF) were detected. Additionally, Myeloperoxidase (MPO) expression was detected by immunohistochemical assay. Lung injury was evaluated with the wet/dry (W/D) ratio and pathologic changes, respectively. Besides, the expression of NLRP3 inflammasome and IkB-α/NF-κB pathway proteins was estimated by immunofluorescence, quantitative real-time PCR (qRT-PCR), and Western blotting assays, respectively. XFZYD pretreatment significantly ameliorated pulmonary ventilation function and reduced the CPB-induced lung histopathological injury, inflammatory cell infiltration in BALF and lung, and the apoptosis of lung cells. Interestingly, XFZYD decreased the CPB-induced NLRP3, ASC, Caspase-1 p20, Pro-GSDMD, GSDMD p30, IL-18, IL-1ß p-P65, and p-IKBα mRNA or protein levels in lung tissues in ALI model rats. In summary, these findings suggest that XFZYD effectively mitigates NLRP3 inflammasome-dependent pyroptosis in CPB-induced ALI model rats, possibly by inhibiting the IkB-α/NF-κB pathway in the lung.

Flow Regulation Performance Analysis of Microfluidic Passive Valve for High Throughput Liquid Delivery.

A microfluidic passive valve (MPV) is important for precise flow control, and it determines the reliability of the microfluidic system. In this paper, a novel MPV capable of delivering a constant flow rate independently of inlet pressure changes is proposed. The flow rate of the MPV is adjusted by the difference between the fluid force on the upper surface of the valve core and the spring force. The constant flow rate of the MPV is maintained by automatically changing the size of the gap channel formed by the groove on the valve core and the baffle on the valve body. The nearly constant flow rate of the MPV is 6.26 mL/min, with a variation of 6.5% under the inlet pressure varied from 1.25 kPa to 3.5 kPa. In addition, the flow characteristics of the MPV are analyzed by numerical simulation. With the increase in the inlet pressure, the maximum velocity gradually increases, while the increment of the maximum velocity decreases. In the movement process of the valve core, the region of pressure drop becomes larger. This work has a certain reference value for the design and research of the MPVs with high throughput liquid delivery.

A self-matching, ultra-fast film forming and washable removal bio-crosslinked hydrogel films for perishable fruits.

Spoilage of food has aggravated the issue of food shortage worldwide. Here, we report a strategy for ultrafast hydrogel film forming within 10 s on fruit surfaces with good self-matching, washable removal and preservative property. This carboxymethyl chitosan (CMCS)/tannic acid (TA) hydrogel film (CTHF) is fabricated by bio-material of CMCS and TA via in-situ rapidly crosslinking with high-density hydrogen bonds. Simply blending TA and CMCS solution at room temperature can form CTHF with different roughness (Ra: ranges from 123 to 1.55 nm) on different fruit surfaces, so as to perfectly match the hydrogel protective layer of pericarp. The CTHF slows down fruit decay by its outstanding antioxidant and antibacterial activity. It is soluble and easily removed (within 3-5 min) by washing without environmental pollution and food safety issues. As natural polymer, CTHF shows high promise as sustainable substitutes for conventional plastics packing because of its non-toxic, edible, biodegradable, and environmentally friendly.

Analysis of changes in circular RNA expression and construction of ceRNA networks in human dilated cardiomyopathy.

Dilated cardiomyopathy (DCM) is a severe life-threatening disease worldwide, and the underlying mechanisms remain unclear. Circular RNAs (circRNAs) have been reported to play important roles in various cardiovascular diseases and can function as competitive endogenous RNAs (ceRNAs). However, their role in human DCM has not been fully elucidated. In the present study, heart samples from DCM patients and healthy controls were used to identify circRNAs by RNA sequencing. Real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was conducted to validate differentially expressed circRNAs and mRNAs. A total of 9585 circRNAs and 22050 mRNAs were detected in the two groups. Overall, 213 circRNAs and 617 mRNAs were significantly up-regulated in the DCM group compared with the control group. Similarly, 85 circRNAs and 1125 mRNAs were significantly down-regulated. According to the ceRNA theory, circRNAs can indirectly interact with mRNAs by directly binding to microRNAs (miRNAs), and circRNAs and mRNAs should be concurrently either up-regulated or down-regulated. Based on this theory, we constructed two circRNA-miRNA-mRNA networks by using the RNA sequencing data and prediction by proprietary software. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to probe the potential functions of differentially expressed circRNAs. In conclusion, this study revealed that the expression of cardiac circRNAs was altered in human DCM and explored the potential functions of circRNAs by constructing ceRNA networks. These findings provide a foundation for future studies of circRNAs in DCM.

Fluid-Structure Interaction Analysis on Membrane Behavior of a Microfluidic Passive Valve.

In this paper, the effect of membrane features on flow characteristics in the microfluidic passive valve (MPV) and the membrane behavior against fluid flow are studied using the fluid-structure interaction (FSI) analysis. Firstly, the microvalve model with different numbers of microholes and pitches of microholes are designed to investigate the flow rate of the MPV. The result shows that the number of microholes on the membrane has a significant impact on the flow rate of the MPV, while the pitch of microholes has little effect on it. The constant flow rate maintained by the microvalve (the number of microholes n = 4) is 5.75 mL/min, and the threshold pressure to achieve the flow rate is 4 kPa. Secondly, the behavior of the membrane against the fluid flow is analyzed. The result shows that as the inlet pressure increases, the flow resistance of the MPV increases rapidly, and the deformation of the membrane gradually becomes stable. Finally, the effect of the membrane material on the flow rate and the deformation of the membrane are studied. The result shows that changes in the material properties of the membrane cause a decrease in the amount of deformation in all stages the all positions of the membrane. This work may provide valuable guidance for the optimization of microfluidic passive valve in microfluidic system.

Inhibitory Effect of Vascular Endothelial Growth Factor on the Slowly Activating Delayed Rectifier Potassium Current in Guinea Pig Ventricular Myocytes.

BACKGROUND: Vascular endothelial growth factor (VEGF) exerts a number of beneficial effects on ischemic myocardium via its angiogenic properties. However, little is known about whether VEGF has a direct effect on the electrical properties of cardiomyocytes. In the present study, we investigated the effects of different concentrations of VEGF on delayed rectifier potassium currents (IK) in guinea pig ventricular myocytes and their effects on action potential (AP) parameters. METHODS AND RESULTS: IK and AP were recorded by the whole-cell patch clamp method in ventricular myocytes. Cells were superfused with control solution or solution containing VEGF at different concentrations for 10 minutes before recording. Some ventricular myocytes were pretreated with a phosphatidylinositol 3-kinase inhibitor for 1 hour before the addition of VEGF. We found that VEGF inhibited the slowly activating delayed rectifier potassium current (IKs) in a concentration-dependent manner (18.13±1.04 versus 12.73±0.34, n=5, P=0.001; 12.73±0.34 versus 9.05±1.20, n=5, P=0.036) and prolonged AP duration (894.5±36.92 versus 746.3±33.71, n=5, P=0.021). Wortmannin, a phosphatidylinositol 3-kinase inhibitor, eliminated these VEGF-induced effects. VEGF had no significant effect on the rapidly activating delayed rectifier potassium current (IKr), resting membrane potential, AP amplitude, or maximal velocity of depolarization. CONCLUSIONS: VEGF inhibited IKs in a concentration-dependent manner through a phosphatidylinositol 3-kinase-mediated signaling pathway, leading to AP prolongation. The results indicate a promising therapeutic potential of VEGF in prevention of ventricular tachyarrhythmias under conditions of high sympathetic activity and ischemia.

Microarray expression profiling and gene ontology analysis of long non-coding RNAs in spontaneously hypertensive rats and their potential roles in the pathogenesis of hypertension.

Long non-coding RNAs (lncRNAs) have been demonstrated to be significant in numerous biological processes. Hypertension is a form of cardiovascular disease with at least one billion cases worldwide. The present study sought to compare the differential expression profiles of lncRNAs in the renal cortex of spontaneously hypertensive rats (SHRs) and normotensive Wistar­Kyoto (WKY) rats. The ipsilateral renal cortex was obtained from 15­week­old SHRs and WKY rats whose blood pressures had been monitored. Total RNA was extracted using TRIzol, and lncRNAs and messenger RNAs were profiled by microarray and validated using fluorescent quantitative reverse transcription­polymerase chain reaction. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to predict the function of differentially expressed genes. Microarray analysis demonstrated that 145 lncRNAs were differentially expressed between SHRs and WKY rats. GO and KEGG pathway analysis indicated that these lncRNAs are involved in numerous biological processes. Thus, lncRNAs may contribute to the pathogenesis of hypertension.

[Telmisartan reduces retina vessel endothelial cell apoptosis via upregulating retinal ACE2-Ang-(1-7)-Mas axis in spontaneous hypertensive rats].

OBJECTIVE: To investigate the effects of angiotensin II (Ang II) antagonist telmisartan on retina vessel endothelial cell apoptosis and its impact on the ACE2-Ang-(1-7)-Mas axis in spontaneous hypertensive rats (SHR). METHODS: Thirty-six SHR 16 week-old were randomly divided into 3 groups (n = 12 each): SHR, SHRT (telmisartan 10 mg · kg-1 · d-1 by gastric gavage) and SHRTA group (telmisartan 10 mg · kg-1 · d-1 by gastric gavage plus intravenous injection of A-779 0.5 mg · kg-1 · d-1), twelve WKY rats served as normotensive control group. Systolic blood pressure was measured at pre-treatment and 8 weeks later. After 8 weeks, rats were sacrificed, the expression of ACE2 and Mas in retina were analyzed by qRT-PCR, Western blot and Immunohistochemistry, the Ang-(1-7) concentration in serum was measured by ELISA. Specimens were obtained and stained by hematoxylin and eosin, and the morphology of retina vessel was observed. Apoptosis of vessel endothelial cells were determined by using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method. RESULTS: The systolic blood pressure of SHR, SHRT and SHRTA groups at baseline were significantly higher than age-matched WKY group (all P < 0.01). Eight weeks later, the systolic blood pressure group was significantly lower in SHRT group than in the SHR group (P < 0.01), this effect was partly reversed in SHRTA group. The retinal ACE2 mRNA and protein expression was significantly lower in SHR group than in WKY and SHRT groups (P < 0.01), which was similar between SHRT group and SHRTA group (P > 0.05). The retinal Mas mRNA and protein expression were significantly lower in SHR group compared to WKY and SHRT groups (all P < 0.01), which was significantly lower in SHRTA group than in the SHRT group (P < 0.05). ELISA results showed that serum Ang-(1-7) protein level was significantly lower in SHR group than in WKY group and SHRT group (both P < 0.05), which was lower in SHRTA group compared to SHRT group. Retinal vessel endothelial cell apoptosis was higher in SHR group than in WKY group, which could be reduced by cotreatment with telmisartan and this beneficial effect could be reversed by A-779. CONCLUSION: Telmisartan can reduce retinal vessel endothelial cell apoptosis via upregulating the ACE2-Ang-(1-7)-Mas axis.