Platycodin D facilitates antiviral immunity through inhibiting cytokine storm via targeting K63-linked TRAF6 ubiquitination.

Influenza virus infection is a worldwide challenge that causes heavy burdens on public health. The mortality rate of severe influenza patients is often associated with hyperactive immunological abnormalities characterized by hypercytokinemia. Due to the continuous mutations and the occurrence of drug-resistant influenza virus strains, the development of host-directed immunoregulatory drugs is urgently required. Platycodon grandiflorum is among the top 10 herbs of traditional Chinese medicine used to treat pulmonary diseases. As one of the major terpenoid saponins extracted from Platycodon grandiflorum, Platycodin D (PD) has been reported to play several roles, including anti-inflammation, analgesia, anti-cancer, hepatoprotection, and immunoregulation. However, the therapeutic roles of PD to treat influenza virus infection remains unknown. Here, we show that PD can protect the body weight loss in severely infected influenza mice, alleviate lung damage, and thus improve the survival rate. More specifically, PD protects flu mice via decreasing the immune cell infiltration into lungs and downregulating the overactivated inflammatory response. Western blot and immunofluorescence assays exhibited that PD could inhibit the activation of TAK1/IKK/NF-κB and MAPK pathways. Besides that, CETSA, SPR and immunoprecipitation assays indicated that PD binds with TRAF6 to decrease its K63 ubiquitination after R837 stimulation. Additionally, siRNA interference experiments exhibited that PD could inhibit the secretion of IL-1ß and TNF-α in TRAF6-dependent manner. Altogether, our results suggested that PD is a promising drug candidate for treating influenza. Our study also offered a scientific explanation for the commonly used Platycodon grandiflorum in many anti-epidemic classic formulas. Due to its host-directed regulatory role, PD may serve as an adjuvant therapeutic drug in conjunction with other antiviral drugs to treat the flu.

Role of SIRT5 in the analgesic effectiveness of moxibustion at ST36 in mice with inflammatory pain.

Sirtuine5 (SIRT5) is an important molecule involved in the pathology of inflammatory diseases. To investigate the impact of SIRT5 on the analgesic effectiveness of moxibustion, we established a complete Freund's adjuvant- (CFA-) induced inflammatory pain in mice model. Moxibustion was applied at the Zusanli (ST36) acupoint in mice with inflammatory pain. The analgesic effectiveness was evaluated by thermal hyperalgesia and mechanical allodynia tests in the right paws after CFA injection. The expression of inflammatory cytokines, including the pro-inflammatory factors IL-1ß and TNF-α, and the anti-inflammatory factors IL-4 and TGF-ß expressions, was evaluated using by ELISA. Furthermore, SIRT5 was evaluated by immunofluorescence and western blotting. The results showed that, compared with the CFA group, both thermal and mechanical pain thresholds increased with moxibustion and the SIRT5 inhibitor MC3482 intervention at ST36. Additionally, compared to the CFA-induced group, the inflammatory mediators, including IL-1ß and TNF-α, decreased, while the anti-inflammatory cytokines IL-4 and TGF-ß increased with moxibustion and MC3482 ST36 acupoint injection. Western blot results showed a decreased expression of SIRT5 at the ST36 site with moxibustion and MC3482 injection, compared to the CFA-induced group. SIRT5 expression in the right paw of mice injected with moxibustion and MC3482 was higher than that in the CFA-induced group. This study revealed that SIRT5 expression is involved in moxibustion analgesia and may be a potential mediator in the regulation of analgesia.

Cable bacteria regulate sedimentary phosphorus release in freshwater sediments.

Previous studies have demonstrated that e-SOx can regulate the sedimentary release of phosphorus (P) in brackish and marine sediments. When e-SOx is active, an iron (Fe) and manganese (Mn) oxide rich layer is formed near the sediment surface, which prevents P release. When e-SOx becomes inactive, the metal oxide layer is reduced via sulfide-mediated dissolution, and P is subsequently released to the water column. Cable bacteria have been shown to also occur in freshwater sediments. In these sediments, sulfide production is limited, and the metal oxide layer would thus dissolve less efficiently, leaving the P trapped at the sediment surface. This lack of an efficient dissolution mechanism implies that e-SOx could play an important role in the regulation of P availability in eutrophied freshwater streams. To test this hypothesis, we incubated sediments from a eutrophic freshwater river to investigate the impact of cable bacteria on sedimentary cycling of Fe, Mn and P. High-resolution depth profiling of pH, O2 and ΣH2S complemented with FISH analysis and high-throughput gene sequencing showed that the development of e-SOx activity was closely linked to the enrichment of cable bacteria in incubated sediments. Cable bacteria activity caused a strong acidification in the suboxic zone, leading to the dissolution of Fe and Mn minerals and consequently a strong release of dissolved Fe2+ and Mn2+ to the porewater. Oxidation of these mobilized ions at the sediment surface led to the formation of a metal oxide layer that trapped dissolved P, as shown by the enrichment of P-bearing metal oxides in the top layer of the sediment and low phosphate in the pore and overlying water. After e-SOx activity declined, the metal oxide layer did not dissolve and P remained trapped at the surface. Overall, our results suggested cable bacteria can play an important role to counteract eutrophication in freshwater systems.

Spatiotemporal differences in riverine nitrogen and phosphorus fluxes and associated drivers across China from 1980 to 2018.

Increases in nutrient loadings to waterways over the past four decades have led to widespread eutrophication and water quality impairments across China. Understanding the spatial, interannual and long-term variations in nutrient loadings and associated drivers at the national scale is crucial for developing effective nutrient reduction strategies. However, the controls on, and spatiotemporal variations in, nutrient fluxes remain a problem from both an academic and management perspective. This study provides spatially extensive and temporally contiguous estimates of changes in riverine total nitrogen (TN), ammonia nitrogen (NH3-N) and total phosphorus (TP) fluxes for continental area of China based on machine learning stack models and empirical modeling over the period from 1980 to 2018. Results reveal considerable spatial, interannual and long-term variability in annual TN, NH3-N and TP fluxes, with spatial variations in average TN and NH3-N fluxes primarily driven by net anthropogenic nitrogen inputs. Interannual variability is dominated by precipitation across continental areas of China. Spatial variability in the estimated average annual TP flux in the undeveloped western and the developed middle east regions of China are primarily controlled by net anthropogenic phosphorus inputs and precipitation, respectively. We found that TN, NH3-N and TP fluxes increased from 1980 to 2018 in watersheds in East China; the national mean annual TN, NH3-N and TP fluxes increased before 2015 and decreased after 2015. This study illustrates the important role of precipitation and temperature variability in controlling the spatial, interannual and long-term variability of nutrient fluxes, and indicates that the influence of the meteorological conditions on annual loadings is needed when designing watershed nutrient reduction or management strategies.

Moxibustion Strategies for Mice.

The field of moxibustion research is expanding, with a rapid increase in publications in recent years. Moxibustion is a therapy that ignites moxa on the skin of humans, with an increase in peripheral skin temperature and localized redness. During this treatment, the recipient must remain still to prevent scalding and expose intervention sites for easy manipulation; however, maintaining a fixed posture during moxibustion is a big challenge for animals. Thus, manipulating moxibustion in small animals, such as mice, can lead to several difficulties for researchers. In addition, an uncomfortable posture for animals can lead to fear and resistance to moxibustion, increased risk of injury, diminished animal welfare, and less valid research data. An efficient, comfortable moxibustion method is needed to protect animal welfare and minimize the adverse effects on experimental results. However, moxibustion methods are highly variable and often have limited efficacy. More importantly, an uncomfortable moxibustion posture might cause a stress response, such as those observed with anxiety, fear, and anger, which could influence the research data. Therefore, strategies for animal moxibustion that inflict the least harm possible during the intervention are required. This protocol introduces a mouse tethering method for moxibustion intervention, minimizing mouse discomfort and improving study efficiency. Essential strategies for tethering mice and application of moxibustion are highlighted, and the structure of the tethering instrument is described.

[Introduction of a new type of mouse forelimb fixator and two types of batch mice moxibustion device].

A newly-developed "Mouse Forelimb Fixator" and two types of "Batch Mice Moxibustion Device" on the basis of our "Mouse Safe and Fast Fixation Board" (developed in 2012) were introduced in the present paper. The Forelimb Fixator inserted into the base part of the apparatus in tenon and mortise style is used to control the mouse's posture with the forelimbs' acupoints fully exposed, and can realize simultaneous fixation of several mice at the same time. By using the mobility of the base of the single-hole moxibustion frame and the magnet, the distance between the acupoint surface and the tip of the ignited moxa stick can be accurately controlled, and several acupoints of different meridians can be simultaneously stimulated at the same time. Utilizing the porous transparent moxibustion board, the Batch Mice Moxibustion Device can meet the requirement of moxibustion at multiple acupoints at the same time. In addition, these devices are convenient in operation, innovative in creativity, save manpower and material resources, and help improve experimental efficiency and research on moxibustion.

The role of STIM1/ORAI1 channel in the analgesic effect of grain-sized moxibustion on inflammatory pain mice model.

The therapeutic effect of grain-sized moxibustion (GS-Moxi) on inflammatory pain has been well recognized clinically, but the mechanism remains unclear. STIM1/ORAI1 is a sensible temperature channel, therefore; this study aimed to investigate the analgesic effect of GS-Moxi and the association with STIM1/ORAI1 expression. CFA-induced inflammatory pain model was established and was treated with GS-Moxi after 3 days of CFA injection. The behavioral test was measured after the GS-Moxi; then, serum was prepared for IL-1ß, IL-6, and TNF-α, and the stimulated skin was used for measuring STIM1 and ORAI1 expression. The results indicated GS-Moxi had an analgesic effect on inflammatory pain and the heat variation was significant for the analgesia. GS-Moxi decreased the expression of IL-1ß, IL-6, and TNF-α. Immunofluorescence and western blot analysis illustrated that heat change was associated with the stimulation of STIM1 and ORAI1. Suggesting that heat variation created by GS-Moxi could be crucial in this therapy and STIM1 and ORAI1 were potential enhancers in regulating analgesia of GS-Moxi.

The efficacy and safety of moxibustion for chemotherapy-induced gastrointestinal adverse reaction: A protocol for systematic review and meta-analysis.

BACKGROUND: Many cancer patients experience gastrointestinal adverse reaction during chemotherapy. Pharmacological interventions are commonly used to treat chemotherapy-induced gastrointestinal side effects but have various limitations. Clinical trials have indicated that moxibustion may alleviate gastrointestinal dysfunction and improve quality of life (QoL) after chemotherapy. This study aims to assess the efficacy and safety of moxibustion for chemotherapy-induced gastrointestinal adverse reaction through a systematic review and meta-analysis. METHODS: All randomized controlled trials (RCTs) related to moxibution targeting chemotherapy-induced gastrointestinal adverse reaction will be searched in online databases, such as PubMed, EMBASE, the Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), the Chinese Scientific Journal Database (VIP Database) and WanFang Database from their inception to May 1, 2020. The primary outcome is the incidence and severity of chemotherapy-related gastrointestinal toxicities (nausea and vomiting, diarrhea and constipation). The secondary outcomes include the quality of life, biological parameters' alteration, and adverse events. Study selection, data extraction, and assessment of risk of bias will be performed independently by 2 researchers. The Cochrane Collaboration's Review Manager (RevMan 5.3) software will be used to conduct the direct meta-analysis. RESULTS: This study will provide a comprehensive review of the available evidence for the treatment of chemotherapy-induced gastrointestinal adverse reaction with moxibustion. CONCLUSION: The conclusion of this study will provide evidence to judge whether moxibustion is an effective and safety therapeutic intervention for chemotherapy-induced gastrointestinal adverse reaction. PROSPERO REGISTRATION NUMBER: CRD42020182990.