How to simulate future scenarios of urban stormwater management? A novel framework coupling climate change, urbanization, and green stormwater infrastructure development.

Climate change, urbanization, and green stormwater infrastructure (GSI) planning policies lead to uncertainties in future urban sustainability. Coupling multiple influencing factors such as climate change, urbanization, and GSI development, this study proposes a novel framework for simulating future scenarios of urban stormwater. Subsequently, the changes in annual surface runoff and runoff pollutants in Shanghai's new and old urban areas were compared and analyzed based on 35 typical future and seven baseline scenarios. The following results were obtained: 1) The runoff control rate of the new urban area was significantly higher than that of the old urban area before GSI construction. After GSI construction, both areas could control stormwater runoff and pollutants, while the decline in efficiency in GSI facilities enormously impacted the old area. 2) Surface runoff in the new urban area was mainly affected by urbanization, while climate change was a major factor in the old urban area; runoff pollutants in new and old urban areas were mainly affected by urbanization, and the change in pollutants in new areas was more pronounced. 3) GSI facilities were unlikely to guarantee the quantity and quality of water resources, especially in scenarios where the efficiency of GSI facilities decreases. In old urban areas, the more extreme climate change and urbanization were, the more significant the effect of improving stormwater management facilities. Our findings showed that future studies on stormwater management should specifically consider the different characteristics of new and old urban regions, pay attention to the maintenance and management of GSI facilities, and build adaptive strategies to cope with climate change, urbanization, and GSI facility destruction.

Induction, Flavonoids Contents, and Bioactivities Analysis of Hairy Roots and True Roots of Tetrastigma hemsleyanum Diels et Gilg.

The flavonoids in Tetrastigma hemsleyanum Diels et Gilg (T. hemsleyanum) have high medicinal value. However, because of slow growth and harsh ecological environments, T. hemsleyanum is currently an endangered species. In light of this, we present a detailed hairy root induction procedure as a promising alternative to true roots with medicinal value. The percentage of explants induced by Agrobacterium rhizogenes (A. rhizogenes) to produce hairy roots out of the total number of explants infected (induction rate 1) was 95.83 ± 7.22%, and the proportion of hairy roots that contained Rol B fragments among all the hairy roots with or without Rol B fragments (positive rate) was 96.57 ± 1.72%. The transformation was further confirmed by the expression of the GUS protein. A high-productive hairy root line was screened for the comparative profiling of six flavonoids with true roots using high-performance liquid chromatography (HPLC). The contents of (+)-catechin, (-)-epicatechin, neochlorogenic acid, luteolin-6-C-glucoside, and orientin were 692.63 ± 127.24, 163.34 ± 31.86, 45.95 ± 3.46, 209.68 ± 6.03, and 56.82 ± 4.75 µg/g dry weight (DW) of 30-day-old hairy roots, respectively, which were higher than those of 3-year-old true roots. Hairy roots have stronger antioxidant activity than true roots. Overall, the hairy roots of T. hemsleyanum could serve as promising alternative sources for the production of flavonoids with medicinal uses.

Phylogenetic Analysis of R2R3-MYB Family Genes in Tetrastigma hemsleyanum Diels et Gilg and Roles of ThMYB4 and ThMYB7 in Flavonoid Biosynthesis.

Tetrastigma hemsleyanum Diels et Gilg (T. hemsleyanum) is an extensively used Chinese folk herb with multiple bioactivities. Among these bioactivities, flavonoids are recognized as the representative active ingredients. We previously found an elevated accumulation of flavonoids in T. hemsleyanum under water stress; however, the mechanism remains unclear. R2R3-MYB transcription factors play vital roles in the plant response to environmental stress and the regulation of secondary metabolites. Herein, a systematic transcriptome identification of R2R3-MYB family genes under water stress in T. hemsleyanum was performed to explore their potential function in the biosynthesis of flavonoids. A total of 26 R2R3-MYB genes were identified, most of which were clustered into functional branches of abiotic stress. ThMYB4 and ThMYB7 were then screened out to be associated with the biosynthesis of flavonoids through a protein-protein interaction prediction. An expression correlation analysis based on RNA-seq further confirmed that ThMYB4 and ThMYB7 were positively related to the flavonoid biosynthetic pathway genes of T. hemsleyanum. In ThMYB4- and ThMYB7-overexpression hairy roots, it was found that the expression of ThCHS and ThCHI was significantly increased, suggesting that ThMYB4 and ThMYB7 may act as regulators in flavonoid biosynthesis. This will shed new light on the promotion of flavonoid production and the medicinal value of T. hemsleyanum by manipulating transcription factors.

A new strategy to improve Ganoderma polysaccharides production by symbiotic fungi elicitors through activating the biosynthetic pathway.

Ganoderma lucidum polysaccharides (GLP) attract growing attention due to their remarkable bioactivities, but the low content in raw materials remains a bottleneck severely restricting their application. We previously found a higher polysaccharides accumulation in Ganoderma lucidum cultured in continuous cropping soil, and soil symbiotic fungi are presumed as the key among many factors. Herein, 33 symbiotic fungi were isolated from the soil, and fungal elicitors were prepared to investigate their biotic eliciting effect on GLP biosynthesis. Most elicitors were found to significantly improve GLP production, among which the NO.16 molecularly identified as Penicillium citrinum, exhibited the optimum eliciting effect with GLP yield increasing by 3.4 times. Differences in the biosynthetic pathway genes expressions and the monosaccharide components of GLP were further analyzed. The transcriptions of the main genes of GLP biosynthetic pathway were up-regulated under PCE treatments, suggesting it improves GLP production by activating transcriptions of the biosynthetic pathway genes. Moreover, PCE eliciting significantly altered the monosaccharide compositions of GLP with Gal, Man, GalA, GlcA, and Fuc increasing by 8.17 %, 5.68 %, 5.41 %, 2.66 %, and 1.51 % respectively, but Glc decreased by 23.43 %, which may result in the activity change. It can serve as a new strategy to improve GLP production.

Lucidenic acid A inhibits the binding of hACE2 receptor with spike protein to prevent SARS-CoV-2 invasion.

High infection caused by mutations of SARS-CoV-2 calls for new prevention strategy. Ganoderma lucidum known as a superior immunoenhancer exhibits various antiviral effects, whether it can resist SARS-CoV-2 remains unclear. Herein, virtual screening combined with in vitro hACE2 inhibition assays were used to investigate its anti SARS-CoV-2 effect. Potential 54 active components, 80 core targets and 20 crucial pathways were identified by the component-target-pathway network. The binding characters of these components to hACE2 and its complexes with spike protein including omicron variant was analyzed by molecular docking. Lucidenic acid A was selected as the top molecule with high affinity to all receptors by forming hydrogen bonds. Molecular dynamics simulation showed it had good binding stability with the receptor proteins. Finally, in vitro FRET test demonstrated it inhibited the hACE2 activity with IC50 2 µmol/mL. Therefore, lucidenic acid A can prevent the virus invasion by blocking hACE2 binding with SARS-CoV-2.

Effect of Continuous Cropping Soil on the Biosynthesis of Bioactive Compounds and the Antioxidant Activity of Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes).

Continuous monoculture can increase plant morbidity and reduce crop yields. However, the Ganoderma lucidum mushroom possesses unique nutrient acquisition methods and displays an altered response to the generation of reactive oxygen species (ROS). The effects of continuous cropping on G. lucidum remain unclear. Here, we investigated the effects of consecutive monoculture soil on the yield, bioactive components, antioxidant properties, nutrient metabolism, and enzyme activity of G. lucidum. The results showed no significant differences in terms of yield between the control check and treatment groups after 2 years. However, the content of polysaccharides and triterpenoids after 2 years of treatment and the content of total phenols and flavonoids after 3 years of treatment significantly increased. Moreover, the antioxidant activity (AOA) of the polysaccharides under consecutive monoculture soil treatments also significantly increased. These differences were caused by changes in nutrient metabolism and the activities of antioxidant and ligninolytic enzymes. Under continuous monoculture soil treatment, the content of reducing sugars, water-soluble sugars, free amino acids, and water-soluble proteins significantly increased. The activities of superoxide dismutase, manganese peroxidase, and lignin peroxidase increased with the time of continuous monoculture, while the activities of peroxidase, catalase, and laccase decreased. The increase in ligninolytic enzymes was postulated to mediate the ROS scavenging ability of G. lucidum. In conclusion, continuous cropping soil could increase the levels of bioactive components and AOAs of G. lucidum.

Anti-inflammatory effects of Ganoderma lucidum sterols via attenuation of the p38 MAPK and NF-κB pathways in LPS-induced RAW 264.7 macrophages.

Ganoderma lucidum exhibits pronounced anti-inflammatory effects, polysaccharides and triterpenoids are regarded as major constituents displaying the anti-inflammatory activities, whether sterols contribute to this activity remains unclear. Herein Ganoderma lucidum sterols (GLS) were innovatively isolated by a single-step procedure, the profile of GLS was characterized by HPLC-ELSD and shown similar to that of sterols separated by a traditional method, but much higher in content. Furthermore, GLS inhibited inflammation in macrophages by significantly attenuating LPS-induced cell polarization as well as releases and mRNA expressions of pro-inflammatory mediators NO, TNF-α, IL-1ß and IL-6. Moreover, the anti-inflammatory activity of GLS was mediated by MAPK and NF-κB pathways, GLS suppressed MAPK pathways by blocking phosphorylation of p38 but not ERK and JNK, which is complementary with inhibitory effects of Ganoderma polysaccharides and triterpenes on JNK and ERK, indicating Ganoderma sterols may exert synergistic anti-inflammatory effect with polysaccharides and triterpenes. GLS also inhibited NF-κB pathways by restraining phosphorylation and degradation of IκB-α and blocking phosphorylation of NF-κB p65. Molecular docking confirmed that sterols of GLS were directly bound to active sites of p38 and p65 to suppress their activation. Therefore, our findings suggest GLS as natural and safe anti-inflammatory agents to prevent and treat inflammatory diseases.

Effects of high levels of nitrogen and phosphorus on perennial ryegrass (Lolium perenne L.) and its potential in bioremediation of highly eutrophic water.

The conservation and management of rain and water in a city environment are a crucial aspect of sustainability. Sponge cities are a solution for the remediation of excessive water consumption, high pollution, and rainstorm waterlogging in urban areas. Perennial ryegrass (Lolium perenne L.) is a common species and promising bioremediation plant for the construction of sponge cities. It could be useful to purify high nitrogen and phosphorus content in wastewater runoff, but its response to high total nitrogen (TN) and total phosphorus (TP) levels has not been studied. The present study aimed to evaluate the effects of high TN + TP level on seed germination, plant growth, antioxidant response, and assimilation rate of N and P in perennial ryegrass. Compared with the control, treatments with 20 mg/L N + 4 mg/L P (20N4P) significantly decreased germination potential at day 3, but the germination rate at day 10 was similar between all treatments and the control, suggesting that a high TN + TP concentration might delay germination, but not prevent it. Treatment with 20N4P also induced significant withering and decreased shoot length, root length, and dry shoot weight of ryegrass seedlings, compared with the control. Treatments with 80N16P significantly decreased levels of chlorophyll a (Chl-a) and chlorophyll b (Chl-b), demonstrating inhibition of photosynthesis. Compared with the control, treatment with 40N8P increased the carotenoid (Car), malondialdehyde (MDA), and glutathione (GSH) content, which might alleviate oxidative stress induced by high TN + TP concentration. Moreover, high levels of TN + TP (160N32P and 320N64P) significantly reduced the removal capacity of N and P by plants. Overall, the present results provide a theoretical foundation for the development of ryegrass for bioremediation of wastewater during construction of sponge cities.

Growth, physiological and transcriptomic analysis of the perennial ryegrass Lolium perenne in response to saline stress.

Salinization of soil is a global environmental concern. To bioremediate or use saline-alkali lands, most studies focused on screening of halophytes and breeding of saline-tolerant non-halophyte vegetables. Seldom studies investigated effects of salinity on general landscape plants, which are important for landscape construction in urban areas. In the present study, effects of salinity on seed germination and seedling growth of the perennial ryegrass Lolium perenne were investigated. The final seed germination rate was not affected at salinity up to 6.4‰. Partial seedlings wilted in all saline treatments and the mortality of L. perenne was positively correlated with salinity. Treatments with salinity equal to or lower than 1.6‰ did not affect length and dry weight of shoot and root. These results suggested that L. perenne could be sowed and then grow well in low-salinity areas. To explore the underlying physiological mechanisms, contents of photosynthetic pigments and antioxidant indices were determined. The results showed that contents of chlorophyll a, b and carotenoid significantly decreased in all saline treatments, in comparison to the control. Similarly, activities of superoxide dismutase and peroxidase decreased and contents of glutathione and malondialdehyde increased in saline treatments. Additionally, transcriptome analysis identified 792 differentially expressed genes (DEGs) in L. perenne shoots between 6.4‰ saline treatment and the control. Compared with the control, genes in relation to iron transportation and amino acid metabolism were downregulated, but genes participating in energy metabolism were upregulated. These changes would inhibit toxicity of ion accumulation and provide more energy for plants to resist saline stress.

Evaluation of antioxidant-associated efficacy of flavonoid extracts from a traditional Chinese medicine Hua Ju Hong (peels of Citrus grandis (L.) Osbeck).

ETHNOPHARMACOLOGICAL RELEVANCE: Hua Ju Hong (HJH, peels of Citrus grandis (L.) Osbeck) is a popularly used traditional Chinese medicine recorded by "Compendium of Materia Medica" (Ben Cao Gang Mu) in Ming Dynasty of China (1578 A.D.). With flavonoid components, HJH possesses hypolipidemic effect associated with antioxidation action. However, no report was found regarding the flavonoid profile and antioxidant activity of HJH. MATERIALS AND METHODS: Five purified flavonoid extracts (TFCA, TFCB, TFCC, TFCD and TFCE.) were obtained from HJH using Ca(OH)2 and HPD-300 macroporous resins, and their total flavonoids and representative flavonoid components were analyzed. In vitro tests of DPPH free radical scavenging activity, reducing power, and total antioxidant activity of each extract were evaluated. The most effective extract was selected for in vivo antioxidative evaluation using a rat hyperlipemia model. RESULTS: The total flavonoid content was varying among each HJH extract and decreased in an order of TFCB>TFCD>TFCC>TFCE>TFCA. TFCB, TFCD, and TFCC contained more than 50% of total flavonoids, the highest content of which was found in TFCB (80.7%). HPLC analysis showed that the contents of three flavonoid components, narirutin, naringin and neohesperidin, displayed a similar trend as that of total flavonoids. In vitro antioxidative tests determined that TFCB at 0.24 to 1.2mg/ml possessed the most significant antioxidant effects among other extracts and was also superior to BHT. In vivo experiment also revealed the significant antioxidant and antihyperlipidemic activities of TFCB started from 50 to 200mg/kg after oral administration to hyperlipemia rats. These results indicate that TFCB with the highest flavonoid contents has the strongest antioxidant-associated activities. CONCLUSION: This is the first report regarding antioxidant-associated activities and relevant flavonoid components of HJH extracts, providing a promising candidate of traditional Chinese medicine for antioxidative treatment.

Antitumor and immunomodulatory activity of polysaccharides from the roots of Actinidia eriantha.

AIM OF THE STUDY: The roots of Actinidia eriantha Benth (Actinidiaceae) have been used for cancers in the Chinese folk medicine. The present study aimed at evaluating the antitumor potentials of the polysaccharides from the roots of Actinidia eriantha and elucidating their immunological mechanisms by determining the effects on the growth of tumor transplanted in mice and the immune response in tumor-bearing mice. MATERIALS AND METHODS: The total polysaccharide AEP and fours purified polysaccharides AEPA, AEPB, AEPC and AEPD were isolated and purified from the roots of Actinidia eriantha by hot water extraction, ethanol precipitation, dialysis and gel filtration. Their effects on the growth of mouse transplantable tumor, splenocyte proliferation, the activity of natural killer (NK) cells and cytotoxic T lymphocytes (CTL), production of cytokines from splenocytes, and serum antigen-specific antibody levels in tumor-bearing mice were measured. RESULTS: AEP and four purified polysaccharides could not only significantly inhibit the growth of mouse transplantable tumor, but also remarkably promote splenocytes proliferation, NK cell and CTL activity, IL-2 and IFN-gamma production from splenocytes, and serum antigen-specific antibody levels in tumor-bearing mice. CONCLUSIONS: The antitumor activity of AEP and four purified polysaccharides might be achieved by improving immune response, and the composition of the monosaccharides, uronic acid contents and molecular weight could affect their antitumor and immunomodulatory activity.

Novel polysaccharide adjuvant from the roots of Actinidia eriantha with dual Th1 and Th2 potentiating activity.

The plant polysaccharides are recognized as an effective biological response modifier with low toxicity. In this study, the water-soluble polysaccharide from the roots of Actinidia eriantha (AEPS) was evaluated for its toxicity and adjuvant potential on the specific cellular and humoral immune responses to ovalbumin (OVA) in mice. AEP did not cause any mortality and side effects when mice were administered subcutaneously twice at the dose up to 5.0mg at intervals of 7 days. The mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 microg dissolved in saline containing Quil A (10 microg) or AEPS (25, 50, or 100 microg) on days 1 and 15. Two weeks later, splenocyte proliferation, natural killer (NK) cell activity, production and mRNA expression of cytokines from splenocytes, and serum OVA-specific antibody titers were measured. The Con A-, LPS-, and OVA-induced splenocyte proliferation and the serum OVA-specific IgG, IgG1, IgG2a, and IgG2b antibody titers in the immunized mice were significantly enhanced by AEPS (P<0.05, P<0.01 or P<0.001). AEPS also significantly promoted the production of Th1 (IL-2 and IFN-gamma) and Th2 (IL-10) cytokines and up-regulated the mRNA expression of IL-2, IFN-gamma, IL-4 and IL-10 cytokines and T-bet and GATA-3 transcription factors in splenocytes from the immunized mice (P<0.05, P<0.01 or P<0.001). Besides, AEPS remarkably increased the killing activities of NK cells from splenocytes in the immunized mice (P<0.01 or P<0.001). The results indicated that AEPS had strong potential to increase both cellular and humoral immune responses and elicit a balanced Th1/Th2 response, and that AEPS may be a safe and efficacious adjuvant candidate suitable for a wide spectrum of prophylactic and therapeutic vaccines.