Ginger polysaccharide alleviates the effects of acute exposure to carbonate in crucian carp (Carassius auratus) by regulating immunity, intestinal microbiota, and intestinal metabolism.

Alkaline stress poses a significant challenge to the healthy growth of fish. Ginger polysaccharide (GP) is one of the main active substances in ginger and has pharmacological effects, such as anti-oxidation and immune regulation. However, the physiological regulatory mechanism of GP addition to diet on alkalinity stress in crucian carp remains unclear. This study aimed to investigate the potential protective effects of dietary GP on antioxidant capacity, gene expression levels, intestinal microbiome, and metabolomics of crucian carp exposed to carbonate (NaHCO3). The CK group (no GP supplementation) and COG group (NaHCO3 stress and no GP supplementation) were set up. The GPCS group (NaHCO3 stress and 0.4% GP supplementation) was stressed for seven days. Based on these data, GP significantly increased the activities of total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX), acid phosphatase (ACP), and alkaline phosphatase (AKP) in carp under alkalinity stress (p < 0.05) and decreased the activity of malon dialdehyde (MDA) (p < 0.05). GP restored the activity of GSH-PX, ACP, and AKP to CK levels. The expression levels of tumor necrosis factor ß (TGF-ß), tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), and interleukin 8 (IL-8) genes were decreased, and the expression levels of determination factor kappa-B (NF-κB) and interleukin 10 (IL-10) genes were increased (p < 0.05). Based on 16 S rRNA high-throughput sequencing, GP improved the changes in the intestinal microbial diversity and structural composition of crucian carp caused by NaHCO3 exposure. In particular, GP increased the relative abundance of Proteobacteria and Bacteroidetes and decreased the relative abundance of Actinobacteria. The metabolic response of GP to NaHCO3 exposed crucian carp guts was studied using LC/MS. Compared to the COG group, the GPCS group had 64 different metabolites and enriched 10 metabolic pathways, including lipid metabolism, nucleotide metabolism, and carbohydrate metabolism. The addition of GP to feed can promote galactose metabolism and provide an energy supply to crucian carp, thus alleviating the damage induced by alkalinity stress. In conclusion, GP can mitigate the effects of NaHCO3 alkalinity stress by regulating immune function, intestinal flora, and intestinal metabolism in crucian carp. These findings provide a novel idea for studying the mechanism of salt-alkali tolerance in crucian carp by adding GP to feed.

Dietary supplementation with Lycium barbarum polysaccharides conducive to maintaining the health of Luciobarbus capito via the enhancement of enzyme activities and the modulation of gut microbiota.

Lycium barbarum polysaccharide (LBP) is the main active component of Lycium barbarum (L. barbarum), which has important medicinal and nutritional value. However, the effect of LBP treatment on Luciobarbus capito (L. capito) still remains unknown. Given this, the current work aims to probe the underlying effect of different levels of LBP treatment (i.e. 0.10, 0.50 and 1.00 g/L) on L. capito in the context of enzymatic activity analysis, histological observations and gut microbiota analysis. Compared with control group, the activities of hepatic antioxidant enzymes, intestinal digestive enzymes and hepatic immune enzyme were found to be significantly increased after 0.10 g/L LBP and 0.50 g/L LBP treatment (P < 0.05). This result indicated that moderate levels of LBP treatment could dramatically enhance the immunity and antioxidant capacity of L. capito. Furthermore, the compositional structures of the gut microbiota in L. capito were found to be greatly shaped after LBP treatment, whereas the diversity and abundance of the gut microbiota were only found to be slightly changed (P > 0.05). No significant changes were screened in the morphologic structures of gut constructions. This work would provide theoretical and experimental basis for future application of LBP as supplement in the culture process of the farmed fish.

Assembly of Celastrol to Zeolitic Imidazolate Framework-8 by Coordination as a Novel Drug Delivery Strategy for Cancer Therapy.

Celastrol (Cel), a compound derived from traditional Chinese medicine Tripterygium wilfordii Hook. F, has attracted considerable attention as an anticancer drug. However, its clinical application is limited due to its low bioavailability and potential toxicity. With the advancement of nanoscale metal organic frameworks (MOF), the nano-delivery of drugs can effectively improve those disadvantages. Nevertheless, hydrophobic drugs apparently cannot be encapsulated by the hydrophilic channels of MOF-based drug delivery systems. To address these issues, a new assembly strategy for hydrophobic Cel was developed by coordinating the deprotonated Cel to zeolitic imidazolate framework-8 (ZIF-8) with the assistance of triethylamine (Cel-ZIF-8). This strategy greatly elevates the assembly efficiency of Cel from less than 1% to ca. 80%. The resulted Cel-ZIF-8 remains stable in the physiological condition while dissociating and releasing Cel after a 45-minute incubation in an acidic tumor microenvironment (pH 5.5). Furthermore, Cel-ZIF-8 is proved to be easily taken up by cancer cells and exhibits a better therapeutic effect on tumor cells than free Cel. Overall, the Cel-ZIF-8 provides a novel assembly strategy for hydrophobic drugs, and the findings are envisaged to facilitate the application of Cel in cancer therapies.

Effects of Dangshen Yuanzhi Powder on learning ability and gut microflora in rats with memory disorder.

ETHNOPHARMACOLOGICAL RELEVANCE: Yuanzhi Powder is a commonly used traditional Chinese medical formulae for its potency in enhancing memory and learning. In clinical practice, Yuanzhi Powder is a classic formula in TCM to treat amnesia of the type "deficiency of Qi, turbid phlegm harasses the head and eyes, and stagnation of phlegm converting into the fire". Our previous study showed that Yuanzhi Power, used together with Codonopsis Radix (Dangshen Yuanzhi Power, DYP), could improve learning and memory ability in animals with memory disorder (MD) and its efficacy is superior or equivalent to that of the Yuanzhi Power. AIM OF STUDY: This study aimed to explore the regulatory mechanism of DYP through the "bacteria-gut-brain axis". MATERIALS AND METHODS: The SD rats were divided randomly into control, model, positive, DYP-L, and DYP-H groups. Except for the control group, the rats were intraperitoneally injected with D-Gal (400 mg/kg) and gavaged with aluminum chloride (200 mg/kg) every day for 50 days. The rats in the DYP group were gavaged with DYP (6.67 and 13.34 g/kg, respectively) from the 15th day, once a day. The rats in the positive group were similarly administrated with piracetam (0.5 g/kg). The rats' bodyweight was recorded from the 16th day. The learning and memory ability of animals was tested by Morris water maze. The levels of MCP-1, NF-L, NSE, and TNF-α in serum were determined by Elisa kit, while the histopathology of duodenum and colon tissues was examined by H & E staining. The diversity of intestinal flora was sequenced and analyzed. In order to reveal the role of intestinal flora in DYP treatment of MD, the intestinal flora composition and the correlation analysis of intestinal flora and the above biochemical indexes were investigated. The intestinal flora function and biological metabolic pathways were predicted and analyzed by the KEGG database. RESULTS: The MD animals' learning and spatial memory ability decreased significantly, compared with the normal group, accompanied by weight increase and intestinal flora disorder. DYP can improve the learning and memory ability of MD animals, and its efficacy may exert through the following ways: (i) callback the abnormal biochemical indexes of MCP-1, NF-L, NSE, and TNF-α; (ii) decreasing the relative ratio of Firmicutes/Bacteroidetes and repairing the pathology of MD animal intestinal mucosa; and (iii) the regulation of DYP on biochemical blood indexes of MD animals was significantly correlated with the regulation of intestinal flora; (iv) DYP rats showed a strong correlation between cognitive ability improvement and bodyweight loss; (v) besides, DYP could also regulate the metabolic pathways of carbohydrate, amino acid, nucleotide, and energy by affecting related biological functions. CONCLUSIONS: The results supported that DYP can improve MD animals' learning and memory ability by restoring the intestinal flora disorder and callback the abnormal biochemical indexes in serum, closely related to the "bacteria-gut-brain axis".

Integrated lipidomics and network pharmacology analysis of the protective effects and mechanism of Yuanzhi San on rats with cognitive impairment.

Cognitive impairment (CI) can seriously affect people's mental and physical health. Yuanzhi San (YZS) is a classic prescription for treating CI, but the mechanisms need further exploration. The aim of this study is to explore the effect of YZS on promoting the learning and memory ability of CI rats induced by d-galactose combined with aluminum chloride. Behavioral experiments had been used to comprehensively evaluate the established CI model. Brain histological morphology and the expressions of calcium ion signaling pathway related factors in serum were used to evaluate the effect of YZS against CI. Lipids in rat serum were analyzed by ultra-performance liquid chromatography-mass spectrometry (UHPLC-MS) and chemical pattern recognition methods. Network pharmacology was used to find potential chemical compounds, targets, and related signaling pathways against CI with treatment of YZS. The integrated lipidomics and network pharmacology analysis were conducted by Cytoscape software. The results showed that YZS could alleviate neurodegenerative impairment. It was verified that model rats had longer latency time, shorter exploration paths, lower new objects recognition indexes, and shorter exercise time and distances compared with the normal rats in behavioral experiments, indicating that the model rats were successfully established. Rats of YZS 6.67 had significant differences in retention time (p < 0.05), number of entrances (p < 0.01), new object recognition indexes (p < 0.05, p < 0.01), exercise time (p < 0.05), and content of Ca [2]+, CAM, APP, CREB (p < 0.01), CAMK2 (p < 0.05). Rats of YZS 6.67 had five cell layers in hippocampus histological morphology. Behavioral experiments results showed that YZS had an active effect on CI rats. From lipidomics analysis, 129 lipids were screened out by conditions of VIP > 1 and p < 0.05, and 17 lipid markers were identified from the databases, which were divided mainly into five types. Pathway analysis indicated that linoleic acid, α-linolenic acid, arachidonic acid, and glycerophospholipid metabolisms were potential target pathways closely involved in the mechanism YZS's effects against CI. Network pharmacology focused on 84 chemical compounds, 130 intersection targets, and 10 hub genes of YZS's effects against CI. Six hub genes and four lipid compounds had intrinsic contact with arachidonic acid metabolism, glycerophospholipid metabolism and linoleate metabolism. The study revealed that YZS could improve animal cognitive behaviors, the expression of factors associated with memory in serum and the histological morphology of hippocampus. Four lipid compounds, three metabolic pathways, and six hub genes of YZS could effectively modulated CI. These results collectively suggest that the main mechanism of YZS in improving CI involves lipid metabolism, which affects biological processes and targets of action in the body.

A Network Pharmacology to Explore the Mechanism of Astragalus Membranaceus in the Treatment of Diabetic Retinopathy.

BACKGROUND: Diabetic retinopathy (DR) includes a series of typical lesions affected by retinal microvascular damage caused by diabetes mellitus (DM), which not only seriously damages the vision, affecting the life's quality of patients, but also brings a considerable burden to the family and society. Astragalus Membranaceus (AM) is a commonly used medicine in clinical therapy of eye disorders in traditional Chinese medicine (TCM). In recent years, it is also used for treating DR, but the specific mechanism is unclear. Therefore, this study explores the potential mechanism of AM in DR treatment by using network pharmacology. METHODS: Based on the oral bioavailability (OB) and drug likeness (DL) of two ADME (absorption, distribution, metabolism, excretion) parameters, Traditional Chinese Medicine Systems Pharmacology Database (TCMSP), Swiss Target Prediction platform, GeneCards, and OMIM database were used to predict and screen the active compounds of AM, the core targets of AM in DR treatment. The Metascape data platform was used to perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis on the core targets. RESULTS: 24 active compounds were obtained, such as quercetin, kaempferol, and astragaloside IV. There were 169 effective targets of AM in DR treatment, and the targets were further screened and finally, 38 core targets were obtained, such as VEGFA, AKT1, and IL-6. EGFR tyrosine kinase inhibitor resistance, AGE-RAGE signaling pathway in diabetic complications, PI3K-Akt signaling pathway, and other metabolic pathways participated in oxidative stress, cell apoptosis, angiogenesis signal transduction, inflammation, and other biological processes. CONCLUSION: AM treats DR through multiple compounds, multiple targets, and multiple pathways. AM may play a role in the treatment of DR by targeting VEGFA, AKT1, and IL-6 and participating in oxidative stress, angiogenesis, and inflammation.

Traditional Chinese medicine Xiaosheng Powder for dry eye disease: A protocol for systematic review and meta analysis.

BACKGROUND: Dry eye disease (DED) has shown a significant increase in recent years, which seriously affects people's work and life. Xiaosheng Powder, a traditional Chinese medicine decoction, has been widely used in treating DED. However, there is no systematic review of the results of the study on this therapeutic effect. The purpose of this review is to evaluate the effectiveness and safety of Xiaosheng Powder in the treatment of DED. METHODS AND ANALYSIS: The electronic databases to be searched will include MEDLINE (PubMed), Cochrane Central Register of Controlled Trials in the Cochrane Library, Excerpta Medica Database, China National Knowledge Infrastructure, China Scientific Journal Database, Wanfang Database and Chinese Biomedical Literature Database. Papers in English or Chinese published from inception to 2020 will be included without any restrictions. Improvement in Ocular Surface Disease Index will be assessed as the primary outcomes. Tear break-up time, Schirmer I test, fluorescent, adverse events, and the recurrence rate after at least 3 months of the treatment will be evaluated as secondary outcomes. We will conduct a meta-analysis of randomized controlled trial if possible. The methodological qualities, including the risk of bias, will be evaluated using the Cochrane risk of bias assessment tool, while confidence in the cumulative evidence will be evaluated using the Grading of Recommendations Assessment, Development, and Evaluation approach. ETHICS AND DISSEMINATION: It is not necessary for a formal ethical approval because the data is not individualized. The results of this review will offer implications for the use of Xiaosheng Powder as a treatment for DED. This knowledge will inform recommendations by ophthalmologist and researchers who are interested in the treatment of DED. The findings of this systematic review will be disseminated through peer-reviewed publications and conference presentations. TRAIL REGISTRATION NUMBER: PROSPERO CRD42020147709.

Quality evaluation of Panax ginseng adventitious roots based on ginsenoside constituents, functional genes, and ferric-reducing antioxidant power.

In the study, six adventitious root lines of Panax ginseng have been successfully established. HPLC-ESI-MS analysis showed that 20 ginsenosides were identified in root lines, notoginsenoside Fa and notoginsenoside R2 were not found in AR lines. In AR lines, the highest accumulation of total ginsenosides was obtained in five-year main AR (24.87 mg/g). Principal component analysis classified root lines into three groups. Five-year ginseng was mostly similar with five-year main AR, five-year rootlet AR, and four-year rootlet AR in ginsenosides composition of group 1. Besides, gene expressions were consistent with the production of total ginsenosides, and correlation analysis revealed that total ginsenosides biosynthesis was significantly positively correlated with the gene expression of dammarenediol synthase. Five-year rootlet AR showed the highest activity on ferric-reducing antioxidant power test among samples. It provides a scientific evidence for the further exploitation and large-scale production of P. ginseng. PRACTICAL APPLICATIONS: This study provides valuable information for the commercial scale culture of ginseng adventitious roots. This report combines morphology, ginsenoside composition and content, gene expression, and ferric-reducing antioxidant power test to evaluate the quality of P. ginseng adventitious root, and combined with principal component analysis to screen out the high yield and stable ginseng adventitious roots. It would be profitable to use adventitious root culture of P. ginseng instead of field cultivation.

Effect of temperature on morphology, ginsenosides biosynthesis, functional genes, and transcriptional factors expression in Panax ginseng adventitious roots.

This study researched the effect of temperature on growth and ginsenosides accumulation in adventitious root cultures of Panax ginseng. Results showed that the ginseng adventitious roots growth and differentiation ability could be affected faced with different incubation temperatures (15, 20, 25, and 30°C for 35 days). Besides, the research also demonstrated that low-temperature stimulation could promote the accumulation of ginsenosides and the content of total ginsenosides increased by 2.53 times at 10°C-7d (10°C for 7 days and then transferred to 25°C for 28 days) compared with that at 25°C. Moreover, the transcriptional levels of functional genes and PgWRKYs were analyzed by this study and the correlation analysis showed that GPS, SS, CYP716A47, CYP716A53v2, UGT74AE2, UGT94Q2, PgWRKY1, PgWRKY3, and PgWRKY8 were significantly correlated with total ginsenosides content. Furthermore, HPLC-ESI-MSn analyzed that Malonyl-Rb1 only existed in 10°C-7d group. PRACTICAL APPLICATIONS: The survey showed that after a certain time of stimulating P. ginseng adventitious roots at low temperature, the accumulation of ginsenosides could be enhanced as their expression of related genes were regulated. It provides a theoretical foundation for the mass production of ginsenosides by controlling the temperature conditions of P. ginseng adventitious roots.

Screening and evaluation of adventitious root lines of Panax notoginseng by morphology, gene expression, and metabolite profiles.

Panax notoginseng is a commonly used Traditional Chinese Medicine (TCM) and has diverse pharmacological activities with triterpenoid saponins as its main active ingredient. In the cultivation of P. notoginseng, continuous cropping is a serious problem, which could induce reduced productivity, low tuber quality, and plant mortality. With unique advantages of easy control, relative stability, high yields, tissue culture is widely used in the protection of TCM resources. In this study, we screened one adventitious root line, multi-branched (MB) root induced from wild-type roots of P. notoginseng, with a high yield of total triterpenoid saponins (17.92 mg/g). The morphology analysis showed that MB root had structure similar to that of wild-type roots, except for the highly branched phenotype. MB root also showed close gene expression levels and metabolite profiles, which were also similar to those of wild-type roots of Demonstration Park (S3Y). Pearson's correlation coefficient analysis confirmed the importance of key gene, 3-hydroxy-3-methylglutaryl CoA reductase (HMGR), and transcription factor, PnERF1, in regulating triterpenoid saponin biosynthesis in P. notoginseng. These results suggested that MB root possesses potential value in the large-scale cultivation of P. notoginseng.

Characterization of UDP-Glycosyltransferase Involved in Biosynthesis of Ginsenosides Rg1 and Rb1 and Identification of Critical Conserved Amino Acid Residues for Its Function.

Ginsenosides attract great attention for their bioactivities. However, their contents are low, and many UDP-glycosyltransferases (UGTs) that play crucial roles in the ginsenoside biosynthesis pathways have not been identified, which hinders the biosynthesis of ginsenosides. In this study, we reported that one UDP-glycosyltransferase, UGTPg71A29, from Panax ginseng could glycosylate C20-OH of Rh1 and transfer a glucose moiety to Rd, producing ginsenosides Rg1 and Rb1, respectively. Ectopic expression of UGTPg71A29 in Saccharomyces cerevisiae stably generated Rg1 and Rb1 under its corresponding substrate. Overexpression of UGTPg71A29 in transgenic cells of P. ginseng could significantly enhance the accumulation of Rg1 and Rb1, with their contents of 3.2- and 3.5-fold higher than those in the control, respectively. Homology modeling, molecular dynamics, and mutational analysis revealed the key catalytic site, Gln283, which provided insights into the catalytic mechanism of UGTPg71A29. These results not only provide an efficient enzymatic tool for the synthesis of glycosides but also help achieve large-scale industrial production of glycosides.

Advances in ginsenoside biosynthesis and metabolic regulation.

In this paper, we reviewed the advances in ginsenoside biosynthesis and metabolic regulation. To begin with, the application of elicitors in the ginsenoside biosynthesis was discussed. Methyl jasmonate (MJ) and analogues have the best effect on accumulation of ginsenoside compared with other elicitors, and few biotic elicitors are applied in Panax genus plants tissue culture. In addition, so far, more than 40 genes encoding ginsenoside biosynthesis related enzymes have been cloned and identified from Panax genus, such as UDP-glycosyltransferases (UGT) genes UDPG, UGTAE2, UGT94Q2, UGTPg100, and UGTPg1. However, the downstream pathway of the ginsenoside biosynthesis is still not clear. Moreover, some methods have been used to increase the expression of functional genes and ginsenoside content in the ginsenoside synthesis pathway, including elicitors, overexpression, RNAi, and transcription factors. The ginsenoside biosynthesis pathway should be revealed so that ginsenoside contents can be regulated.

Therapeutic effects of lactosyl derivative Gu-4 in a collagen-induced arthritis rat model.

Rheumatoid arthritis (RA) is an inflammatory disorder that is characterized by persistent recurrence of joint inflammation leading to cartilage and bone destruction. The present anti-arthritis therapies failed to achieve satisfactory remission in all patients; therefore, it is still necessary to develop novel approaches to fulfill the demand in clinic. Here, we reported the therapeutic effects of lactosyl derivative Gu-4, a synthetic compound that was previously identified as a selective inhibitor against leukocyte integrin CD11b, in a bovine type II collagen induced arthritis (CIA) rat model. First, prophylactic administration of Gu-4 (1.2728 mg/kg) to rats by intraperitoneal injection every 2 days from the first day of collagen immunization significantly decreased the incidence of CIA, diminished the mean paw volume increase, and reduced the number of swollen paws. Second, administration of Gu-4 (1.2728 mg/kg) to rats at early-onset stage of CIA prevented the progression of the pathological process of RA, accelerated the remission of paw edema, and declined the arthritis score; after 5 weeks treatment, X-ray and histological examinations were carried out, the ankle joint of hind limb of Gu-4 treated CIA rats exhibited slighter bone erosion and much less inflammatory cell infiltration compared to those of saline treated animals; furthermore, Gu-4 remarkably attenuated the production of rheumatoid factor (RF) in the serum of CIA rats as determined by ELISA. Moreover, we performed in vitro lymphocyte proliferation assay and found that Gu-4 significantly inhibited the proliferation of splenic lymphocytes isolated from CIA rats in a dose-dependent manner. Our results suggest that Gu-4 can effectively ameliorate CIA and might be an alternative option for the treatment of RA.

[Growth comparison among Psammosilene tunicoides populations in tissue culture].

The comparison between the growth of eight populations from Psammosilene tunicoides at Yunnan Province was made by the tissue culture. The initial results showed out two populations from Yunshanping (Lijiang) and Xiaomoyu (Kunming) was dominant than orthers. It would be regard as one of fine germplasm resources for the culture of Psammosilence tunicoides.