Neuroprotection and Mechanism of Gas-miR36-5p from Gastrodia elata in an Alzheimer's Disease Model by Regulating Glycogen Synthase Kinase-3ß.

Alzheimer's disease (AD) is currently the most common neurodegenerative disease. Glycogen synthase kinase 3ß (GSK-3ß) is a pivotal factor in AD pathogenesis. Recent research has demonstrated that plant miRNAs exert cross-kingdom regulation on the target genes in animals. Gastrodia elata (G. elata) is a valuable traditional Chinese medicine that has significant pharmacological activity against diseases of the central nervous system (CNS). Our previous studies have indicated that G. elata-specific miRNA plays a cross-kingdom regulatory role for the NF-κB signaling pathway in mice. In this study, further bioinformatics analysis suggested that Gas-miR36-5p targets GSK-3ß. Through western blot, RT-qPCR, and assessments of T-AOC, SOD, and MDA levels, Gas-miR36-5p demonstrated its neuroprotective effects in an AD cell model. Furthermore, Gas-miR36-5p was detected in the murine brain tissues. The results of the Morris water maze test and western blot analysis provided positive evidence for reversing the learning deficits and hyperphosphorylation of Tau in AD mice, elucidating significant neuroprotective effects in an AD model following G. elata RNA administration. Our research emphasizes Gas-miR36-5p as a novel G. elata-specific miRNA with neuroprotective properties in Alzheimer's disease by targeting GSK-3ß. Consequently, our findings provide valuable insights into the cross-kingdom regulatory mechanisms underlying G. elata-specific miRNA, presenting a novel perspective for the treatment of Alzheimer's disease.

Gestational Vitamin E Status and Gestational Diabetes Mellitus: A Retrospective Cohort Study.

OBJECTIVES: To examine the association between vitamin E (VE) status and gestational diabetes mellitus (GDM). METHODS: A retrospective cohort study was conducted by using data of 52,791 women at 137 hospitals across 22 provinces of China. A fasting plasma glucose (FPG) level of ≥5.1 mmol/L between the 24th and 40th weeks of gestation was used as the criteria for the diagnosis of GDM. Mean FPG level and GDM rate were calculated within each combination of the first-trimester VE concentration categories and gestational change categories. The associations of the first-trimester VE concentrations and gestational VE change with FPG and GDM were examined by employing generalized additive models (GAMs). RESULTS: 7162 (13.57%) cases were diagnosed with GDM. The GDM rate was 22.44%, 11.50%, 13.41%, 12.87%, 13.17%, 13.44%, 12.64%, and 14.24% among women with the first-trimester VE concentrations of <7.2, 7.2-7.9, 8.0-9.3, 9.4-11.0, 11.1-13.2, 13.3-15.8, 15.9-17.7, and 17.8-35.9 mg/L, respectively. The GDM rate was 15.96%, 13.10%, 13.64%, and 12.87% among women with gestational VE change of <0, 0-0.19, 0.20-0.29, ≥0.30 mg/L per week, respectively. Multivariable adjusted GAM analyses found that the first-trimester VE concentration was associated with the FPG levels and GDM risk in an L-shaped pattern; the FPG levels and GDM risk decreased sharply to a threshold (around 7 mg/L), and then were keep flat. Gestational VE decreases when the first-trimester VE level was less than 11 mg/L were related to increased FPG levels and GDM risk. CONCLUSIONS: Both low first-trimester VE levels and subsequent gestational VE decrease were related with increased risk of GDM. The findings suggest the necessity of having VE-rich foods and appropriate VE supplementation to prevent GDM for pregnant women with low baseline VE levels.

Daphnetin inhibits spinal glial activation via Nrf2/HO-1/NF-κB signaling pathway and attenuates CFA-induced inflammatory pain.

Daphnetin (7, 8-dihydroxycoumarin, DAPH), a coumarin derivative isolated from Daphne odora var., recently draws much more attention as a promising drug candidate to treat neuroinflammatory diseases due to its protective effects against neuroinflammation. However, itscontribution to chronic inflammatory pain is largely unknown. In the current work, we investigated the effects of DAPH in a murine model of inflammatory pain induced by complete Freund's adjuvant (CFA) and its possible underlying mechanisms. Our results showed that DAPH treatment significantly attenuated mechanical allodynia provoked by CFA. A profound inhibition of spinal glial activation, followed by attenuated expression levels of spinal pro-inflammatory cytokines, was observed in DAPH-treated inflammatory pain mice. Further study demonstrated that DAPH mediated negative regulation of spinal NF-κB pathway, as well as its preferential activation of Nrf2/HO-1 signaling pathway in inflammatory pain mice. This study, for the first time, indicated that DAPH might preventthe development of mechanical allodynia in mice with inflammatory pain. And more importantly, these data provide evidence for the potential application of DAPH in the treatment of chronic inflammatory pain.

Structural and functional organization of the MYC transcriptional factors in Camellia sinensis.

MAIN CONCLUSION: Genome-wide identification, expression analysis of the MYC family in Camellia sinensis, and potential functional characterization of CsMYC2.1 have laid a solid foundation for further research on CsMYC2.1 in jasmonate (JA)-mediated response. Myelocytomatosis (MYC) of basic helix-loop-helix (bHLH) plays a major role in JA-mediated plant growth and developmental processes through specifically binding to the G-box in the promoters of their target genes. In Camellia sinensis, studies on the MYC gene family are limited. Here, we identified 14 C. sinensis MYC (CsMYC) genes, and further analyzed the evolutionary relationship, gene structure, and motif pattern among them. The expression patterns of these CsMYC genes in different tissues suggested their important roles in diverse function in tea plant. Four MYC transcription factors with the highest homology to MYC2 in Arabidopsis were localized in the nucleus. Two of them, named CsMYC2.1 and CsMYC2.2, exhibited transcriptional self-activating activity, and, therefore, could significantly activate the promoter containing G-box motif, whereas CsJAM1.1 and CsJAM1.2 lack the transcriptional self-activating activity, indirectly mediating the JA pathway through interacting with CsMYC2.1 and CsMYC2.2. Furthermore, Yeast Two-Hybrid (Y2H) and Bimolecular Fluorescent Complimentary (BiFC) assays showed that CsMYC2.1 could interact with CsJAZ3/7/8 proteins. Genetically, the complementation of CsMYC2.1 in myc2 mutants conferred the ability to restore the sensitivity to JA signals. The results provide a comprehensive characterization of the 14 CsMYCs in C. sinensis, establishing a solid foundation for further research on CsMYCs in JA-mediated response.

Identification and Investigation of miRNAs From Gastrodia elata Blume and Their Potential Function.

Gastrodia elata Blume (G. elata) is a valuable traditional Chinese medicine with neuroprotection, anti-inflammatory, and immune regulatory functions. MicroRNAs (miRNA) is a kind of endogenous noncoding small RNAs that plays distinctly important roles for gene regulation of organisms. So far, the research on G. elata is mainly focused on the pharmacological functions of the natural chemical ingredients, and the function of G. elata miRNA remains unknown. In this study, 5,718 known miRNAs and 38 novel miRNAs were identified by high-throughput sequencing from G. elata. Based on GO and KEGG analysis, we found that the human genes possibly regulated by G. elata miRNAs were related to the cell cycle, immune regulation, intercellular communication, etc. Furthermore, two novel miRNAs as Gas-miR01 and Gas-miR02 have stable and high expression in the medicinal tissues of G. elata. Further bioinformatics prediction showed that both Gas-miR01 and Gas-miR02 could target Homo sapiens A20 gene, furthermore, the dual-luciferase reporter gene assay and Western Blotting verified the interaction of Gas-miR01 or Gas-miR02 with A20. These evidences suggested that G. elata-unique miRNAs might be involved in certain physiological processes. The animal experiment showed that Gas-miR01 and Gas-miR02 could be detected in some tissues of mice by intragastric administration; meanwhile, the A20 expression in some tissues of mice was downregulated. These results supported for the functional study of G. elata miRNAs.

Core Microbiome of Medicinal Plant Salvia miltiorrhiza Seed: A Rich Reservoir of Beneficial Microbes for Secondary Metabolism?

Seed microbiome includes special endophytic or epiphytic microbial taxa associated with seeds, which affects seed germination, plant growth, and health. Here, we analyzed the core microbiome of 21 Salvia miltiorrhiza seeds from seven different geographic origins using 16S rDNA and ITS amplicon sequencing, followed by bioinformatics analysis. The whole bacterial microbiome was classified into 17 microbial phyla and 39 classes. Gammaproteobacteria (67.6%), Alphaproteobacteria (15.6%), Betaproteobacteria (2.6%), Sphingobacteria (5.0%), Bacilli (4.6%), and Actinobacteria (2.9%) belonged to the core bacterial microbiome. Dothideomycetes comprised 94% of core fungal microbiome in S. miltiorrhiza seeds, and another two dominant classes were Leotiomycetes (3.0%) and Tremellomycetes (2.0%). We found that terpenoid backbone biosynthesis, degradation of limonene, pinene, and geraniol, and prenyltransferases, were overrepresented in the core bacterial microbiome using phylogenetic examination of communities by reconstruction of unobserved states (PICRUSt) software. We also found that the bacterial genera Pantoea, Pseudomonas, and Sphingomonas were enriched core taxa and overlapped among S. miltiorrhiza, maize, bean, and rice, while a fungal genus, Alternaria, was shared within S. miltiorrhiza, bean, and Brassicaceae families. These findings highlight that seed-associated microbiomeis an important component of plant microbiomes, which may be a gene reservoir for secondary metabolism in medicinal plants.

Comparative study on the composition of free amino acids and derivatives in the two botanical origins of an edible Chinese herb "Xiebai", i.e., Allium chinense G. Don and Allium macrostemon Bunge species.

Xiebai is an edible Chinese herb with various health and therapeutic benefits. To evaluate its nutritional and health values, the free amino acids and derivatives of its two botanical origins (i.e., Allium chinense G. Don and Allium macrostemon Bunge) were isolated using a solvent extraction method and analyzed using automatic amino acid analysis and ultra-performance liquid chromatography-quadrupole-time of flight (UPLC-Q-TOF) mass spectrometry. Our data show that both plants contain abundant free amino acids, and the amount of total free amino acids in A. chinense G. Don is higher than that in A. macrostemon Bunge. The free amino acid compositions in the two plants are qualitatively similar, including nineteen proteinogenic and four non-proteinogenic amino acids. The identified proteinogenic amino acids include eight essential amino acids and five semi-essential amino acids. The sum of essential and semi-essential amino acids accounts for 64.9% and 69.7% of the total free amino acids of the two plants, respectively. The principal amino acids of both plants, from highest concentration to lowest concentration, are arginine, glutamine, glutamic acid, asparagine and serine. A. chinense G. Don is also rich in citrulline and lysine. In addition, two amino acid derivatives were identified from the two plants, i.e., the proline analog N­methyl­proline and the dipeptide H-Glu-Tyr-OH. For the first time, the presence of N­methyl­proline in the plants of the Allium genus and the presence of H-Glu-Tyr-OH in unprocessed food sources are reported. The influences of the identified substances on the flavor, nutrition and health values of Xiebai are discussed.

[Teaching research of Molecular Pharmacognosy for talent development in industry of Chinese medicinal materials under new situation].

The industry of Chinese medicinal materials is going through another high-level development stage with some important files issued by Chinese government in the past months, such as "the protection and development plans of Chinese medicinal materials (2015-2020)" and "the strategic development plans of Chinese medicine (2016-2030)". In addition, the effect of "TU Youyou" will not only improve the industry development, but also indicates the increasing international competition intensely. Therefore, one of the core problems of the sustainable-development industry is the training of senior talents under the "New Situation" with opportunity and intense competition. As one of the forefront courses of Chinese Pharmacology, Molecular Pharmacognosy (MP) is a new interdisciplinary science, which integrates the pharmacognosy and molecular biology, and combines many discipline theories and technological systems. MP not only inherits the traditional concepts,but also makes up for the shortages of pharmacognosy, and improves the development of pharmacognosy. Thus, these are importance of MP for cultivation of senior talents, and also the difficult teaching points of MP with no unified teaching mode. We will, in this paper, discuss the possible teaching modes through several aspects for talent cultivation and meeting the needs of social and industry development, such as teaching state of MP, the education of undergraduate and graduate students, social identity, and self renewal of curriculum theories and practice.

[Effects of different salt-alkaline stress on seed germination and physiological characteristics of Hedysarum polybotrys].

OBJECTIVE: In order to get the method for improving the salt resistance of Hedysarum polybotrys seeds and seedlings under different salt-alkaline stress, the seed germination and physiological characteristics of H. polybotrys seedlings were studied. METHOD: Several physiological indexes of H. polybotrys seeds under different salt-alkaline stress, such as the germination vigor, germination rate, relative germination rate, relative salt damage rate were measured. And others indexes of the seedlings like chlorophyll contents, soluble protein contents, the permeability of plasmalemma, the activities of POD and SOD were also measured. RESULT: Different salt-alkaline stress decreased the germination rate, vigor of germinate, germination index, while relative salt damage rate increased. With the increased salt-alkaline concentration, the adverse effects became more obvious. The strength of the salts: Na2CO3 > Na2SO4 > NaCl. With the increase of the salt-alkaline concentration, the chlorophyll contents and the soluble protein contents decreased, but the permeability of plasmalemma increased. The change trend of SOD and POD activity was similar, it is increased firstly, and then decreased as the stress intensity extended, the most significant increase of Na2SO4 and Na2CO3 in the concentration of salt-alkaline was 25 mmol x L(-1), but NaCl was 50 mmol x L(-1). CONCLUSION: The seeds and seedlings inhibition of the salts was Na2CO3 > Na2SO4 > NaCl.

[Study on liver injury of Oncomelania hupensis caused by Eomecon chinanthe sanguinarine].

OBJECTIVE: To analyze the effects of Eomecon chinanthe sanguinarine (SAN) on glucogen, enzyme activity and lipid peroxidation of Oncomelania hupensis liver so as to explore the mechanism of SAN against Oncomelania hupensis. METHODS: SAN was extracted and purified from the dry powder of Eomecon chionantha. Oncomelania hupensis were immersed in 5 mg/L sanguinarine (50 Oncomelania hupensis per 500 ml solution) or clean water at 25 degrees C for 36 h, the livers were isolated from live snails. Total glucogen content, malondialdehyde (MDA) level, activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), acid phosphatase (ACP), alkaline phosphatase (AKP), superoxide dismutase (SOD), peroxidase (POD) were determined respectively and the data were analyzed by independent t test. RESULTS: The glucogen content of snail livers in the SAN group and the control group were (12.151 +/- 0.204) and (18.113 +/- 0.163) mg/g respectively, the difference between the two groups was significant (P < 0.05); the MDA levels of the two groups were (5.298 +/- 0.441) and (4.351 +/- 0.197) nmol/mgprot respectively, and the difference was not significant (P > 0.05); the activities of ALT, AST, ACP, AKP, SOD in the SAN group were (2.760 +/- 0.076) U/mgprot, (68.723 +/- 2.295) U/mgprot, (407.949 +/-19.868) U/gprot, (191.287 +/- 0.771) U/ gprot and (48.452 +/- 0.193) U/mgprot respectively, the activities of these enzymes in the control group were (1.104 +/- 0.000) U/mgprot, (49.448 +/- 1.626) U/mgprot, (344.475 +/- 30.186) U/gprot, (121.905 +/- 3.127) U/gprot and (38.814 +/- 2.765) U/mgprot respectively, the activities of ALT, AST, ACP, AKP and SOD were significantly increased after immersed in 5 mg/L SAN for 36 h, the differences were significant (All P values < 0.05); yet the difference of POD between the SAN group [(22.170 +/- 0.018) U/mgprot] and the control group [(21.747 +/- 0.264) U/mgprot] was not significant (P > 0.05). CONCLUSION: SAN can destroy liver functions of Oncomelania hupensis through decreasing glucogen content and changing activities of some important enzymes in snail liver.

The chemical composition of human milk from Inner Mongolia of China.

In Inner Mongolia, China, the chemical composition of 66 breast milk samples at three lactation stages was analysed. Except for total nitrogen content, the contents of total solid, fat, NPN, lactose and ash were not significantly different between colostral, transitional and mature milk. Fatty acids did not vary over the three lactation stages, while unsaturated fatty acids accounted for 59.95-63.22% of the total fatty acids. Relatively low contents of vitamins were in the milk because the volunteer mothers did not take any vitamin supplementation over the entire lactation period. Besides sodium and phosphate, the concentrations of most minerals in the breast milk remained fairly constant across the three lactation stages.

[Determination of lead in microemulsified rapeseed oil and bio-diesel oil by GFAAS].

Bio-diesel oil has attracted much attention as a substitutable energy sources for its renewable and eco-friendly property. However, problems of lead contamination in fuel are also emphasized increasingly at present. So it was of quite significance to determine the contents of lead in bio-diesel oil and its raw material rapeseed oil. An effective method was developed for the rapid determination of lead in rapeseed oil and bio-diesel oil by graphite furnace atomic absorption spectrometry (GFAAS) after their stabilization as microemulsions. In this research work, polyethyleneglycol octyl phenyl ether and n-butanol were used for emulsifier and auxiliary emulsifying agent, respectively. For Pb, efficient thermal stabilization was obtained using NH4H2PO4 as matrix modifier. Sample stabilization was necessary because of evident analyte losses that occurred immediately after sampling. Excellent long-term sample stabilization and the influence of the microemulsion composition on the GFAAS response were observed by mixing different organic solvents. The ashing and atomization temperature and ramp rate influenced the sensitivity obtained for Ph. Take this into account, the optimum conditions of the graphite furnace atomic absorption spectrometric determination of Pb in rapeseed oil and bio-diesel oil samples were investigated. The results showed that the microemulsion was quite stable when the value of V(20% polyethyleneglycol octyl phenyl ether), V(n-butanol), V(oil) and V(water) was 0.1: 8.9: 0.5: 0.5, without matrix interference effect. The determination limit of the proposed method was 126.2 microg x L(-1) for Pb, comfortably below the values found in the analyzed samples. The recoveries were from 81.8% to 109.0%, which performed using the addition of different concentrations of lead to bio-diesel oil, rapeseed oil and petrochemical diesel samples. The relative standard deviation of determination was 5.84%. This work showed the great efficiency of the microemulsion, indicating that it is possible to extract lead from the oil phase. The method was applied to the determination of lead in oil samples with satisfactory results.