Properties and Fungal Communities of Different Soils for Growth of the Medicinal Asian Water Plantain, Alisma orientale, in Fujian, China.

The Asian water plantain, Alisma orientale (Sam.) Juzep, is a traditional Chinese medicinal plant. The dried tubers of the Alisma orientale, commonly referred to as Alismatis rhizome (AR), have long been used in traditional Chinese medicine to treat a variety of diseases. Soil properties and the soil microbial composition are known to affect the quality and bioactivity of plants. Here, we sought to identify variations in soil fungal communities and soil properties to determine which would be optimal for cultivation of A. orietale. Soil properties, heavy metal content, and pesticide residues were determined from soils derived from four different agricultural regions around Shaowu City, Fujian, China, that had previously been cultivated with various crops, namely, Shui Dao Tu (SDT, rice), Guo Shu Tu (GST, pecan), Cha Shu Tu (CST, tea trees), and Sang Shen Tu (SST, mulberry). As fungi can either positively or negatively impact plant growth, the fungal communities in the different soils were characterized using long-read PacBio sequencing. Finally, we examined the quality of A. orientale grown in the different soils. Our results show that fungal community diversity of the GST soil was the highest with saprotrophs the main functional modes in these and SDT soils. Our data show that GST and SDT soils were most suitable for A. orientale growth, with the quality of the AR tubers harvested from GST soil being the highest. These data provide a systematic approach at soil properties of agricultural lands in need of replacement and/or rotating crops. Based on our findings, GST was identified as the optimal soil for planting A. orientale, providing a new resource for local farmers.

Design of an ultrabroadband infrared metamaterial absorber in the atmospheric transparent window.

In order to meet the demand for broadband absorbers in the infrared transparent window of the atmosphere, we designed an ultrabroadband metamaterial absorber (MA), which is composed of a germanium-zinc sulfide-chromium-zinc sulfide (Ge-ZnS-Cr-ZnS) disk periodic array and a chromium (Cr) bottom layer. In the infrared transparent window of the atmosphere, the average absorption of the MA is as high as 99.1%, and ultrabroadband near-perfect absorption is realized. Moreover, the absorber is polarization independent and insensitive to the angle of incidence. The ultrabroadband and high-absorption metamaterial absorber has broad application prospects in solar cells, photodetectors, thermophotovoltaics, and thermal emitters.

Hypolipidemic effect of Alisma orientale (Sam.) Juzep on gut microecology and liver transcriptome in diabetic rats.

Alisma orientale (Sam.) Juzep (A. orientale) is a traditional herb that is often used to treat disease including edema and hyperlipidemia. However, the molecular mechanism by which Alisma orientale (Sam.) Juzep exerts its hypolipidemic effects remains unclear. In this study, a diabetic rat model was established by feeding a high-fat and high-sugar diet combined with a low-dose streptozotocin injection (HFS). Then the rats were treated with an A. orientale water extract (AOW), an A. orientale ethanolic extract (AOE) or metform (MET). The gut microflora and liver transcriptome were analyzed by high-throughput next-generation sequencing. Ultra-performance liquid chromatography-triple quadrupole-mass spectrometry was employed to analyze the major compounds in the AOE. The results showed that the serum total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) levels in rats of the AOE group (2.10 g/kg/day, 14 days) were significantly lower than those in the HFS group (p<0.01). Moreover, AOE treatment altered the gut microecology, particularly modulating the relative abundance of gut microflora involved in lipid metabolism compared with the HFS group. Furthermore, compared with the HFS group, the mRNA expression levels of Fam13a, Mapk7, Mpp7, Chac1, Insig1, Mcpt10, Noct, Greb1l, Fabp12 and Hba-a3 were upregulated after the administration of AOE. In contrast, the mRNA expression levels of Lox, Mybl1, Arrdc3, Cyp4a2, Krt20, Vxn, Ggt1, Nr1d1 and S100a9 were downregulated. Moreover, AOE treatment for two weeks markedly promoted the relative abundance of Lachnospiraceae (p = 0.0013). The triterpenoids contents in AOE were alisol A, alisol A 24-acetate, alisol B, alisol B 23-acetate, alisol C 23-acetate, alisol F, alisol F 24-acetate, and alisol G. Our findings above illustrated that the hypolipidemic effect of the triterpenoids of A. orientale is mediated mainly through alteration of the gut microecology and the regulation of genes involved in cholesterol metabolism, especially Insig1.

Construction and Analysis of circRNA-miRNA-mRNA Molecular Regulatory Networks During Herba Gelsemium elegans Intoxication.

Gelsemium elegans (Gardner & Champ.) Benth. (GE) has therapeutic effects for pain and malignant tumors but also has high toxicity. Its mechanism of toxicity has not yet been fully clarified, thus limiting its application. Meanwhile, evidence has shown that circRNAs are closely related to the progression of disease. However, very little is known about their expression profiles during intoxication. In this paper, circRNA/mRNA microarrays were respectively performed to detect their expression profiles in mice with acute GE intoxication versus normal controls. CircRNAs were verified by qRT-PCR in subsequent experiments. A regulation pattern of circRNA→miRNA→mRNA was deduced based on intersection analysis of circRNA/mRNA microarrays. The results revealed circRNAs (143) and mRNAs (1,921) were significantly expressed during intoxication. Most of the circRNAs were exonic, and most distributions in chromosomes were transcribed from chr1, chr2, chr7, and chr11. Furthermore, dysregulated expression of mmu-circRNA-013703 and mmu-circRNA-010022 was verified. Then a circRNA-targeted miRNA-mRNA co-expression network was constructed. The network map contained 2 circRNAs, 52 miRNAs, and 752 mRNAs. GO & KEGG analysis further predicted that mmu-circRNA-013703 and mmu-circRNA-010022 may participate in cellular survival/demise-related, neuron/synapse-related, and channel-related pathways. Based on functional modules analysis, a new network was formed, in which mmu-circRNA-013703 VS mmu-miR-361-3p linked to most mRNAs. Most of these mRNAs were known to be involved in the aforementioned functional module. This indicated that mmu-circRNA-013703 functioned as a sponge of miRNAs to regulate the more comprehensive circRNA-miRNA-mRNA co-expression network. Our approach revealed a landscape of dysregulated circRNA-miRNA-mRNA and may be valuable for the identification of new biomarkers during intoxication.

Comparative transcriptome and metabolite profiling of four tissues from Alisma orientale (Sam.) Juzep reveals its inflorescence developmental and medicinal characteristics.

Alisma orientale (Sam.) Juzep (A. orientale) is an important medicinal plant in traditional Chinese medicine. In this study, de novo RNA-seq of A. orientale was performed based on the cDNA libraries from four different tissues, roots, leaves, scapes and inflorescences. A total of 41,685 unigenes were assembled, 25,024 unigene functional annotations were obtained by searching against the five public sequence databases, and 3,411 simple sequence repeats in A. orientale were reported for the first time. 15,402 differentially expressed genes were analysed. The morphological characteristics showed that compared to the other tissues, the leaves had more chlorophyll, the scapes had more vascular bundles, and the inflorescences contained more starch granules and protein. In addition, the metabolic profiles of eight kinds of alisols metabolite profiling, which were measured by ultra-Performance liquid chromatography-triple quadrupole-mass spectrometry showed that alisol B 23-acetate and alisol B were the major components of the four tissues at amounts of 0.068~0.350 mg/g and 0.046~0.587 mg/g, respectively. In addition, qRT-PCR validated that farnesyl pyrophosphate synthase and 3-hydroxy-3-methylglutaryl-CoA reductase should be considered the critical candidate genes involved in alisol biosynthesis. These transcriptome and metabolic profiles of A. orientale may help clarify the molecular mechanisms underlying the medicinal characteristics of A. orientale.

[Correlation study between accumulation of triterpenoids and expression of relative genes in Alisma orientale].

To research the correlation between accumulation of triterpenoids and expression of key enzymes genes in triterpenoid biosynthesis of Alisma orientale,the study utilized UPLC-MS/MS method to detect eight triterpenoids content in the tuber of A. orientale from different growth stages,including alisol A,alisol A 24 acetate,alisol B,alisol B 23 acetate,alisol C 23 acetate,alisol F,alisol F 24 acetate and alisol G,and then the Real time quantitative PCR was used to analyze the expression of key enzymes genes HMGR and FPPS in triterpenoid biosynthesis. Correlation analysis showed that there was a significant positive relation between the total growth of these eight triterpenoids and the average relative expression of HMGR and FPPS(HMGR: r = 0. 998,P<0. 01; FPPS: r = 0. 957,P<0. 05),respectively. Therefore,the study preliminarily determined that HMGR and FPPS genes could regulate the biosynthesis of triterpenoids in A. orientale,which laid a foundation for further research on the biosynthesis and regulation mechanism of triterpenoids in A. orientale.

Distributive differences of P2Xs between the forelimb and hind limb of adjuvant arthritis rats and intervention by Notopterygh rhizoma et radix.

CONTEXT: Notopterygium incisum Ting ex H. T. Chang (Umbelliferae) (NI) specializes in treatment of upper limb rheumatoid arthritis (RA), but the exact mechanism is unclear. P2Xs are useful targets for inflammatory pain therapy. It led us to hypothesize that NI may preferentially act on particular P2Xs and these receptors may be unevenly distributed in the upper/lower limb. OBJECTIVE: To investigate P2Xs distribution in the upper/lower limb and NI's targets in upper limb RA. MATERIALS AND METHODS: The SD rats were randomized into 11 groups of 10 animals each. Eight experimental groups were established by the injection of 0.1 mL FCA into the plantar surface of rat paw. Three control groups suffered the same volume of saline. The articular cavities were then taken on the seventh day to detect P2Xs expression. NI (3 g/kg) and prednisone (10 mg/kg) were respectively given by oral gavage once daily for 14 d. The swelling degree and P2Xs were evaluated individually. RESULTS: In normal rats, the expressions of P2X3 and P2X6 in forelimb were markedly higher than that of in hind limb (P < 0.05). After induced by FCA, P2X1, P2X3, P2X4, P2X5 and P2X7 were increased significantly (P < 0.01). The biggest difference was P2X3. In NI treatment rats, swelling degree of the 7th/14th day in forelimb was 68.24%/38.89%, whereas that of in hind limb was 88.72%/79.92%. P2X3 mRNA and protein expression was significantly reduced as contrasted with the control group (P < 0.05). CONCLUSIONS: P2X3 receptor was predominantly expressed in the forelimb RA rat. NI relieved the FCA-induced RA by inhibiting upper limb's P2X3 receptor.

Herba Gelsemii elegantis is detoxified by ramulus et folium Mussaendae pubescentis extract by modulating hepatic cytochrome P450 and glutathione S-transferase enzymes in rats.

Herba Gelsemii elegantis (GE) has been frequently used as a Chinese folk medicine but has high acute toxicity. In Traditional Chinese Medicine, it may be detoxified by Ramulus et Folium Mussaendae pubescentis (MP), but the detoxification mechanism has remained elusive. The present study aimed to evaluate the detoxification mechanisms by which MP modulates the effect of GE in rats, including the inhibition of hepatic cytochrome P (CYP)450 and glutathione S-transferase (GST) enzymes. Male Sprague Dawley rats were orally administered GE at three doses (0.36, 0.43 or 0.54 g/kg) alone and, at the highest dose, in combination with MP (21.6 g/kg) every day for 7 consecutive days. The control group of animals received the same volume of saline. The mRNA and protein expression of hepatic CYPs representative of two subfamilies (CYP2E1 and CYP1A2) were separately assessed by reverse-transcription quantitative polymerase chain reaction (RT-qPCR), western blot and immunohistochemistry assays. The mRNA and protein expression as well as enzyme activity of hepatic GST were assessed by RT-qPCR, western blot and colorimetric assays, respectively. The results indicated that GE significantly inhibited CYP2E1 mRNA and protein expression in a dose-dependent manner. Co-administration of MP increased CYP2E1 mRNA and protein expression as compared with the high GE dose alone. Cells expressing CYP2E1, located around the hepatic vascular plexus under a clear background, were identified by immunohistochemical staining. The results for CYP1A2 were similar to those for CYP2E1. At all concentrations used, GE significantly inhibited GST mu 1 (GSTm1) mRNA and protein expression in a dose-dependent manner, as compared with the control. Combination of GE and MP increased the mRNA and protein expression of GSTm1 as compared with the high dose of GE. However, the differences in GST-pi mRNA and protein expression between the GE and GE + MP groups were not significant. Of note, rats co-treated with MP were significantly protected from the decrease in GST activity produced by GE. The present study indicated that co-administration of GE and MP upregulated the activities of CYP450 and GST enzymes when compared with GE alone. This modulation may explain for the effect of MP in reducing the acute toxicity of GE.

Construction of a heterologous gene expression system in the banana rhizobacterium strain GW-3 and its colonization ability.

Rhizobacteria inhabiting the rhizosphere are beneficial to their host plants, and can potentially serve as biocontrol agents to control plant diseases. We isolated the rhizobacterium strain GW-3, which was the dominant bacterium in the rhizosphere soils of healthy banana plants. Then, we constructed an expression system with a kanamycin resistance gene to express a heterologous protein in GW-3. Using the green fluorescent protein gene as the reporter, we monitored expression of the heterologous protein by detecting fluorescence intensity and conducting western blot analyses. The standard fluorescence intensity of the recombinant strain reached 1,482 ± 3.49 RFU. To study the colonization ability of GW-3, we inoculated this bacterium into sterilized and unsterilized rhizosphere soils and monitored the bacterial population over 25 days. The populations of GW-3 in rhizosphere soils first increased, then decreased, and finally reached a balance. Laser scanning confocal microscope analyses of fluorescence in banana roots after inoculation with GW-3 confirmed that the recombinant GW-3 strain stably colonized banana root surfaces. Analyses of the bacterial population in unsterilized rhizosphere soils showed that the recombinant GW-3 strain was still the dominant bacterium in banana rhizosphere soils at 25 days after inoculation. Together, these results showed that this expression system can be used to express a heterologous protein at high levels in a dominant rhizobacterium. By incorporating relevant resistance genes into the expression system, this method could be used to genetically engineer GW-3 to control banana wilt disease.

VP5-deficient mutant virus induced protection against challenge with very virulent infectious bursal disease virus of chickens.

A mutant infectious bursal disease virus (IBDV) deficient in expressing VP5, rGx-F9VP2DeltaVP5, was generated using reverse genetics technology. In comparison to the characteristics of rGx-F9VP2 virus in vitro, the mutant virus demonstrated lower viral titer and cytopathogenicity. To understand the role of VP5 in the pathogenicity of IBDV in vivo, animal experiments were conducted. rGx-F9VP2DeltaVP5 caused reduced bursal lesion of SPF chickens compared to rGx-F9VP2. Although rGx-F9VP2DeltaVP5 induced lower serum antibody than rGx-F9VP2 did, both inoculated groups were fully protected against vvIBDV challenge 4 weeks post-inoculation. In addition, immunosuppression induced by VP5-deficient virus was studied in 2-week-old SPF chickens immunized with AIV inactivated vaccine. And there was reduced immunosuppression as shown in our experimental results. The results showed that AIV hemagglutination inhibition (HI) antibodies of the rGx-F9VP2DeltaVP5 inoculated group were similar to those of the mock-inoculated group, however, they were higher than those of the rGx-F9VP2 inoculated group, indicating that deficiency of VP5 decreased the immunosuppression of rGx-F9VP2DeltaVP5 in chickens. All data indicated that VP5 played an important role in viral replication and pathogenesis both in vitro and in vivo. The VP5-deficient mutant virus could be a good candidate as a marked vaccine.

Synthesis of bisdesmosidic kryptogenyl saponins using the 'random glycosylation' strategy and evaluation of their antitumor activity.

A bisdesmosidic steroidal saponins library, composed of 16 novel kryptogenin glycosides, was set up via six random glycosylation procedures, wherein two compounds showed their antitumor activity against HeLa cell in the preliminary pharmacological research.