Preliminary Analysis of Transcriptome Response of Dioryctria sylvestrella (Lepidoptera: Pyralidae) Larvae Infected with Beauveria bassiana under Short-Term Starvation.

The Dioryctria genus contains several destructive borer pests that are found in coniferous forests in the Northern Hemisphere. Beauveria bassiana spore powder was tested as a new method of pest control. In this study, Dioryctria sylvestrella (Lepidoptera: Pyralidae) was used as the object. A transcriptome analysis was performed on a freshly caught group, a fasting treatment control group, and a treatment group inoculated with a wild B. bassiana strain, SBM-03. Under the conditions of 72-h fasting and a low temperature of 16 ± 1 °C, (i) in the control group, 13,135 of 16,969 genes were downregulated. However, in the treatment group, 14,558 of 16,665 genes were upregulated. (ii) In the control group, the expression of most genes in the upstream and midstream of the Toll and IMD pathways was downregulated, but 13 of the 21 antimicrobial peptides were still upregulated. In the treatment group, the gene expression of almost all antimicrobial peptides was increased. Several AMPs, including cecropin, gloverin, and gallerimycin, may have a specific inhibitory effect on B. bassiana. (iii) In the treatment group, one gene in the glutathione S-transferase system and four genes in the cytochrome P450 enzyme family were upregulated, with a sharp rise in those that were upregulated significantly. In addition, most genes of the peroxidase and catalase families, but none of the superoxide dismutase family were upregulated significantly. Through innovative fasting and lower temperature control, we have a certain understanding of the specific defense mechanism by which D. sylvestrella larvae may resist B. bassiana in the pre-wintering period. This study paves the way for improving the toxicity of B. bassiana to Dioryctria spp.

Characteristics of Carbapenem-resistant Klebsiella pneumoniae in sewage from a tertiary hospital in Jilin Province, China.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) infection is a serious problem in hospitals worldwide. We monitored a tertiary hospital in Changchun, Jilin Province, China, and found that CRKP was the major species among the carbapenem-resistant isolates in sewage. Subsequently, we evaluated the drug susceptibility, resistance genes, virulence genes, outer pore membrane protein-related genes (OmpK35 & OmpK 36), multi-locus sequence typing and replicons, biofilm formation capabilities, and resistance to chlorine-containing disinfectants among KP isolates. Identification of drug sensitivity, multiple resistance profiles were observed including 77 (82.80%) multidrug resistant (MDR), 16 (17.20%) extensive drug resistant (XDR). Some antibiotic resistance genes were detected, the most prevalent carbapenemase gene was blaKPC, and 16 resistance genes were associated with other antibiotics. In addition, 3 (3.23%) CRKP isolates demonstrated loss of OmpK-35 and 2 (2.15%) demonstrated loss of OmpK-36. In the detection of multi-locus sequence typing (MLST), 11 ST11 isolates carried virulence genes. The most common replicon type was IncFII. Biofilm-forming capabilities were demonstrated by 68.8% of the isolates, all of which were resistant to chlorine-containing disinfectants. The results of the study showed that antibiotic-resistant isolates, especially CRKP, could resist disinfectants in hospital wastewater, and improper treatment of hospital wastewater may lead to the spread of drug-resistant bacteria and their genes. Thus, these bacteria must be eliminated before being discharged into the municipal sewage system.

Propolis Ethanolic Extract Attenuates D-gal-induced C2C12 Cell Injury by Modulating Nrf2/HO-1 and p38/p53 Signaling Pathways.

Previous study has shown that propolis ethanolic extract (PEE) has a protective effect on aging skeletal muscle atrophy. However, the exact molecular mechanism remains unclear. This study aimed to investigate the effect of PEE on D-galactose (D-gal)-induced damage in mouse C2C12 cells. The results revealed that PEE increased the viability of senescent C2C12 cells, decreased the number of senescence-associated ß-galactosidase (SA-ß-Gal)-positive cells and promoted the differentiation of C2C12 cells. PEE resisted oxidative stress caused by D-gal by activating the Nrf2/HO-1 signaling pathway and maintained the differentiation ability of C2C12 cells. PEE inhibited apoptosis by suppressing p38 phosphorylation and reducing p53 expression. In summary, our findings reveal the molecular mechanism by which PEE protects D-gal-induced C2C12 cells, providing a theoretical basis for the development of PEE for the alleviation of muscle atrophy.

The effect of spraying bacterial and fungal solutions on Korean pine Pinus koraiensis Sieb. et Zucc. cone development and seed quality when sprayed during the flowering phase.

Korean pine is an economically essential afforestation species limited by the unreasonable collection of cones, indiscriminate use of chemical pesticides and pest damage. This study aimed to determine whether spraying bacterial or fungal solutions affected insect pests, cone development, and the seed quality of Korean pine Pinus koraiensis Sieb. et Zucc. The experiment was conducted in a forest plantation in Linkou County (Heilongjiang, China) in 2019. Four fungal strains and one bacterial strain were applied during the flowering phase of Korean pine. The results after a year and a half of study indicated that a high concentration of Bacillus thuringiensis 223176 promoted cone development, increased seed weight, and reduced the proportion of damaged cones. Under this treatment, there were 15.873% damaged cones; the seed weight reached 0.829 g, and there were 82.738% fully developed cones. Trees treated with the second most effective strain, Beauveria bassiana 122077, had 30.556% damaged cones and an average seed weight of 0.810 g. Leucanicillium antillanum 01 performed the worst in this study. The seed weight was only 0.775 g, and the damaged and fully developed cones were 52.444 and 41.773%, respectively. In summary, spraying bacterial or fungal solutions during the flowering stage of Korean pine positively impacted seed quality and effectively decreased damage by the lepidopteran species that feed on the cones and seeds in this study.

Systematic Review and Bioinformatic Analysis of microRNA Expression in Autism Spectrum Disorder Identifies Pathways Associated With Cancer, Metabolism, Cell Signaling, and Cell Adhesion.

Background: Previous studies have identified differentially expressed microRNAs in autism spectrum disorder (ASD), however, results are discrepant. We aimed to systematically review this topic and perform bioinformatic analysis to identify genes and pathways associated with ASD miRNAs. Methods: Following the Preferred Reporting Items for Systematic reviews and Meta-Analyses, we searched the Web of Science, PubMed, Embase, Scopus, and OVID databases to identify all studies comparing microRNA expressions between ASD persons and non-ASD controls on May 11, 2020. We obtained ASD miRNA targets validated by experimental assays from miRTarBase and performed pathway enrichment analysis using Metascape and DIANA-miRPath v3. 0. Results: Thirty-four studies were included in the systematic review. Among 285 altered miRNAs reported in these studies, 15 were consistently upregulated, 14 were consistently downregulated, and 39 were inconsistently dysregulated. The most frequently altered miRNAs including miR-23a-3p, miR-106b-5p, miR-146a-5p, miR-7-5p, miR-27a-3p, miR-181b-5p, miR-486-3p, and miR-451a. Subgroup analysis of tissues showed that miR-146a-5p, miR-155-5p, miR-1277-3p, miR-21-3p, miR-106b-5p, and miR-451a were consistently upregulated in brain tissues, while miR-4742-3p was consistently downregulated; miR-23b-3p, miR-483-5p, and miR-23a-3p were consistently upregulated in blood samples, while miR-15a-5p, miR-193a-5p, miR-20a-5p, miR-574-3p, miR-92a-3p, miR-3135a, and miR-103a-3p were consistently downregulated; miR-7-5p was consistently upregulated in saliva, miR-23a-3p and miR-32-5p were consistently downregulated. The altered ASD miRNAs identified in at least two independent studies were validated to target many autism risk genes. TNRC6B, PTEN, AGO1, SKI, and SMAD4 were the most frequent targets, and miR-92a-3p had the most target autism risk genes. Pathway enrichment analysis showed that ASD miRNAs are significantly involved in pathways associated with cancer, metabolism (notably Steroid biosynthesis, Fatty acid metabolism, Fatty acid biosynthesis, Lysine degradation, Biotin metabolism), cell cycle, cell signaling (especially Hippo, FoxO, TGF-beta, p53, Thyroid hormone, and Estrogen signaling pathway), adherens junction, extracellular matrix-receptor interaction, and Prion diseases. Conclusions: Altered miRNAs in ASD target autism risk genes and are involved in various ASD-related pathways, some of which are understudied and require further investigation.

Genes involved in the Type I pheromone biosynthesis pathway and chemoreception from the sex pheromone gland transcriptome of Dioryctria abietella.

Dioryctria abietella is a coniferous seed orchard pest that can damage a series of host plants and cause huge losses to the forest economy. Sex pheromones play an important role in lepidopteran sex communication for reproduction and can be used as biological control agents to monitor and trap pests. However, the genes involved in the biosynthesis, transportation, and degradation of D. abietella sex pheromones have not been studied extensively. Transcriptome analysis of female D. abietella sex pheromone glands (PGs) revealed that 210 candidate genes might be involved in sex pheromone biosynthesis (139 genes) and chemoreception systems (71 genes). The gene expression patterns exhibited four desaturase genes (DabiDES4-7) and one fatty acid reductase gene (DabiFAR6), which were more highly expressed in sex pheromone glands than in other tissues, suggesting that these enzymes play an important role in D. abietella sex pheromone synthesis. In addition, most DabiOBPs showed high expression in antennae, but only DabiOBP4 exhibited specific expression in sex pheromone glands, suggesting that they may play many physiological roles in D. abietella. We put forth a reasonable hypothesis about type I pheromone biosynthesis pathways based on these genes identified in the D. abietella sex pheromone gland transcriptome. Our findings lay a foundation for population monitoring, mating disruption, mass trapping, and the development of ecologically acceptable management strategies.

Study on Hypoglycemic Effect of the Drug Pair of Astragalus Radix and Dioscoreae Rhizoma in T2DM Rats by Network Pharmacology and Metabonomics.

Type 2 diabetes mellitus (T2DM) is a metabolic disease accompanied by a series of diseases such as diabetic nephropathy. The drug pair (HS) of Astragalus Radix (HQ) and Dioscoreae Rhizoma (SY) was designed by Dr. Shi Jinmo to improve the treatment of T2DM. However, the exact mechanism involved requires further clarification. In this work, 1H-NMR-based metabonomics and network pharmacology were adopted. Metabolic profiling indicated that the metabolic perturbation was reduced after HS treatment. The results found 21 biomarkers. According to the network pharmacology, we found that the regulation of T2DM was primarily associated with 18 active compounds in HS. These active compounds mainly had an effect on 135 targets. Subsequently, combining network pharmacology and metabonomics, we found four target proteins, which indicated that HS has potential hypoglycemic effects through regulating monoamine oxidases B (MAOB), acetyl-CoA carboxylase 1 (ACACA), carbonic anhydrase 2 (CA2), and catalase (CAT). In conclusion, the result showed that these four targets might be the most relevant targets for the treatment of T2DM with HS. This study clarified the mechanism of HS in the treatment of T2DM and also confirmed the feasibility of combining metabonomics and network pharmacology to study the mechanisms of traditional Chinese medicine (TCM). In the future, this approach may be a potentially powerful tool to discovery active components of traditional Chinese medicines and elucidate their mechanisms.

Effect of seedling grade standard on improving the quality of licorice (Glycyrrhiza uralensis F.): changes in the seven bioactive components and root biomass during two-year growth.

Licorice cultivated is one of the most popular herbal medicines, while its quality is unstable. The aim of present study is to investigate the effect of licorice seedling grade standard on improving its quality. One-year-old Glycyrrhiza uralensis seedlings were classified into three grades 1, 2, and 3 by weight per plant. The major root biomass indexes (root fresh weight, root dry weight and taproot diameter) and contents of 7 bioactive components (glycyrrhizin, liquiritin, liquiritin apioside, liquiritigenin, isoliquiritin, isoliquiritin apioside, and isoliquiritigenin) varied in different grades seedlings. Further, the contents of 7 investigated compounds of 3-year-old licorice produced by grade 1 seedlings were 1.5-2 times as much as those produced by grade 2 and 3. Additionally, the contents of liquiritin apioside and isoliquiritin apioside were positively correlated with licorice root biomass. These results indicated that establishing licorice seedling grade standard is an effective way to improve and control its quality.

Phytotoxicity of umbelliferone and its analogs: Structure-activity relationships and action mechanisms.

Two coumarins, umbelliferone and daphnoretin, were isolated from roots of Stellera chamaejasme L; the former had been identified as one of the main allelochemicals in our previous studies. Both of them have the skeleton of 7-hydroxycoumarin, but showed different phytotoxic effects. Umbelliferone and its analogs were then prepared to investigate the structure-activity relationship of hydroxycoumarins and screened for phytotoxicity. The inhibitory effects varied observably in response to the coumarin derivatives, especially umbelliferone (1), 7-hydroxy-4-methylcoumarin (3) and coumarin (10) displayed strong inhibition of lettuce and two field weeds, Setaria viridis and Amaranthus retroflexus, and compounds 11 and 12 also exhibited phytotoxic activity with species specificity. The number and location of hydroxyl groups were importantly responsible for the phytotoxicity. A C7 hydroxyl group was considered to be a potentially active site and methyl substitution at the C4 position contributed significantly to the activity. The phytotoxic mechanism was briefly studied with umbelliferone by evaluating the reactive oxygen species (ROS) and chlorophylls level in lettuce seedlings. The results showed that umbelliferone induced the accumulation of ROS in the root tip and significantly decreased the chlorophyll content in the leaves. Thus, a ROS-mediated regulation pathway and the inhibition of photosynthesis were definitely involved in the phytotoxicity of umbelliferone.

Phytotoxic flavonoids from roots of Stellera chamaejasme L. (Thymelaeaceae).

Allelopathy, the negative effect on plants of chemicals released to the surroundings by a neighboring plant, is an important factor which contributes to the spread of some weeds in plant communities. In this field, Stellera chamaejasme L. (Thymelaeaceae) is one of the most toxic and ecologically-threatening weeds in some of the grasslands of north and west China. Bioassay-guided fractionation of root extracts of this plant led to the isolation of eight flavonoids 1-8, whose structures were elucidated by spectroscopic analysis. All compounds obtained, except 7-methoxylneochaejasmin A (4) and (+)-epiafzelechin (5), showed strong phytotoxic activity against Arabidopsis thaliana seedlings. Seedling growth was reduced by neochamaejasmin B (1), mesoneochamaejasmin A (2), chamaejasmenin C (3), genkwanol A (6), daphnodorin B (7) and dihydrodaphnodorin B (8) with IC50 values of 6.9, 12.1, 43.2, 74.8, 7.1 and 27.3µg/mL, respectively, and all of these compounds disrupted root development. Endogenous auxin levels at the root tips of the A. thaliana DR5::GUS transgenic line were largely reduced by compounds 1, 2 and 6-8, and were increased by compound 4. Moreover, the inhibition rate of A. thaliana auxin transport mutants pin2 and aux1-7 by compounds 1-8 were all lower than the wild type (Col-0). The influence of these compounds on endogenous auxin distribution is thus proposed as a critical factor for the phytotoxic effect. Compounds 1, 2, 4 and 8 were found in soils associated with S. chamaejasme, and these flavonoids also showed phytotoxicity to Clinelymus nutans L., an associated weed of S. chamaejasme. These results indicated that some phytotoxic compounds from roots of S. chamaejasme may be involved in the potential allelopathic behavior of this widespread weed.

Allelochemicals in the rhizosphere soil of Euphorbia himalayensis.

Weed infestation has been known to cause considerable reductions in crop yields, thereby hindering sustainable agriculture. Many plants in genus Euphorbia affect neighboring plants and other organisms by releasing chemicals into the environment. In view of the serious threat of weeds to agriculture, the allelochemicals of Euphorbia himalayensis and their allelopathic effects were investigated. The extract of root exudates from rhizosphere soil exhibited allelopathic activities against crops (wheat, rape, and lettuce) and grasses (Poa annua, Festuca rubra, and red clover). Bioassay-guided fractionation and isolation from the root extract of E. himalayensis led to the characterization of two ellagic acid derivatives and a jatrophane diterpene, which observably showed phytotoxic activities against lettuce, Festuca arundinacea, and F. rubra. They were further confirmed by ultra-performance liquid chromatography-tandem mass spectrometry to have concentrations of 3.6, 3.8, and 8.99 nmol/g in the rhizospere soil, respectively. Bioassay indicated that the combination of the allelochemicals could be selective plant growth regulator in agriculture.

[Application and prospect of near infrared reflectance spectroscopy in forage analysis].

Forage was the material basis of animal husbandry production, and its quality is directly related to the quality of animal products. It was very important to control the forage quality and detect the composition of forage raw materials in forage production. Predication of forage quality was often completed by the traditional and classical methods in the past, which were complex, time consuming and expensive, and could not acquire the nutritional value of forage timely. Near infrared reflectance spectroscopy was a highly efficient and rapid modern analysis technique developed in 1970's. It comprehensively applied the latest research results of computer technique, spectroscopy and chemometrics, and has been widely used in various fields owing to its unique advantages such as being timely, less expensive, non-destructive, and so on. Near infrared reflectance spectroscopy has gained more and more importance though its application to forage analysis was very late. Presently, not only conventional composition (such as moisture, dry matter, crude protein, crude fiber, crude fat, crude ash neutral detergent fiber, acid detergent fiber, etc.), but also non-conventional composition (including minerals, trace elements, enzyme and anti-nutritional factors etc. ) and anti-nutritional factors in forage were determined by means of near infrared reflectance spectroscopy. Testing and analyzing the conventional composition in forage was the traditional applied field of near infrared reflectance spectroscopy, a lot of studies of which were done and it has already been one of the standard methods of testing the conventional composition. Forage bioavaibility was also evaluated by near infrared reflectance spectroscopy, so as to assess the utilization rate and nutritional value of forage. Moreover, near infrared spectroscopy could be used successfully to predict the botanical composition in grassland and leaf/stem ratios. Near infrared spectroscopy technique and its application and prospect in forage analysis were reviewed in the present paper.

[Allelopathy of different plants on wheat, cucumber and radish seedlings].

By means of bioassay in laboratory and field, this paper studied the allelopathy of 18 kinds of plants in Gansu Province on the seedlings of wheat, cucumber and radish. The results showed that the aqueous extract of the stems and leaves of Artemisia annua, Solanum nigrum and Datura stramonium had the strongest allelopathy on test receptor plants, and their synthetic inhibitory effect (SE) was 47.66%, 32.89% and 26.63%, respectively. The SE of Xanthium sibiricum, Portulaca oleraca, Cephalanoplos segetum, and Chenopodium album was 21.71%, 20.93%, 20.83% and 20.2%, respectively, while Vicia amoena (SE 3.5%), Setaria viridis (SE 2.2%), and Cymamchum chinense (SE 1.97%) had a weaker allelopathy. Chenopodium ambrosioides (SE - 1.03%), Polygonum caespitosum (SE - 1.63%) and Avena fatua (SE 5.33%) had no evident allelopathy, but Artemisia annua affected the seedling height and fresh weight of radish, cucumber, wheat and maize, with the SE being 54.07%, 38.46%, 33.35% and 20.88%, respectively. Artemisia annua had a 44.70% of SE on wheat growth, and thus, had a certain value to develop and use.