Magnetic nanoparticles immobilized thrombin ligand fishing to screen thrombin inhibitors in natural products.

In this study, thrombin was immobilized with magnetic particles modified by glutaraldehyde. The changes in secondary structures of immobilized enzyme revealed an increment in conformational rigidity and stability, which can be reflected in temperature and pH stability as well as the tolerance of organic reagents. The optimal reutilization times of magnetic particle immobilized thrombin were 7 times, and the half-life of enzyme activity preserved at room temperature was 5 days, which was 2.5 times higher than that of free enzyme. Ligusticum chuanxiong and Anemarrhenae Rhizoma with high enzyme inhibitory activity were selected for primary screening, and six potential inhibitors of thrombin were identified by HPLC/MS. The results showed that three compounds in Anemarrhenae Rhizoma had better predictive thrombin inhibitory activity. Through the in vitro thrombin activity inhibition experiment, it was also verified that mangiferin and neo-mangiferin had an ideal thrombin activity inhibition effect, which was consistent with the results of molecular docking.

Identification of milk-related genes and regulatory networks in Bactrian camel either supplemented or under grazing.

Using gene co-expression networks to understand dynamic characterizations in lactating animals becomes a common method. However, there are rarely reporters focusing on milk traits in Bactrian camel by high-throughput sequencing. We used RNA-seq to generate the camel transcriptome from the blood of 16 lactating Alxa Bactrian camel in different feeding groups. In total, we obtained 1185 milk-related genes correlated with milk yield, milk protein, milk fat, and milk lactose across the WGCNA analysis. Moreover, 364 milk-related genes were differentially expressed between supplementation and grazing feeding groups. The differential expression-camel milk-related genes CMRGs (DE-CMRGs) in supplement direct an intensive gene co-expression network to improve milk performance in lactating camels. This study provides a non-invasive method to identify the camel milk-related genes in camel blood for four primary milk traits and valuable theoretical basis and research ideas for the study of the milk performance regulation mechanism of camelid animals.

Bitter Almond Albumin ACE-Inhibitory Peptides: Purification, Screening, and Characterization In Silico, Action Mechanisms, Antihypertensive Effect In Vivo, and Stability.

Almond expeller is an undeveloped reservoir of bioactive peptides. In the current study, a zinc ion ligand Arg-Pro-Pro-Ser-Glu-Asp-Glu-Asp-Gln-Glu (RPPSEDEDQE) offering a noncompetitive inhibitory effect on ACE (IC50: 205.50 µmol·L-1) was identified from almond albumin hydrolysates via papain and thermolysin hydrolysis, subsequent chromatographic separation, and UPLC-Q-TOF-MS/MS analysis. Molecular docking simulated the binding modes of RPPSEDEDQE to ACE and showed the formation of hydrogen bonds between RPPSEDEDQE and seven active residues of ACE. Moreover, RPPSEDEDQE could bind to fifteen active sites of ACE by hydrophobic interactions, and link with the His387 and zinc ions of the zinc tetrahedral coordination. Ultraviolet wavelength scanning and Fourier-transformed infrared spectroscopy analysis revealed that RPPSEDEDQE can provide multiple binding sites for zinc ions. However, RPPSEDEDQE cannot bind with any central pocket of ACE, which was evidenced by an inhibition kinetics experiment. Additionally, the zinc-chelating capacity and inhibiting ability against ACE of RPPSEDEDQE were both not significantly reduced by the hydrolysis of gastrointestinal enzymes. A moderate to high dose of RPPSEDEDQE (100-150 mg·kg bw-1) significantly reduced the systolic and diastolic blood pressure of spontaneous hypertensive rats, but chelation with zinc ions decreased its antihypertensive efficiency. These results indicate that bitter almond albumin peptides may be used for lowering blood pressure.

Stimulation of primed carbon under climate change corresponds with phosphorus mineralization in the rhizosphere of soybean.

Elevated CO2 and temperature likely alter photosynthetic carbon inputs to soils, which may stimulate soil microbial activity to accelerate the decomposition of soil organic carbon (SOC), liberating more phosphorus (P) into the soil solution. However, this hypothesis on the association of SOC decomposition and P transformation in the plant rhizosphere requires robust soil biochemical evidence, which is critical to nutrient management for the mitigation of soil quality against climate change. This study investigated the microbial functional genes relevant to P mineralization together with priming processes of SOC in the rhizosphere of soybean grown under climate change. Soybean plants were grown under elevated CO2 (eCO2, 700 ppm) combined with warming (+ 2 °C above ambient temperature) in open-top chambers. Photosynthetic carbon flow in the plant-soil continuum was traced with 13CO2 labeling. The eCO2 plus warming treatment increased the primed carbon (C) by 43 % but decreased the NaHCO3-extratable organic P by 33 %. Furthermore, NaHCO3-Po was negatively correlated with phosphatase activity and microbial biomass C. Elevated CO2 increased the abundances of C degradation genes, such as abfA and ManB, and P mineralization genes, such as gcd, phoC and phnK. The results suggested that increased photosynthetic carbon inputs to the rhizosphere of plants under eCO2 plus warming stimulated the microbial population and metabolic functions of both SOC and organic P mineralization. There is a positive relationship between the rhizosphere priming effect and P mineralization. The response of microorganisms to plant-C flow is decisive for coupled C and P cycles, which are likely accelerated under climate change.

External sodium acetate improved Cr(VI) stabilization in a Cr-spiked soil during chemical-microbial reduction processes: Insights into Cr(VI) reduction performance, microbial community and metabolic functions.

Interest combined chemical and microbial reduction for Cr(VI) remediation in contaminated sites has greatly increased. However, the effect of external carbon sources on Cr(VI) reduction during chemical-microbial reduction processes has not been studied. Therefore, in this study, the role of external sodium acetate (SA) in improving Cr(VI) reduction and stabilization in a representative Cr(VI)-spiked soils was systemically investigated. The results of batch experiments suggested that the soil Cr(VI) content declined from 1000 mg/kg to 2.6-5.1 mg/kg at 1-5 g C/kg SA supplemented within 15 days of reaction. The external addition of SA resulted in a significant increase in the relative abundances of Cr(VI)-reducing microorganisms, such as Tissierella, Proteiniclasticum and Proteiniclasticum. The relative abundance of Tissierella increased from 9.1% to 29.8% with the SA treatment at 5 g C/kg soil, which was the main contributors to microbial Cr(VI) reduction. Redundancy analysis indicated that pH and SA were the predominant factors affecting the microbial community in the SA treatments at 2 g C/kg soil and 5 g C/kg soil. Functional prediction suggested that the addition of SA had a positive effect on the metabolism of key substances involved in Cr(VI) microbial reduction. This work provides new insightful guidance on Cr(VI) remediation in contaminated soils.

Effects of 24-Form Tai Chi on Cardio-Pulmonary Functions, Exercise Performances, and Cognitive Functions of the Aged.

Background: The study aimed to discuss the influences of 24-form Tai Chi on cardio-pulmonary functions, exercise performances, and cognitive functions of the aged. Methods: Forty old people in Pingdingshan of China were selected through convenient sampling method from 2020 to 2021. They were randomly divided into the observation group (n=20) and the control group (n=20). The observation group exercised 24-form Tai Chi for 8 weeks, three times per week, 45 min per session. The respondents were evaluated by cardiopulmonary exercise testing, sitting-standing test, 6-min walking test, and the Berg balance scale. The P300 test was performed to evaluate the cognitive functions of respondents. Results: After the intervention, the observation group was significantly (P<0.05) superior to the control group concerning maximal oxygen consumption, anaerobic threshold, vital capacity, stroke volume (SV), cardiac output (CO), left ventricular end-diastolic diameter (LVEDD), left end-diastolic velocity (EDV), sitting-standing test, 6-min walking test, and the Breg balance function scores, showing statistically significant differences (P<0.05). The content of body fat and body mass index (BMI) of the observation group decreased significantly, whereas muscle mass and fat-free mass increased dramatically, which were however significantly lower than those of the controlled group (P<0.05). Moreover, the P3b amplitude of the observation group increased significantly (P<0.05) and was significantly higher than that of the control group (P<0.05). Conclusion: The 24-form Tai Chi not only improves cardio-pulmonary functions and exercise performances of the aged but also increases cognitive functions of the aged.

Dormancy-release, germination and seedling growth of Paeonia ostii 'Fengdan' seeds under measures of physical and chemical treatment and sowing.

Paeonia ostii 'Fengdan', a woody oleaginous plant native from China, is considered an oil crop with economic potential. However, a low germination rate was still a restriction for Paeonia ostii 'Fengdan' production. The present research evaluated the germination, rooting and physiological characteristics of seedlings of Paeonia ostii 'Fengdan' in response to different physical treatments and the application of exogenous chemicals. Results indicated that seeds stored in sand at room temperature, and soaked in water for 3 days prior to planting, had a beneficial effect on hypocotyl dormancy-breaking. The rate of rooting and root growth of Paeonia ostii 'Fengdan' were significantly improved with 5 cm sowing depth in 15-20℃ soils. Compared with other sowing depths, the rooting percentage was significantly increased by 1.19% (2.5 cm), 0.98% (7.5 cm) and 1.47% (10 cm), respectively. Epicotyl dormancy was relieved when taproot length reached 50 mm. Soaking seeds in 0.76 mmol/L 5-aminolevulinic acid for 48 hours had the greatest beneficial effect on seed germination and seedling growth, the germination percentage was significantly increased by 4.25% (24 h) and 5.08% (72 h) compared with other treatments. While seed soaked in 10 mmol/L sodium nitroprusside for 48 hours also exhibited enhanced seedling growth, and the germination percentage was significantly increased by 4.36% (24 h) and 7.40% (72 h). Those results benefited seed germination and seedling growth of Paeonia ostii 'Fengdan' which could suggest the promotion of its industrial values and productive potentials. The mechanism of seed breaking dormancy and germination of Paeonia ostii 'Fengdan' needs further study.

Molecular and biochemical characterization of two 4-coumarate: CoA ligase genes in tea plant (Camellia sinensis).

KEY MESSAGE: Two 4-coumarate: CoA ligase genes in tea plant involved in phenylpropanoids biosynthesis and response to environmental stresses. Tea plant is rich in flavonoids benefiting human health. Lignin is essential for tea plant growth. Both flavonoids and lignin defend plants from stresses. The biosynthesis of lignin and flavonoids shares a key intermediate, 4-coumaroyl-CoA, which is formed from 4-coumaric acid catalyzed by 4-coumaric acid: CoA ligase (4CL). Herein, we report two 4CL paralogs from tea plant, Cs4CL1 and Cs4CL2, which are a member of class I and II of this gene family, respectively. Cs4CL1 was mainly expressed in roots and stems, while Cs4CL2 was mainly expressed in leaves. The promoter of Cs4CL1 had AC, nine types of light sensitive (LSE), four types of stress-inducible (SIE), and two types of meristem-specific elements (MSE). The promoter of Cs4CL2 also had AC and nine types of LSEs, but only had two types of SIEs and did not have MSEs. In addition, the LSEs varied in the two promoters. Based on the different features of regulatory elements, three stress treatments were tested to understand their expression responses to different conditions. The resulting data indicated that the expression of Cs4CL1 was sensitive to mechanical wounding, while the expression of Cs4CL2 was UV-B-inducible. Enzymatic assays showed that both recombinant Cs4CL1 and Cs4CL2 transformed 4-coumaric acid (CM), ferulic acid (FR), and caffeic acid (CF) to their corresponding CoA ethers. Kinetic analysis indicated that the recombinant Cs4CL1 preferred to catalyze CF, while the recombinant Cs4CL2 favored to catalyze CM. The overexpression of both Cs4CL1 and Cs4CL2 increased the levels of chlorogenic acid and total lignin in transgenic tobacco seedlings. In addition, the overexpression of Cs4CL2 consistently increased the levels of three flavonoid compounds. These findings indicate the differences of Cs4CL1 and Cs4CL2 in the phenylpropanoid metabolism.

Identification of differentially expressed miRNAs and their target genes in response to brassinolide treatment on flowering of tree peony (Paeonia ostii).

Tree peony is a famous flower plant in China, but the short and concentrated flowering period limits its ornamental value and economic value. Brassinolide (BR) plays an important role in plant growth and development including flowering. There have been a large number of reports on the molecular aspects of the flowering process, but the genetic mechanism that was responsible for miRNA-guided regulation of tree peony is almost unclear. In this study, the leaves of tree peony cultivar, 'Feng Dan', were sprayed with different concentrations of BR, and the obvious bloom delay was found at the treatment with BR 50 µg/L. The small RNA sequencing and transcriptome sequencing were performed on the petals of tree peony under an untreated control (CK) and the treatment with BR 50 µg/L during four consecutive flowering development stages. A total of 22 known miRNAs belonging to 12 families were identified and 84 novel miRNAs were predicted. Combined with transcriptome data, a total of 376 target genes were predicted for the 18 differentially expressed known miRNAs and 177 target genes were predicted for the 23 differentially expressed novel miRNAs. Additionally, the potential miRNAs and their target genes were identified, including miR156b targeting SPL, miR172a_4 targeting AP2 and four novel miRNAs targeting SPA1, and revealed that they might affect the flowering time in tree peony. Collectively, these results would provide a theoretical basis for further analysis of miRNA-guided regulation on flowering period in tree peony.

Thiosulfatihalobacter marinus gen. nov. sp. nov., a novel member of the family Roseobacteraceae, isolated from the West Pacific Ocean.

Two strains (GL-11-2T and ZH2-Y79) were isolated from the seawater collected from the West Pacific Ocean and the East China Sea, respectively. Cells were Gram-stain-negative, strictly aerobic, non-motile and rod-shaped. Cells grew in the medium containing 0.5-7.5 % NaCl (w/v, optimum, 1.0-3.0 %), at pH 6.0-8.0 (optimum, pH 6.5-7.0) and at 4-40 °C (optimum, 30 °C). H2S production occurred in marine broth supplemented with sodium thiosulphate. The almost-complete 16S rRNA gene sequences of the two isolates were identical, and exhibited the highest similarity to Pseudoruegeria aquimaris JCM 13603T (97.5 %), followed by Ruegeria conchae TW15T (97.2%), Shimia aestuarii DSM 15283T (97.1 %) and Ruegeria lacuscaerulensis ITI-1157T (97.0 %). Phylogenetic analysis revealed that the isolates were affiliated with the family Roseobacteraceae and represented an independent lineage. The sole isoprenoid quinone was ubiquinone 10. The principal fatty acids were summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and cyclo-C19 : 0 ω8c. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and diphosphatidylglycerol. The DNA G+C content was 62.3 mol%. The orthologous average nucleotide identity, in silico DNA-DNA hybridization and average amino acid identity values among the genomes of strain GL-11-2T and the reference strains were 73.2-79.0, 20.3-22.5 and 66.0-80.8 %, respectively. Strains GL-11-2ᵀ and ZH2-Y79 possessed complete metabolic pathways for thiosulphate oxidation, dissimilatory nitrate reduction and denitrification. Phylogenetic distinctiveness, chemotaxonomic differences and phenotypic properties revealed that the isolates represent a novel genus and species of the family Roseobacteraceae, belonging to the class Alphaproteobacteria, for which the name Thiosulfatihalobacter marinus gen. nov., sp. nov. (type strain, GL-11-2T=KCTC 82723T=MCCC M20691T) is proposed.

Plant phosphorus acquisition links to phosphorus transformation in the rhizospheres of soybean and rice grown under CO2 and temperature co-elevation.

Climate change is likely to influence the reservoir of soil phosphorus (P) as plants adaptably respond to climate change in the perspective of P acquisition capability via root proliferation and mediating biochemical properties in the rhizosphere to access various soil P fractions. It is particularly important in cropping soils where P fertilizer plus soil P is required to synchronize crop P demand for the production sustainability under climate change. However, few studies have examined the effect of CO2 and temperature co-elevation on plant P acquisition, P fractions and relevant functional genes in the rhizosphere of different crops. Thus, the present study investigated the effect of elevated CO2 and warming on P uptake of soybean and rice grown in Mollisols, and soil P fractions and relevant biochemical properties and microbial functions in the rhizosphere with or without P application. Open-top chambers were used to achieve elevated CO2 of 700 ppm combined with warming (+ 2 °C above ambient temperature). CO2 and temperature co-elevation increased P uptake in soybean by 23% and 28% under the no-P and P application treatments, respectively; and in rice, by 34% and 13%, respectively. CO2 and temperature co-elevation depleted organic P in the rhizosphere of soybean, but increased in the rhizosphere of rice. The phosphatase activity negatively correlated with organic P in the highland soil while positively in the paddy soil. The P mineralization likely occurs in soybean-grown soils under climate change, while the P immobilization in paddy soils. CO2 and temperature co-elevation increased the copy numbers of P functional genes including phoD, phoC, pstS and phnX, in soils with P application. These results indicate that the P application would be requested to satisfy the increased P demand in soybean under climate change, but not in rice in paddy soils where soil P availability is sufficient. Therefore, elevated CO2 and temperature facilitated the crop P uptake via biochemical and microbial pathways, and P functional genes played an essential role in the conversion of P.

Traditional Chinese medicine residue-derived micropore-rich porous carbon frameworks as efficient sulfur hosts for high-performance lithium-sulfur batteries.

Biomass-derived carbon-based energy materials are receiving extensive attention nowadays. With the widespread use of traditional Chinese medicines in the treatment of diseases and health care, a great deal of herb residues are thrown away after the unique decoction process. Here, through hydrothermal carbonization combined with KOH activation, a micropore-rich and nitrogen-doped porous carbon framework (MRNCF) is prepared from the waste roots of a kind of well-known and widely used traditional Chinese medicine, Acanthopanax senticosus. Compared with ordinary carbon-based sulfur host materials, the MRNCFs can effectively hinder the shuttling effect and dissolution of polysulfides through the synergistic action of physical confinement in micropores and chemical anchoring for nitrogen doping, and the lithium-sulfur batteries using MRNCF as the host present superior electrochemical performance. In a high sulfur content of over 75%, the as-prepared electrodes exhibit a highly reversible specific capacity of 540.4 mA h g-1 at a current density of 0.5C after 150 cycles and an excellent rate capability at different current densities.

Quantitative selenium speciation in feed by enzymatic probe sonication and ion chromatography-inductively coupled plasma mass spectrometry.

A rapid, sensitive and species preservative analytical method for the simultaneous determination of six selenium (Se) species has been developed. Enzymatic probe sonication (EPS) was investigated as a novel and alternative technology for the extraction of Se species from feed matrices and the results were compared with the conventional hot water extraction, enzymatic hydrolysis and sequential extraction. The critical parameters of EPS such as enzyme types, extraction time, temperature, ultrasonic power and sample/enzyme ratio were varied with control. The Se species were separated and quantitatively determined by ion chromatography-inductively coupled plasma mass spectrometry (IC-ICP-MS). Under current optimised conditions, six inorganic and organic Se species were completely separated within 15 min in a single chromatographic run. The spectral interferences from the argon plasma 40Ar2, 40Ar37Cl or 1H79Br were effectively removed by employing the kinetic energy discrimination (KED) mode. Quantitative extraction for total Se (>94.8%) and more than 89.0% for the sum of different Se chemical forms without species transformation were obtained in only 60 s by applying the EPS treatment using aqueous protease XIV. The limits of detection (LODs) and quantification (LOQs) for Se species were in the ranges of 0.21-0.56 µg kg-1 and 0.69-1.87 µg kg-1, respectively. The proposed method was successfully applied to the speciation of Se in several reference materials and feed samples collected from the markets and local farms.

Transcriptome analysis reveals mechanism of early ripening in Kyoho grape with hydrogen peroxide treatment.

BACKGROUND: In a previous study, the early ripening of Kyoho grape following H2O2 treatment was explored at the physiological level, but the mechanism by which H2O2 promotes ripening at the molecular level is unclear. To reveal the molecular mechanism, RNA-sequencing analysis was conducted on the different developmental stages of Kyoho berry treated with H2O2. RESULTS: In the comparison of treatment and control groups, 406 genes were up-regulated and 683 were down-regulated. Time course sequencing (TCseq) analysis showed that the expression patterns of most of the genes were similar between the treatment and control, except for some genes related to chlorophyll binding and photosynthesis. Differential expression analysis and the weighted gene co-expression network were used to screen significantly differentially expressed genes and hub genes associated with oxidative stress (heat shock protein, HSP), cell wall deacetylation (GDSL esterase/lipase, GDSL), cell wall degradation (xyloglucan endotransglucosylase/ hydrolase, XTH), and photosynthesis (chlorophyll a-b binding protein, CAB1). Gene expression was verified with RT-qPCR, and the results were largely consistent with those of RNA sequencing. CONCLUSIONS: The RNA-sequencing analysis indicated that H2O2 treatment promoted the early ripening of Kyoho berry by affecting the expression levels of HSP, GDSL, XTH, and CAB1 and- photosynthesis- pathways.

Transcriptional Analysis of the Early Ripening of 'Kyoho' Grape in Response to the Treatment of Riboflavin.

Previous study has demonstrated that the riboflavin treatment promoted the early ripening of the 'Kyoho' grape berry. However, the molecular mechanism causing this was unclear. In order to reveal the regulation mechanism of riboflavin treatment on grape berry development and ripening, the different berry developmental stages of the 'Kyoho' berry treated with 0.5 mmol/L of riboflavin was sampled for transcriptome profiling. RNA-seq revealed that 1526 and 430 genes were up-regulated and down-regulated, respectively, for the comparisons of the treatment to the control. TCseq analysis showed that the expression patterns of most of the genes were similar between the treatment and the control, except for some genes that were related to the chlorophyll metabolism, photosynthesis-antenna proteins, and photosynthesis, which were revealed by the enrichment analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). The differentially expressed genes and weighted gene co-expression network analysis (WGCNA) analysis identified some significantly differentially expressed genes and some hub genes, including up-regulation of the photosynthesis-related ELIP1 and growth and development-related GDSL; and down-regulation of the oxidative stress-related ATHSP22 and berry softening-related XTH32 and GH9B15. The results suggested that the riboflavin treatment resulted in the variations of the expression levels of these genes, and then led to the early ripening of the 'Kyoho' berry.

Preparation, optimization and cellular uptake study of tanshinone I nanoemulsion modified with lactoferrin for brain drug delivery.

Tanshinone I (TSI) is one of the bioactive compound obtained from the root of Salvia miltiorrhiza which is a well-known traditional Chinese medicine (TCM) used for the treatment of various diseases. Although TSI possesses several pharmacological effects, it has poor water solubility, blood-brain barrier (BBB) permeability and brain bioavailability. Therefore, in the present study, we developed TSI nanoemulsion (TSI-NE) modified with a brain targeting ligand (Lactoferrin (Lf)) to improve the BBB permeability. Pseudo-ternary phase diagrams were used to optimize the formulation. The optimal TSI-NE and TSI-Lf-NE were prepared and characterized. Finally, the uptake of TSI-Lf-NE by mouse brain microvascular endothelial cell line (bEnd.3 cells) was assessed using Coumarin-6 as a fluorescent probe. The results of the study showed that the stable optimal formulation of O/W nanoemulsion was successfully developed and modified with Lf. The cellular uptake study has shown that the fluorescence intensity (FI) increased with time over the incubation period. The FI at all time intervals increased in the following order: Coumarin-6-Solution＜Coumarin-6-NE＜Coumarin-6-Lf-NE. The results suggest that the BBB permeability of Coumarin-6-Lf-NE was better than those of Coumarin-6-NE and Coumarin-6 solution. Lf modified nanoemulsion has great potential for improving the brain delivery of TSI.

Effects of the stems and leaves of Astragalus membranaceus on growth performance, immunological parameters, antioxidant status, and intestinal bacteria of quail.

This study was designed to evaluate the potential application of the stems and leaves of Astragalus membranaceus (AMSL) in the poultry industry. Quails were divided into four groups and fed daily with an AMSL-free diet (control) or with 1%, 3%, or 5% (w/w) AMSL-incorporated diets for 35 days. The results showed that supplementing AMSL in the diet, especially at a concentration of 3%, increased daily gain and feed intake during the entire experiment (p < 0.05). The immune organ development of the thymus and bursa of Fabricius was promoted, and the immune system was enhanced by increasing the quantities of IgA and complements C3 and C4 (p < 0.05). The total antioxidant capacity and the activities of glutathione peroxidase and catalase were increased (p < 0.05). Moreover, the 3%-5% AMSL groups regulated the intestinal flora by promoting the proliferation of lactic acid bacteria and inhibiting the growth of coliform bacteria (p < 0.05). In conclusion, feeding incorporated diets with appropriate AMSL levels significantly increased growth performance, strengthened the immune system, improved antioxidative status, and regulated the intestinal microflora of quails, suggesting that AMSL has the potential to serve as a feed additive in the poultry industry.

Transcriptome Analysis on Maternal Separation Rats With Depression-Related Manifestations Ameliorated by Electroacupuncture.

Maternal separation (MS), a stressful event in early life, has been linked to neuropsychiatric disorders later in life, especially depression. In this study we investigated whether treatment with electroacupuncture (EA) could ameliorate depression-related manifestations in adult animals that had adverse early life experiences. We demonstrated depression-like behavior deficiencies in a sucrose preference test and a forced swimming test in a rat model with neonatal MS. Repeated EA treatment at the acupoints Baihui (GV20) and Yintang (GV29) during adulthood was shown to be remarkably attenuated above behavioral deficits. Using unbiased genome-wide RNA sequencing to investigate alterations in the transcriptome of the prefrontal cortex (PFC), we explored the altered gene sets involved in circadian rhythm and neurotransmitter transporter activity in MS rats, and their expression tended to be reversed after EA treatment. In addition, we analyzed the interaction network of differentiated lncRNA- or circRNA-miRNA-mRNA by using the principle of competitive endogenous RNA (ceRNA). These results suggest that EA at GV20 and GV29 ameliorates depression-related manifestations by regulating the expression of multiple genes.

Electroacupuncture Ameliorates Cognitive Deficit and Improves Hippocampal Synaptic Plasticity in Adult Rat with Neonatal Maternal Separation.

Exposure to adverse early-life events is thought to be the risk factors for the development of psychiatric and altered cognitive function in adulthood. The purpose of this study was to investigate whether electroacupuncture (EA) treatment in young adult rat would improve impaired cognitive function and synaptic plasticity in adult rat with neonatal maternal separation (MS). Wistar rats were randomly divided into four groups: control group, MS group, MS with EA treatment (MS + EA) group, and MS with Sham-EA treatment (MS + Sham-EA) group. We evaluated the cognitive function by using Morris water maze and fear conditioning tests. Electrophysiology experiment used in vivo long-term potentiation (LTP) at Schaffer Collateral-CA1 synapses was detected to assess extent of synaptic plasticity. Repeated EA stimulation at Baihui (GV 20) and Yintang (GV 29) during postnatal 9 to 11 weeks was identified to significantly ameliorate poor performance in behavior tests and improve the impaired LTP induction detected at Schaffer Collateral-CA1 synapse in hippocampus. Collectively, the findings suggested that early-life stress due to MS may induce adult cognitive deficit associated with hippocampus, and EA in young adult demonstrated that its therapeutic efficacy may be via ameliorating deficit of hippocampal synaptic plasticity.

[Key techniques for precision cultivation of nitrogenous fertilizer of pollution-free ginseng].

Planting pollution-free farmland is the main mode of industrialization of ginseng cultivation, fine management of nitrogen fertilizer ginseng pollution-free farmland cultivation technology system is one of the key factors. In order to investigate the effect of nitrogenous fertilizer on the accumulation of ginseng biomass and saponins synthesis in vegetative growth stage, two-years-old ginsengs were used as test materials in this study. The test materials were cultivated by Hoagland medium with different nitrogen concentration (0，10，20，40 mg·L⁻¹) for 40 days. During the cultivation, photosynthetic rate was measured four times. After 40 days cultivation, chlorophyll content, stem diameter and the spatiotemporal expression of saponin synthesis related genes PgHMGR and PgSQE were tested. The results showed that there were significant differences in the photosynthetic rate and chlorophyll content among different nitrogen concentrations. The relative expression level of PgHMGR gene and PgSQE gene in root, stem and leaves of ginseng were different. Ginseng seedlings cultivated by 20 mg·L⁻¹ nitrogen possess the highest photosynthetic rate and chlorophyll content, while PgHMGR and PgSE showed the highest gene expression level. The optimal nitrogen concentration for the growth of 2-years-old ginseng might be 20 mg·L⁻¹ with 57.14 g ammonium nitrate each plant or pure 20.00 mg nitrogen each plant. It is concluded that this concentration is the most suitable concentration for the ginsenoside synthesis. Pollution-free ginseng with fine nitrogen fertilizer cultivation is conducive to the production of high quality and efficient ginseng medicinal materials. It lays a theoretical foundation for the rational fertilization and environment-friendly sustainable ecological ginseng planting industry.