High-throughput visualization mutation screening technology to enhance the specificity of CadR based whole-cell cadmium biosensor.

As a heavy metal pollutant, Cd2+ often enters the human body through the food chain causing great harm to human health. Whole-cell biosensor is an emerging technology for rapid on-site detection of heavy metals with the advantages of inexpensive, fast to mass-produce, and strong in anti-interference resistance, but suffering from insatisfactory specificity. In this study, a strategy of Adjacent Site Saturation Mutation (ASSM) was designed to improve the specificity of transcription factor CadR, which acted as the recognition element and determined the specificity of whole cell Cd2+ biosensors. A specific saturated library was constructed using the strategy of adjacent mutation. After two rounds of high-throughput visual screening, a whole-cell biosensor with good response to Cd2+, and with significant weakened Hg2+ interference was obtained. The optimized whole-cell biosensor showed a linear dynamic concentration range from 500 nM to 100 µM, a detection limit of 0.079 µM, and has satisfactory specificity and anti-interference. The ASSM strategy proposed in this study can provide a new method for the application of synthetic biology in food safety detection, indicating the importance of whole-cell biosensors for the detection of heavy metals.

Ammonia removal mitigates white plague type II in the coral Pocillopora damicornis.

White Plague Type II (WPL II) is a disease increasingly affecting scleractinian coral species and progresses rapidly. However, the etiological pathogen and remedy remain elusive. In this study, transmission experiments demonstrated that Aureimonas altamirensis and Aurantimonas coralicida, representing the WPL II pathogens, could infect Pocillopora damicorni. The infection produced selected pathological symptoms, including bleaching, tissue loss, and decolorization. Furthermore, ammonia degradation significantly reduced the severity of infection by these pathogens, indicating that ammonia may be a virulence factor for WPL II. Coral microbiome analysis suggested that ammonia degradation mediates the anti-white plague effect by maintaining the density of Symbiodiniaceae and stabilizing the core and symbiotic bacteria. Aureimonas altamirensis and Aurantimonas coralicida have been shown to cause diseases of P. damicornis, with ammonia acting as a virulence factor, and ammoniac degradation may be a promising and innovative approach to mitigate coral mortality suffering from increasing diseases.

Microbial community and transcriptional responses to V. coralliilyticus stress in coral Favites halicora and Pocillopora damicornis holobiont.

Variability in coral hosts susceptibility to Vibrio coralliilyticus is well-documented; however, the comprehensive understanding of tolerance of response to pathogen among coral species is lacked. Herein, we investigated the microbial communities and transcriptome dynamics of two corals in response to Vibrio coralliilyticus. Favites halicora displayed greater resistance to Vibrio coralliilyticus challenge than Pocillopora damicornis. Furthermore, the relative abundances of Flavobacteriaceae, Vibrionacea, Rhodobacteraceae, and Roseobacteraceae increased significantly in Favites halicora following pathogen stress, whereas that of Akkermansiaceae increased significantly in Pocillopora damicornis, leading to bacterial community imbalance. In contrast to the previous results, pathogen infection did not have much effect on the community structures of Symbiodiniaceae and fungi, but led to a decrease in the density of Symbiodiniaceae. Transcriptome analysis indicated that Vibrio infection triggered a coral immune response, resulting in higher expression of immune-related genes, which appeared to have higher transcriptional plasticity in Favites halicora than in Pocillopora damicornis. Specifically, the upregulated genes of Favites halicora were predominantly involved in the apoptosis pathway, whereas Pocillopora damicornis were significantly enriched in the nucleotide excision repair and base excision repair pathways. These findings suggest that coral holobionts activate different mechanisms across species in response to pathogens through shifts in microbial communities and transcriptomes, which provides novel insight into assessing the future coral assemblages suffering from disease outbreaks.

Replacing manual operation with bio-automation: A high-throughput evolution strategy to construct an integrated whole-cell biosensor for the simultaneous detection of methylmercury and mercury ions without manual sample digestion.

Methylmercury is primarily responsible for most food mercury pollution cases. However, most biosensors developed for mercury pollution analysis can only detect mercury ions. Although oxidative strong-acid digestion or microwave-assisted digestion can convert methylmercury into mercury ions, it is unsuitable for on-site detection. This study designed a bio-digestion gene circuit and integrated it into a mercury ion whole-cell biosensor，creating a novel on-site methylmercury detection method. Five alkyl mercury lyases from different bacterial genomes were screened via bioinformatics analysis, of which goMerB from Gordonia otitis showed the highest catalytic biological digestion efficiency. The goMerB site-specific saturation and random mutation libraries were constructed. After two rounds of high-throughput visualization screening, the catalytic activity of the mutant increased 2.5-fold. The distance between the three crucial amino acid sites and methylmercury changed in the mutant, which likely contributed to the enhanced catalytic efficiency. The optimized whole-cell biosensor showed a linear dynamic concentration range of 100 nM to 100 µM (R2 =0.991), satisfactory specificity, and interference resistance. The detection limit of the goMerBt6-MerR-RFP biosensor was 0.015 µM, while the limit of quantitation was 0.049 µM. This study demonstrated the application of synthetic biology for food safety detection and highlighted the future potential of "Lab in a Cell" for hazard analysis.

Bacterial carbonic anhydrase-induced carbonates mitigate soil erosion in biological soil crusts.

Soil erosion is a significant environmental issue worldwide, particularly in island regions where land resources are exceedingly scarce. Biological soil crusts play a crucial role in mitigating soil erosion, yet the precise effect and mechanism of biological soil crusts against erosion remain ambiguous. In this study, biological soil crusts at various developmental stages from a tropical coral island in the South China Sea were chosen to investigate the role of carbonic anhydrase in mitigating erosion. A cohesive strength meter, real-time quantitative PCR, and 16S rRNA gene high-throughput sequencing were employed to assess variations in soil antiscouribility as well as bacterial abundance and composition during the formation and development of biological soil crusts. Scanning electron microscopy was utilized to detect carbonates induced by bacterial carbonic anhydrase and elucidate their role in the solidification of sand particles. The findings indicate that the formation and development of biological soil crusts significantly enhance anti-scouribility. Comparison to those of bare coral sand, the shear stress increased from 0.35 to 1.11 N/m2 in the dark biocrusts. Moreover, significantly elevated carbonic anhydrase activity was observed in biological soil crusts, demonstrating a positive correlation with antiscouribility. In addition, there was a significant increase in bacterial abundance within the biological soil crusts. The enrichment of Cyanobacteriales and Chloroflexales potentially contributed to the increased carbonic anhydrase activity and antiscouribility. Furthermore, three cyanobacterial strains with carbonic anhydrase activity were isolated from biological soil crusts and subsequently confirmed to enhance sand solidification through microbial carbonate precipitation. This study presents initial evidence for the role of microbial carbonic anhydrase in enhancing the antiscouribility of biological soil crusts during their formation and development. These findings offer novel insights into the functional and mechanistic dimensions underlying the mitigation of soil erosion facilitated by biological soil crusts, which are valuable for implementing sustainable biorestoration and environmental management technologies to prevent soil erosion.

m6A-regulated tumor glycolysis: new advances in epigenetics and metabolism.

Glycolytic reprogramming is one of the most important features of cancer and plays an integral role in the progression of cancer. In cancer cells, changes in glucose metabolism meet the needs of self-proliferation, angiogenesis and lymphangiogenesis, metastasis, and also affect the immune escape, prognosis evaluation and therapeutic effect of cancer. The n6-methyladenosine (m6A) modification of RNA is widespread in eukaryotic cells. Dynamic and reversible m6A modifications are widely involved in the regulation of cancer stem cell renewal and differentiation, tumor therapy resistance, tumor microenvironment, tumor immune escape, and tumor metabolism. Lately, more and more evidences show that m6A modification can affect the glycolysis process of tumors in a variety of ways to regulate the biological behavior of tumors. In this review, we discussed the role of glycolysis in tumor genesis and development, and elaborated in detail the profound impact of m6A modification on different tumor by regulating glycolysis. We believe that m6A modified glycolysis has great significance and potential for tumor treatment.

Advances in Synthetic-Biology-Based Whole-Cell Biosensors: Principles, Genetic Modules, and Applications in Food Safety.

A whole-cell biosensor based on synthetic biology provides a promising new method for the on-site detection of food contaminants. The basic components of whole-cell biosensors include the sensing elements, such as transcription factors and riboswitches, and reporting elements, such as fluorescence, gas, etc. The sensing and reporting elements are coupled through gene expression regulation to form a simple gene circuit for the detection of target substances. Additionally, a more complex gene circuit can involve other functional elements or modules such as signal amplification, multiple detection, and delay reporting. With the help of synthetic biology, whole-cell biosensors are becoming more versatile and integrated, that is, integrating pre-detection sample processing, detection processes, and post-detection signal calculation and storage processes into cells. Due to the relative stability of the intracellular environment, whole-cell biosensors are highly resistant to interference without the need of complex sample preprocessing. Due to the reproduction of chassis cells, whole-cell biosensors replicate all elements automatically without the need for purification processing. Therefore, whole-cell biosensors are easy to operate and simple to produce. Based on the above advantages, whole-cell biosensors are more suitable for on-site detection than other rapid detection methods. Whole-cell biosensors have been applied in various forms such as test strips and kits, with the latest reported forms being wearable devices such as masks, hand rings, and clothing. This paper examines the composition, construction methods, and types of the fundamental components of synthetic biological whole-cell biosensors. We also introduce the prospect and development trend of whole-cell biosensors in commercial applications.

Applications of the Whole-Cell System in the Efficient Biosynthesis of Heme.

Heme has a variety of functions, from electronic reactions to binding gases, which makes it useful in medical treatments, dietary supplements, and food processing. In recent years, whole-cell system-based heme biosynthesis methods have been continuously explored and optimized as an alternative to the low-yield, lasting, and adverse ecological environment of chemical synthesis methods. This method relies on two biosynthetic pathways of microbial precursor 5-aminolevulinic acid (C4, C5) and three known downstream biosynthetic pathways of heme. This paper reviews the genetic and metabolic engineering strategies for heme production in recent years by optimizing culture conditions and techniques from different microorganisms. Specifically, we summarized and analyzed the possibility of using biosensors to explore new strategies for the biosynthesis of heme from the perspective of synthetic biology, providing a new direction for future exploration.

A selectivity-enhanced fluorescence imprinted sensor based on yellow-emission peptide nanodots for sensitive and visual smart detection of λ-cyhalothrin.

The development of precise and efficient detection technologies to recognize λ-cyhalothrin (LC) in agricultural products has attracted attention worldwide due to its widespread use and notable toxic effects on humans. Herein, a novel fluorescence biomimetic nanosensor was elaborately designed based on Zn(II)-doped cyclo-ditryptophan (c-WW)-type peptide nanodots and incorporating molecularly imprinted polymer (c-WW/Zn-PNs@MIP) for LC assays. C-WW/Zn-PNs obtained by self-assembly with aromatic cyclic dipeptides as basic building blocks and coordination with Zn(II) have low-toxicity, photostability, and bright yellow fluorescence emission, as a sensitive signal transducer. High-affinity imprinting sites further endow c-WW/Zn-PNs@MIP with superior selectivity and reusability. Based on prominent merits, c-WW/Zn-PNs@MIP demonstrated a good linear range (1-360 µg/L) with a low limit of detection (LOD) (0.93 µg/L), fast kinetics in target capture (10 min), and strong practicability in the capture of LC from real samples (spiked recovery of 81.0-107.7%). Additionally, to attain onsite profiling of LC, a visual platform was developed by integrating c-WW/Zn-PNs@MIP with a smartphone-assisted optical device. This smart evaluation system can capture concentration-dependent fluorescent images and accurately digitize them, enabling quantitative analysis of LC. This study developed a fluorescent c-WW/Zn-PNs@MIP-based smart evaluation system as a novel platform for LC monitoring applications, which not only has enormous economic value but also great environmental health significance.

Magnetically Controlled Nanorobots Based on Red Emissive Peptide Dots and Artificial Antibodies for Specific Recognition and Smart Scavenging of Nε-(Carboxymethyl)lysine in Dairy Products.

Food-borne advanced glycation end products (AGEs) are highly related to various irreversible diseases, and Nε-(carboxymethyl)lysine (CML) is the typical hazardous AGE. The development of feasible strategies to monitor and reduce CML exposure has become desirable to address the problems. In this work, we proposed magnetically controlled nanorobots by integrating an optosensing platform with specific recognition and binding capability, realizing specific anchoring and accurate determination as well as efficient scavenging of CML in dairy products. The artificial antibodies offered CML imprinted cavities for highly selective absorption, and the optosensing strategy was designed based on electron transfer from red emissive self-assembling peptide dots (r-SAPDs) to CML, which was responsible for the identity, response, and loading process. The r-SAPDs overcame the interference from autofluorescence, and the limit of detection was 0.29 µg L-1, which bestowed accuracy and reliability for in situ monitoring. The selective binding process was accomplished within 20 min with an adsorption capacity of 23.2 mg g-1. Through an external magnetic field, CML-loaded nanorobots were oriented, moved, and separated from the matrix, which enabled their scavenging effects and reusability. The fast stimuli-responsive performance and recyclability of the nanorobots provided a versatility strategy for effective detection and control of hazards in food.

Targets and mechanisms of Alpinia oxyphylla Miquel fruits in treating neurodegenerative dementia.

The dried and ripe fruits of Alpinia oxyphylla and ripe fruits of Alpinia oxyphylla Miquel (AO) have the effects of tonifying kidney-essence and nourishing intelligence and thus have been widely used in treating dementia. Alzheimer's disease (AD) is a typical form of neurodegenerative dementia with kidney-essence deficiency in Traditional Chinese Medicine (TCM). So far, there is a lack of systematic studies on the biological basis of tonifying kidney-essence and nourishing intelligence and the corresponding phytochemicals. In this study, we investigated the targets of AO in tonifying kidney-essence and nourishing intelligence based on the key pathophysiological processes of neurodegenerative dementia. According to ultra-high-performance liquid chromatography with triple quadrupole mass spectrometry data and Lipinski's rule of five, 49 bioactive phytochemicals from AO were identified, and 26 of them were found to target 168 key molecules in the treatment of neurodegenerative dementia. Nine phytochemicals of AO were shown to target acetylcholinesterase (ACHE), and 19 phytochemicals were shown to target butyrylcholinesterase (BCHE). A database of neurodegenerative dementia with kidney-essence deficiency involving 731 genes was constructed. Furthermore, yakuchinone B, 5-hydroxy-1,7-bis (4-hydroxy-3-methoxyphenyl) heptan-3-one (5-HYD), oxyhylladiketone, oxyphyllacinol, butyl-ß-D-fructopyranoside, dibutyl phthalate, chrysin, yakuchinone A, rhamnetin, and rhamnocitrin were identified as the key phytochemicals from AO that regulate the pathogenesis of neurodegenerative dementia in a multitargeted manner. The approach of studying the pharmacological mechanism underlying the effects of medicinal plants and the biological basis of TCM syndrome may be helpful in studying the translation of TCM.

Significant Shifts in Microbial Communities Associated with Scleractinian Corals in Response to Algae Overgrowth.

Microbes play a key role in reef dynamics, mediating the competition between scleractinian corals and benthic algae; however, major shifts in bacterial communities among coral species in response to increases in the abundance of algae are not well understood. We investigated the taxonomic composition of coral-associated microbial communities under algae-overgrowth conditions using 16S rRNA gene sequencing. The results showed that non-algal (i.e., healthy) tissue (HH) had lower bacterial abundance and diversity than tissue collected from the coral-algae interface boundary (HA) and areas of algae growth (AA). Specifically, the HA and AA samples had higher relative abundances of Saprospiraceae, Rhodobacteraceae, and Alteromonadaceae. Compared with Platygyra sp. and Montipora sp., the physiological response of Pocillopora sp. was more intense under algae-induced stress based on microbial gene function prediction. Our results indicate that algal pressure can significantly alter the microbial community structure and function of coral ecosystems. Our data thus provide new insight into the relationship between corals and their microbiome under environmental stress.

A Tau Pathogenesis-Based Network Pharmacology Approach for Exploring the Protections of Chuanxiong Rhizoma in Alzheimer's Disease.

Alzheimer's disease (AD) is the most common cause of neurodegenerative dementia and one of the top medical concerns worldwide. Currently, the approved drugs to treat AD are effective only in treating the symptoms, but do not cure or prevent AD. Although the exact causes of AD are not understood, it is recognized that tau aggregation in neurons plays a key role. Chuanxiong Rhizoma (CR) has been widely reported as effective for brain diseases such as dementia. Thus, we explored the protections of CR in AD by a tau pathogenesis-based network pharmacology approach. According to ultra-HPLC with triple quadrupole mass spectrometry data and Lipinski's rule of five, 18 bioactive phytochemicals of CR were screened out. They were shown corresponding to 127 tau pathogenesis-related targets, among which VEGFA, IL1B, CTNNB1, JUN, ESR1, STAT3, APP, BCL2L1, PTGS2, and PPARG were identified as the core ones. We further analyzed the specific actions of CR-active phytochemicals on tau pathogenesis from the aspects of tau aggregation and tau-mediated toxicities. It was shown that neocnidilide, ferulic acid, coniferyl ferulate, levistilide A, Z-ligustilide, butylidenephthalide, and caffeic acid can be effective in reversing tau hyperphosphorylation. Neocnidilide, senkyunolide A, butylphthalide, butylidenephthalide, Z-ligustilide, and L-tryptophan may be effective in promoting lysosome-associated degradation of tau, and levistilide A, neocnidilide, ferulic acid, L-tryptophan, senkyunolide A, Z-ligustilide, and butylidenephthalide may antagonize tau-mediated impairments of intracellular transport, axon and synaptic damages, and neuron death (especially apoptosis). The present study suggests that acting on tau aggregation and tau-mediated toxicities is part of the therapeutic mechanism of CR against AD.

Description of Prasinibacter corallicola gen. nov., sp. nov., a zeaxanthin-producing bacterium isolated from stony coral Porites lutea.

Thermal stress is considered one of the main causes of mass scleractinian coral degradation; however, it is still unknown how corals can adapt to future global warming. In this study, 11 strains of coral-associated Flavobacteria were shown to produce zeaxanthin, a carotenoid antioxidant, which may help coral holobionts to alleviate thermal stress. In addition, a novel zeaxanthin-producing Flavobacterium, designated R38T, was identified using polyphasic taxonomy. Although strain R38T shared a maximum 16S rRNA gene sequence similarity of 93% with Mesoflavibacter aestuarii KYW614T, phylogenetic analyses based on whole genome and 16S rRNA gene sequences revealed that strain R38T forms a distinct branch in a robust cluster composed of strain R38T and Leptobacterium flavescens KCTC 22160T under the family Flavobacteriaceae. Strain R38T exhibited average nucleotide identities of 70.2% and 72.5% for M. aestuarii KYW614T and L. flavescens KCTC 22160T, respectively. The only detected respiratory quinone was menaquinone 6 (MK-6). The genomic DNA G + C content was 33.2 mol%. The major polar lipids were phosphatidylmethylethanolamine, phosphatidylethanolamine, one unidentified ninhydrin phospholipid, three unidentified ninhydrin-positive lipids, and three unidentified lipids. The major cellular fatty acids were iso - C15: 0, iso - C15: 0 ω6c, C16:2 DMA, and C13:1 ω3c. The distinct biochemical, chemotaxonomic, phylogenetic, and phylogenomic differences from validly published taxa suggest that strain R38T represents a new species of a new genus, for which Prasinibacter corallicola gen. nov., sp. nov. is proposed. The type strain R38T (= MCCC 1K03889T = KCTC 72444T).

Comparative Genomics Reveal the Animal-Associated Features of the Acanthopleuribacteraceae Bacteria, and Description of Sulfidibacter corallicola gen. nov., sp., nov.

Members of the phylum Acidobacteria are ubiquitous in various environments. Soil acidobacteria have been reported to present a variety of strategies for their success in terrestrial environments. However, owing to lack of pure culture, information on animal-associated acidobacteria are limited, except for those obtained from 16S rRNA genes. To date, only two acidobacteria have been isolated from animals, namely strain M133T obtained from coral Porites lutea and Acanthopleuribacter pedis KCTC 12899T isolated from chiton. Genomics and physiological characteristics of strain M133T and A. pedis KCTC 12899T were compared with 19 other isolates (one strain from each genus) in the phylum Acidobacteria. The results revealed that strain M133T represents a new species in a new genus in the family Acanthopleuribacteraceae. To date, these two Acanthopleuribacteraceae isolates have the largest genomes (10.85-11.79 Mb) in the phylum Acidobacteria. Horizontal gene transfer and gene duplication influenced the structure and plasticity of these large genomes. Dissimilatory nitrate reduction and abundant secondary metabolite biosynthetic gene clusters (including eicosapentaenoic acid de novo biosynthesis) are two distinct features of the Acanthopleuribacteraceae bacteria in the phylum Acidobacteria. The absence of glycoside hydrolases involved in plant polysaccharide degradation and presence of animal disease-related peptidases indicate that these bacteria have evolved to adapt to the animal hosts. In addition to low- and high-affinity respiratory oxygen reductases, enzymes for nitrate to nitrogen, and sulfhydrogenase were also detected in strain M133T, suggesting the capacity and flexibility to grow in aerobic and anaerobic environments. This study highlighted the differences in genome structure, carbohydrate and protein utilization, respiration, and secondary metabolism between animal-associated acidobacteria and other acidobacteria, especially the soil acidobacteria, displaying flexibility and versatility of the animal-associated acidobacteria in environmental adaption.

Use of a purified ß-glucosidase from coral-associated microorganisms to enhance wine aroma.

BACKGROUND: ß-Glucosidases (3.2.1.21) play essential roles in the removal of nonreducing terminal glucosyl residues from saccharides and glycosides. However, the full potential and different applications of recombinant high-yield microbial ß-glucosidase-producing systems remain to be tackled. RESULTS: A ß-glucosidase gene designated as Mg132 was isolated from a coral microorganism by high-throughput sequencing and functional screening. The deduced amino acid sequences of Mg132 showed a highest identity of 97% with ß-glucosidase predicted in the GenBank database. This gene was cloned and overexpressed in Escherichia coli BL21 (DE3) for the first time. The optimal pH and temperature of purified recombinant Mg132 were 8.0 and 50 °C respectively. It exhibited a high level of stability at high concentration of glucose and ethanol, and glucose concentrations below 300 mmol L-1 distinctly stimulated p-nitrophenyl-ß-d-glucopyranoside hydrolysis, reaching 200% at 15% ethanol. The Km and Vmax values were 0.293 mmol L-1 and 320 µmol min-1  mg-1 respectively while using p-nitrophenyl-ß-d-glucopyranoside as a substrate. Wine treated with Mg132 had an obvious positive catalytic specificity for glycosides, which give a pleasant flavor of temperate fruity and floral aromas. The total concentration of fermentative volatiles was 201.42 ± 10.22 µg L-1 following Mg132 treatment and 99.21 ± 7.72 µg L-1 in control samples. CONCLUSION: Good tolerance of winemaking and aroma fermentative properties suggest that Mg132 has potential application in aroma enhancement in wine and warrants further study. © 2021 Society of Chemical Industry.

A Novel Neutral and Mesophilic ß-Glucosidase from Coral Microorganisms for Efficient Preparation of Gentiooligosaccharides.

ß-glucosidases can produce gentiooligosaccharides that are lucrative and promising for the prebiotic and alternative food industries. However, the commercial production of gentiooligosaccharides using ß-glucosidase is challenging, as this process is limited by the need for high thermal energy and increasing demand for the enzyme. Here, a putative ß-glucosidase gene, selected from the coral microbial metagenome, was expressed in Escherichia coli. Reverse hydrolysis of glucose by Blg163 at pH 7.0 and 40 °C achieved a gentiooligosaccharide yield of 43.02 ± 3.20 g·L-1 at a conversion rate of 5.38 ± 0.40%. Transglycosylation of mixed substrates, glucose and cellobiose, by Blg163 consumed 21.6 U/0.5 g glucose/g cellobiose, achieving a gentiooligosaccharide yield of 70.34 ± 2.20 g·L-1 at a conversion rate of 15.63%, which is close to the highest yield reported in previous findings. Blg163-mediated synthesis of gentiooligosaccharides is the mildest reaction and the lowest ß-glucosidase consumption reported to date.

Cross-Linked Regulation of Coral-Associated Dinoflagellates and Bacteria in Pocillopora sp. during High-Temperature Stress and Recovery.

As the problem of ocean warming worsens, the environmental adaptation potential of symbiotic Symbiodiniaceae and bacteria is directly related to the future and fate of corals. This study aimed to analyse the comprehensive community dynamics and physiology of these two groups of organisms in the coral Pocillopora sp. through indoor simulations of heat stress (which involved manually adjusting the temperature between both 26 °C and 34 °C). Heat treatment (≥30 °C) significantly reduced the abundance of Symbiodiniaceae and bacteria by more than 70%. After the temperature was returned to 26 °C for one month, the Symbiodiniaceae density was still low, while the absolute number of bacteria quickly recovered to 55% of that of the control. At this time point, the Fv/Fm value rose to 91% of the pretemperature value. The content of chlorophyll b associated with Cyanobacteria increased by 50% compared with that under the control conditions. Moreover, analysis of the Symbiodiniaceae subclade composition suggested that the relative abundance of C1c.C45, C1, and C1ca increased during heat treatment, indicating that they might constitute heat-resistant subgroups. We suggest that the increase in the absolute number of bacteria during the recovery period could be an important indicator of coral holobiont recovery after heat stress. This study provides insight into the cross-linked regulation of key symbiotic microbes in the coral Pocillopora sp. during high-temperature stress and recovery and provides a scientific basis for exploring the mechanism underlying coral adaptation to global warming.

Protective effects of Da-cheng-qi decoction in rats with intracerebral hemorrhage.

BACKGROUND: Intracerebral hemorrhage (ICH), the most fatal subtype of stroke, has no disease-modifying treatment. Da-cheng-qi decoction (DCQ), composed of rhubarb, is one of the most commonly used Chinese traditional decoctions in ICH treatment. But the mechanism is not clear. Emodin is an active compound found in rhubarb. PURPOSE: To study the protective effects of DCQ on ICH and its possible mechanisms of action. METHODS: The ICH model was reproduced by injecting collagenase-VII into the left caudate putamen (CPu) of rats. DCQ and emodin were used to treat the ICH rats for 7 days. Behavior tests, proteomic analysis, morphological studies, and western blotting were performed. RESULTS: The neurological deficits in the ICH rats recovered with DCQ and emodin on the 14th day after ICH. The proteomics data revealed that DCQ significantly corrected the pathological signals in the CPu and hippocampus after ICH. The numbers of amoebic microglia in the CPu and M2 microglia in both CPu and hippocampus were significantly increased after DCQ and emodin treatment. The increase in GluN2B-containing NMDA receptor (NR2B) and postsynaptic density protein-95, activation of mitogen-activated protein kinase (MAPK) signals in the CPu, and secondary neurodegeneration (SND) in the hippocampus were significantly recovered in DCQ-treated rats. Inhibition of MAPK p38 (p38) in the hippocampus was observed after DCQ and emodin treatment. CONCLUSION: The protective effects of DCQ on ICH were confirmed in this study, and its mechanism may be related to the inhibition of MAPK and activation of M2 microglia. These results are beneficial to the development of ICH therapeutic targets.

Mechanistic insights into the anti-depressant effect of emodin: an integrated systems pharmacology study and experimental validation.

Depression is a complex neuropsychiatric disease involved multiple targets and signaling pathways. Systems pharmacology studies could potentially present a comprehensive molecular mechanism to delineate the anti-depressant effect of emodin (EMO). In this study, we investigated the anti-depressant effects of EMO in the chronic unpredictable mild stress (CUMS) rat model of depression and gained insights into the underlying mechanisms using systems pharmacology and molecular simulation analysis. Forty-three potential targets of EMO for treatment of depression were obtained. GO biological process analysis suggested that the biological functions of these targets mainly involve the regulation of reactive oxygen species metabolic process, response to lipopolysaccharide, regulation of inflammatory response, etc. KEGG pathway enrichment analysis showed that the PI3K-Akt signaling pathway, insulin resistance, IL-17 signaling pathway were the most significantly enriched signaling pathways. The molecular docking analysis revealed that EMO might have a strong combination with ESR1, AKT1 and GSK3B. Immunohistochemical staining and Western blotting showed that 2 weeks' EMO treatment (80 mg/kg/day) reduced depression related microglial activation, neuroinflammation and altered PI3K-Akt signaling pathway. Our findings provide a systemic pharmacology basis for the anti-depressant effects of EMO.