Comparative Transcriptome Analysis Revealed Candidate Gene Modules Involved in Salt Stress Response in Sweet Basil and Overexpression of ObWRKY16 and ObPAL2 Enhanced Salt Tolerance of Transgenic Arabidopsis.

Sweet basil (Ocimum basilicum L.) is an important aromatic plant with high edibility and economic value, widely distributed in many regions of the tropics including the south of China. In recent years, environmental problems, especially soil salinization, have seriously restricted the planting and spread of sweet basil. However, the molecular mechanism of the salt stress response in sweet basil is still largely unknown. In this study, seed germination, seedling growth, and chlorophyll synthesis in sweet basil were inhibited under salt stress conditions. Through comparative transcriptome analysis, the gene modules involved in the metabolic processes, oxidative response, phytohormone signaling, cytoskeleton, and photosynthesis were screened out. In addition, the landscape of transcription factors during salt treatment in sweet basil was displayed as well. Moreover, the overexpression of the WRKY transcription factor-encoding gene, ObWRKY16, and the phenylalanine ammonia-lyase-encoding gene, ObPAL2, enhanced the seed germination, seedling growth, and survival rate, respectively, of transgenic Arabidopsis, suggesting that they might be important candidates for the creation of salt-tolerant sweet basil cultivars. Our data enrich the study on salt responses in sweet basil and provide essential gene resources for genetic improvements in sweet basil in the future.

Preliminary Study on the Pathogenic Mechanism of Jujube Flower Disease in Honeybees (Apis mellifera ligustica) Based on Midgut Transcriptomics.

Honeybees are prone to poisoning, also known as jujube flower disease, after collecting nectar from jujube flowers, resulting in the tumultuous demise of foragers. The prevalence of jujube flower disease has become one of the main factors affecting the development of the jujube and beekeeping industries in Northern China. However, the pathogenic mechanisms underlying jujube flower disease in honeybees are poorly understood. Herein, we first conducted morphological observations of the midgut using HE-staining and found that jujube flower disease-affected honeybees displayed midgut damage with peritrophic membrane detachment. Jujube flower disease was found to increase the activity of chitinase and carboxylesterase (CarE) and decrease the activity of superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and the content of CYP450 in the honeybee midgut. Transcriptomic data identified 119 differentially expressed genes in the midgut of diseased and healthy honeybees, including CYP6a13, CYP6a17, CYP304a1, CYP6a14, AADC, and AGXT2, which are associated with oxidoreductase activity and vitamin binding. In summary, collecting jujube flower nectar could reduce antioxidant and detoxification capacities of the honeybee midgut and, in more severe cases, damage the intestinal structure, suggesting that intestinal damage might be the main cause of honeybee death due to jujube nectar. This study provides new insights into the pathogenesis of jujube flower disease in honeybees.

Liposomal STAT3-Degrading PROTAC Prodrugs Promote Anti-Hepatocellular Carcinoma Immunity via Chemically Reprogramming Cancer Stem Cells.

Cancer stem cells (CSCs) with hyperactivated signal transducer and activator of transcription 3 (STAT3) are a major driver of hepatocellular carcinoma (HCC). Herein, we report a nanointegrative proteolysis-targeting chimera (PROTAC)-based STAT3 degradation strategy that enables efficient chemical reprogramming of HCC-associated CSCs, which potently inhibits CSC growth while evoking anti-HCC immune responses. The PROTAC prodrug was synthesized by conjugating the STAT3 binding domain (inS3) with a thioketal-caged E3 ligase ligand (VL-TK) via an oligo(ethylene glycol) linker (OEG) with tuned length and flexibility and encapsulating it in cRGD-modified cationic liposomes for CSC-targeted delivery while facilitating their lysosomal escape. The PROTAC prodrugs were activated by the upregulated ROS levels in CSCs and efficiently degraded STAT3 for chemical reprogramming, which would not only impair their stemness features but also remodel the immunosuppressive TME into an immunosupportive state to boost anti-HCC immunity. This strategy provides an approach for improving HCC treatment in clinics.

[Effect of Suanzaoren Decoction on expression of ionotropic glutamate receptors and synaptic plasticity in hippocampus of anxiety rats].

This study investigated the effect of Suanzaoren Decoction on the expression of N-methyl-D-aspartate receptors(NMDAR) and α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptors(AMPAR) in the hippocampus and synaptic plasticity in rats with conditioned fear-induced anxiety. The effect of Suanzaoren Decoction on rat behaviors were evaluated through open field experiment, elevated plus maze experiment, and light/dark box experiment. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of glutamate(Glu) and γ-aminobutyric acid(GABA) in the rat hippocampus. Real-time fluorescence quantitative PCR(qRT-PCR) and Western blot were employed to assess the gene and protein expression of ionotropic glutamate receptors in the hippocampal region. Transmission electron microscopy was utilized to observe the changes in the ultrastructure of synaptic neurons in the hippocampal region. Long-term potentiation(LTP) detection technique was employed to record the changes in population spike(PS) amplitude in the hippocampal region of mice in each group. The behavioral results showed that compared with the model group, the Suanzaoren Decoction group effectively increased the number of entries into open arms, time spent in open arms, percentage of time spent in open arms out of total movement time, number of entries into open arms out of total entries into both arms(P<0.01), and significantly increased the time spent in the light box and the number of shuttle crossings(P<0.01). There was an increasing trend in the number of grid crossings, entries into the center grid, and time spent in the center grid, indicating a significant anxiolytic effect. ELISA results showed that compared with the model group, the Suanzaoren Decoction group exhibited significantly reduced levels of Glu, Glu/GABA ratio(P<0.01), and significantly increased levels of GABA(P<0.01) in the rat hippocampus. Furthermore, Suanzaoren Decoction significantly decreased the gene and protein expression of NMDAR(GluN2B and GluN2A) and AMPAR(GluA1 and GluA2) compared with the model group. Transmission electron microscopy results demonstrated improvements in synapses, neuronal cells, and organelles in the hippocampal region of the Suanzaoren Decoction group compared with the model group. LTP detection results showed a significant increase in the PS amplitude changes in the hippocampal region of Suanzaoren Decoction group from 5 to 35 min compared with the model group(P<0.05, P<0.01). In conclusion, Suanzaoren Decoction exhibits significant anxiolytic effects, which may be attributed to the reduction in NMDAR and AMPAR expression levels and the improvement of synaptic plasticity.

Inhibition of glycolysis-driven immunosuppression with a nano-assembly enhances response to immune checkpoint blockade therapy in triple negative breast cancer.

Immune-checkpoint inhibitors (ICI) are promising modalities for treating triple negative breast cancer (TNBC). However, hyperglycolysis, a hallmark of TNBC cells, may drive tumor-intrinsic PD-L1 glycosylation and boost regulatory T cell function to impair ICI efficacy. Herein, we report a tumor microenvironment-activatable nanoassembly based on self-assembled aptamer-polymer conjugates for the targeted delivery of glucose transporter 1 inhibitor BAY-876 (DNA-PAE@BAY-876), which remodels the immunosuppressive TME to enhance ICI response. Poly ß-amino ester (PAE)-modified PD-L1 and CTLA-4-antagonizing aptamers (aptPD-L1 and aptCTLA-4) are synthesized and co-assembled into supramolecular nanoassemblies for carrying BAY-876. The acidic tumor microenvironment causes PAE protonation and triggers nanoassembly dissociation to initiate BAY-876 and aptamer release. BAY-876 selectively inhibits TNBC glycolysis to deprive uridine diphosphate N-acetylglucosamine and downregulate PD-L1 N-linked glycosylation, thus facilitating PD-L1 recognition of aptPD-L1 to boost anti-PD-L1 therapy. Meanwhile, BAY-876 treatment also elevates glucose supply to tumor-residing regulatory T cells (Tregs) for metabolically rewiring them into an immunostimulatory state, thus cooperating with aptCTLA-4-mediated immune-checkpoint inhibition to abolish Treg-mediated immunosuppression. DNA-PAE@BAY-876 effectively reprograms the immunosuppressive microenvironment in preclinical models of TNBC in female mice and provides a distinct approach for TNBC immunotherapy in the clinics.

Comparative transcriptomic analysis provides key genetic resources in clove basil (Ocimum gratissimum) under cadmium stress.

Planting aromatic plant might be a promising strategy for safely utilizing heavy metal (HM)-contaminated soils, as HMs in essential oil could be completely excluded using some special technologies with ease. Clove basil (Ocimum gratissimum L.) is an important aromatic plant used in essential oil production. Improving cadmium (Cd) tolerance in clove basil can increase its production and improve the utilization efficiency of Cd-contaminated soils. However, the lack of genomic information on clove basil greatly restricts molecular studies and applications in phytoremediation. In this study, we demonstrated that high levels of Cd treatments (0.8, 1.6 and 6.5 mg/L) significantly impacted the growth and physiological attributes of clove basil. Cd contents in clove basil tissues increased with treatment concentrations. To identify Cd stress-responsive genes, we conducted a comparative transcriptomic analysis using seedlings cultured in the Hoagland's solution without Cd ion (control) or containing 1.6 mg/L CdCl2 (a moderate concentration of Cd stress for clove basil seedlings). A total of 104.38 Gb clean data with high-quality were generated in clove basil under Cd stress through Illumina sequencing. More than 1,800 differential expressed genes (DEGs) were identified after Cd treatment. The reliability and reproducibility of the transcriptomic data were validated through qRT-PCR analysis and Sanger sequencing. KEGG classification analysis identified the "MAPK signaling pathway," "plant hormone signal transduction" and "plant-pathogen interaction" as the top three pathways. DEGs were divided into five clusters based on their expression patterns during Cd stress. The functional annotation of DEGs indicated that downregulated DEGs were mainly involved in the "photosynthesis system," whereas upregulated DEGs were significantly assigned to the "MAPK signaling pathway" and "plant-pathogen interaction pathway." Furthermore, we identified a total of 78 transcription factors (TFs), including members of bHLH, WRKY, AP2/ERF, and MYB family. The expression of six bHLH genes, one WRKY and one ERF genes were significantly induced by Cd stress, suggesting that these TFs might play essential roles in regulating Cd stress responses. Overall, our study provides key genetic resources and new insights into Cd adaption mechanisms in clove basil.

A Multi-Scale Tool Orientation Generation Method for Freeform Surface Machining with Bull-Nose Tool.

Free-form surface parts are widely used in industries, and they consist of intricate 3D surfaces such as molds, impellers, and turbine blades that possess complex geometrical contours and demand high precision. Proper tool orientation is crucial for ensuring the efficiency and accuracy of five-axis computer numerical control (CNC) machining. Multi-scale methods have received much attention and have been widely used in various fields. They have been proven to be instrumental and can obtain fruitful outcomes. Ongoing research on multi-scale tool orientation generation methods, which aim to acquire tool orientations that satisfy both macro- and micro-scale requirements, is significantly important for improving the machining quality of workpiece surfaces. This paper proposes a multi-scale tool orientation generation method that considers both the machining strip width and roughness scales. This method also ensures a smooth tool orientation and avoids interference in the machining process. First, the correlation between the tool orientation and rotational axis is analyzed, and feasible area calculation and tool orientation adjustment methods are introduced. Then, the paper introduces the calculation method for machining strip widths on the macro-scale and the roughness calculation method on the micro-scale. Besides, tool orientation adjustment methods for both scales are proposed. Next, a multi-scale tool orientation generation method is developed to generate tool orientations that meet the macro- and micro-scale requirements. Finally, to verify the effectiveness of the proposed multi-scale tool orientation generation method, it is applied to the machining of a free-form surface. Experimental verification results have shown that the tool orientation generated by the proposed method can obtain the expected machining strip width and roughness, meeting both macro- and micro-scale requirements. Therefore, this method has significant potential for engineering applications.

The molecular mechanism of cardiac injury in SARS-CoV-2 infection: Focus on mitochondrial dysfunction.

BACKGROUND: Coronavirus disease 2019(COVID-19) caused a large number of infections worldwide. Although some patients recovered from the disease, some of the other problems that accompanied it, such as cardiac injury, could affect the patient's subsequent quality of life and prognosis. OBJECTIVES: To clarify the molecular mechanism of cardiac injury in SARS-CoV-2 Infection. METHODS: The RNA-Seq dataset (GSE184715) comparing expression profiling of Mock human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and SARS-CoV-2-infected hiPSC-CMs was downloaded from Gene Expression Omnibus (GEO). Differentially expressed genes(DEGs) were performed by the R software. Degs were analyzed by enrichment analysis to clarify the affected pathways. Hub genes were screened out by a PPI network constructed from Degs. Finally, Connectivity Map was used to screen for the treatment of COVID-19 induced cardiac injury. RESULTS: 2705 differentially expressed genes were identified. Enrichment analysis confirmed that mitochondrial dysfunction was caused by SARS-CoV-2, meanwhile, cardiac muscle contraction was suppressed and NF-κB was activated. Based on the PPI network, 15 hub genes were identified. These 15 down-regulated hub genes were mainly involved in the reduced activity of complexes in the mitochondrial respiratory chain associated with mitochondrial dysfunction. Moreover, 5 candidate drugs were identified to treat cardiac injury. CONCLUSION: In conclusion, SARS-CoV-2 infection of cardiomyocytes causes mitochondrial dysfunction, including reduced mitochondrial respiratory chain complex activity and decreased ATP synthesis, leading to cardiomyocyte apoptosis, while the activated NF-κB also induced cytokine storms, ultimately resulting in cardiac injury.

Low-level cadmium exposure induced hormesis in peppermint young plant by constantly activating antioxidant activity based on physiological and transcriptomic analyses.

As one of the most toxic environmental pollutants, cadmium (Cd) has lastingly been considered to have negative influences on plant growth and productivity. Recently, increasing studies have shown that low level of Cd exposure could induce hormetic effect which benefits to plants. However, the underlying mechanisms of Cd-triggered hormesis are poorly understood. In this study, we found that Cd stress treatment showed a hormetic effect on peppermint and Cd treatment with 1.6 mg L-1 concertation manifested best stimulative effects. To explore the hormesis mechanisms of Cd treatment, comparative transcriptome analysis of peppermint young plants under low (1.6 mg L-1) and high (6.5 mg L-1) level of Cd exposure at 0 h, 24 h and 72 h were conducted. Twelve of differentially expressed genes (DEGs) were selected for qRT-PCR validation, and the expression results confirmed the credibility of transcriptome data. KEGG analysis of DEGs showed that the phenylpropanoid biosynthesis and photosynthesis were important under both low and high level of Cd treatments. Interestingly, GO and KEGG analysis of 99 DEGs specifically induced by low level of Cd treatment at 72 h indicated that these DEGs were mainly involved in the pathway of phenylpropanoid biosynthesis and their functions were associated with antioxidant activity. The expression pattern of those genes in the phenylpropanoid biosynthesis pathway and encoding antioxidant enzymes during 72 h of Cd exposure showed that low level of Cd treatment induced a continuation in the upward trend but high level of Cd treatment caused an inverted V-shape. The changes of physiological parameters during Cd exposure were highly consistent with gene expression pattern. These results strongly demonstrate that low level of Cd exposure constantly enhanced antioxidant activity of peppermint to avoid oxidative damages caused by Cd ion, while high level of Cd stress just induced a temporary increase in antioxidant activity which was insufficient to cope with lasting Cd toxicity. Overall, the results presented in this study shed a light on the underlying mechanisms of the Cd-mediated hormesis in plant. Moreover, our study provided a safe method for the efficient utilization of mild Cd-contaminated soil as peppermint is an important cash plant.

Full length transcriptomes analysis of cold-resistance of Apis cerana in Changbai Mountain during overwintering period.

Apis cerana in Changbai Mountain is an ecological type of Apis cerana, which is an excellent breeding material with cold-resistant developed by long-term natural selection under the ecological conditions. However, the physiological and molecular mechanisms of Changbai Mountain population under cold stress are still unclear. In this study, the Nanopore sequencing was carried out for the transcriptome of Apis cerana in Changbai Mountain in the coldest period of overwintering, which will provide a reference to the cold-resistant mechanism. We determined 5,941 complete ORF sequences, 1,193 lncRNAs, 619 TFs, 10,866 SSRs and functional annotations of 11,599 new transcripts. Our results showed that the myosin family and the C2H2 zinc finger protein transcription factor family possibly have significant impacts on the response mechanism of cold stress during overwintering. In addition, the cold environment alters genes expression profiles in honeybees via different AS and APA mechanisms. These altered genes in Hippo, Foxo, and MARK pathways help them counter the stress of cold in overwinter period. Our results might provide clues about the response of eastern honeybees to extreme cold, and reflect the possible genetic basis of physiological changes.

Effects of melatonin treatment on browning alleviation of fresh-cut foods.

Enzymatic browning is the main quality issue of fresh-cut foods. This study investigated the effects of different concentrations (0.05, 0.1, and 0.2 mM) of melatonin (MT) treatment on the enzymatic browning of four fresh-cut foods, including apples (Malus domestica), pears (Pyrus spp.), potatoes (Solanum tuberosum), and taros (Colocasia esculenta), and found that only 0.05 mM MT was most significant at enhancing the L* values and reducing the browning index (BI) in all four foods. This suggests that 0.05 mM MT might be a universal concentration for the browning alleviation of fresh-cut foods. MT treatment increased the total phenolic contents and PAL activities but reduced the activities of POD, PPO, and LOX in fresh-cut taros. Furthermore, MT significantly affected the expression of sixteen browning-related genes in fresh-cut taros during storage. These results suggest that MT reduced fresh-cut food browning by regulating the activities of browning-related enzymes. However, MT did not significantly influence the browning degree or PPO activity of the taro mash in vitro, suggesting that MT might regulate enzyme activity in an indirect manner. Overall, these results indicate that MT might be a promising anti-browning agent to alleviate the browning of fresh-cut foods. PRACTICAL APPLICATIONS: Melatonin (MT) is an endogenously produced indoleamine. Previous studies have reported that MT displays protective activities in many agricultural products. However, the reported protective MT concentrations vary between different products. To extend this agent to the fresh-cut industry, it is necessary to determine the universal dosage of MT needed for application efficiency. In this study, the effects of three concentrations (0.05, 0.1, and 0.2 mM) of MT on four fresh-cut foods, including apples, pears, potatoes, and taros, were investigated. The results showed that MT treatment alleviated browning development in the slices of these four foods and that 0.05 mM MT was the most effective treatment. Moreover, 0.05 mM MT significantly affected the activities of browning-related enzymes. These results suggest that 0.05 mM MT is a universal dosage for reducing surface browning in fresh-cut foods. This study provides a foundation for the application of MT in the processing of fresh-cut foods, especially fresh-cut taros.

The complete chloroplast genome sequence of Artocarpus hypargyreus.

The complete chloroplast genome sequences of Artocarpus hypargyreus was reported in this study. The length of the sequence was 160,952 bp in length with a large single copy (LSC) region of 89,476 bp, the small single copy (SSC) region of 20,070 bp and two inverted repeat (IR) regions of 25,703 bp. The complete genome contains 129 genes including 84 protein-coding genes, 8 rRNA genes and 37 tRNA genes. Phylogenetic analysis of the Moraceae based on 8 plastome sequence shows that A. hypargyreus is most related to Morus cathayana.

Application of citronella and rose hydrosols reduced enzymatic browning of fresh-cut taro.

Hydrosols are byproducts produced by steam distillation of aromatic plant materials. Previous studies reported hydrosols owned effectively inhibitory effects on microbials, but few focused on the effects in reducing enzymatic browning of fresh-cut products. Here, application of citronella hydrosol (CH) and rose hydrosol (RH) obviously reduced values of L*, a* and b* in surfaces of fresh-cut taros, suggesting these hydrosols could reduce browning severity. About 500 ml/L of RH and CH reduced total phenol contents, and activities of phenylalanine ammonia lyase (PAL), peroxidase (POD) and polyphenol oxidase (PPO), suggesting hydrosols suppressed browning development by reducing PAL, POD and PPO activities. Studies reported terpenoids had potential inhibition on PPO activity. Chemical composition analysis showed that RH and CH extracts contained abundant terpenoids. These imply that terpenoids contained in two hydrosols might have contributed to the reduction of enzyme activity. This study indicates that hydrosols might be promising natural anti-browning agents for fresh-cut foods. PRACTICAL APPLICATIONS: Hydrosols are the byproducts of essential oil from steam distillation. The work described here shows that application of citronella and rose hydrosols significantly reduced the browning of fresh-cut taros. Moreover, aromatic hydrosols also present inhibitory activity against microorganisms and are generally considered safe for human health. These suggested that aromatic hydrosols are the natural and useful anti-browning agents to preserve the quality of fresh cut foods, and can be well integrated into industrial procedures.

Genetic diversity of Aoluguya Reindeer based on D-loop region of mtDNA and its conservation implications.

Aoluguya Reindeer is the only reindeer population in China. In recent years, habitat loss and inbreeding have led to population decline, and population growth has been slow, maintaining a thousand or so. To better protect the Aoluguya Reindeer and improve its fecundity, we have introduced reindeer from Finland, crossbreeding help us to reach this goal. However, it is lacking in the study of genetic diversity of reindeer in China and Finland. Therefore, we used the partial sequences of the D-loop region of mitochondrial DNA to analyze the genetic diversity of Chinese reindeer (Aoluguya Reindeer) and the introduced Finnish reindeer, and identified twenty-six haplotypes, including nineteen in China, five in Finland, and two in Russia. There is no shared haplotype among them. The nucleotide diversity of Aoluguya Reindeer is 0.00752, which is significantly lower than that of reindeer in Finland and other countries. The haplotype and phylogenetic analysis show that reindeer from different geographical origins are not clustered completely according to geographical distribution. Aoluguya Reindeer populations and the introduced reindeer herds from Finland are all closely related to the reindeer from Russia. AMOVA analysis showed that there was significant differentiation between reindeer populations in China and Finland, and mismatch analysis showed that both populations had not experienced expansion. In this study, we identified the genetic diversity of Aoluguya Reindeer and the introduced reindeer, and provided a scientific basis for the conservation and breeding of Aoluguya Reindeer resources.

Is GPR39 the natural receptor of obestatin?

GPR39, an orphan receptor belonging to the family of G protein-coupled receptors, was originally reported to be the receptor of obestatin. However recently, numerous reports have questioned this conclusion. In mammals, GPR39 was reported to be involved in the regulation of gastrointestinal and the metabolic functions. In this article, a latest and brief review on the receptor family, structure, distribution and physiological functions of GPR39 has been reported.

Effect of phytase supplementation to barley-canola meal and barley-soybean meal diets on phosphorus and calcium balance in growing pigs

Se realizaron dos experimentos metabólicos para determinar el efecto de adicionar una fitasa microbiana a dietas cebada pasta de canola y cebada-pasta de soya en el balance de P y Ca de cerdos en crecimiento. En el experimento 1, seis cerdos (29,6kg peso vivo inicial) se alimentaron con dietas elaboradas con cebada y pasta de canola, sin o con fitasa (500 unidades.kg-¹ dieta) durante 24 días, en un diseño de sobrecambio. Las dietas proveyeron niveles de P inferior es al óptimo. En el experimento 2, doce cerdos (53,5kg peso vivo inicial) recibieron 4 dietas cebada-pasta de soya, en un diseño de sobrecambio con dos periodos experimentales. Las dietas 2, 3 y 4 contenían niveles de P inferiores al óptimo. La dieta 1 se adicionó con P y Ca inorgánico para cubrir los requerimientos; las dietas 2, 3 y 4 contenían niveles de P inferiores al óptimo, acondicionándose la dieta 3 con fitasa y la dieta 4 con fitasa y una mezcla de xilanasa y ß-glucanasa. La adición de fitasa incrementó (p<0,05) la digestibilidad y la retención de P, así como la digestibilidad de Ca. La adición conjunta de fitasa y mezcla de xylanasa y ß-glucanasa a la dieta 2 produjo valores de retención de P y digestibilidades de P y Ca similares (P> 0,05) a los de las dietas 1 y 3. En conclusión, la adición de fitasa a dietas cebada-pasta de canola y cebada-pasta de soya mejoró la utilización de P y la digestibilidad de Ca, pero no se obtuvo efecto adicional con la adición de xylanasa y ß-glucanasa