Programming Intracellular Clustering of Spiky Nanoparticles via Liposome Encapsulation.

The intracellular clustering of anisotropic nanoparticles is crucial to the improvement of the localized surface plasmon resonance (LSPR) for phototherapy applications. Herein, we programmed the intracellular clustering process of spiky nanoparticles (SNPs) by encapsulating them into an anionic liposome via a frame-guided self-assembly approach. The liposome-encapsulated SNPs (lipo-SNPs) exhibited distinct and enhanced lysosome-triggered aggregation behavior while maintaining excellent monodispersity, even in acidic or protein-rich environments. We explored the enhancement of the photothermal therapy performance for SNPs as a proof of concept. The photothermal conversion efficiency of lipo-SNPs clusters significantly increased 15 times compared to that of single lipo-SNPs. Upon accumulation in lysosomes with a 2.4-fold increase in clustering, lipo-SNPs resulted in an increase in cell-killing efficiency to 45% from 12% at 24 µg/mL. These findings indicated that liposome encapsulation provides a promising approach to programing nanoparticle clustering at the target site, which facilitates advances in the development of smart nanomedicine with programmable enhancement in LSPR.

In vitro drug screening models derived from different PC12 cell lines for exploring Parkinson's disease based on electrochemical signals of catecholamine neurotransmitters.

Gold nanostructures and a Nafion modified screen-printed carbon electrode (Nafion/AuNS/SPCE) were developed to assess the cell viability of Parkinson's disease (PD) cell models. The electrochemical measurement of cell viability was reflected by catecholamine neurotransmitter (represented by dopamine) secretion capacity, followed by a traditional tetrazolium-based colorimetric assay for confirmation. Due to the  capacity to synthesize, store, and release catecholamines as well as their unlimited homogeneous proliferation, and ease of manipulation, pheochromocytoma (PC12) cells were used for PD cell modeling. Commercial low-differentiated and highly-differentiated PC12 cells, and home-made nerve growth factor (NGF) induced low-differentiated PC12 cells (NGF-differentiated PC12 cells) were included in the modeling. This approach achieved sensitive and rapid determination of cellular modeling and intervention states. Notably, among the three cell lines, NGF-differentiated PC12 cells displayed the enhanced neurotransmitter secretion level accompanied with attenuated growth rate, incremental dendrites in number and length that were highly resemble with neurons. Therefore, it was selected as the PD-tailorable modeling cell line. In short, the electrochemical sensor can be used to sensitively determine the biological function of neuron-like PC12 cells with negligible destruction and to explore the protective and regenerative impact of various substances on nerve cell model.

Assembly and comparative analysis of the complete mitochondrial genome of Pinellia ternata.

Pinellia ternata is an important natural medicinal herb in China. However, it is susceptible to withering when exposed to high temperatures during growth, which limits its tuber production. Mitochondria usually function in stress response. The P . ternata mitochondrial (mt) genome has yet to be explored. Therefore, we integrated PacBio and Illumina sequencing reads to assemble and annotate the mt genome of P . ternata . The circular mt genome of P . ternata is 876 608bp in length and contains 38 protein-coding genes (PCGs), 20 tRNA genes and three rRNA genes. Codon usage, sequence repeats, RNA editing and gene migration from chloroplast (cp) to mt were also examined. Phylogenetic analysis based on the mt genomes of P . ternata and 36 other taxa revealed the taxonomic and evolutionary status of P . ternata . Furthermore, we investigated the mt genome size and GC content by comparing P . ternata with the other 35 species. An evaluation of non-synonymous substitutions and synonymous substitutions indicated that most PCGs in the mt genome underwent negative selection. Our results provide comprehensive information on the P . ternata mt genome, which may facilitate future research on the high-temperature response of P . ternata and provide new molecular insights on the Araceae family.

Analysis of codon usage patterns in Bupleurum falcatum chloroplast genome.

Objective: In order to distinguish the traditional Chinese medicine Bupleurum falcatum and its adulterants effectively and develop a better understanding of the factors affecting synonymous codon usage, codon usage patterns of chloroplast genome, we determine the complete chloroplast (cp) genome of B. falcatum and clarify the main factors that influence codon usage patterns of 78 genes in B. falcatum chloroplast genome. Methods: The total genomic DNA of fresh leaves from a single individual of B. falcatum was extracted with EASYspin plus Total DNA Isolation Kit and 2 µg genome DNA was sequenced using Illumina Hiseq 2500 Sequencing Platform. The cp genome of B. falcatum was reconstructed with MITObim v1.8 and annotated in the program CPGAVAS2 with default parameters. Python script and Codon W were used to calculate the codon usage bias parameters. Results: The full length of B. falcatum cp genome was 155 851 bp, 132 different genes were annotated in this cp genome containing 80 protein-coding genes, 30 tRNA genes, and four rRNA genes. The codon usage models tended to use A/T-ending codons. The neutrality plot, ENC plot, PR2-Bias plot and correspondence analysis showed that both compositional constraint under selection and mutation could affect the codon usage models in B. falcatum cp genome. Furthermore, three optimal codons were identified and most of these three optimal codons ended with G/U. Conclusion: The cp genome of B. falcatum has been characterized and the codon usage bias in B. falcatum cp genome is influenced by natural selection, mutation pressure and nucleotide composition. The results will provide much more barcode information for species discrimination and lay a foundation for future research on codon optimization of exogenous genes, genetic engineering and molecular evolution in B. falcatum.

Nanotechnology-Based Drug Delivery System for Anticancer Active Ingredients from Traditional Chinese Medicines: A Review.

The variable dosage forms of most traditional Chinese medicines (TCMs) could be disadvantaged by low selectivity, poor biological distribution, limited bioavailability with low efficacy, and some adverse effects. These issues limit the control of clinical pharmacodynamics of the antitumor active components. With the progress of science and technology, many new polymer materials and new technologies have emerged, such as nanotechnology, cyclodextrin inclusion, solid dispersion, microcapsule and microsphere technologies. These new technologies provide a good basis for exploring novel TCM dosage forms for overcoming the shortcomings. The increased numbers of new technologies have been used to study TCM dosage forms with remarkable achievements. In this review paper, we will provide a systematic overview of the new dosage forms of nano-formulations and co-medications in relation to nano-delivery systems in an attempt to provide useful references for practical application of active antitumor ingredients from the TCMs.

Urolithin A alleviates acute kidney injury induced by renal ischemia reperfusion through the p62-Keap1-Nrf2 signaling pathway.

Acute kidney injury (AKI) induced by renal ischemia reperfusion (RIR) is typically observed in renal surgeries and is a leading cause of renal failure. However, there is still an unmet medical need currently in terms of clinical treatments. Herein, we report the effect of Urolithin A (UA) in a mouse RIR model, wherein we demonstrated its underlying mechanism both in vitro and in vivo. The expression levels of p62 and Keap1 significantly decreased, while that of nuclear Nrf2 increased in vitro in a hypoxia cell model after UA treatment. Furthermore, the apoptosis of tubular cells was attenuated and the reactive oxygen species (ROS) levels were reduced in the kidneys in a mouse RIR model after UA administration. In this study, we demonstrated that UA can alleviate oxidative stress and promote autophagy by activating the p62-Keap1-Nrf2 signaling pathway, which could protect the kidneys from ischemia reperfusion injury.

Protective effects of lotus plumule ethanol extracts on bleomycin-induced pulmonary fibrosis in mice.

Pulmonary fibrosis (PF) is a progressive fibrosing disease, characterized by excessive accumulation of extracellular matrix (ECM) and remodeling of the lung architecture, which finally result in respiratory failure. Currently, there is no satisfactory treatment for PF， therefore, the development of effective agents is urgently needed. Lotus plumule, the green embryo of Nelumbo nucifera Gaertn., a plant of the Nymphaeaceae family, is a traditional Chinese food with exceptional nutritional value and its extracts exert prominent anti-inflammatory and anti-fibrotic effects. The aim of the present study was to investigate the inhibitory effects of lotus plumule extracts (LPEs) on bleomycin (BLM)-induced PF in mice. Therefore, enzyme-linked immunosorbent assay, RT-PCR, and western blot analysis were performed. The histopathological examination demonstrated that LPEs could obviously decrease the degree of alveolitis, deposition of ECM and the production of collagen I (Col-I) in the pulmonary interstitium. In addition, the results showed that LPEs markedly alleviated the expression of interleukin (IL)-6, IL-17, transforming growth factor (TGF)-ß, and α-smooth muscle actin (α-SMA). Additionally, the content of Col-I and hydroxyproline (HYP) was also attenuated. In conclusion, LPEs could ameliorate the BLM-induced lung fibrosis, thus suggesting that LPEs could serve as a potential therapeutic approach for PF.

The study of schizogenous formation of secretory ducts in Ferula ferulaeoides (Steud.) Korov.

The secretory ducts of Ferula ferulaeoides (Steud.) Korov. are the main tissue of synthesis, secretion, and accumulation of resin. The formation of secretory ducts is closely related to the harvest and quality of resin, but the lumen formation mode and corresponding mechanism have not been thoroughly studied. This study of F. ferulaeoides investigated the microstructure and ultrastructure of the secretory ducts from a developmental point of view. Stem samples were analyzed by light microscopy, transmission electron microscopy, and fluorescence microscopy. The data results showed (1) the walls of secretory cells were intact during the development of secretory ducts in F. ferulaeoides; (2) the plastids and endoplasmic reticulum of secretory cells participated in the synthesis of resin; (3) pectinase was involved in the degradation of the middle lamella; and (4) no features of programmed cell death during the formation of secretory ducts. The results suggested that the formation of F. ferulaeoides' secretory ducts was schizogenous, and pectinase was involved in its formation. These data may be beneficial to further explore the formation of secretory duct in other species of Ferula L. and the formation mechanism of schizogenous secretory structures.

Berberine inhibits pancreatic intraepithelial neoplasia by inhibiting glycolysis via the adenosine monophosphate -activated protein kinase pathway.

Most cases of pancreatic cancer develop in patients with chronic pancreatitis (CP). Berberine is natural product that exhibits anti-tumor effects in various types of cancer and is used in traditional Chinese medicine. In this study, we demonstrated that berberine inhibited the development of pancreatic intraepithelial neoplasia (PanIN) in an in vivo CP model and an in vitro acinar-to-ductal metaplasia model. As berberine may inhibit glycolysis during the development of PanIN, we measured indicators of glycolysis. Quantitative reverse transcription polymerase chain reaction and western blotting assays revealed that berberine activated the adenosine monophosphate-activated protein kinase (AMPK) pathway. This demonstrated that berberine suppressed glycolysis by targeting AMPK, a key metabolic sensor. Furthermore, berberine acted via the AMPK-hypoxia-inducible factor 1 alpha pathway to achieve suppression of PanIN. These findings show that berberine is a potential therapeutic candidate for preventing the progression of CP to PanIN.

Simultaneous process optimization of ultrasound-assisted extraction of polyphenols and ellagic acid from pomegranate (Punica granatum L.) flowers and its biological activities.

This study was designed to optimize the extraction rate of total polyphenols and ellagic acid from pomegranate flowers. Single factors were investigated for liquid-to-material ratio (5-25), ethanol concentration (20%-60%), sonication time (5-60 min), and sonication power (150-500 W). The level range of the Box-Bokhen design was determined with respect to the single-factor results. The components of each index were normalized using the entropy weighting method for obtaining the comprehensive evaluation value. Under the actual conditions, the final optimization results were 17 for liquid-to-material ratio, 43% for ethanol concentration, 10 min for ultrasonic time, and 300 W for ultrasonic power. The extracts obtained under optimal conditions were tested for the inhibition of Streptococcus mutans and its biofilm, and results showed that pomegranate flowers exerted some inhibitory effects on the bacterium. Phosphomolybdenum and FRAP assays were used, and DPPH, ABTS, and O2- radical scavenging tests were conducted, indicating that pomegranate flower extracts have good antioxidant capacity.

Oral absorption mechanism of the polysaccharides from Gastrodia elata Blume base on fluorescence labeling.

The mechanisms of action of polysaccharides in vivo have been widely elucidated. However, the systematic research of its absorption and transport mechanisms remains unclear. Herein, we extracted a polysaccharide fraction (GEP) from Gastrodia elata by water extraction and alcohol precipitation and aimed to reveal its oral absorption processes through animal models and Caco-2 cells monolayer models. Our research data showed that GEP-Cy5.5 could be absorbed through the small intestine and the main absorption intestinal segment was the ileum (the absorption rate constant [Ka]: (3.64 ± 0.70) × 10-4 cm/s; the effective apparent permeability [Papp value]: (4.88 ± 1.02) × 10-5 cm/s). The ligated intestinal loops also revealed that GEP-Cy5.5 could pass through the villi of the small intestine and the mucosal barrier into the submucosa. Furthermore, GEP-Cy5.5 was readily absorbed into the blood through the gastrointestinal tract, then distributed in the liver and the kidney. The Papp value of in vitro transport study was (1.29 ± 0.08) × 10-6 cm/s, which was a time-dependent process. Notably, GEP-Cy5.5 was transported through the endocytosis process mediated by clathrin and macropinocytosis. The underlying absorptive mechanisms of GEP in vivo and in vitro were clarified, which provided the guidance for clinical medicine administration and could deepen the biological understanding of oral polysaccharides.

The adsorption efficiency of nitrogen and phosphorus by in-situ remediation of modified sediment composite material.

Dredged sediment can occupy a large amount of land area, resulting in waste of land resources, and high disposal costs. In response to the problem, this work calcinates and modified the sediment and compounds it with the modified water purification plant sludge, zeolite powder, and bentonite. This is used as a covering material to inhibit the release of nitrogen (N) and phosphorus (P) in the sediment. The results showed that sediment modified composite material covering effectively reduces the release of nitrogen (N) and phosphorus (P) in the sediment, especially the release of P. When the thickness of the covering layer is 3 cm, the reduction rate of total N, NH4+-N, and total P in the overlying water by the modified composite material of sediment is 61.58, 79.59, and 70.34%, respectively. It can be seen that the covering material has a significant effect on the control of the release of N and P in the sediment. Additionally, the reduction of nutrients in the overlying water can overcome the negative effects of temperature rise in controlling the release of N and P in the sediment.

Analysis of synonymous codon usage of transcriptome database in Rheum palmatum.

BACKGROUND: Rheum palmatum is an endangered and important medicinal plant in Asian countries, especially in China. However, there is little knowledge about the codon usage bias for R. palmatum CDSs. In this project, codon usage bias was determined based on the R. palmatum 2,626 predicted CDSs from R. palmatum transcriptome. METHODS: In this study, all codon usage bias parameters and nucleotide compositions were calculated by Python script, Codon W, DNA Star, CUSP of EMBOSS. RESULTS: The average GC and GC3 content are 46.57% and 46.6%, respectively, the results suggested that there exists a little more AT than GC in the R. palmatum genes, and the codon bias of R. palmatum genes preferred to end with A/T. We concluded that the codon bias in R. palmatum was affect by nucleotide composition, mutation pressure, natural selection, gene expression levels, and the mutation pressure is the prominent factor. In addition, we figured out 28 optimal codons and most of them ended with A or U. The project here can offer important information for further studies on enhancing the gene expression using codon optimization in heterogeneous expression system, predicting the genetic and evolutionary mechanisms in R. palmatum.

Microscopic anatomy and ultrastructure of the resin ducts of Ferula ferulaeoides (Steud.) Korov. in Xinjiang.

Ferula ferulaeoides (Steud.) Korov. is a perennial herb that belongs to Umbelliferae (Apiaceae). Its resin and roots have extensive commercial and medicinal value in the Xinjiang region. However, the resin-secreting resin ducts (RDs) of F. ferulaeoides have not been studied in detail. This study used optical and transmission electron microscopy to explore the anatomical features, including the distribution, size, and structure, of the RDs among different organs of F. ferulaeoides. The microstructure data revealed that the RDs consisted of a round lumen, a layer of secretory cells, and multiple layers of sheath cells. Notably, the RDs in stem were arranged alternatively in a multilayered ring with vascular bundles of three distinct sizes. The ultrastructural analysis revealed that organelles in the secretory cells potentially play important roles in resin secretion. Those data may be of great significance to understanding the anatomy of the RDs in Ferula L. and Umbelliferae.

Mining genes associated with furanocoumarin biosynthesis in an endangered medicinal plant, Glehnia littoralis.

The endangered medicinal plant Glehnia littoralis is one of the important natural source of furanocoumarin, which has been used as mucolytic, antitussive, antitumour and antibacterial. However, the genetic information of furanocoumarin biosynthesis in G. littoralis is scarce at present. The objective of this study was to mine the putative candidate genes involved in the biosynthesis pathwayof furanocoumarin and provide references for gene identification, and functional genomics of G. littoralis. We carried out the transcriptome analysis of leaves and roots in G. littoralis, which provided a dataset for gene mining. Psoralen, imperatorin and isoimperatorin were detected in G. littoralis by high performance liquid chromatography analysis. Candidate key genes were mined based on the annotations and local BLAST with homologous sequences using BioEdit software. The relative expression of genes was analysed using quantitative real-time polymerase chain reaction. Further, the CYP450 genes were mined using phylogenetic analyses using MEGA 6.0 software. Atotal of 156,949 unigenes were generated, of which 9021 were differentially-expressed between leaves and roots. A total of 82 unigenes encoding eight enzymes in furanocoumarin biosynthetic pathway were first obtained. Seven genes that encoded key enzymes in the downstream furanocoumarin biosynthetic pathway and expressed more in roots than leaves were screened. Twenty-six candidate CYP450 unigenes expressed abundantly in roots and were chiefly concentrated in CYP71, CYP85 and CYP72 clans. Finally, we filtered 102 differentially expressed transcription factors (TFs) unigenes. The transcriptome of G. littoralis was characterized which would help to elucidate the furanocoumarin biosynthetic pathway in G. littoralis and provide an invaluable resource for further study of furanocoumarin.

Characteristics and Diversity of Endophytic Bacteria in Endangered Chinese Herb Glehnia littoralis Based on Illumina Sequencing.

Glehnia littoralis is an endangered medicinal plant growing in the coastal ecological environment and plays an important role in coastal ecosystems. The endophytes in the plant have a significant role in promoting plant growth and enhancing plant stress resistance. However, the endophytic bacterial structure associated with halophyte G. littoralis is still not revealed. In this project, the construction and diversity of endophytic bacterial consortium associated with different tissues of G. littoralis were illustrated with high throughput sequencing of the V3-V4 region of the bacterial 16S rRNA. The results resolved that the diversity and richness of endophytic bacteria were significantly higher in root than in leaf and stem. The operational taxonomic units (OTU) analysis demonstrated that the Actinobacteria and Proteobacteria were dominant in all the samples at the phylum level, and Pseudomonas, Bacillus, Rhizobium were the dominant genera. Our results unraveled that the bacterial communities differed among different tissues of G. littoralis. Endophytic bacterial communities in leaf and stem shared more similarity than that in the root. Furthermore, the difference of bacteria community and structure among different tissues were also detected by principal coordinate analysis. Taken altogether, we can conclude that the bacterial communities of different tissues are unique, which could facilitate understanding the diversity of endophytic bacteria in G. littoralis.

The Chinese Herbal Formula Shenzhu Tiaopi Granule Results in Metabolic Improvement in Type 2 Diabetic Rats by Modulating the Gut Microbiota.

OBJECTIVE: The aim of this study is to investigate the implication of the Chinese herbal formula (CHF) Shenzhu tiaopi Granule (STG) in type 2 diabetes mellitus (T2DM) and discuss the mechanisms by which STG regulates the gut microbiota. METHOD: Goto-Kakizaki (GK) rats and age-matched Wistar (W) rats were acclimatized for 1 week. The GK rats were randomly divided into 3 groups and orally gavaged with saline (model group, M), acarbose (acarbose group, A), and STG (granule of CHF group, G; the component of this formula includes Codonopsis pilosula, Rhizoma Atractylodis, Pinellia, Poria cocos, Pericarpium Citri Reticulatae, Coptis chinensis Franch, and Pueraria). The W rats were orally gavaged with saline (control group, C). The observation time was 8 weeks. The weight, fasting blood glucose (FBG) level, and blood lipid levels were tested. The 16S rRNA genes in the V3-V4 region were sequenced, and the structure of the gut microbiota was analysed. RESULTS: Compared to C, M displayed significant differences in blood glucose, gut microbiota, etc. (P<0.05; P<0.01). Compared to M, A and G showed a similar reduction in the FBG gain and a shift in the structure of the gut microbiota (P<0.05; P<0.01). Compared with A, G exhibited a significant decrease in weight, FBG level, and total cholesterol (P<0.05). The gut microbiota, Bacteroidetes, the Firmicutes/Bacteroidetes ratio, Allobaculum, and Desulfovibrionaceae were significantly decreased in response to the STG treatment, while Lactobacillus was significantly enriched (P<0.05; P<0.01). The community composition also differed at the phylum and genus levels based on the linear discriminant analysis effect size and heat map. CONCLUSION: Our findings suggest that the composition of the gut microbiota was significantly changed in the diabetic GK rats compared with that in the normal W rats. STG treatment can improve glucose and lipid levels and modulate the gut microbiota in T2DM rats.

Astragaloside IV prevents high glucose­induced cell apoptosis and inflammatory reactions through inhibition of the JNK pathway in human umbilical vein endothelial cells.

Endothelial dysfunction is a key pathophysiological step in early stage diabetes mellitus (DM) macrovascular complications and is also crucial in the inflammatory mechanisms of macrovascular complications. However, there is currently no effective intervention to improve endothelial dysfunction associated with DM macrovascular complications. Astragaloside IV (AS­IV), which can be extracted from the traditional Chinese medicine Astragalus membranaceus, has potential therapeutic effects on DM and its complications. The present study evaluated the effect of AS­IV on high glucose­induced human umbilical vein endothelial cell (HUVEC) injury and its possible mechanism. The result indicated that AS­IV has a significant protective effect on high glucose­induced HUVEC injury. AS­IV could significantly promote cell proliferation, reduce apoptosis and decrease the protein and mRNA expression levels of tumor necrosis factor­α and interleukin­1ß in HUVECs. Furthermore, AS­IV could decrease the expression of phosphorylated c­Jun NH2­terminal kinase (JNK) phosphorylated apoptosis signal­regulating kinase 1, cytochrome c, cleaved­caspase­9, cleaved­caspase­3 and the relative ratio of B­cell lymphoma­2 associated X protein/B­cell lymphoma­2 in HUVECs. In conclusion, the present study demonstrated that AS­IV could suppress apoptosis and inflammatory reactions promoted by high glucose conditions in HUVECs by inhibiting the JNK signaling pathway. These findings suggest that AS­IV could inhibit the process of endothelial dysfunction in diabetic macrovascular complications.

Characterization of the complete chloroplast genome of medicinal plant Rheum officinale (Polygonaceae).

The complete chloroplast (cp) genome of Rheum officinale Baill. was determined based on the Illumina Sequencing data. The cp genome is 161,563 bp in length. The overall G + C content of the cp genome was 37.31%. The R. officinale cp genome contains 129 genes, including 84 protein-coding genes, eight rRNA genes (four rRNA species), and 37 tRNA genes (20 tRNA species). A neighbour-joining phylogenetic tree clarified that the cp genome of R. officinale was closely related to that of R. palmatum in Polygonaceae.

The complete chloroplast genome sequence of Rheum tanguticum, an endangered Chinese medicinal plant (Polygonaceae).

Rheum tanguticum is a valuable medicinal plant endemic to the Qinghai-Tibetan Plateau. It has been listed classified under the IUCN Red List categories of Vulnerable due to the low reproductive rate and heavy exploitation. In this study, the complete chloroplast (cp) genome of R. tanguticum has been assembled using data from the whole-genome Illumina sequencing. The cp genome is 161,515 bp in size and contains two inverted repeat regions of 30,823 bp each, which is separated by a large single-copy region of 86,675 bp and a small single-copy region of 13,194 bp. The cp genome contains 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Phylogenetic analysis revealed that the cp genome of R. tanguticum was closely related to that of the R. palmatum.