Preparation of Tetrandrine Nanocrystals by Microfluidic Method and Its In Vitro and In Vivo Evaluation.

The anti-hepatocellular carcinoma effects of TET are acknowledged, but its application is hindered by its poor water solubility and low bioavailability. Conventional methods for nanocrystal preparation are laborious and lack control. To address these limitations, we propose employing the microfluidic method in the preparation of TET nanocrystals, aiming to enhance the aforementioned constraints. The objectives of this study were to prepare TET nanocrystals (TET-NC@GL) using a Y-microfluidic method with glycyrrhetinic acid (GL) as a stabilizer. The optimal preparation prescription was determined through a single-factor test and Box-Behnken response surface method. Additionally, the nanocrystals prepared with the commonly used stabilizer polyvinylpyrrolidone K30 (PVPK30), known as TET-NC@PVPK30, were characterized and evaluated for their toxicity to HepG2 cells. Hybridized nanocrystals (TET-HNC@GL and TET-HNC@PVPK30) were synthesized using a water-soluble aggregation-induced emission (AIE) fluorescent probe (TVP). Qualitative and quantitative cellular uptake experiments were conducted using these hybridized nanocrystals. Conducting in vivo pharmacokinetic assays evaluates the relative bioavailability of nanocrystals. The results indicated that TET-NC@GL, optimized using the response surface method, had a particle size of 136.47 ± 3.31 nm and a PDI of 0.219 ± 0.002. The administration of TET-NC@GL significantly enhanced the cell inhibition rate compared to the TET group and the TET-NC@PVPK30 group (P < 0.01). Moreover, the qualitative and quantitative cellular uptake results revealed a significant enhancement in cellular uptake in the TET-HNC@GL administration group compared to the TET-HNC@PVPK30 group (P < 0.01). In vivo pharmacokinetic results showed that the bioavailability of TET-NC@GL group was 3.5 times higher than that of the TET group. The results demonstrate the successful preparation of TET-NC@GL nanocrystals.

Efficient degradation of atrazine through in-situ anchoring NiCo2O4 nanosheets on biochar to activate sulfite under neutral condition.

Sulfite (S(IV)) is a promising substitute for sulfate radical-based advanced oxidation processes. Here, a composite of in-situ anchoring NiCo2O4 nanosheets on biochar (BC) was firstly employed as a heterogeneous activator for sulfite (NiCo2O4@BC-sulfite) to degrade atrazine (ATZ) in the neutral environment. The synergistic coupling of BC and NiCo2O4 endows the resulting composite excellent catalytic activity. 82% of the degradation ratio of ATZ (1 mg/L) could be achieved within 10 min at initial concentrations of 0.6 g/L NiCo2O4@BC, 3.0 mmol/L sulfite in neutral environment. When further supplementing sulfite into the system at 20 min (considering the depletion of sulfite), outstanding degradation efficiency (∼ 100%) were achieved in the next 10 min without any other energy input by the NiCo2O4@BC-sulfite system. The features of the prepared catalysts and the effects of some key parameters on ATZ degradation were systematically examined. A strong inner-sphere complexation (Co2+/Ni2+-SO32-) was explored between sulfite and the metal sites on the NiCo2O4@BC surface. The redox cycle of the surface metal efficiently mediated sulfite activation and triggered the series radical chain reactions. The generated radicals, in particular the surface-bound radicals were involved in ATZ degradation. High performance liquid chromatography-tandem mass spectrometry (LC-MS) technique was used to detect the degradation intermediates. Density functional theory (DFT) calculations were performed to illustrate the possible degradation pathways of ATZ. Finally, an underlying mechanism for ATZ removal was proposed. The present study offered a low-cost and sustainable catalyst for sulfite activation to remove ATZ in an environmentally friendly manner from wastewater.

Nitrate promoted defluorination of perfluorooctanoic acid in UV/sulfite system: Coupling hydrated electron/reactive nitrogen species-mediated reduction and oxidation.

A significantly accelerated defluorination of recalcitrant perfluorooctanoic acid (PFOA) was explored with the co-present nitrate (20 mg L-1) by UV/sulfite treatment (UV/sulfite-nitrate). The deep defluorination of PFOA and complete denitrification of nitrate were simultaneously achieved in UV/sulfite-nitrate system. At the initial 30 min, PFOA defluorination exhibited an induction period, exactly corresponding to the removal of the co-existed nitrate. Upon the induction period passed, an accelerated removal of PFOA (5 mg L-1) occurred, nearly 100% defluorination ratio reached within 2 h. Compared with those in UV/sulfite, the kinetics of PFOA decay, defluorination, and transformation product formations were greatly enhanced in UV/sulfite-nitrate system. Reactive nitrogen species (RNS) generated from eaq--induced reduction of nitrate were found to play significant roles on the promoted defluorination apart from eaq--mediated reductive defluorination. The investigations on solution pH (7.0-11.0) confirmed that the reductive defluorination of PFOA was more efficient under alkaline conditions, however, the presence of nitrate can promote the defluorination even under neutral pH. Theoretical calculations of Fukui function demonstrated that RNS could easily launch electrophilic attack toward H-rich moieties of fluorotelomer carboxylates (FTCAs, CnF2n+1-(CH2)m-COO-), more persistent intermediates (formed via H/F exchange), and convert FTCAs into shorter-chain perfluorinated carboxylic acids, thus facilitating the deep defluorination. Along with the analysis on the denitrification products, the liberation of fluoride ions and generated intermediates, possible decomposition pathways were proposed. This work highlights the indispensable synergy from eaq-/RNS with integrated reduction and oxidation on PFOA defluorination and will advance remediation technologies of perfluorinated compound contaminated water.

Efficient removal of PFOA with an In2O3/persulfate system under solar light via the combined process of surface radicals and photogenerated holes.

The environmental persistence, high toxicity and wide spread presence of perfluorooctanoic acid (PFOA) in aquatic environment urgently necessitate the development of advanced technologies to eliminate PFOA. Here, the simultaneous application of a heterogeneous In2O3 photocatalyst and homogeneous persulfate oxidation (In2O3/PS) was demonstrated for PFOA degradation under solar light irradiation. The synergistic effect of direct hole oxidation and in-situ generated radicals, especially surface radicals, was found to contribute significantly to PFOA defluorination. Fourier infrared transform (FTIR) spectroscopy, Raman, electrochemical scanning microscope (SECM) tests and density functional theory (DFT) calculation showed that the pre-adsorption of PFOA and PS onto In2O3 surface were dramatically critical steps, which could efficiently facilitate the direct hole oxidation of PFOA, and boost PS activation to yield high surface-confined radicals, thus prompting PFOA degradation. Response surface methodology (RSM) was applied to regulate the operation parameters for PFOA defluorination. Outstanding PFOA decomposition (98.6%) and near-stoichiometric equivalents of fluorides release were achieved within illumination 10 h. An underlying mechanism for PFOA destruction was proposed via a stepwise losing CF2 unit. The In2O3/PS remediation system under solar light provides an economical, sustainable and environmentally friendly approach for complete mineralization of PFOA, displaying a promising potential for treatment of PFOA-containing water.

The Phoebe genome sheds light on the evolution of magnoliids.

Lauraceae includes the genus Phoebe, and the family is linked to the evolution of magnoliids. We sequenced the genome of Phoebe bournei Nanmu. The assembled genome size was 989.19 Mb, with a contig N50 value of 2.05 Mb. A total of 28,198 protein-coding genes were annotated in P. bournei. Whole-genome duplication (WGD) analysis showed that Lauraceae has experienced two WGD events; the older WGD event occurred just before the divergence of Lauraceae and Magnoliales, and the more recent WGD was shared by all lineages of Lauraceae. The phylogenetic tree showed that magnoliids form a sister clade to monocots and eudicots. We also identified 63 MADS-box genes, including AGL12-like genes that may be related to the regulation of P. bournei roots and FIN219-like genes encoding GH3 proteins, which are involved in photomorphogenesis. SAUR50-like genes involved in light signal-mediated pedicel or stem development were also identified. Four ATMYB46- and three PtrEPSP-homologous genes related to lignin biosynthesis were identified. These genes may be associated with the formation of straight trunks in P. bournei. Overall, the P. bournei reference genome provides insight into the origin, evolution, and diversification of Phoebe and other magnoliids.

Smart pH-Regulated Switchable Nanoprobes for Photoelectrochemical Multiplex Detection of Antibiotic Resistance Genes.

Antibiotic resistance, encoded via particular genes, has become a major global health threat and substantial burden on healthcare. Hence, the facile, low-cost, and precise detection of antibiotic resistance genes (ARGs) is crucial in the realm of human health and safety, especially multiplex sensing assays. Here, a smart pH-regulated switchable photoelectrochemical (PEC) bioassay has been created for ultrasensitive detection of two typical subtypes of penicillin resistance genes bla-CTX-M-1 (target 1, labeled as TDNA1) and bla-TEM (target 2, labeled as TDNA2), whereby pH-responsive antimony tartrate (SbT) complex-grafted silica nanospheres are ingeniously adopted as signal DNA1 tags (labeled as SDNA1-SbT@SiO2NSs). The operations of the PEC bioassay depend on the switchable dissociation of the pH-responsive SDNA1-SbT@SiO2NSs complex under the external pH stimuli, thus initiating the pH-regulated release of ions pre-embedded in sandwich-type DNA nanoassemblies. At acidic conditions, the dissociation of SDNA1 tags (ON state) triggers the release of the embedded SbO+. Under alkaline conditions, the dissociation of SDNA1 tags is inhibited (OFF state). The detection of TDNA2 was achieved via DNA hybridization-triggered metal ion release. The unwinding of the introduced hairpin T-Hg2+-T fragment, hybridized with the second anchored signal DNA (SDNA2), ignites the release of Hg2+. The released SbO+ or Hg2+ ions would trigger the formation of Sb2S3/ZnS or HgS/ZnS heterostructure through ion-exchange with the photosensitive ZnS layer, giving rise to the amplified photocurrents and eventually realizing the ultrasensitive detection of penicillin resistance genes subtypes, bla-CTX-M-1 and bla-TEM. The as-fabricated pH-regulated PEC bioassay, smartly integrating the pH-responsive intelligent unit as SDNA tags, pH-regulated release of embedded ions, and the subsequent ion-exchange-based signal amplification strategy, exhibits high sensitivity, specificity, low-cost, and ease of use for multiplex detection of ARGs. It can be successfully used for measuring bla-CTX-M-1 and bla-TEM in real E. coli plasmids, demonstrating great promise for developing a new class of genetic point-of-care devices.

Rapid photochemical decomposition of perfluorooctanoic acid mediated by a comprehensive effect of nitrogen dioxide radicals and Fe3+/Fe2+ redox cycle.

Developing efficient methods to degrade perfluorochemicals (PFCs), an emerging class of highly recalcitrant contaminants, are urgently needed in recent years, due to their persistence, high toxicity, and resistance to most regular treatment procedures. Here, a UV-photolysis system is reported for efficient mineralization of perfluorooctanoic acid (PFOA) via irradiation of ferric nitrate aqueous solution, where in-situ generating •NO2 and the effective Fe3+/Fe2+ redox cycle synergistically play great roles on rapidly mediating the mineralization of PFOA. A fast PFOA removal kinetics with first-order kinetic constants of 2.262 h-1 is observed at initial PFOA concentration of 5 ppm (50 mL volume), reaching ∼ 92 % removal efficiency within only 0.5-h irradiation. Near-stoichiometric fluoride ions liberation and high total organic carbon (TOC) removal efficiency (∼100 %) further validated the capability for completely destructive removal of PFOA. A tentative pathway for PFOA destruction is proposed. This work, by UV photolysis of abundant existing iron/nitrate-based systems in natural environment, provides an economical, sustainable and highly efficient approach for complete mineralization of perfluorinated chemicals.

Less-invasive chromosome screening of embryos and embryo assessment by genetic studies of DNA in embryo culture medium.

PURPOSE: To perform a preliminary exploration of a new embryo rank in clinical practice by combining the embryo chromosome copy number and mitochondrial copy number analysis of DNA extracted from embryo culture medium and blastocoel fluid. METHOD: Eighty-three ICSI embryos from day 2 or day 3 were cultured to day 5 or day 6. Thirty-two blastocysts of 3 cc or above were obtained. Culture medium and blastocoel fluid were collected at 24 h before blastocyst formation. The genomic DNA and mitochondrial DNA (mtDNA) from the culture medium combined with blastocoel fluid and the whole blastocyst were amplified and sequenced by MALBAC-NGS. We compared the chromosomal information generated by the new protocol from the culture medium and the information employed by the whole embryo method. A multivariable linear regression was performed to study the impact of the blastocyst morphological score, chromosomal abnormality, embryo mtDNA copy number, and female age on the culture medium mtDNA copy number. RESULTS: (1) The DNA from 31 blastocysts was successfully amplified, and the successful amplification rate was 96.9% (31/32). The success rate of the amplification of genomic DNA extracted from the culture medium was 87.5% (28/32). (2) There were 18 blastocysts in which the less invasive method and the whole embryo method revealed the same results. The consistency rate was 66.7% (18/27). (3) The culture medium mitochondrial DNA copy number (MCN) had a significantly positive correlation with the blastocyst mitochondrial DNA copy number (P = 0.001), female age (P = 0.012), and blastocyst score (P = 0.014), but there was no obvious correlation with blastocyst chromosome (P = 0.138). CONCLUSIONS: The preliminary exploration result of the less invasive approach for having an embryo rank was not satisfying, which still awaits further long-term evaluation.

Self-assembly of supra-amphiphiles building block fabricated by ß-cyclodextrin and adamantane-based ionic liquid.

A new adamantane-based ionic liquid, (11-(((-adamantane-1-carbonyl)oxy)undecyl)-1-methylimidazol-3-ium bromide (AD-C11im), was synthesized from 1-adamantanecarboxylic acid and observed that it can aggregate into micelles in aqueous solution. A number of experiments were conducted to understand the self-assembly of supra-amphiphiles building block fabricated by ß-cyclodextrin (ß-CD) and adamantane-based ionic liquid at diverse molar ratios. Studies revealed that host-guest interaction between the adamantane group and ß-CD occurred and AD-C11im@1ß-CD building block formed when same amount ß-CD was added. Then the micelles aggregates formed by AD-C11im only turned into spherical vesicles, which was confirmed by AFM, DLS and TEM. Besides, according to the results of AFM, it can be confirmed that the vesicles were monolayer structure. When double amount ß-CD was added, both the adamantane group and the hydrophobic chain were encapsulated by ß-CD and AD-C11im@2ß-CD building block formed. Thus the aggregations changed from vesicles to net-like nanofiber, which was observed by TEM. When the ß-CD concentration increased to 40 mM, the formation of light blue hydrogel was observed during the self-assembling process of AD-C11im@2ß-CD building block.

Chloroplast characterizations and phylogenetic position of an endangered orchid, Vanda coerulea (Orchidaceae).

Vanda coerulea possess a high ornamental value and medical effect against glaucoma and cataract. The whole complete chloroplast (cp) genome of V. coerulea and the phylogenetic position based on the cp sequences remain unclear. Herein, we report the complete chloroplast genome of V. coerulea. The chloroplast genome was 149,376 bp in length, including a large single-copy (LSC) region of 86,100 bp, a small single-copy (SSC) region of 11,702 bp, and two inverted repeat (IRs) regions of 25,787 bp. A total of 129 genes were characterized, including 74 protein-coding genes, 36 tRNA genes, and 8 rRNA genes. The overall GC content was 36.6%, and GC percentages range from 27.9% to 43.2% throughout LSC, IRs, and SSC regions. Phylogenetic analysis based on 20 chloroplast genomes of Orchidaceae indicated that V. coerulea is closely related to V. brunnea. Our study provides a valuable resource for the identification and distinction of Vanda genus, and will lay a foundation for further research and conservation measures of V. coerulea.

Whole genome and transcriptome analysis reveal adaptive strategies and pathogenesis of Calonectria pseudoreteaudii to Eucalyptus.

BACKGROUND: Leaf blight caused by Calonectria spp. is one of the most destructive diseases to affect Eucalyptus nurseries and plantations. These pathogens mainly attack Eucalyptus, a tree with a diversity of secondary metabolites employed as defense-related phytoalexins. To unravel the fungal adaptive mechanisms to various phytoalexins, we examined the genome of C. pseudoreteaudii, which is one of the most aggressive pathogens in southeast Asia. RESULTS: A 63.7 Mb genome with 14,355 coding genes of C. pseudoreteaudii were assembled. Genomic comparisons identified 1785 species-specific gene families in C. pseudoreteaudii. Most of them were not annotated and those annotated genes were enriched in peptidase activity, pathogenesis, oxidoreductase activity, etc. RNA-seq showed that 4425 genes were differentially expressed on the eucalyptus(the resistant cultivar E. grandis×E.camaldulensis M1) tissue induced medium. The annotation of GO term and KEGG pathway indicated that some of the differential expression genes were involved in detoxification and transportation, such as genes encoding ABC transporters, degrading enzymes of aromatic compounds and so on. CONCLUSIONS: Potential genomic determinants of phytoalexin detoxification were identified in C. pseudoreteaudii by comparison with 13 other fungi. This pathogen seems to employ membrane transporters and degradation enzymes to detoxify Eucalyptus phytoalexins. Remarkably, the Calonectria genome possesses a surprising number of secondary metabolism backbone enzyme genes involving toxin biosynthesis. It is also especially suited for cutin and lignin degradation. This indicates that toxin and cell wall degrading enzymes may act important roles in the establishment of Calonectria leaf blight. This study provides further understanding on the mechanism of pathogenesis in Calonectria.

Transcriptomic Analysis of Calonectria pseudoreteaudii during Various Stages of Eucalyptus Infection.

Eucalyptus leaf blight caused by Calonectria spp. is a serious disease in Eucalyptus seedling and plantations. However, the molecular mechanisms of the infection process and pathogenesis of Calonectria to Eucalyptus is not well-studied. In this study, we analyzed the transcriptomes of C. pseudoreteaudii at three stages of Eucalyptus leaf infection, and in mycelium grown in potato dextrose broth using Illumina RNA-Seq technology. We identified 161 differentially expressed genes between C. pseudoreteaudii from leaf and mycelium grown in potato dextrose broth. GO and KEGG enrichment analyses of these genes suggested that they were mainly involved in oxidoreductase activity, hydrolase activity, and transmembrane transporter activity. Most of the differentially expressed genes at the early infection stage were upregulated. These upregulated genes were mainly involved in cell wall hydrolysis and toxin synthesis, suggesting a role for toxin and cell wall hydrolases in the establishment of Calonectria leaf blight. Genes related to detoxification of phytoalexins were continually upregulated during infection. The candidate effectors and putative pathogenicity determinants identified in this study will help in the functional analysis of C. pseudoreteaudii virulence and pathogenicity.

Data for transcriptome and proteome analysis of Eucalyptus infected with Calonectria pseudoreteaudii.

Cylindrocladium leaf blight is one of the most important diseases in Eucalyptus plantations. We investigated the proteome and transcriptome of Eucalyptus infected or not infected with Calonectria pseudoreteaudii. Here we provide the information about the processing of raw data obtained by RNA-seq and iTRAQ technologies. The data are related to [1].

Transcriptome and proteome analysis of Eucalyptus infected with Calonectria pseudoreteaudii.

Cylindrocladium leaf blight is one of the most severe diseases in Eucalyptus plantations and nurseries. There are Eucalyptus cultivars with resistance to the disease. However, little is known about the defense mechanism of resistant cultivars. Here, we investigated the transcriptome and proteome of Eucalyptus leaves (E. urophylla×E. tereticornis M1), infected or not with Calonectria pseudoreteaudii. A total of 8585 differentially expressed genes (|log2 ratio| ≥1, FDR ≤0.001) at 12 and 24hours post-inoculation were detected using RNA-seq. Transcriptional changes for five genes were further confirmed by qRT-PCR. A total of 3680 proteins at the two time points were identified using iTRAQ technique.The combined transcriptome and proteome analysis revealed that the shikimate/phenylpropanoid pathway, terpenoid biosynthesis, signalling pathway (jasmonic acid and sugar) were activated. The data also showed that some proteins (WRKY33 and PR proteins) which have been reported to involve in plant defense response were up-regulated. However, photosynthesis, nucleic acid metabolism and protein metabolism were impaired by the infection of C. pseudoreteaudii. This work will facilitate the identification of defense related genes and provide insights into Eucalyptus defense responses to Cylindrocladium leaf blight. BIOLOGICAL SIGNIFICANCE: In this study, a total of 130 proteins and genes involved in the shikimate/phenylpropanoid pathway, terpenoid biosynthesis, signalling pathway, cell transport, carbohydrate and energy metabolism, nucleic acid metabolism and protein metabolism in Eucalyptus leaves after infected with C. pseudoreteaudii were identified. This is the first report of a comprehensive transcriptomic and proteomic analysis of Eucalyptus in response to Calonectria sp.