Polyethyleneimine-mediated assembly of DNA nanotubes for KRAS siRNA delivery in lung adenocarcinoma therapy.

Self-assembled DNA nanostructures hold great promise in biosensing, drug delivery and nanomedicine. Nevertheless, challenges like instability and inefficiency in cellular uptake of DNA nanostructures under physiological conditions limit their practical use. To tackle these obstacles, this study proposes a novel approach that integrates the cationic polymer polyethyleneimine (PEI) with DNA self-assembly. The hypothesis is that the positively charged linear PEI can facilitate the self-assembly of DNA nanostructures, safeguard them against harsh conditions and impart them with the cellular penetration characteristic of PEI. As a demonstration, a DNA nanotube (PNT) was successfully synthesized through PEI mediation, and it exhibited significantly enhanced stability and cellular uptake efficiency compared to conventional Mg2+-assembled DNA nanotubes. The internalization mechanism was further found to be both clathrin-mediated and caveolin-mediated endocytosis, influenced by both PEI and DNA. To showcase the applicability of this hybrid nanostructure for biomedical settings, the KRAS siRNA-loaded PNT was efficiently delivered into lung adenocarcinoma cells, leading to excellent anticancer effects in vitro. These findings suggest that the PEI-mediated DNA assembly could become a valuable tool for future biomedical applications.

An ultrasensitive solid-state ECL biosensor based on synergistic effect between Zn-NGQDs and porphyrin-based MOF as "on-off-on" platform.

To develop an ultra-sensitive solid-state electrochemiluminescence (ECL) biosensor for detection of miRNA 24, three different forms of porphyrin metal-organic framework (MOF) nanomaterials with good biocompatibility were synthesized through small molecule ligand modulation. We investigated various properties of synthesized MOFs in the presence of different small molecule ligands. The as-obtained 2D MOF nanodisk exhibited high ECL intensity and outstanding stability in the presence of a co-reactant at low concentrations. We also synthesized zinc-based quantum dots (Zn-NGQDs) with excellent photovoltaic properties by doping zinc dithiothreitol (DTT-Zn) into quantum dots. Accordingly, an enzyme-free solid-state ECL biosensor for miRNA 24 based on the "on-off-on" signal conversion strategy was created. Dependent on the synergy between the luminophor 2D MOF and Zn-NGQDs, the biosensor achieves a wide linear range from 1.00 × 10-16 to 1.00 × 10-10 mol·L-1 and an exceedingly low detection limit of 0.03 fM. Furthermore, the ECL biosensor exhibits outstanding selectivity, repeatability, and stability. The method has great potential for investigating sensitive detection models for various biomolecules and the design of highly efficient MOF luminescent materials.

Transcriptome modulation by endophyte drives rice seedlings response to Pb stress.

This study investigated the growth, SPAD value, chlorophyll fluorescence and transcriptome response of endophyte uninoculated and inoculated rice seedlings under Pb stress after treatment of 1 d and 5 d. Inoculation of endophytes significantly improved the plant height, SPAD value, Fv/F0, Fv/Fm and PIABS by 1.29, 1.73, 0.16, 1.25 and 1.90 times on the 1 d, by 1.07, 2.45, 0.11, 1.59 and 7.90 times on the 5 d, respectively, however, decreased the root length by 1.11 and 1.65 times on the 1 d and 5 d, respectively under Pb stress. Analysis of rice seedlings leaves by RNA-seq, there were 574 down-regulated and 918 up-regulated genes after treatment of 1 d, 205 down-regulated and 127 up-regulated genes after treatment of 5 d, of which 20 genes (11 up-regulated and 9 down-regulated) exhibited the same changing pattern after treatment of 1 d and 5 d. Using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) to annotate these DEGs, and it was found that many of DEGs involved in photosynthesis, oxidative detoxification, hormone synthesis and signal transduction, protein phosphorylation/kinase and transcription factors. These findings provide new insights into the molecular mechanism of interaction between endophyte and plants under heavy metal stress, and contribute to agricultural production in limited environments.

Neuroprotective Effects of Sodium Butyrate by Restoring Gut Microbiota and Inhibiting TLR4 Signaling in Mice with MPTP-Induced Parkinson's Disease.

Parkinson's disease (PD) is a prevalent type of neurodegenerative disease. There is mounting evidence that the gut microbiota is involved in the pathogenesis of PD. Sodium butyrate (NaB) can regulate gut microbiota and improve brain functioning in neurological disorders. Hence, we examined whether the neuroprotective function of NaB on PD was mediated by the modulation of gut microbial dysbiosis and revealed its possible mechanisms. Mice were administered 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 7 consecutive days to construct the PD model. NaB gavage was given 2 h after the daily MPTP injections for 21 days. NaB improved the motor functioning of PD mice, increased striatal neurotransmitter levels, and reduced the death of dopaminergic neurons. The 16S rRNA sequencing analysis revealed that NaB restored the gut microbial dysbiosis. NaB also attenuated the intestinal barrier's disruption and reduced serum, colon, and striatal pro-inflammatory cytokines, along with inhibiting the overactivation of glial cells, suggesting an inhibitory effect on inflammation from NaB throughout the gut-brain axis of the PD mice. Mechanistic studies revealed that NaB treatment suppressed the TLR4/MyD88/NF-kB pathway in the colon and striatum. In summary, NaB had a neuroprotective impact on the PD mice, likely linked to its regulation of gut microbiota to inhibit gut-brain axis inflammation.

Metabolomics analysis reveal the molecular responses of high CO2 concentration improve resistance to Pb stress of Oryza sativa L. seedlings.

Rice seedlings were exposed to two CO2 concentrations (400 ± 20 and 800 ± 20 µmol mol-1) and three PbNO3 concentrations (0, 50 and 100 µmol L-1) for 10 days to explore the regulatory mechanisms of elevated CO2 for Pb stress resistance. Electrical conductivity, MDA content, SOD, POD, CAT activities and metabolomics changes were studied. Results showed that: Pb stress damaged cell membrane system, electrical conductivity and MDA content increased 49.34 % and 73.27 %, respectively, and some antioxidant enzymes activities increased. Sugar, polyol, amino acid metabolism and fatty acid ß-oxidation were all enhanced to improve osmotic adjustments, maintain cell membrane stability, supply energy, nitrogen assimilates and antioxidant capacity; Under composite treatments, cell membrane damage was reduced, activities of protective enzymes increased compared with only Pb stress, POD activity increased the most (49.14 %) under severe Pb composite treatment. High CO2 caused the enhance of cells antioxidant capacity, TCA cycle intermediate products contents and fatty acid desaturation under mild Pb stress. Many sugars, polyols and amino acids contents were increased as osmotic regulatory substances by high CO2 under severe Pb stress; Secondary metabolites played an important role under Pb stress and composite treatments. The object of this study is to provide a possible molecular mechanism of rice response to Pb stress under high CO2 in the future.

Application of RNA sequencing to understand the response of rice seedlings to salt-alkali stress.

BACKGROUND: Salt-alkali stress represents one of the most stressful events with deleterious consequences for plant growth and crop productivity. Despite studies focusing on the effects of salt-alkali stress on morphology and physiology, its molecular mechanisms remain unclear. Here, we employed RNA-sequencing (RNA-seq) to understand how Na2CO3 stress inhibits rice seedling growth. RESULTS: Na2CO3 stress significantly inhibited the growth of rice seedlings. Through RNA-seq, many differentially expressed genes (DEGs) were shown to be potentially involved in the rice seedling response to salt-alkali stress. After 1-day and 5-day treatments, RNA-seq identified 1780 and 2315 DEGs in the Na2CO3-treated versus -untreated rice seedling shoots, respectively. According to the gene ontology enrichment and the Kyoto Encylopedia of Genes and Genomes annotation of DEGs, the growth-inhibition processes associated with salt-alkali stress involve a myriad of molecular events, including biosynthesis and metabolism, enzyme activity, and binding, etc. CONCLUSION: Collectively, the transcriptome analyses in the present work revealed several potential key regulators of plant response to salt-alkali stress, and might pave a way to improve salt-alkali stress tolerance in rice.

A novel sensor for visual and selective detection of Hg2+ based on functionalized doped quantum dots.

In this paper, a novel analytical platform for the visual, sensitive and reliable analysis of mercury ions (Hg2+) is fabricated based on functionalized doped quantum dots. We synthesized a new specific nano-material, zinc dithiothreitol combined with graphene quantum dots (ZnNCs-NGQDs), by a simple and convenient method which, as an efficient luminophore, was then applied to construct an electrochemiluminescence (ECL) system for the first time. Under optimized conditions, the ECL sensor showed an excellent response for Hg2+ in the linear range of 1.0 mM to 10 pM, with a low detection limit of 3 pM. Moreover, the proposed method demonstrated satisfactory selectivity, stability and acceptable reproducibility for the detection of Hg2+. The recovery of tap water and lake water samples ranged from 96% to 105%, indicating the potential applicability of the proposed method for monitoring environmental water samples. Meanwhile, visual attempts for mercury ion detection by using doped quantum dots have also obtained satisfactory results. Importantly, our research revealed a viable method for improving the sensitivity and convenience of target studies in sensing fields derived from functional material design.

Toward efficient multiple-site incorporation of unnatural amino acids using cell-free translation system.

Amber suppression has been widely used to incorporate unnatural amino acids (UNAAs) with unique structures or functional side-chain groups into specific sites of the target protein, which expands the scope of protein-coding chemistry. However, this traditional strategy does not allow multiple-site incorporation of different UNAAs into a single protein, which limits the development of unnatural proteins. To address this challenge, the suppression method using multiple termination codons (TAG, TAA or TGA) was proposed, and cell-free unnatural protein synthesis (CFUPS) system was employed. By the analysis of incorporating 3 different UNAAs (p-propargyloxy-l-phenylalanine, p-azyl-phenylalanine and L-4-Iodophenylalanine) and mass spectrometry, the simultaneous usage of the codons TAG and TAA were suggested for better multiple-site UNAA incorporation. The CFUPS conditions were further optimized for better UNAA incorporation efficiency, including the orthogonal translation system (OTS) components, magnesium ions, and the redox environment. This study established a CFUPS approach based on multiple termination codon suppression to achieve efficient and precise incorporation of different types of UNAAs, thereby synthesizing unnatural proteins with novel physicochemical functions.

Whole genome sequencing analysis of a dexamethasone-degrading Burkholderia strain CQ001.

This study is to analyze the functional genes and metabolic pathways of dexamethasone degradation in Burkholderia through genome sequencing.A new Burkholderia sp. CQQ001 (B. CQ001) with dexamethasone degrading activity was isolated from the hospital wastewater and sequenced using Illumina Hiseq4000 combined with the third-generation sequencing technology. The genomes were assembled, annotated, and genomically mapped. Compared with six Burkholderia strains with typical features and four Burkholderia strains with special metabolic ability, the functional genes and metabolic pathways of dexamethasone degradation were analyzed and confirmed by RT-qPCR.Genome of B. CQ001 was 7,660,596 bp long with 6 ring chromosomes. The genes related to material metabolism accounted for 80.15%. These metabolism related genes could participate in 117 metabolic pathways and cover various microbial metabolic pathways in different environments and decomposition pathways of secondary metabolites, especially the degradation of aromatic compounds. The steroidal metabolic pathway containing 1 ABC transporter and 9 key metabolic enzymes related genes were scattered in the genome. Among them, the ABC transporter, KshA, and KshB increased significantly under the culture conditions of dexamethasone sodium phosphate as carbon source.B. CQ001 is a bacterium with strong metabolic function and rich metabolic pathways. It has the potential to degrade aromatics and other exogenous chemicals and contains genes for steroid metabolism. Our study enriches the genetic information of Burkholderia and provides information for the application of Burkholderia in bioremediation and steroid medicine production.

Endophyte infection enhances accumulation of organic acids and minerals in rice under Pb2+ stress conditions.

This study aimed to assess the effects of endophyte infection on Pb accumulation, organic acids (OAs) and mineral element contents by comparing endophyte-uninfected (E-) and endophyte-infected (E+) rice exposed to 0, 10.36, 20.72, 31.08 and 41.44 mg/L Pb2+ for 10 days. Pb2+ stress decreased growth, dry weight, and chlorophyll (Chl) content in E- and E+ in a concentration-dependent manner. Pb2+ accumulation was similar in E- and E+. Under Pb2+ stress, E+ accumulated more malate and fumarate in the leaves compared to E-, as well as more tartrate, malate, succinate and fumarate in the roots. Furthermore, E+ secreted more malate and lactate under 41.44 mg/L Pb2+ stress. Malate accumulation was induced by endophyte infection under Pb2+ exposure suggesting that malate is the most obvious candidate ligand for Pb2+. Endophyte infection increased Ca, Mg, P, Fe and Ni contents in the leaves and Ni contents in the roots under Pb2+ stress, but reduced Fe content in the roots under high Pb2+ stress. Under same Pb2+ concentration, endophyte infection significantly increased plant height, the dry weight of the shoots, and Chl content. The effects of endophyte infection might be due to changes in OAs accumulation and exudation and improvements in mineral uptake under Pb2+ stress.

ß-Caryophyllene/Hydroxypropyl-ß-Cyclodextrin Inclusion Complex Improves Cognitive Deficits in Rats with Vascular Dementia through the Cannabinoid Receptor Type 2 -Mediated Pathway.

This work was conducted to prepare ß-caryophyllene-hydroxypropyl-ß-cyclodextrin inclusion complex (HPßCD/BCP) and investigate its effects and mechanisms on cognitive deficits in vascular dementia (VD) rats. First, HPßCD/BCP was prepared, optimized, characterized, and evaluated. HPßCD/BCP and AM630 were then administered to VD rats to upregulate and downregulate the cannabinoid receptor type 2 (CB2). Results showed that HPßCD/BCP can significantly increase the bioavailability of BCP. Through the Morris water maze test, HPßCD/BCP can attenuate learning and memory deficits in rats. Cerebral blood flow (CBF) monitoring results indicated that HPßCD/BCP can promote the recovery of CBF. Moreover, molecular biology experiments showed that HPßCD/BCP can increase the expression levels of CB2 in brain tissues, particularly the hippocampus and white matter tissues, as well as the expression levels of PI3K and Akt. Overall, the findings demonstrated the protective effects of HPßCD/BCP against cognitive deficits induced by chronic cerebral ischemia and suggested the potential of HPßCD/BCP in the therapy of vascular dementia in the future.

Degradation of dexamethasone by acclimated strain of Pseudomonas Alcaligenes.

This study is to investigate the use of microbial remediation technology for degradation of dexamethasone in polluted water. A strain of Pseudomonas Alcaligenes with the ability of dexamethasone degradation was isolated from hospital polluted water. This strain was further acclimated into a bacterial strain that could highly degrade dexamethasone. Domesticated bacterial proteins were separated by osmotic shock method and were analyzed using SDS-PAGE. Enzyme activity of dexamethasone degradation was detected by high performance liquid chromatography. Protein bands with different molecular weight were found in all regions of the bacteria and a band with molecular weight of about 100 kDa was most obvious. In intracellular and periplasmic liquid, there was a band with molecular weight of about 41 kDa. Enzyme activity mainly existed in intracellular liquid. The 41 kDa protease was purified using ammonium sulfate precipitation, DEAE-52 ion exchange column and Sephadex G-100 column. Dexamethasone and dexamethasone sodium phosphate degrading rates of the purified enzyme were 36% and 95%, respectively. The 100 kDa protein had a 19% coverage rate to TonB receptor dependent protein, with 11 peptides matching. The 41 kDa protein had a 56% coverage rate to isovaleryl coenzyme A dehydrogenase, with 5 peptides matching. The 41 kDa protein had good degradation between the temperature of 25-40°C and PH value of 6.5-8.5. The enzyme kinetics equation was Ct = C0 e(-0.1769t), in accordance with the first-order kinetic equation. This study laid the foundation for further preparation of bioremediation agents for clearance of dexamethasone pollution in water.

[Studies on screening of matrix and permeation enhancer of sustained-release patches of duliang].

OBJECTIVE: To screen the matrix and permeation enhancer of Duliang Patches. METHODS: Based on L9 (3(4)) orthogonal experimental design, the content of imperatorin of the release rate and transdermal osmolality were regarded as evaluation indexes to optimize the matrix and permeation enhancer of patches suing of Drug dissolution tester and Franz diffusion cell. RESULTS: The best prescriplion of sustained-release patch of Duliang was: the quality percentage content of the starch, sodium carboxymethyl cellulose, glycerin, azone, propylene glycol and PEG400 was 6%, 2%, 30%, 1%, 15% and 2.5% respectively. The release behavior of sustained-release patches of Duliang tallied with Higuchi equation and the effect of sustained-release was apparent. CONCLUSION: The sustained-release patches of Duliang have good property of sustained-release and transdermal in vitro and the stability of patches is also sound while the release in vivo awaits further inspection.

Growth, photosynthesis and antioxidant responses of endophyte infected and non-infected rice under lead stress conditions.

An endophytic fungus was tested in rice (Oryza sativa L.) exposed to four levels of lead (Pb) stress (0, 50, 100 and 200 µM) to assess effects on plant growth, photosynthesis and antioxidant enzyme activity. Under Pb stress conditions, endophyte-infected seedlings had greater shoot length but lower root length compared to non-infected controls, and endophyte-infected seedlings had greater dry weight in the 50 and 100 µM Pb treatments. Under Pb stress conditions, chlorophyll and carotenoid levels were significantly higher in the endophyte-infected seedlings. Net photosynthetic rate, transpiration rate and water use efficiency were significantly higher in endophyte-infected seedlings in the 50 and 100 µM Pb treatments. In addition, chlorophyll fluorescence parameters Fv/Fm and Fv/Fo were higher in the infected seedlings compared to the non-infected seedlings under Pb stress. Malondialdehyde accumulation was induced by Pb stress, and it was present in higher concentration in non-infected seedlings under higher concentrations of Pb (100 and 200 µM). Antioxidant activity was either higher or unchanged in the infected seedlings due to responses to the different Pb concentrations. These results suggest that the endophytic fungus improved rice growth under moderate Pb levels by enhancing photosynthesis and antioxidant activity relative to non-infected rice.

Alleviation of exogenous oligochitosan on wheat seedlings growth under salt stress.

Hydroponic experiments were carried out to study the role of oligochitosan in enhancing wheat (Triticum aestivum L.) resistance to salt stress. Data were collected on plant biomass, chlorophyll content, photosynthetic rate (P (n)), stomatal conductance (g (s)), proline content, antioxidant enzyme activities, and malondialdehyde (MDA) content. Under 150 mM salt stress, plant growth was significantly inhibited. Shoot length, root length, and dry weight were sharply reduced by 26%, 31%, and 20%, respectively, of the control. Superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) were increased by 28%, 13%, and 26%, respectively, of the control and MDA content largely accumulated, which was 1.5-fold of the control. However, 0.0625% oligochitosan pretreatment alleviated the adverse effects of salt stress, which was reflected by increasing root length, shoot length, dry weight, chlorophyll content, P (n,) and g (s). Furthermore, it also showed that oligochitosan pretreatment significantly increased antioxidant enzyme (SOD, CAT and POD) activities, and reduced MDA content in leaves. Meanwhile, the accumulation of proline was markedly accelerated. The results indicated that oligochitosan pretreatment ameliorated the adverse effects and partially protected the seedlings from salt stress during the following growth period.

Effects of Na2CO3 stress on photosynthesis and antioxidative enzymes in endophyte infected and non-infected rice.

Endophyte infected and uninfected seedlings of rice (Oryza sativa L.) were subjected to five different levels of Na(2)CO(3) for 2 weeks. Under both Na(2)CO(3) stress as well as no stress, endophyte-infected plants were higher for above-ground dry weight and shoot length, but lower for root length compared to non-infected controls, and there was no significant difference for below-ground dry weight. Chlorophylls and carotenoid contents, net photosynetic rate, transpiration rate, catalase and peroxidase activities increased, but malondialdehyde content declined in the infected plants compared to non-infected controls under Na(2)CO(3) stress. Compared to non-infected controls, water use efficiency, stomatal conductance, chlorophyll fluorescence parameters (F(v)/F(m), F(v)/F(o)) and superoxide dismutase activity in infected plants increased under high concentrations of Na(2)CO(3) while there was no significant difference under low concentrations of Na(2)CO(3). Endophyte infection was concluded to be beneficial to the growth and antioxidative mechanisms in Oryza sativa under Na(2)CO(3) stress.

Elevated carbon dioxide and/or ozone concentrations induce hormonal changes in Pinus tabulaeformis.

We investigated endogenous plant hormones and needle growth in Pinus tabulaeformis plants grown in open-top chambers and exposed to ambient or elevated concentrations of carbon dioxide (CO(2)) and/or ozone (O(3)). Exposure to elevated CO(2) for 100 days significantly increased the change in fresh needle weight, indole-3-acetic acid (IAA), isopentenyl-adenosine (iPA), and dihydrozeatin riboside (DHZR) content. Abscisic acid (ABA) content decreased, and no effect was observed on zeatin riboside (ZR) content or changes in needle dry weight. The ratios of IAA/ABA and total cytokinins (CKs)/ABA [Formula: see text] were increased. Elevated O(3) significantly decreased IAA and ZR, and decreased the ratios of IAA/ABA and CKs/ABA. Ozone treatment increased ABA content but did not change iPA or DHZR content or change fresh or dry needle weights. The combination treatment significantly increased ABA content and the IAA/ABA ratio but decreased the total CKs/ABA ratio and had no effect on CKs or IAA content or change in fresh and dry needle weights. The results indicate that elevated CO(2) ameliorated the effects of elevated O(3) on tree growth.

[Hydrolysis technology optimization of phorbol esters by orthogonal experiment].

OBJECTIVE: To establish the best hydrolytic conditions from phorbol esters. METHOD: The orthogonal experiment was used to optimize 4 factors, which were reaction time, ratio of solid-to-liquid, hydrolytic times, and temperature. Diamonsil C18 column (4. 6 mm x 250 mm, 5 microm) was used and the mobile phase was consisted of acetonitrile and water for HPLC detection. The detection wavelength was set at 234 nm, the flow rate was 1 mL x min(-1), and the column temperature was 25 degrees C. RESULT: The optimum conditions were 10 h of reaction time, 1:6 of solid-to-liquid (BaOH/MeOH) ratio, 25 degrees C of temperature, and one time of hydrolysis. There was a good linear relationship of phorbol in the range of 4.28-107 mg x L(-1) (r = 0.999 9), and the average recovery was 97.89%, with RSD 0.78%. CONCLUSION: The method is steady, reliable and reproducible, and it provides a mean for future study.