Redox-sensitive self-assembling polymer micelles based on oleanolic modified hydroxyethyl starch: Synthesis, characterisation, and oleanolic release.

Our study aimed at developing polymer micelles that possess redox sensitivity and excellent controlled release properties. 3,3'-dithiodipropionic acid (DTDPA, Abbreviation in synthetic polymers: SS) was introduced as ROS (Reactive oxygen species)response bond and connecting arm to couple hydroxyethyl starch (HES) with oleanolic acid (OA), resulting in the synthesis of four distinct grafting ratios of HES-SS-OA. FTIR (Fourier Transform infrared spectroscopy) and 1H NMR (1H Nuclear magnetic resonance spectra) were used to verify the triumphant combination of HES-SS-OA. Polymer micelles were found to encapsulate OA in an amorphous form, as indicated by the results of XRD (X-ray diffraction) and DSC (Differential scanning calorimetry). When the OA grafting rate on HES increased from 7.72 % to 11.75 %, the particle size decreased from 297.79 nm to 201.39 nm as the polymer micelles became compact due to enhanced hydrophobicity. In addition, the zeta potential changed from -16.42 mv to -25.78 mv, the PDI (polydispersity index) decreased from 0.3649 to 0.2435, and the critical micelle concentration (CMC) decreased from 0.0955 mg/mL to 0.0123 mg/mL. Results of erythrocyte hemolysis, cytotoxicity and cellular uptake illustrated that HES-SS-OA had excellent biocompatibility and minimal cytotoxicity for AML-12 cells. Disulfide bond breakage of HES-SS-OA in the presence of H2O2 and GSH confirmed the redox sensitivity of the HES-SS-OA micelles and their excellent controlled release properties for OA. These findings suggest that HES-SS-OA can be potentially used in the future as a healthcare drug and medicine for the prevention or adjuvant treatment of inflammation.

Lemon essential oil nanoemulsions: Potential natural inhibitors against Escherichia coli.

Lemon essential oil (LEO) is a common natural antibacterial substance, and encapsulating LEO into nanoemulsions (NEs) can improve their stability and broaden its application. Our study aimed to investigate the bacterial inhibitory effect of LEO-NEs against Escherichia coli (E. coli). Results showed that the minimum inhibitory concentration (MIC) of LEO-NEs was 6.25 mg/mL, and the time-kill curve showed that E. coli were significantly killed by LEO-NEs after 5 h of treatment at 1MIC. Flow-cytometry analysis showed that LEO-NEs adversely affected the cell-membrane depolarisation, cell-membrane integrity, and efflux pump function of E. coli. Confocal laser scanning microscopy demonstrated that 8MIC of LEO-NEs induced changes in the cell-membrane permeability and cell-wall integrity of E. coli. Proteomic results suggested that the mode of action LEO-NEs against E. coli was to enhance bacterial chemotaxis and significantly inhibit ribosomal assembly. They may also affect butyric acid, ascorbic acid and aldehyde metabolism, and sulphur-relay system pathways. In conclusion, LEO-NEs had potential application as a natural antibacterial agent for the control of E. coli in the food industry.

pH-Responsive doxorubicin-loaded magnetosomes for magnetic resonance-guided focused ultrasound real-time monitoring and ablation of breast cancer.

MR-guided focused ultrasound surgery (MRgFUS) is driving a new direction in non-invasive thermal ablation therapy with spatial specificity and real-time temperature monitoring. Although widely used in clinical practice, it remains challenging to completely ablate the tumor margin due to fear of damaging the surrounding tissues, thus leading to low efficacy and a series of complications. Herein, we have developed novel pH-responsive drug-loading magnetosomes (STPSD nanoplatform) for increasing the T2-contrast and improved the ablation efficiency with a clinical MRgFUS system. Specifically, this STPSD nanoplatform is functionalized by pH-responsive peptides (STP-TPE), encapsulating superparamagnetic iron oxide (SPIO) and doxorubicin (DOX), which can cause drug release and SPIO deposition at the tumor site triggered by acidity and MRgFUS. Under MRgFUS treatment, the increased vascular permeability caused by hyperthermia can improve the uptake of SPIO and DOX by tumor cells, so as to enhance ultrasound energy absorption and further enhance the efficacy of chemotherapy to completely ablate tumor margins. Moreover, we demonstrated that a series of MR sequences including T2-weighted imaging (T2WI), contrast-enhanced T1WI imaging (T1WI C+), maximum intensity projection (MIP), volume rendering (VR) and ADC mapping can be further utilized to monitor the MRgFUS ablation effect in rat models. Overall, this smart nanoplatform has the capacity to be a powerful tool to promote the therapeutic MRgFUS effect and minimize the side effects to surrounding tissues.

Development of Citrus-Based Functional Jelly and an Investigation of Its Anti-Obesity and Antioxidant Properties.

Intervention with natural products is becoming a promising obesity control strategy as healthy eating becomes increasingly popular. The present study aimed to prepare a citrus-based functional jelly (CFJ) from citrus by-products and investigate its bioactive effects in mice. The results of the CFJ preparation showed that the optimal formula of CFJ was 29.12%, 20%, and 3.61% for chenpi, orange juice, and pectin, respectively. The optimized CFJ can be personalized and designed with jelly shapes using 3D food printing technology. The evaluation of the biological activity of the CFJ showed that it was low in calories, with a total phenolic content of 12.44 ± 0.26 mg GAE/g. Moreover, the CFJ has a good free radical scavenging ability for ABTS. The results of the mouse experiments showed that the CFJ significantly suppressed the body weight gain and fat deposits with a dose-dependent effect, compared with the control group (p < 0.05). In addition, the activities of the antioxidant-related enzymes (CAT and SOD) of the mice were also enhanced after a supplementation with the CFJ. In short, the CFJ is a functional snack enriched in phenolic substances with low-calorie, antioxidant and anti-obesity properties. This work promotes the utilization of citrus by-products and the healthy development of its processing industry.

Membrane-camouflaged supramolecular nanoparticles for co-delivery of chemotherapeutic and molecular-targeted drugs with siRNA against patient-derived pancreatic carcinoma.

Pancreatic cancer remains one of the most lethal malignancies worldwide. The combination of the first-line standard agent gemcitabine (GEM) with the molecular-targeted drug erlotinib (Er) has emerged as a promising strategy for pancreatic cancer treatment. However, the clinical benefit from this combination is still far from satisfactory due to the unfavorable drug antagonism and the fibrotic tumor microenvironment. Herein, we propose a membrane-camouflaged dual stimuli-responsive delivery system for the co-delivery of GEM and Er into pancreatic cancer cells and tissues to block the antagonism, as well as reshapes profibrotic tumor microenvironment via simultaneous delivery of small interference RNA (siRNA) for synergistic pancreatic cancer treatment. This "all-in-one" delivery system exhibits sensitive GSH and pH-dependent drug release profiles and enhances the inhibitory effects on the proliferation and migration of tumor cells in vitro. Excitingly, the systemic injection of such a biomimetic drug co-delivery system not only resulted in superior inhibitory effects against orthotopic pancreatic tumor and patient-derived tumor (PDX), but also greatly extended the survival rate of tumor-bearing mice. Our findings provide a promising therapeutic strategy against pancreatic cancer through the enhanced synergistic effect of target therapy, chemotherapy and anti-fibrotic therapy, which represents an appealing way for pancreatic cancer treatment.

Development of Functional Kiwifruit Jelly with chenpi (FKJ) by 3D Food Printing Technology and Its Anti-Obesity and Antioxidant Potentials.

With the growing popularity of the concept of healthy diet, modern obesity treatment is gradually shifting from surgical or pharmacological treatment to nutritional intervention. As a safe and effective measure, natural product interventions are a potential strategy of obesity management. The present study aimed to develop a kind of functional food rich in bioactive compounds (chenpi, kiwifruit, and pectin as raw materials) and investigate their bioactive effects on a mouse model. For development of functional kiwifruit jelly with chenpi (FKJ), the results of single-factor and response surface experiments showed that the optimized formulation was composed of a 30.26% addition of chenpi, 35% addition of kiwifruit juice, and 2.88% addition of pectin. The FKJ obtained with the optimal formulation could be used as a 3D printing raw material to print the desired food shapes successfully. For bioactivity evaluation of FKJ, the results with a mouse model showed that the food intake, liver weight, and adipose tissue weight were significantly decreased after administration of FKJ with dose-dependent effect compared to the CON group (p < 0.05). Meanwhile, the serum levels of several inflammatory factors (TG, IL-6, and TNF-α) were decreased and the activities of several antioxidant-related enzymes (SOD, GSH-PX, and CAT) were increased. In short, a functional kiwifruit jelly with chenpi was developed in this study. It is a functional snack food rich in active phenolic compounds, low in calories, with antioxidant and anti-inflammatory activity, and prevents fat accumulation. FKJ could well meet the needs of modern people for nutrition and health and also promote the processing and utilization of natural products, and has good development prospects in the functional food industry.

Preparation, Characterization, and Antioxidant Activity of Nanoemulsions Incorporating Lemon Essential Oil.

Lemon essential oil (LEO) is a kind of citrus essential oil with antioxidant, anti-inflammatory, and antimicrobial activities, but low water solubility and biological instability hinder its industrial application. In this study, LEO was nanoemulsified to solve these problems. The preparation procedure of lemon essential oil nanoemulsions (LEO-NEs) was optimized, and the physicochemical characterization and antioxidant activities were explored. Single-factor experiments (SFEs) and response surface methodology (RSM) were conducted for the effects on the mean droplet size of LEO-NEs. Five factors of SFE which may influence the droplet size were identified: HLB value, concentration of essential oil, concentration of surfactant, ultrasonic power, and ultrasonic time. On the basis of the SFE, the RSM approach was used to optimize the preparation procedure to obtain LEO-NEs with the smallest droplet size. LEO-NEs exhibited good antioxidant activity when the HLB value was 13, content of surfactant was 0.157 g/mL, ultrasonic time was 23.50 min, and ultrasonic power was 761.65 W. In conclusion, these results can provide a good theoretical basis for the industrial application of lemon essential oil.

A Duplex CRISPR-Cas9 Ribonucleoprotein Nanomedicine for Colorectal Cancer Gene Therapy.

Based on the high frequency of concurrent adenomatous polyposis coli (APC) and KRAS mutations and their strong cooperative interaction in human colorectal cancer (CRC) promotion, we herein develop a CRISPR-Cas9-based genome-editing nanomedicine to target both APC and KRAS mutations for the treatment of CRC. To this end, a hyaluronic acid (HA)-decorated phenylboronic dendrimer (HAPD) was designed for the targeted delivery of Cas9 ribonucleoprotein (RNP), by which both APC and KRAS genetic mutations harboring in CRC cells can be synergistically disrupted. Systemic administration of Cas9 RNP targeting APC and KRAS enabled by HAPD significantly inhibits tumor growth on xenografted and orthotopic CRC mouse models and also greatly prevents CRC-induced liver metastasis and lung metastasis. Thus, this duplex genome-editing system provides a promising gene therapy strategy for the treatment of human CRC and can be extended to other types of cancers with activated Wnt/ß-catenin and RAS/extracellular signal-regulated kinase (ERK) pathways.

Global transcriptomic response of Listeria monocytogenes exposed to Fingered Citron (Citrus medica L. var. sarcodactylis Swingle) essential oil.

Listeria monocytogenes, which could cause severe disease of listeriosis, is one of the most concerned foodborne pathogens worldwide. Citrus medica L. var. sarcodactylis Swingle (Fingered Citron) is one of the citrus species cultivated in south China. Here, we investigated the efficacy of Fingered Citron essential oil (FCEO) against L. monocytogenes and explored the response of L. monocytogenes in the presence of FCEO using genome-wide transcriptome analysis. FCEO exhibited strong anti-listeria activity and obvious alterations of cell morphology were observed by scanning electron microscopy and transmission electron microscopy. Moreover, GO analysis demonstrated many potential cell responses, including metabolic process, cellular process, single-organism process, cell part, membrane, catalytic activity, binding, and transporter activity. KEGG analysis suggests that L. monocytogenes respond and adapt by (1) increasing motility through the enhancement of flagella rotation; (2) promoting cell tumbles and re-orientating to escape from FCEO; (3) enhancing the uptake of carbohydrates from environment to gain more energy; (4) changing the uptake of several metallic cations, including iron, zinc, cobalt, and nickel. Our research contributes to the understanding of the adaptive responses of L. monocytogenes exposed to FCEO and provides novel insights for finding new targets of anti-listeria therapy.

Changes in the Volatile Components of Candied Kumquats in Different Processing Methodologies with Headspace-Gas Chromatography-Ion Mobility Spectrometry.

The effects of two different processing methods on the volatile components of candied kumquats were investigated via headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS). The characteristic volatile fingerprints of fresh kumquats (FKs), vacuum sugaring osmosis combined with hot-air drying kumquats (VS-ADKs), and atmospheric pressure sugaring osmosis combined with hot-air drying kumquats (AS-ADKs) were established using 3D topographic plots. From the fingerprints, 40 signal peaks for 22 compounds were confirmed and quantified in all types of kumquats, namely, two terpenes, four esters, seven aldehydes, three ketones, and six alcohols. 3-Pentanone was identified as the major component of FKs; followed by 1-hexanol and the Z-3-hexen-1-ol dimer. The hexanal dimer, 2-hexen-1-ol, and the ethyl acetate dimer were the major markers of VS-ADKs. Benzaldehyde and furfurol were the prominent constituent parts of AS-ADKs. Compared with that in FKs, the pentanal and dimethyl ketone contents of VS-ADKs and AS-ADKs exhibited a dramatic increase (p < 0.05). By contrast, the change in ethanol dimer tended to decrease (p < 0.05). Principal component analysis (PCA) clearly showed that the samples, which were distributed in a separate space could be well-distinguished. Furthermore, the similarity of different processed kumquats and their corresponding volatile components was demonstrated via heat map clustering analysis. The results confirmed the potential of HS-GC-IMS-based approaches to evaluate processed kumquats with various volatile profiles.

Ganoderma lucidum Polysaccharides Prevent Palmitic Acid-Evoked Apoptosis and Autophagy in Intestinal Porcine Epithelial Cell Line via Restoration of Mitochondrial Function and Regulation of MAPK and AMPK/Akt/mTOR Signaling Pathway.

Ganoderma lucidum polysaccharide (GLP) extracted from Ganoderma lucidum (Leyss. ex Fr.) Karst, a traditional Chinese medicine, is a biologically active substance reported to possess anti-oxidative, anti-apoptotic, and neurological protection. However, it is unknown whether GLP have any protective effect against high-fat constituents-induced epithelial cell injury. The aim of this study was to investigate the protection and molecular mechanism of GLP on injury induced by palmitic acid (PA) in the intestinal porcine epithelial cell line (IPEC-J2). First, we tested whether the treatment of GLP attenuate PA-induced IPEC-J2 cell death. GLP markedly blocked PA-caused cytotoxicity and apoptosis in IPEC-J2 cells. Moreover, GLP recovered the decreased mitochondrial function and inhibited activation of caspase-dependent apoptotic pathway. Interestingly, PA promoted cell apoptosis and autophagy through stimulation of phosphorylation of mitogen-activated protein kinases (MAPKs), AMP-activated protein kinase (AMPK), and inhibition of phosphorylation of Akt and mammalian target of rapamycin (mTOR), which was reversed by GLP. Taken together, this study revealed a protective effect of GLP against PA-evoked IPEC-J2 cell death through anti-apoptotic and anti-autophagic properties.