Lipid-Based Nanocarriers Enabled Oral Delivery of Oleanolic Acid Derivative DKS26 for Diabetes Management.

Oleanolic acid derivative DKS26 has hypolipidemic, islet, and hepatoprotective effects. However, high lipophilicity and low water solubility led to DKS26 extremely low oral bioavailability. Herein, lipid-based nanocarriers, including lipid nanodiscs (sND/DKS26) and liposomes (sLip/DKS26), are prepared to improve DKS26 oral absorption. In comparison to free DKS26 (5.81%), the absolute oral bioavailabilities are significantly increased to 29.47% (sND/DKS26) and 37.25% (sLip/DKS26) without detectable toxicity or immunogenicity even after repeated administrations. Both sND/DKS26 and sLip/DKS26 significantly reduce the feeding glucose level and the AUC of OGTT in db/db diabetic mice. Aiding by the newly developed scFv-based nanocarrier separation methods, no intact nanocarriers are detected in blood circulation after oral administration, suggesting that both formulations are unable to penetrate the intestinal epithelium. They enhance DKS26 absorption mainly by improving intestinal cell uptake and rapid intracellular release of the payload. Since pre-existing anti-PEG is widely detected in humans, the present oral absorption pathway of both nanocarriers successfully avoids unfavorable immunological responses after interaction with anti-PEG antibodies. The application of lipid-based nanocarriers paves an efficient and safe avenue for the clinical translation and application of poorly soluble therapeutics derived from traditional Chinese medicine.

De novo full length transcriptome analysis and gene expression profiling to identify genes involved in phenylethanol glycosides biosynthesis in Cistanche tubulosa.

BACKGROUND: The dried stem of Cistanche, is a famous Chinese traditional medicine. The main active pharmacodynamic components are phenylethanol glycosides (PhGs). Cistanche tubulosa produces higher level of PhGs in its stems than that of Cistanche deserticola. However, the key genes in the PhGs biosynthesis pathway is not clear in C. tubulosa. RESULTS: In this study, we performed the full-length transcriptome sequencing and gene expression profiling of C. tubulosa using PacBio combined with BGISEQ-500 RNA-seq technology. Totally, 237,772 unique transcripts were obtained, ranging from 199 bp to 31,857 bp. Among the unique transcripts, 188,135 (79.12%) transcripts were annotated. Interestingly, 1080 transcripts were annotated as 22 enzymes related to PhGs biosynthesis. We measured the content of echinacoside, acteoside and total PhGs at two development stages, and found that the content of PhGs was 46.74% of dry matter in young fleshy stem (YS1) and then decreased to 31.22% at the harvest stage (HS2). To compare with YS1, 13,631 genes were up-regulated, and 15,521 genes were down regulated in HS2. Many differentially expressed genes (DEGs) were identified to be involved in phenylpropanoid biosynthesis pathway, phenylalanine metabolism pathway, and tyrosine metabolism pathway. CONCLUSIONS: This is the first report of transcriptome study of C. tubulosa which provided the foundation for understanding of PhGs biosynthesis. Based on these results, we proposed a potential model for PhGs biosynthesis in C. tubulosa.

Mechanisms of straw biochar's improvement of phosphorus bioavailability in soda saline-alkali soil.

High pH and exchangeable sodium percentage, structural deterioration due to alkalinity, and nutrient deficiencies are typical characteristics of soda saline-alkali soil. In addition, phosphorus is typically the main limiting nutrient. Thus, there have been intense efforts to counter the salinity and improve the phosphorus availability of these soils (which cover large and growing areas). A promising approach is long-term application of straw biochar, which can significantly reduce soil salinity and promote the transformation of soil phosphorus. However, the mechanisms involved remain unclear. Thus, major aims of this review are to systematically address the mechanisms whereby biochar improves phosphorus bioavailability in soda saline-alkali soil through changes in the soil's physico-chemical properties, aggregate stability, contents of organic acids, enzyme activities, key functional genes, and microbial community structure. Another is to provide theoretical foundations for establishing effective methods for applying straw biochar to improve soda saline-alkali land and optimize phosphorus fertilizer applications.

Dietary curcumin supplementation effects on blood immunological profile and liver enzymatic activity of laying hens after exposure to high temperature conditions.

Various environmental factors affect livestock production but heat stress is a major challenge in the poultry farming. Poultry exposes to high temperature alters blood immunological parameters and liver enzymatic function which in turn, suppress the immunity and disease resistance of chickens. Thus, the purpose of present study was to explore the effect of dietary curcumin supplementation on blood immunological biomarker and liver enzymatic activity of laying hens under heat stress conditions. Experimental groups contained two control groups (normal temperature control (NC) and heat stress control (HC) and 3 heat stress curcumin treatment groups (HT100, HT200 and HT300). Hens in HC group with basal diet and heat stress curcumin treatment groups were exposed 6 h/day heat stress (32 ± 1 °C) from 10:00 a.m. to 16:00 p.m. for 9 week. The results of present study showed that heat stressed curcumin treatment group had improved liver weight, WBC values and immunoglobulin level as compared to untreated HC and NC groups. The available results also indicated that laying hens supplemented with curcumin under high temperature conditions had reduced H/L ratio, serum corticosterone levels, inflammatory cytokines response and liver enzymatic activity (ALT) which enhanced the immunity of laying hens under hot climatic conditions. Therefore, it is concluded that curcumin has ability to combat harsh environmental conditions which can be used as anti-inflammatory and immune booster feed additive in the poultry nutrition.

Effect of diacylglycerol interfacial crystallization on the physical stability of water-in-oil emulsions.

The influence of diacylglycerol (DAG) combined with polyglycerol polyricinoleate (PGPR) on the stability of water-in-oil (W/O) emulsions containing hydrogenated palm oil (HPO) was studied. Polarized light microscope revealed that DAG promoted HPO to crystallize at the water-oil interface, providing the combination of Pickering and network stabilization effects. It was proposed that the molecular compatibility of fatty acids in DAG with HPO accounted for the promotional effect. The interfacial crystallization of DAG together with the surface activity of PGPR led to the formation of emulsions with uniform small droplets and high freeze-thaw stability. Further exploration of physical properties indicated that the combination of DAG and PGPR dramatically improved the emulsion's viscoelasticity and obtained a larger deformation yield. Water droplets in DAG-based emulsions acted as active fillers to improve the network rigidity. Therefore, DAG is a promising material to be used as emulsifier to enhance the physical stability of W/O emulsions.

Simultaneous Unlocking Optoelectronic and Interfacial Properties of C60 for Ultrasensitive Immunosensing by Coupling to Metal-Organic Framework.

Due to exceptional electron-accepting ability, light-absorption, and a delocalized conjugated structure, buckminsterfullerene (C60) has attracted fascinating interest in the field of organic solar cells. However, poor delocalization and accumulation of electrons for pristine C60 in physiological aqueous solution and difficulties in conjugation with biomolecules limit its extended photovoltaic applications in bioassay. Herein, we reported the noncovalent coupling of C60 to an electronically complementary porphyrin-derived metal-organic framework (PCN-224) with carboxyl-group terminals. Such assembly not only offered a friendly interface for bioconjugation but also resulted in a long-range ordering C60@PCN-224 donor-acceptor system that demonstrated an unprecedented photocurrent enhancement up to 10 times with respect to each component. As an example, by further cooperating with Nanobodies, the as-prepared C60@PCN-224 was applied to a photoelectrochemical (PEC) immunosensor for S100 calcium-binding protein B with by far the most promising detection activities. This work may open a new venue to unlock the great potential of C60 in PEC biosensing with excellent performances.

Effect of dietary curcumin on the antioxidant status of laying hens under high- temperature condition.

Heat stress induces oxidative stress, and reduces antioxidant defenses of birds, which may affect poultry-production performance. Dietary antioxidants may protect against heat stress. We evaluated the effect of increasing concentrations of dietary curcumin on antioxidant parameters of hens under high-temperature conditions for nine weeks. Roman laying hens (n = 336, 22 weeks old, 1420 g weight) were divided into three treatment groups. The first group served as a thermo-neutral control (kept at 25 ±â€¯1 °C). The second group was exposed to high temperatures (32 ±â€¯1 °C, 6 h/day), and fed a basal diet. The third group was further divided into five groups, and all were exposed to high temperatures (32 ±â€¯1 °C, 6 h/day) and provided a basal diet supplemented with 100, 150, 200, 250, 300 mg/kg curcumin (H1, H2, H3, H4, H5). All treatments included four replicates of 12 hens. Total superoxide dismutase (SOD) activity was significantly higher in H2 and H3 groups, and total antioxidant capacity (T-AOC) was higher in H2, H3, and H5 groups. Catalase (CAT) and glutathione peroxidase (GSH-Px) activities were significantly higher in the H3 group. Malondialdehyde concentrations were lower in curcumin supplemented hens compared to control groups hens. Hens in all curcumin treatment groups had slightly (but non-significantly) higher activities of CAT, SOD, GSH-Px, and T-AOC in liver, heart, and lung tissues, compared to heat stressed control group. It is concluded that dietary curcumin given to laying hens under heat stress may enhance their antioxidant status, and ameliorate stressful environmental conditions.

Effect of curcumin supplementation on TLR4 mediated non-specific immune responses in liver of laying hens under high-temperature conditions.

The liver performs a significant role in innate and adaptive immunity. Heat stress causes oxidative stress in liver tissues and reduces the immune responses of laying hens which can cause several diseases affecting poultry-production performance. Hepatic inflammation is a common trigger of liver disease, which is reflected by hepatic tissue damage leading to fibrogenesis and hepatocellular carcinoma. Dietary manipulation of curcumin has been proposed to ameliorate the immune status of chickens under heat stress. Thus, this study aimed to investigate the effect of curcumin supplementation on TLR4 mediated non-specific immune response in liver of laying hens under high-temperature conditions. Experimental groups contained two controls groups (high temperature and thermo-neutral control (HC and NC) fed basal diet) and three high-temperature curcumin treatments groups (HT100, HT200 and HT300). Laying hens in HC and HT groups exposed 6 h/day heat stress (32 ±â€¯1 °C). The results of present study showed that heat stress curcumin treatment group had reduced inflammatory responses (IL-6, IL-1ß, TNF-α) as compared to HC and NC group. Pathological lesions and DNA damage of immune tissues were decreased in heat stress curcumin supplementation as compared to HC and NC group. Furthermore, PCNA, TLR4 and its downstream gene expression as well as protein expression (TLR4, NF-κB and PCNA) were significantly down regulated in heat stress curcumin supplemented group as compared to HC and NC group. Therefore, it is concluded that heat stressed hens supplemented with dietary curcumin enhance the immunity of laying hens and combat stressful environmental conditions.

Antimicrobial Activity of Punicalagin Against Staphylococcus aureus and Its Effect on Biofilm Formation.

Punicalagin, one of the main active compounds in pomegranate peel, has been reported to possess many properties, including antioxidant, antimicrobial, antiviral, and immunosuppressive activities. The aim of this study was to investigate the antibacterial effect of punicalagin against Staphylococcus aureus and possible mode of action. Growth inhibition activity was examined by the agar diffusion method. Then agar dilution method was adopted to determine the minimum inhibitory concentration (MIC). The effects of punicalagin on cell membrane were assessed by measuring potassium efflux. Morphological changes of S. aureus were assessed by scanning and transmission electron microscopy. Crystal violet assay was applied to investigate antibiofilm activity of punicalagin. Punicalagin exhibited good antistaphylococcal effect with an MIC of 0.25 mg/mL. An increase of potassium efflux was observed when cells were treated with punicalagin at 2 × MIC. Punicalagin induced morphological damages to the cell membrane. Moreover, punicalagin exerted a remarkable inhibitory effect on biofilm formation of S. aureus. These findings suggest that punicalagin has antimicrobial and antibiofilm activities against S. aureus and may have potential application to control S. aureus contamination in food industry.

Tribological behavior of a Ni-free Zr-based bulk metallic glass with potential for biomedical applications.

In this study, the tribological behavior of a Ni-free Zr53Al16Co23·25Ag7.75 bulk metallic glass (BMG) was investigated in dry-sliding and simulated physiological media using ball-on-disk reciprocating friction. The effects of sliding load, speed, media and counterpart materials on the wear resistance of the Zr-Al-Co-Ag BMG were illustrated. Under dry-sliding in air, wear resistance of the Zr-based BMG decreases with increasing sliding load, and wear deterioration is controlled by oxidation and abrasive wear. With increasing sliding velocity, larger plastic deformation occurs on the surface of BMG due to the frictional heat. The BMG exhibits decreased wear resistance in 0.9% NaCl and phosphate buffer saline (PBS) solutions in comparison with that in air and deionized water, which is probably associated with tribocorrosion controlled by synergistic effects of abrasive and corrosive wear. The wear resistance of the Zr-based BMG against Si3N4 counterpart material is inferior to that against ZrO2, whereas the case is contrary to that against Al2O3. The effect of ceramic counterpart materials on the wear resistance of BMG is discussed based on their Young's modulus and fracture toughness.

Protective effect of oleanolic acid on oxidized-low density lipoprotein induced endothelial cell apoptosis.

Oleanolic acid (3ß-hydroxyolean-12-en-28-oic acid, OA) is a naturally-occurring triterpenoid with various promising pharmacological properties. The present study was conducted to determine the protective effects of OA against oxidized low-density lipoprotein (ox-LDL) induced endothelial cell apoptosis and the possible underlying mechanisms. Our results showed that ox-LDL significantly decreased cell viability and induced apoptosis in human umbilical vein endothelial cells (HUVECs). OA in the co-treatment showed a protective effect against ox-LDL induced loss in cell viability and an increase in apoptosis, which was associated with the modulating effect of OA on ox-LDL induced hypoxia-inducible factor 1α(HIF-1α) expression. Moreover, our results showed that the modulating effect of OA against ox-LDL induced HIF-1α expression was obtained via inhibition of lipoprotein receptor 1 (LOX-1)/reactive oxygen species (ROS) signaling. Collectively, we suggested that the protective effect of OA against ox-LDL induced HUVEC apoptosis might, at least in part, be obtained via inhibition of the LOX-1/ROS/HIF-1α signaling pathway.

The anti-infective activity of punicalagin against Salmonella enterica subsp. enterica serovar typhimurium in mice.

Punicalagin, a major bioactive component of pomegranate peel, has been proven to have antioxidant, antiviral, anti-apoptosis, and hepatoprotective properties. The aim of this study was to investigate the anti-infective activity of punicalagin in a mouse model. C57BL/6 mice were initially challenged with Salmonella enterica subsp. enterica serovar typhimurium (S. typhimurium) and then treated with punicalagin. Food and water consumption and body weight were recorded daily. On day 8 post infection, the mice were sacrificed to examine pathogen counts in tissues, hematological parameters, cytokine levels, and histological changes. Compared to mice only infected with S. typhimurium, punicalagin-treated mice had more food consumption and less weight loss. A higher survival rate and lower counts of viable S. typhimurium in feces, liver, spleen, and kidney were found in the punicalagin-treated mice. The enzyme linked immunosorbent assay showed that the levels of IL-6, IL-10, and IFN-γ in serum and the spleen and TNF-α in serum, the spleen and the liver were reduced by punicalagin. Moreover, more neutrophils and higher neutrophil-to-mononuclear cell ratios in the punicalagin-treated mice were observed. Histological examination showed that punicalagin protected cells in the liver and spleen from hemorrhagic necrosis. It is concluded that punicalagin has a beneficial effect against S. typhimurium infection in mice. The anti-infective properties, together with other nutritionally beneficial effects, make punicalagin a promising supplement in human food or animal feeds to prevent disease associated with S. typhimurium.

Tannin-rich pomegranate rind extracts reduce adhesion to and invasion of Caco-2 Cells by Listeria monocytogenes and decrease its expression of virulence genes.

Pomegranate rind is rich in tannins that have remarkable antimicrobial activities. This study aimed to evaluate the effects of a tannin-rich fraction from pomegranate rind (TFPR) on Listeria monocytogenes virulence gene expression and on the pathogen's interaction with human epithelial cells. Growth curves were monitored to determine the effect of TFPR on L. monocytogenes growth. The 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide and fluorescence staining assays were used to examine the cytotoxicity of TFPR. The effects of TFPR on L. monocytogenes adhesion to and invasion of epithelial cells were investigated using Caco-2 cells. Real-time quantitative PCR analysis was conducted to quantify mRNA levels of three virulence genes in L. monocytogenes. Results showed that a MIC of TFPR against L. monocytogenes was 5 mg/ml in this study. TFPR exhibited cytotoxicity against Caco-2 cells when the concentration was 2.5 mg/ml. Subinhibitory concentrations of TFPR significantly reduced, in a dose-dependent manner, adhesion to and invasion of Caco-2 cells by L. monocytogenes. When L. monocytogenes was grown in the presence of 2.5 mg/ml TFPR, the transcriptional levels of prfA, inlA, and hly decreased by 17-, 34-, and 28-fold, respectively.

Bactrian camel nanobody-based immunoassay for specific and sensitive detection of Cry1Fa toxin.

The variable domain of the heavy-chain-only antibody (VHH) or nanobody (Nb), derived from camelids, begins to play an important role on the detection of protein markers. In this study, we constructed a phage-displayed library of VHHs against Cry1Fa by immunizing a healthy Bactrian camel with Cry1Fa toxin. After a series of bio-panning and screening by phage display technology, three anti-Cry1Fa nanobodies (Nbs) with great difference in complementarity determining region 3 (CDR3) were obtained and they were highly specific to Cry1Fa as well as showed full of activity when exposed to 70 °C for 3 h. Through modifying Nbs with Horseradish Peroxidase (HRP) and biotin, two Nbs which can recognize the different epitopes of Cry1Fa were determined and they were used to establish a novel sandwich immune ELISA based on biotin-SA interaction for Cry1Fa detection. The immunoassay exhibited a linear range from 1 to 100 ng/mL with a detection limit of 0.88 ng/mL. The recoveries from spiked corn and soybean samples were ranged from 83.33 to 117.17%, with a coefficient of variation (C.V) less than 6.0%. All together, the proposed immunoassay will be a promising way for sensitive and accurate determination of Cry1Fa toxin.

Tannin-rich fraction from pomegranate rind damages membrane of Listeria monocytogenes.

Pomegranate rind has been reported to inhibit several foodborne pathogens, and its antimicrobial activity has been attributed mainly to its tannin fraction. This study aimed to investigate the antimicrobial activity of the tannin-rich fraction from pomegranate rind (TFPR) against Listeria monocytogenes and its mechanism of action. The tannin-related components of TFPR were analyzed by high-performance liquid chromatography and liquid chromatography-mass spectrometry, and the minimum inhibitory concentration (MIC) of TFPR was determined using the agar dilution method. Extracellular potassium concentration, the release of cell constituents, intra- and extracellular ATP concentrations, membrane potential, and intracellular pH (pHin) were measured to elucidate a possible antibacterial mechanism. Punicalagin (64.2%, g/g) and ellagic acid (3.1%, g/g) were detected in TFPR, and the MICs of TFPR were determined to be 1.25-5.0 mg/mL for different L. monocytogenes strains. Treatment with TFPR induced a decrease of the intracellular ATP concentration, an increase of the extracellular concentrations of potassium and ATP, and the release of cell constituents. A reduction of pHin and cell membrane hyperpolarization were observed after treatment. Electron microscopic observations showed that the cell membrane structures of L. monocytogenes were apparently impaired by TFPR. It is concluded that TFPR could destroy the integrity of the cell membrane of L. monocytogenes, leading to a loss of cell homeostasis. These findings indicate that TFPR has the potential to be used as a food preservative in order to control L. monocytogenes contamination in food and reduce the risk of listeriosis.

Berberine inhibits acute radiation intestinal syndrome in human with abdomen radiotherapy.

Radiation-induced acute intestinal symptoms (RIAISs) are the most relevant complication of abdominal or pelvic radiation. Considering the negative impact of RIAIS on patients' daily activities, the preventive effects of berberine on RIAIS in patients were investigated. Thirty-six patients with seminoma or lymphomas were randomized to receive berberine oral (n = 18) or not (n = 18). Forty-two patients with cervical cancer were randomized to a trial group (n = 21) and control group (n = 21). Radiotherapy used a parallel opposed anterior and posterior. 300-mg berberine was administered orally three times daily in trial groups. Eight patients with RIAIS were treated with 300-mg berberine three times daily from the third to the fifth week. Toxicities, such as fatigue, anorexia/nausea, etc., were graded weekly according to CTC version 2.0. Patients with abdominal/pelvic radiation in the control group showed grade 1 fatigue, anorexia/nausea, colitis, vomiting, proctitis, weight loss, diarrhea and grade 2 anorexia/nausea, fatigue. Only grade 1 colitis, anorexia/nausea, and fatigue were seen in patients of abdominal radiation treated with berberine. Grade 1 fatigue, colitis, anorexia/nausea, and proctitis occurred in patients of pelvic radiotherapy treated with berberine. Pretreatment with berberine significantly decreased the incidence and severity of RIAIS in patients with abdominal/pelvic radiotherapy when compared with the patients of the control group (P < 0.05). RIAIS were reduced in patients with abdominal radiotherapy/pelvic radiation after receiving berberine treatment. Berberine significantly reduced the incidence and severity of RIAIS and postponed the occurrence of RIAIS in patients with abdominal or whole pelvic radiation.

[Effects of Yunnan Baiyao on peri-operative bleeding of patients undergoing cervical open-door laminoplasty: a multi-center randomized double-blind placebo-control trial].

OBJECTIVE: To investigate the effects of Yunnan Baiyao on peri-operative bleeding of the patients undergoing cervical open-door laminoplasty. METHODS: 197 patients undergoing cervical open-door laminoplasty were randomly divided into 2 groups Yunnan Baiyao group (receiving Yunnan Baiyao 500 mg three times daily for 5 days) and placebo group. The amounts of intra-operative and post-operative bleeding were compared, and the side effect of Yunnan Baiyao was also analyzed. RESULTS: Statistical analysis showed that the amount of intra-operative bleeding of the Yunnan Baiyao was 350 ml +/- 190 ml, significantly lower than that of the placebo group (443 ml +/- 266 ml, P < 0.05). There were no statistical differences in the amount of postoperative bleeding and side effect rate between the two groups. CONCLUSION: Yunnan Baiyao is effective and safe in reducing the amount of intra-operative bleeding of cervical open-door laminoplasty.