Pharmacokinetics and antioxidant activity of dihydrocaffeic acid grafted chitosan nanomicelles loaded with chicoric acid in broilers.

Chicoric acid (CA) is a natural nutrient found in plants, showcasing diverse biological activities, including anti-inflammatory and antioxidant properties. Despite its valuable properties, CA faces limitations in bioavailability and susceptibility to oxidative breakdown during utilization. Previous research introduced synthesized dihydrocaffeic acid grafted chitosan self-assembled nanomicelles (DA-g-CS), demonstrating its potential to enhance CA absorption. This study aims to investigate the pharmacokinetics, tissue distribution, and antioxidant activity of both CA and DA-g-CS loaded CA (DA-g-CS/CA) in broilers. An IPEC-J2 cell model was established and evaluated to delve deeper into the transport mechanism and antioxidant potential. The in vivo pharmacokinetic analysis in broilers highlighted a substantial difference: the maximum plasma concentration (Cmax) of DA-g-CS/CA exceeded CA by 2.6-fold, yielding a notable increased relative bioavailability to 214%. This evidence underscores the significant enhancement in CA's oral absorption, facilitated by DA-g-CS. The collective evaluation outcomes affirm the successful development of the cell model, indicating its suitability for drug transporter experiments. The findings from the intestinal transit analysis revealed that both CA and DA-g-CS/CA underwent passive entry into IPEC-J2 cells. Notably, the cellular uptake rate of DA-g-CS loaded with CA was significantly amplified, reaching 2.1 times higher than that of CA alone. Intracellular transport mechanisms involved microtubules, lysosomes, and the endoplasmic reticulum, with an additional pathway involving the endoplasmic reticulum observed specifically for DA-g-CS/CA, distinguishing it from CA. Moreover, the results from both in vivo and in vitro antioxidant assessments highlight the potent antioxidant activity of DA-g-CS/CA, showcasing its efficacy in preventing and treating cellular damage induced by oxidative stress. In summary, these findings underscore the significant enhancement of CA's efficacy facilitated by DA-g-CS, establishing a robust theoretical foundation for the prospective application of CA within livestock and poultry farming.

Preparation of caffeic acid grafted chitosan self-assembled micelles to enhance oral bioavailability and antibacterial activity of quercetin.

Quercetin (QR) is a naturally occurring flavonoid organic compound that has poor solubility in water and highly unstable in alkaline conditions, resulting in limited absorption in poultry. Consequently, in our experiment, QR was employed as a model compound, encapsulated within the caffeic acid graft chitosan copolymer (CA-g-CS) self-assembled micelles to enhance its solubility, stability and exhibit a synergistic antibacterial effect. The optimization of the formula was carried out using a combination of single-factor experimentation and the response surface method. The in vitro release rate and stability of CA-g-CS-loaded QR micelles (CA-g-CS/QR) in various pH media were studied and the pharmacokinetics in white feather broiler chickens was evaluated in vivo. Additionally, the antibacterial activity was investigated using Escherichia coliCMCC44102 and Escherichia coli of chicken origin as the test strain. The results showed the optimized formula for the self-assembled micelles were 4 mL water, 0.02 mg/mL graft copolymer, and 1 mg QR, stirring at room temperature. The encapsulation efficiency was 72.09%. The resulting CA-g-CS/QR was uniform in size with an average diameter of 375.6 ± 5.9 nm. The release pattern was consistent with the Ritger-Peppas model. CA-g-CS/QR also significantly improved the stability of QR in alkaline condition. The relative bioavailability of CA-g-CS/QR was found to be 1.67-fold that of the reference drug, indicating a substantial increase in the absorption of QR in the broiler. Compared to the original drug, the antibacterial activity of CA-g-CS/QR was significantly enhanced, as evidenced by a reduction of half in the MIC and MBC values. These results suggest that CA-g-CS/QR improves the bioavailability and antibacterial activity of QR, making it a promising candidate for clinical use.

Dihydrocaffeic acid grafted chitosan self-assembled nanomicelles with enhanced intestinal transport and antioxidant properties of chicoric acid.

Chicoric acid (CA) plays a crucial role as a functional factor within the realm of foods, showcasing a wide array of bioactivities. Nevertheless, its oral bioavailability is significantly limited. To optimize the intestinal absorption and bolster the antioxidant capacity of CA, a water-soluble dihydrocaffeic acid grafted chitosan copolymer (DA-g-CS) was synthesized using a conventional free radicals system, and subsequently utilized for the encapsulation of CA within self-assembled nanomicelles (DA-g-CS/CA). The average particle size of DA-g-CS/CA was 203.3 nm, while the critical micelle concentration was 3.98 × 10-4 mg/mL. Intestinal transport studies revealed that DA-g-CS/CA penetrated cells via the macropinocytosis pathway, exhibiting the cellular uptake rate 1.64 times higher than that of CA. This substantial enhancement in the intestinal transport of CA underscores the significant improvements achieved through DA-g-CS/CA delivery. The pharmacokinetic results demonstrated that DA-g-CS/CA exhibited a remarkable bioavailability 2.24 times that of CA. Furthermore, the antioxidant assessment demonstrated that DA-g-CS/CA exhibited exceptional antioxidant properties in comparison to CA. It demonstrated enhanced protective and mitigating effects in the H2O2-induced oxidative damage model, while also displaying a stronger emphasis on protective effects rather than attenuating effects. These findings aim to establish a solid theoretical foundation for the advancement of CA in terms of its oral absorption and the development of functional food products.

Spatial-temporal Evolution and Influencing Factors of Digital Financial Inclusion: County-level Evidence from China.

The vigorous development of information and communications technology has accelerated reshaping of the financial industry. The COVID-19 pandemic has further catalyzed the demand for digital financial services. Digital financial inclusion relies on information technology to overcome spatial limitations. In this case, the research question is whether it adheres to the spatial laws governing conventional financial activities. This study uses exploratory spatial data analysis and a geographical detector to elucidate the spatiotemporal characteristics and factors influencing digital financial inclusion at the county level in China (Data don't include that of Hong Kong, Macao and Taiwan of China) from 2014 to 2020. The research findings indicate: first, China's county-level digital financial inclusion is generally increasing and exhibits significant spatial autocorrelation. Second, population density, level of traditional financial development, government regulation, and education level are key determinants of China's county-level digital financial inclusion. Third, policies should be differentiated by region to narrow the spatial gap in digital financial inclusion. The results provide a reference for other developing countries on using digital technology to develop financial inclusion.

A strategy for the improvement of the bioavailability and antiosteoporosis activity of BCS IV flavonoid glycosides through the formulation of their lipophilic aglycone into nanocrystals.

Epimedium-derived flavonoid glycosides are widely used for the prevention of osteoporosis, but these compounds generally exhibit poor membrane permeability and oral absorption. To address these limitations, the bioactive lipophilic aglycone icaritin (ICT) was selected and successfully developed into nanocrystals (ICTN) through the antisolvent-precipitation method. After the parameters in the preparation of ICTN were optimized, the morphology, crystallinity, adsorption of the stabilizers on the ICT surface, and the dissolution of the resulting nanocrystals were characterized. The pharmacokinetics in rat and the in vitro antiosteoporosis activity of serum withdrawn after the oral administration of ICTN to rats on mouse osteoblastic cells were evaluated. Consistent with its good performance in stabilizing the ICT nanosuspension, atomic force microscopy showed that hydroxypropyl methylcellulose (HPMC) exhibits better adsorption on the ICT surface compared with other stabilizers. Needle-shaped crystals (∼ 220 nm in diameter) with a high drug loading (∼ 90%) were generated when 0.16 mL of the ICT acetone solution (10 mg/mL) was injected quickly into 2 mL of the HPMC solution (0.02%, w/w) under ultrasonication for 10 s at room temperature. The thermal analysis demonstrated that the majority of the particles are in their crystalline forms, similarly to the unformulated ICT. After oral administration, ICTN exhibited a faster dissolution rate and significantly faster absorption, as supported by the increased AUC0-36h and Cmax and the reduced Tmax of these nanocrystals compared with the raw suspension (p < 0.05). Compared with blank serum, enhanced proliferation and differentiation activities were observed when serum withdrawn after the oral administration of ICTN in rat was incubated with osteoblast MC3T3-E1 cells. The present delivery system could provide a new promising strategy for BCS IV glycoside of flavonoids or other natural products by formulation of their bioactive lipophilic aglycone forms to enhance oral absorption and in vivo bioactivity.

Oral absorption and excretion of icaritin, an aglycone and also active metabolite of prenylflavonoids from the Chinese medicine Herba Epimedii in rats.

Icaritin (ICT) is a main aglycone and also active intestinal metabolite of prenylflavonoids from the Chinese medicine Herba Epimedii. In the present study, the oral absorption and excretion of this compound was investigated using rats for exploring its fate in the body, so as to better understanding its in vivo pharmacological activities. The free (parent) and total (parent plus conjugated metabolites) ICT concentrations in rat plasma, urine and bile, after intravenous (i.v.) and oral administration both at 5mg/kg, were determined before and after enzymatic hydrolysis with ß-glucuronidase/sulphatase, respectively, by a HPLC-UV method. The results showed that free ICT plasma concentration after i.v. dose was rapidly decreased with average t(1/2, λ) of 0.43 h, while the total ICT concentration was decreased slowly with t(1/2, λ) of 6.86 h. The area under the curve of ICT conjugated metabolites was about 11-fold higher than that of free ICT. The majority of ICT in the body was excreted from the bile with 68.05% of dose over 8 h after i.v. dosing, in which only 0.15% was in parent form. While very little amount of ICT was excreted from the urine with 3.01% of dose over 24 h, in which the parent form was 0.62%. After oral administration, very little amount of parent ICT was detected only in 0.5, 1 or 2 h plasma samples with the concentration less than LOQ, however, its total plasma concentration after enzymatic hydrolysis treatment was at relative high level with average maximum concentration of 0.49 µg/ml achieved at 1h post dose. The oral bioavailability of ICT was 35% of dose, estimated by its total plasma drug concentrations. It is concluded that ICT can be easily absorbed into the body, and then rapidly conversed to its conjugated metabolites, and finally removed from the body mainly by biliary excretion.

An LC-MS/MS method for determination of forsythiaside in rat plasma and application to a pharmacokinetic study.

A highly sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of forsythiaside in rat plasma using epicatechin as internal standard. The analytes were extracted by solid-phase extraction and chromatographied on a C(18) column eluted with a gradient mobile phase of acetonitrile and water both containing 0.2% formic acid. The detection was performed by negative ion electrospray ionization in multiple reaction monitoring mode, monitoring the transitions m/z 623-->161 and m/z 289-->109 for forsythiaside and epicatechin, respectively. The assay was linear over the concentration ranges of 2.0-50.0 and 50.0-5000.0ng/mL with limits of detection and quantification of 0.2 and 1.0ng/mL, respectively. The precision was <10.8% and the accuracy was >91.9%, and extraction recovery ranged from 81.3% to 85.0%. This method was successfully applied to a pharmacokinetic study of forsythiaside in rats after intravenous (20mg/kg) and oral (100mg/kg) administration, and the result showed that the compound was poorly absorbed with an absolute bioavailability being approximately 0.5%.

An LC-MS/MS method for determination of 3,6'-disinapoylsucrose in rat plasma and its application to a pharmacokinetic study.

3,6'-Disinapoylsucrose (DSS), a major active component of traditional Chinese medicine Yuan-Zhi (the roots of Polygala tenuifolia), has significant effects for neuroprotection and improving learning memory. In order to explore the pharmacokinetic properties of DSS so as to further understand its in vivo activities, a sensitive LC-MS/MS method was developed for determination of DSS in rat plasma and applied to a pharmacokinetic study in the present study. After treatment by protein precipitation, the plasma sample was separated on a C(18) HPLC column and analyzed by a mass spectrometry under positive electrospray ionization. Multiple-reaction monitoring was employed to measure the ion transition at m/z 777.4 --> 409.2 for DSS and m/z 557.2 --> 309.1 for forsythin as internal standard. The method was linear over the studied concentration range of 0.5-1000.0 ng/mL. The precision and accuracy ranged from 1.4 to 18.4%, and from -3.7 to -9.5%, respectively, for within-day and between-day assay. Extraction recovery was higher than 86.6%. The limits of detection and quantification were 0.3 and 0.5 ng/mL, respectively. The present method was successfully applied to a pharmacokinetic study. DSS was found to have poor oral absorption with only about 0.5% bioavailability.