Synthesis of cationic ß-cyclodextrin functionalized silver nanoparticles and their drug-loading applications.

Silver nanoparticles have attracted great attention owing to their distinct physicochemical properties, which inspire the development of their synthesis methodology and their potential biomedical applications. In this study, a novel cationic ß-cyclodextrin (C-ß-CD) containing a quaternary ammonium group and amino group was applied as a reducing agent as well as a stabilizing agent to prepare C-ß-CD modified silver nanoparticles (CßCD-AgNPs). Besides, based on the inclusion complexation between drug molecules and C-ß-CD, the application of CßCD-AgNPs in drug loading was explored by the inclusion interaction with thymol. The formation of AgNPs was confirmed by ultraviolet-visible spectroscopy (UV-vis) and X-ray diffraction spectroscopy (XRD). Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed the prepared CßCD-AgNPs were well dispersed with particle sizes between 3-13 nm, and the zeta potential measurement result suggested that the C-ß-CD played a role in preventing their aggregation in solution. 1H Nuclear magnetic resonance spectroscopy (1H-NMR) and Fourier transform infrared spectroscopy (FT-IR) revealed the encapsulation and reduction of AgNPs by C-ß-CD. The drug-loading action of CßCD-AgNPs was demonstrated by UV-vis and headspace solid-phase microextraction gas chromatography mass spectrometry (HS-SPME-GC-MS), and the results of TEM images showed the size increase of nanoparticles after drug loading.

Screening and identification of tyrosinase inhibitors in edible plant materials by on-line UPLC-enzyme reactor coupled with UHPLC-FTMS.

Tyrosinase plays a primary role in melanin biosynthesis and enzymatic browning of freshly cut fruits and vegetables. Herein, an on-line ultraperformance liquid chromatography diode array detector biochemical detection (UPLC-DAD-BCD) method was established to identify trace amount potent tyrosinase inhibitors and antibrowning agents in complex mixtures. The tyrosinase inhibition activities of some representative compounds were evaluated by using the established method and their chromatography-activity relationships were obtained. Then the proposed UPLC-DAD-BCD method was applied to screen tyrosinase inhibitors in edible herbal extracts and identified two tyrosinase inhibitors in green tea and three in cinnamon. The above active ingredients were determined by ultra-high-performance liquid chromatography linear ion trap/orbitrap high resolution mass spectrometry (UHPLC-FTMS). The on-line UPLC-DAD-BCD in combination with UHPLC-FTMS was confirmed to be a powerful technique to screen and elucidate the active ingredients in complex matrixes and could be applied to evaluate the integrated effects of multiple ingredients against corresponding targets.

Pharmacokinetic and tissue distribution study of eight volatile constituents in rats orally administrated with the essential oil of Artemisiae argyi Folium by GC-MS/MS.

Artemisia argyi is commonly used as a remedy for gynecological and respiratory disease in traditional Chinese medicine. The essential oil is considered as the major active ingredients of A. argyi, mainly composed of eucalyptol, α-thujone, camphor, borneol, bornyl acetate, eugenol, ß-caryophyllene, and caryophyllene oxide, while limited study addresses the in vivo disposition of these volatile ingredients. In present study, a rapid, sensitive and selective GC-MS/MS method has been developed and validated for the quantification of the eight volatile constituents in rat plasma and tissues after orally dosing with the essential oil of Artemisiae Argyi Folium (AAEO) using naphthalene as an internal standard (IS). The analytes were extracted from biosamples by liquid-liquid extraction with hexane/ethyl acetate. The GC separation was achieved on a TG-5SILMS column (30 m × 0.25 mm, 0.25 µm film thickness) and MS detection was performed on selective reaction monitoring (SRM) mode. The assay had a lower limit of quantification (LLOQ) less than 2 ng/ml for the analytes with good linearity (r ≥ 0.9907). Their disposition profile in rat plasma and tissues was characterized after orally giving AAEO, and the data revealed the analytes underwent rapid absorption from GI tract and were mainly transferred to the liver, heart, kidney, lung, and spleen with prompt elimination. The results provided a meaningful basis for guiding the pharmacodynamic study and clinical applications of this herbal medicine.

Huganbuzure Granule Attenuates Concanavalin-A-Induced Immune Liver Injury in Mice via Regulating the Balance of Th1/Th2/Th17/Treg Cells and Inhibiting Apoptosis.

In Uygur medicine, Huganbuzure granule (HBG) is one of the classical prescriptions for liver protection. However, its role in immune liver injury remains unknown. This study evaluates the effect of HBG on concanavalin-A- (ConA-) induced immune liver injury and investigates its protective underlying mechanism. BALB/c mice were randomly divided into five groups (n = 24 mice per group): control, ConA, 1.6 g/kg HBG + ConA, 3.2 g/kg HBG + ConA, and 6 mg/kg prednisolone + ConA. HBG was intragastrically administrated once daily for ten consecutive days, prior to ConA (20 mg/kg) injection. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), superoxide dismutase (SOD), and malondialdehyde (MDA) in mouse serum were measured after ConA injection. Moreover, liver-related mRNA levels were evaluated by qPCR. The detection of liver-related proteins was assessed by immunohistochemistry and western blot analysis. Compared with the ConA group, HBG reduced the mRNA expression of IL-17A and IFN-γ and the protein expression of T-bet and ROR-γt. In addition, HBG increased the mRNA expression of IL-4 and TGF-ß and protein expression of GATA3 and Foxp3, indicating that HBG regulated the balance of Th1/Th2 and Th17/Treg. Furthermore, HBG alleviated immune liver injury by reducing oxidative stress, inhibiting apoptosis, and decreasing the expression of p-JNK, p-ERK, p-p38, p-JAK1, p-STAT1, p-STAT3, and IRF1. Our data suggested that HBG attenuated ConA-induced immune liver injury by regulating the immune balance and inhibiting JAK1/STATs/IRF1 signaling, thereby reducing apoptosis induced by JNK activation. The findings indicate that HBG may be a promising drug for immune liver injury.

Simultaneous determination of phenolic acids and flavonoids in Artemisiae Argyi Folium by HPLC-MS/MS and discovery of antioxidant ingredients based on relevance analysis.

A rapid and reliable LC-MS/MS method was developed for the quantitation of major components in Folium Artemisiae Argyi (mugwort), a widely used traditional Chinese herbal medicine. A total of 5 phenolic acids and 17 flavonoids were separated and simultaneously determined by using a Shiseido C18 column (150 × 3.0 mm, 3 µm) and gradient elution of acetonitrile-aqueous formic acid (100:0.1, v/v) at a 0.5 mL min-1 flow rate, via multiple reaction monitor (MRM) in polarity switching mode. The quantitative method was validated in terms of sensitivity, linearity, precision, accuracy and stability, which proved to be sensitive, accurate and reproducible. Then 65 samples collected from different areas were selected for component analysis by LC-MS/MS and assessment of antioxidant activity using DPPH, ABTS, FRAP, O2- and OH scavenging assays. Grey relational analysis and partial least square regression were used to evaluate the relevance between chemicals and bioactivities, and the results indicated chlorogenic acid, isochlorogenic acid B, A, C, eriodictyol, jaceosidin and eupatilin made the key contribution to antioxidant activity. The present study combines chemical analysis and bioassay to identify bioactive markers, which possesses potential value for the activity-oriented quality control of mugwort.

Network Pharmacology Deciphering Mechanisms of Volatiles of Wendan Granule for the Treatment of Alzheimer's Disease.

OBJECTIVE: To explore the mechanisms of the volatiles of Wendan granule (WDG) for the treatment of Alzheimer's disease, network pharmacology method integrating absorption, distribution, metabolism, and excretion (ADME) screening, target fishing, network constructing, pathway analysing, and correlated diseases prediction was applied. METHODS: Twelve small molecular compounds of WDG were selected as the objects from 74 volatiles with the relative abundances above 2 %, and their ADME parameters were collected from Traditional Chinese Medicine Systems Pharmacology platform (TCMSP), and the corresponding targets, genes, pathways, and diseases were predicted according to the data provided by TCMSP, DrugBank, Uniport, and the Database for Annotation, Visualization, and Integrated Discovery (DAVID). Then the related pathways and correlation analysis were explored by the Kyoto Encyclopedia and Genomes (KEGG) database. Finally, the networks of compound target, target pathway, and pathway disease of WDG were constructed by Cytoscape software. RESULTS: Twelve compounds interacted with 49 targets, of which top three targets were gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1), prostaglandin G/H synthase 2 (PGHS-2), and sodium-dependent noradrenaline transporter. Interestingly, these targets were highly associated with depression, insomnia, and Alzheimer's disease that mainly corresponded to mental and emotional illnesses. CONCLUSION: The integrated network pharmacology method provides precise probe to illuminate the molecular mechanisms of the main volatiles of WDG for relieving senile dementia related syndromes, which will also facilitate the application of traditional Chinese medicine as an alternative or supplementary to conventional treatments of AD, as well as follow-up studies such as upgrading the quality standard of clinically applied herbal medicine and novel drug development.

Effect of Poria cocos hydroethanolic extract on treating adriamycin-induced rat model of nephrotic syndrome.

OBJECTIVE: To evaluate the effect of Poria cocos (Schw.) Wolf hydroethanolic extract (PHE) against nephrotic syndrome (NS) in rats and to identify the potential active components from PHE. METHODS: The high content compounds were isolated and purified by using column chromatography followed by preparative highperformance liquid chromatography (p-HPLC). Forty male Wistar rats with adriamycin (ADR)-induced NS were randomly divided into 5 groups, 8 in each group: model control group, positive control group (with prednisone treatment), PHE low-dose group, PHE middle-dose group and PHE high-dose group. Another 8 rats were recruited as vehicle control group. All rats received the intragastric administration of corresponding drugs or saline for 30 days. During the experimental period, rats' behavior and appearance were observed and recorded daily, and their body weights were recorded weekly. After treatment, 24-h urine samples were collected to evaluate the urine protein and urine creatinine (Ucr); then the rats were sacrificed to collect carotid blood and to determine the levels of serum total protein (TP), albumin (Alb), globulin (Glo), total cholesterol (TC) and cytokine interlukin-4 (IL-4). RESULTS: Six acidic components were isolated and identified from the PHE section: pachymic acid, 15α-hydroxydehydrotumulosic acid, trametenolic acid, dehydropachymic acid, 3ß-hydroxy-lanosta-7,9(11), 24-trien-21-oic-acid and dehydroeburicoic acid. Compared with the model control group, the urine protein content were significantly decreased in the PHE treatment groups and positive control group (P<0.05), especially PHE middle-dose group (P<0.01). The Ucr values and serum levels of TP, Glo, TC and IL-4 in PHE low- and middle-dose groups were also presented obvious recover tendency as compared with the model control group (P<0.05 or P<0.01). However, positive control group and all PHE groups indicated no significant therapeutic effect on raising Alb value, although PHE low- and middle-dose treatment groups showed better outcomes than positive control group (P>0.05). CONCLUSIONS: PHE showed an encouraging therapeutic effect against ADR-induced NS in a rat model. PHE might be a group of effective substances for the treatment of NS.

A comparison of lateral fixation versus dual plating for simple bicondylar fractures.

BACKGROUND: The best fixation method for bicondylar tibial plateau fracture is debated. The aim of this study was to compare the effect of a lateral locking plate with that of dual plates in the treatment of tibial plateau fracture with a relatively intact medial condyle fragment. METHODS: Eighty-six patients diagnosed with a bicondylar tibial plateau fracture with a relatively intact medial condyle were assigned to groups treated with either dual buttress plates (DP group) or a lateral locking plate (LP group). Preoperative characteristics, surgical details, and postoperative complications were recorded and compared between groups. Hospital for Special Surgery Knee Scoring System score was used to evaluate clinical outcome. RESULTS: Both groups displayed satisfactory clinical and radiographic results. There were no significant differences in associated preoperative conditions between the DP and LP groups. Durations of hospital stay and operation were significantly shorter, and blood loss significantly less, in the LP group than in the DP group (P=0.045;0.038;0.031). The rate of delayed-union was significantly lower in the LP group than in the DP group (P=0.023). CONCLUSION: Both dual-plate and lateral locking-plate fixation can provide satisfactory treatment of bicondylar tibial plateau fractures if properly used. When the medial tibial condyle is relatively intact, a lateral locking plate can provide stability similar to that of dual plate while decreasing operative duration and soft-tissue complications, which can assist fracture healing and shorten hospital stay. LEVEL OF EVIDENCE: I - Randomized controlled trial.

Functional outcomes of bicondylar tibial plateau fractures treated with dual buttress plates and risk factors: a case series.

OBJECTIVE: Although there are different ways to treat bicondylar tibial plateau fractures, treatment remains as a challenge to most orthopaedic surgeons. The purpose of this study was to assess the outcomes of dual buttress plates fixation in treatment of bicondylar tibial plateau fractures and to determine risk factors for outcome. METHODS: Clinical data for 74 patients with bicondylar tibial plateau fracture were retrospectively reviewed. Dual buttress plates were placed through two incisions. All patients were evaluated both radiographically and clinically according to Rasmussen scores and Hospital for Special Surgery (HSS) scores during the follow-up period. The data were analyzed using univariate analysis and multivariate logistic regression analysis to identify independent risk factors for functional outcome (Rasmussen functional score). RESULTS: Rasmussen anatomic score was excellent in 34, good in 23, and fair in 17 patients. Sixty-nine patients were followed for a mean of 27 months. Functional results of excellent and good were obtained in 56(81.2%) patients by Rasmussen functional score and in 57 (82.6%) patients by HSS score. Univariate analysis showed Arbeitsgemeinschaft für Osteosynthesefragen (AO) fracture type and reduction quality were significantly related to functional results. Multivariate analysis showed that AO fracture type and reduction quality were independent risk factors for outcome. CONCLUSION: Dual buttress plates technique provided a favourable quality of fracture reduction and satisfactory clinical function. Appropriate selection for operation chance, limited soft tissue stripping and filling of osseous defects may lessen or prevent serious complications. Fracture reduction quality and AO fracture type were independent risk factors for clinical function.

Headspace, solid-phase micro-extraction, gas chromatographic-mass spectrometric analysis of terpenoids in the latex of Euphorbia species.

The volatile and semi-volatile terpenoids in the latex of Euphorbia amygdaloides, E. exigua, E. helioscopia, and E. peplus were analyzed by headspace, solid-phase micro-extraction (HS-SPME), coupled with gas chromatography-mass spectrometry. The volatiles were extracted using a 100 microm polydimethylsiloxane SPME fiber under optimized extraction conditions. The compounds detected encompassed a range of chemical classes, but only terpenoids were evaluated. Only sesquiterpene hydrocarbons were detected in the tested samples of E. exigua, E. helioscopia, and E. peplus, with beta-caryophyllene being the major one, but were never recorded in latex samples of E. amygdaloides, in which only the diterpene hydrocarbon kaur-16-ene was detected. Alpha-Humulene was consistently found in samples of E. helioscopia, and E. peplus, but never in those of the other two species. These preliminary results show that the developed procedure is suitable for the analysis of small samples of Euphorbia latex and that, for each individual species, there is very little qualitative difference between samples, regardless of either place or date of collection.