Preparation and characterization of calcium carbonate microspheres and their potential application as drug carriers.

The influence of lecithin from egg yolk (LE) on calcium carbonate (CaCO3) biomineralization was investigated. In the present study, spherical CaCO3 particles were synthesized via coprecipitation in the presence of LE. LE multilamellar liposomes were first tuned by sonication to provide better control over the nucleation of CaCO3. Subsequently, monodisperse microspheres ~2 µm in size were generated by controlling the aggregation and growth of CaCO3 under appropriate concentrations of LE. In contrast to unstable vaterite, the microspheres generated in aqueous solution remained stable for at least 10 days without transforming into calcite, due to the strong interaction between the LE and calcium ions. The microspheres as drug carriers of doxorubicin (DOX) were assessed and were observed to have a good encapsulation efficiency, sustained drug release without a burst release and notable pH sensitivity. In addition, in vivo tumor inhibition examination demonstrated that DOX­loaded CaCO3 microspheres formulation had more superior efficacy to significantly restrain tumor growth. These novel LE/CaCO3 hybrids may provide novel options for various biomedical applications.

Metabonomic study on the plasma of streptozotocin-induced diabetic rats treated with Ge Gen Qin Lian Decoction by ultra high performance liquid chromatography-mass spectrometry.

Changes in endogenous metabolites in the plasma of streptozotocin (STZ)-induced diabetic rats treated with Ge Gen Qin Lian Decoction (GGQLD) were studied. The endogenous compounds in plasma were detected using ultra high performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS). Rats were divided into three groups: control, model, and administration (4.95g crude drug/kg body weight). After the final administration, plasma samples from the three groups were analyzed using metabonomics. The three sample groups could be clearly distinguished. The administration group exhibited a distinct return to the levels of phytosphingosine and dihydrosphingosine of the control group according to the principal component analysis score, and the corresponding biomarkers were defined. Significant changes in endogenous metabolites, such as dihydrosphingosine, phytosphingosine, cholylglycine, and pantothenic acid, were identified in STZ-induced diabetic rats. These biochemical changes are associated with the metabolism of sphingolipids, fats, and acetyl coenzyme-A, which could be useful to further investigate the characteristics of STZ-induced diabetes mellitus and the therapeutic mechanism of action of GGQLD. This metabonomic analysis could provide a useful starting point toelucidate the therapeutic effects and mechanism of action of GGQLD in diabetes mellitus.

Novel B,O-chelated fluorescent probe for nitric oxide imaging in Raw 264.7 macrophages and onion tissues.

A novel fluorescent probe based on B,O-chelated dipyrromethene chromophore in far-visible and near-infrared spectral region (600-900 nm), boron chelated 8-(3,4-diaminophenyl)-3,5-bis(2-hydroxyphenyl)-4-bora-3a,4a-diaza-s-indancene (BOPB), has been first developed for nitric oxide (NO) imaging. BOPB, a turn-on fluorescent probe, can react with NO rapidly under physiological condition. The reaction product of BOPB with NO, BOPB-T, emits bright red fluorescence at 643 nm when excited at 622 nm. Meanwhile, BOPB-T displays high fluorescent quantum yield of 0.21 and good photostability. The selectivity for NO over other reactive oxygen/nitrogen species and ascorbic acid has been investigated and BOPB has good specificity for the detection of NO. MTT assay shows that the toxicity of BOPB (below 10 µM) to living cells can be neglected. Based on these investigations, BOPB has been used for NO imaging in Raw 264.7 cells and onion tissues. Meanwhile, mechanical injury to onion tissues results in a brighter fluorescence around the wound, which indicates that more NO has been produced in plant tissues in response to external stimuli. Our studies illustrate that BOPB has advantages of high sensitivity, low background interference and little photo damage on fluorescence imaging of NO.

Determination of fatty acids in bio-samples based on the pre-column fluorescence derivatization with 1,3,5,7-tetramethyl-8-butyrethylenediamine-difluoroboradiaza-s-indacene by high performance liquid chromatography.

1,3,5,7-Tetramethyl-8-butyrethylenediamine-difluoroboradiaza-s-indacene (TMBB-EDAN) has been designed and synthesized as a highly fluorescent labeling reagent for carboxylic acids. By using TMBB-EDAN, a sensitive and rapid method based on high performance liquid chromatography-fluorescence detection for the determination of twelve fatty acids (FAs) in bio-samples has been developed. Under optimized conditions, these FAs were tagged with TMBB-EDAN in the presence of 1-ethyl-3-(3-dimethyla-minopropyl) carbodiamide at 20°C for 30min and then the baseline separation was achieved on a C18 column with a linear gradient elution in 26min. With fluorescence detection at λex/λem=490nm/510nm, the linear ranges of FAs were from 3.0 to 300nM and the detection limits with a signal-to-noise ratio of 3 were in the 0.2-0.4nM range. The proposed method offers advantages of milder derivatization condition and much better sensitivity for the determination of FAs, when compared to the reported fluorescence derivatization-based methods.

Ultrasensitive determination of jasmonic acid in plant tissues using high-performance liquid chromatography with fluorescence detection.

An ultrasensitive and selective high-performance liquid chromatographic method for the volatile signaling hormone, jasmonic acid, has been developed based on precolumn derivatization with 1,3,5,7-tetramethyl-8-aminozide-difluoroboradiaza-s-indacene (BODIPY-aminozide). The derivatization reaction was carried out at 60 °C for 30 min in the presence of phosphoric acid. The formed jasmonic acid derivative was eluted using a mobile phase of methanol/pH 6.50 ammonium formate buffer/tetrahydrofuran (67:30:3, v/v/v) in 10 min on a C(18) column and detected with fluorescence detection at excitation and emission wavelengths of 495 and 505 nm, respectively. The detection limit (signal-to-noise ratio = 4) reached 1.14 × 10(-10) M or 2.29 fmol per injection (20 µL), which is the lowest of the existing methods. The proposed method has been successfully applied to the direct determination of trace jasmonic acid in the crude extracts of soybean leaves from soybean mosaic virus-infected and normal plants with recoveries of 95-104%.

Simultaneous determination of phytohormones containing carboxyl in crude extracts of fruit samples based on chemical derivatization by capillary electrophoresis with laser-induced fluorescence detection.

An efficient and sensitive capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) method has been developed for the simultaneous determination of phytohormones containing carboxyl group, including gibberellic acid, indole-3-acetic acid, abscisic acid, jasmonic acid, indole butyric acid, 1-naphthalene acetic acid and 2,4-dichloro-phenoxy acetic acid, based on the chemical derivatization with 6-oxy-(acetypiperazine) fluorescein. Using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide as the condensing reagent, the derivatization reaction completed at 60°C in 60 min and the derivatization limits could reach 20 nmol L(-1). The formed derivatives of seven phytohormones have been separated and quantified within 20 min. The linearity was found in the range of 0.01-1 µmol L(-1) and the limits of detection were 1.6-6.7 nmol L(-1) (S/N=3). The proposed method has been applied to analyze the crude extract of 0.5 g banana samples directly without further purification and the recoveries varying from 90.7 to 106.1%.

Determination of phytohormones in plant samples based on the precolumn fluorescent derivatization with 1,3,5,7-tetramethyl-8-aminozide-difluoroboradiaza-s-indacene by HPLC for routine use.

Six phytohormones including indole butyric acid (IBA), naphthalene acetic acid (NAA), 2,4-dichloro-phenoxy acetic acid (2,4-D), indole-3-acetic acid (IAA), abscisic acid (ABA), and salicylic acid (SA) in crude plant extractions have been quantitated by means of high-performance liquid chromatography (HPLC) with fluorescence detection based on the precolumn derivatization using 1,3,5,7-tetramethyl-8-aminozide-difluoroboradiaza-s-indacene (BODIPY-aminozide), a fluorescent reagent synthesized in our lab recently. The optimization of derivatization conditions was carefully studied by an L(25) (5(6)) orthogonal array design (OAD) with five factors at five levels that are important influence parameters in the improvement of derivatization efficiency. The separation conditions were also studied in detail. Under the optimal conditions, the detection limits (S/N=3) of the six phytohormones were found from 0.12 to 0.75 nM. The proposed method was the first investigation of aminozide for the analysis of phytohormones and has been successfully applied to the determination of phytohormones in plant samples such as cucumber and tomato with recoveries of 94-105%.

[Primary study on the hepatotoxicity and nephrotoxicity of rats induced by three kinds of nanomaterials].

OBJECTIVE: To investigate the effects of hepatotoxicity and nephrotoxicity on rats induced by Nano-SiO2 Nano-Fe3O4 and SWCNTs. METHODS: 42 Wistar rats were randomly divided into the control group, low and high dose groups of three kinds of nanoparticles. The rats were killed when rats were exposed by oral gavage for 4 weeks. The ratios of liver and kidney to body weight, the pathological changes of liver and kidney, serum certain biochemically indexes were detected. RESULTS: The ratios of liver and kidney to body weight in experimental groups were not significantly difference in comparison to the control. The ALT and AST level in serum of rats in part experiment groups were more higher than those of the control group. The results of pathological analysis indicated that there were small amounts inflammatory cell infiltration and hepatic cell adipose degeneration in the liver of rats in high-dose group, the change of kidney were not found. CONCLUSION: Three kinds of nanoparticles could induce hepatotoxicity of the rats in the condition of this experiment.