Detection of the Adulteration of Dendrobium Huoshanense with Dendrobium Henanense by UV-Vis-Shortwave Near-Infrared Diffuse Reflectance Spectroscopy Combined with Chemometrics.

BACKGROUND: Dendrobium huoshanense (DHS) is a classic traditional Chinese medicine (TCM) with distinctive medicinal benefits and great economic worth; nevertheless, because of similar tastes and looks, it is simple to adulterate with less expensive substitutes (such as Dendrobium henanense [DHN]). OBJECTIVE: This work aimed to develop a reliable tool to detect and quantify the adulteration of DHS with DHN by using UV-Vis-shortwave near-infrared diffuse reflectance spectroscopy (UV-Vis-SWNIR DRS) combined with chemometrics. METHODS: Adulterated samples prepared in varying concentrations (0-100%, w/w) were analyzed with UV-Vis-SWNIR DRS methods. Partial least-square-discriminant analysis (PLS-DA) and partial least-squares (PLS) regression techniques were used for the differentiation of adulterated DHN from pure DHS and the prediction of adulteration levels. RESULTS: The PLS-DA classification models successfully differentiated adulterated and nonadulterated DHS with an over 100% correct classification rate. UV-Vis-SWNIR DRS data were also successfully used to predict adulteration levels with a high coefficient of determination for calibration (0.9924) and prediction (0.9906) models and low error values for calibration (3.863%) and prediction (5.067%). CONCLUSION: UV-Vis-SWNIR DRS, as a fast and environmentally friendly tool, has great potential for both the identification and quantification of adulteration practices involving herbal medicines and foods. HIGHLIGHTS: UV-Vis-SWNIR DRS combined with chemometrics can be applied to identify and quantify the adulteration of herbal medicines and foods.

Development of a joint derivatization protocol for the unequivocal identification of the monosaccharide composition in four dendrobium polysaccharides and free monosaccharide by GC-MS.

The determination of monosaccharides is crucial for studying the structure of polysaccharides and the composition of free monosaccharides in living organisms. Based on previous derivatization gas chromatography-mass spectrometry (GC-MS) methods, we aimed to develop a novel analytical protocol for better quantifying monosaccharides. In this study, sugar alcohol acetylation, saccharonitrile acetylation, silylation and a combination of sugar alcohols acetylation and saccharonitrile acetylation were compared. The optimal method was verified with the monosaccharide determination of four polysaccharides and four free monosaccharides from Dendrobium. The results showed that the novel combined derivatization method was superior to the other three methods in terms of content analysis of monosaccharides. Furthermore, it possessed good linearity (all calibration curves showed relative coefficients ≥ 0.999), sensitivity, precision (relative standard deviation < 2%), and accuracy (recovery, 95.7-105%). Finally, the novel method established in this study was successfully employed in determining the monosaccharide composition of four polysaccharides and four free monosaccharide samples from Dendrobium.

Simultaneous Quantification of 11 Phenolic Compounds and Consistency Evaluation in Four Dendrobium Species Used as Ingredients of the Traditional Chinese Medicine Shihu.

The interchangeable use of different herbs to prepare the same formulation is a common practice in Traditional Chinese Medicine (TCM). However, this practice would require the component herbs to share similar compositions, at least in terms of the bioactive agents, to ensure they can replace each other in drug preparation. In this study, we developed an effective and comprehensive high-performance liquid chromatography-diode array detector (HPLC-DAD) method for simultaneous analysis of 11 phenolic compounds in the methanol extracts of Dendrobium huoshanense, Dendrobium nobile (D. nobile), Dendrobium chrysotoxum (D. chrysotoxum), and Dendrobium fimbriatum (D. fimbriatum), which have been identified as interchangeable ingredients for the same TCM preparation "Shihu" in the Chinese pharmacopeia (ChP). The consistency of the four Dendrobium species was evaluated on the basis of the presence of the 11 investigated compounds and the HPLC fingerprints of the methanol extracts of the plants. When gradient elution was performed with a solvent system of acetonitrile and water on a Zorbax Eclipse XDB-C18 (150 mm × 4.6 mm, 5 µm) with monitoring at 220 nm, all 11 investigated compounds were isolated at the baseline. The established HPLC method showed excellent linearity (all analytical curves showed relative coefficients [R2] > 0.999), sensitivity, precision (relative standard deviation [RSD] < 2%), and accuracy (recovery, 90.65-99.17%). These findings confirmed that the method we constructed was reliable. Quantification analysis showed significant differences in the contents of the investigated polyphenols in the four Dendrobium species. Evaluations of consistency revealed that the similarities among the four species were 0.299-0.906 in assessments based on the 11 polyphenols and 0.685-0.968 in assessments based on HPLC fingerprints. Thus, the components of the four Dendrobium species may be significantly different, and more experiments are required to determine whether they can be used interchangeably in the same amounts for preparing the formulation according to ChP.

Simultaneous analysis of 20 free amino acids by a single marker combined with an HPLC fingerprint evaluation of Dendrobium huoshanense.

A phenylhexyl isothiocyanate (PITC) precolumn derivatization quantitative analysis of multicomponents by a single marker (QAMS) strategy for the simultaneous analysis of 20 free amino acids (FAA) in Dendrobium huoshanense is proposed. The method was validated by the linearity, limit of detection (LDO), and limit of quantitation (LOQ), recovery, precision, and stability. The results showed that when applying the established method, the LOQ of the FFAs was lower than 1 ng/ml except threonine (1.32 ng) and cysteine (1.16 ng). The QAMS investigation revealed that, using any one of the 20 FAAs as the reference internal standard, no significant differences were observed between the external standard method and the QAMS method for the quantification of FAAs in D. huoshanense by PITC precolumn derivatization [The relative standard deviation (RSD, %) by QAMS and ESM were all below 5%]. HPLC fingerprint investigation combined with similar analysis (the similarity values for S1-S25 were >0.875) and quality fluctuation analysis showed that the cultivation environment might have a great effect on the accumulation of FAAs in D. huoshanense. Overall, our study showed that we might increase the accuracy and scope of the simultaneous quantification of multicomponents using the QAMS technique by being derivatized with a strong UV absorbing group, and QAMS combined with chromatographic fingerprinting can be considered good quality criteria for the quality control of D. huoshanense and may provide analytical technical support for research on Maillard Reaction during the further processing of D. huoshanense.

Combination and efficiency: preparation of dissolving microneedles array loaded with two active ingredients and its anti-pigmentation effects on guinea pigs.

Hyperpigmentation is a common skin disorder caused by excessive melanogenesis and uneven dispersion of melanin in the skin. To combine multiple active agents with an efficient transdermal drug delivery system is an effective strategy to combat UV induced skin pigmentation. In this work, Arbutin (Arb) and Vitamin C (Vc) mixed in 1:1 were found to have the greatest inhibition effects on melanogenesis and tyrosinase activity in B16 murine melanoma cells. And hyaluronic acid (HA) based dissolving microneedles array (DMNA) was employed to overcome the skin barriers for improved topical drug delivery, which exhibited the most desirable features, including morphology, mechanical properties, dissolving ability, and the highest drug loading. Furthermore, DMNA could greatly increase the stability of Vc during storage without adding any antioxidant which is an important issue for Vc administration. Pharmacodynamics study showed that DMNA loaded with Arb and Vc could synergistically suppress UVB-induced hyperpigmentation in guinea pig skin. This work provides a promising treatment strategy and solution for skin pigmentation and other skin problems.

A flavone, Wogonin from Scutellaria baicalensis inhibits the proliferation of human colorectal cancer cells by inducing of autophagy, apoptosis and G2/M cell cycle arrest via modulating the PI3K/AKT and STAT3 signalling pathways.

PURPOSE: The purpose of this study was to examine the anticancer effects of a flavone from Scutellaria baicalensis wogonin against a panel of colorectal cancer cells. METHODS: The SW1417, SW48, DLD-1, HCT-15, LS-180 and CCD-18Co cell lines were used for the evaluation of the anticancer effects of wogonin. WST-1 and colony formation assays were used for cell viability assessment. Cell cycle analysis was assessed by flow cytometry. Autophagy was detected by electron microscopy. Apoptosis was detected by acridine orange (AO)/ethidium bromide (EB) staining. Cell protein expression was checked by western blotting. RESULTS: The cytotoxic effects of wogonin were comparatively negligible against the normal CCD-18Co cells with an IC50 of >100 µM. Investigation of the mechanism of action revealed that wogonin exerts growth inhibitory effects on the SW48 colorectal cancer cells by autophagic and apoptotic cell death. This was also accompanied with upregulation of autophagic proteins such as LC3II and Beclin 1 as well as the apoptotic proteins such as caspase 3, 8 and 9 and Bax expressions. Wogonin also induced arrest of the SW48 cells at the G2/M check point of the cell cycle. In addition, wogonin could also inhibit the PI3K/AKT and STAT3 signal transduction pathways. CONCLUSION: These results suggest that wogonin exerts potent anticancer effects on colorectal cancer cells and may prove essential in the management of colorectal cancer.

[Modern research and application of Longgu].

Longgu is the fossil of ancient mammals which was used as a common kind of mineral medicine. Longgu is always used to treat neurological diseases. Currently, the quality standard of Longgu is incomplete. Moreover, because of the non-renewable nature of the resource and the increase of national protection of fossils, the clinical application of Longgu is facing a series of problems. As the discovery of the ingredient and the development of forging technology researchers launched to search the substitutes of Longgu. The article summarizes the usage and the study of Longgu, in order that we can discuss the modern usage and substitutability of Longgu.

Significantly inhibitory effects of low molecular weight heparin (Fraxiparine) on the motility of lung cancer cells and its related mechanism.

Low molecular weight heparin (LMWH) improving the cancer survival has been attracting attention for many years. Our previous study found that LMWH (Fraxiparine) strongly downregulated the invasive, migratory, and adhesive ability of human lung adenocarcinoma A549 cells. Here, we aimed to further identify the antitumor effects and possible mechanisms of Fraxiparine on A549 cells and human highly metastatic lung cancer 95D cells. The ability of cell invasion, migration, and adhesion were measured by Transwell, Millicell, and MTT assays. FITC-labeled phalloidin was used to detect F-actin bundles in cells. Chemotactic migration was analyzed in a modified Transwell assay. Measurement of protein expression and phosphorylation activity of PI3K, Akt, and mTOR was performed with Western blot. Our studies found that Fraxiparine significantly inhibited the invasive, migratory, and adhesive characteristics of A549 and 95D cells after 24 h incubation and showed a dose-dependent manner. Fraxiparine influenced the actin cytoskeleton rearrangement of A549 and 95D cells by preventing F-actin polymerization. Moreover, Fraxiparine could significantly inhibit CXCL12-mediated chemotactic migration of A549 and 95D cells in a concentration-dependent manner. Furthermore, Fraxiparine might destroy the interaction between CXCL12-CXCR4 axis, then suppress the PI3K-Akt-mTOR signaling pathway in lung cancer cells. For the first time, our data indicated that Fraxiparine could significantly inhibit the motility of lung cancer cells by restraining the actin cytoskeleton reorganization, and its related mechanism might be through inhibiting PI3K-Akt-mTOR signaling pathway mediated by CXCL12-CXCR4 axis. Therefore, Fraxiparine would be a potential drug for lung cancer metastasis therapy.

TACN-containing cationic lipids with ester bond: preparation and application in gene delivery.

A series of novel cationic lipids based on 1,4,7-triazacyclononane (TACN) with different hydrophobic chains were synthesized via the formation of a biodegradable ester bond. These lipids were found to have good buffering capacity at the pH range of 5.0-6.5, which is similar to that of the acidic endosomal compartments. The liposomes formed from these lipids and DOPE could condense DNA into nanoparticles with proper sizes. In vitro experiments showed moderate to good gene transfection efficiency of the formed lipoplexes. The structure-activity relationships of this type of lipids were discussed.

Cationic lipids containing protonated cyclen and different hydrophobic groups linked by uracil-PNA monomer: synthesis and application for gene delivery.

In this report, as candidates for non-viral gene vectors, cationic lipids L1, L2 and L3 based on protonated cyclen and different hydrophobic groups (cholesterol, dodecanol or diosgenin) linked by PNA monomer were designed and synthesized. Their liposomes were easily prepared by mixing the synthesized lipids with dioleoylphosphatidyl ethanolamine (DOPE) under appropriate mole ratios. Agarose gel retardation and fluorescent titration by ethidium bromide (EB) showed the strong DNA-binding ability with the K(sv) values of 1.21 × 10(7), 3.76 × 10(6) and 2.90 × 10(6) M(-1) for the liposomes formed from L1-L3, respectively. These liposomes could retard pDNA at an N/P ratio of 3 and form lipoplexes with sizes around 200-300 nm and zeta-potential values of +20-50 mV at N/P ratio from 4 to 10. Besides, the cytotoxicity of the three lipoplexes assayed by MTT is quite different. The results from in vitro transfection in HEK 293T and A549 cell lines showed that the transfection efficiency of L3/DOPE/DNA lipoplex at an N/P ratio of 6 and lipid/DOPE mole ratio of 1:2 is slightly higher than that of Lipofectamine 2000™, indicating that the title PNA monomer-based cationic lipids have great potential to be efficient non-viral gene vector.

Biodegradable cross-linked poly(amino alcohol esters) based on LMW PEI for gene delivery.

Polyethylenimine (PEI, especially with M(w) of 25,000) has been known as an efficient gene carrier and a gold standard of gene transfection due to its high transfection efficiency (TE). However, high concomitant cytotoxicity limited the application of PEI. In this report, several cationic polymers derived from low molecular weight (LMW) PEI (M(w) 600) linked with diglycidyl adipate (DA-PEI) or its analogs (diglycidyl succinate, DS-PEI and diglycidyl oxalate, DO-PEI; D-PEIs for all 3 polymers) were prepared and characterized. GPC gave M(w)s of DA-PEI, DS-PEI and DO-PEI as 6861, 16,015 and 35,281, respectively. Moreover, degradation of the ester-containing DS-PEI was also confirmed by GPC. In addition, hydroxyls in these polymers could improve their water solubility. These polymers exhibited good ability to condense plasmid DNA into nanoparticles with the size of 120-250 nm. ζ-potentials of the polyplexes were found to be around +10-20 mV under weight ratios (polymer/DNA) from 0.5 to 32. Agarose gel retardation showed that DNA could be released from the polyplexes after being pre-incubated for 30 h. In vitro experiments were carried out and it was found that DS-PEI showed about 5 times of TE compared to that of the PEI/DNA polyplex under a weight ratio of 1 in A549 cells. Meanwhile, the cytotoxicity of D-PEIs assayed by MTT is lower than that of 25 kDa PEI in HEK293 cells. These results suggested that this series of PEI derivatives would be promising non-viral biodegradable vectors for gene delivery.