[Identification of genes involved in biosynthesis of paeoniflorin in Paeonia lactiflora based on transcriptome analysis].

Paeoniflorin, a representative pinane monoterpene glycoside, is the main active component and quality index of Paeoniae Radix Alba and Paeoniae Radix Rubra.The possible biosynthesis of paeoniflorin is as follows: GPP is derived from mevalonate(MVA) and/or 2-C-methyl-D-erythritol 4-phosphate(MEP) pathway(s) followed by the catalysis with terpene synthase, cytochrome P450(CYP450), UDP-glucuronosyltransferase(UGT), and acyltransferase(AT), respectively.This study aims to explore the genes rela-ted to the biosynthesis of paeoniflorin.To be specific, the cDNA libraries for flowers, leaves, and roots of Paeonia lactiflora were established and sequenced.A total of 30 609 open reading frames(ORFs) were yielded.Through functional annotation and expression analysis of all CYP450 genes in the transcriptome, 11 CYP450 genes belonging to CYP71 A and CYP71 D subfamilies and showing expression trend consistent with monoterpene synthase PlPIN that may be involved in paeoniflorin biosynthesis were screened out.Subsequently, 7 UGT genes and 9 AT genes demonstrating the expression trend consistent with PlPIN which were possibly involved in paeoniflorin biosynthesis were further screened by functional annotation analysis, full-length sequence analysis, expression analysis, and phylogeny analysis.This study provided a systematic screening method with smaller number of candidate genes, thus reducing the workload of functional gene verification.The result laid a foundation for analyzing the biosynthesis pathway of paeoniflorin and the formation mechanism.

Engineering Gadolinium-Integrated Tellurium Nanorods for Theory-Oriented Photonic Hyperthermia in the NIR-II Biowindow.

Near-infrared (NIR) light-triggered hyperthermia has exhibited promising prospects in oncology therapy due to the unique merits including minimal invasiveness, monitorable, excellent therapeutic effect, and negligible side effects. Especially, the second NIR biowindow (NIR-II, 1000-1700 nm) with less absorbance and scattering by skin tissue, and deep tissue penetration, has received extensive attention for photonic hyperthermia. Unfortunately, the dissatisfactory photothermal conversion efficiency (PCE) and cumbersome preparation process of photo-driven heat conversion nanomaterials seriously hamper the future clinical application. To combat the aforementioned challenges, high imaging performance and desired therapeutic outcome 1D nanorods are constructed based on gadolinium-integrated tellurium nanorods (Te-Gd). In this system, magnetic resonance (MR) imaging and X-ray computed tomography (CT) imaging-guided photonic hyperthermia can be easily implemented in cooperation with Te-Gd. Importantly, Te-Gd possesses high PCE (41%) in the NIR-II biowindow because the transition of the excited electron can easily occur from the valence band (VB) to the conduction band (CB) on (1 0 1) and (1 0 2) crystal planes. Furthermore, the distinctive photostability, high tumor accumulation, as well as low systemic adverse effects of Te-Gd guarantee the potential in the clinic.

A highly active (102) surface-induced rapid degradation of a CuS nanotheranostic platform for in situ T1-weighted magnetic resonance imaging-guided synergistic therapy.

Polyvinylpyrrolidone-modified CuS nanocrystals (CuS NCs) with high photothermal conversion efficiency (46%) and pH and near-infrared (NIR) light-triggered degradation properties are a promising nanotheranostic platform for in situ magnetic resonance imaging (MRI)-guided synergistic photothermal and photodynamic therapy. On the one hand, the (102) surface of CuS NCs has a small bandgap based on density functional theory, which leads to high photothermal conversion efficiency. On the other hand, the S vacancy formation energy of the (102) surface is favourable. On entry into tumor cells through endocytosis, the S2- ions on the (102) surface of CuS NCs can be easily oxidized under the tumor microenvironment and 808 nm laser irradiation; then, a large amount of Cu+ ions can be released from CuS NCs and accelerate the degradation of nanocrystals. Cu+ ions can generate reactive oxygen species (ROS) under the tumor microenvironment and 808 nm laser irradiation. Meanwhile, the oxidation product Cu2+ ions can be generated from the oxidized Cu+ ions and applied for in situ T1-weighted magnetic resonance imaging. Moreover, the biodegradable CuS NCs possess a high tumor uptake and can be rapidly excreted with a low long-term retention/toxicity. Therefore, degradable and multifunctional CuS NCs are a safe and efficient candidate for the diagnosis and treatment of cancer.

Epimedium sagittatum inhibits TLR4/MD-2 mediated NF-κB signaling pathway with anti-inflammatory activity.

BACKGROUND: Epimedium sagittatum (Sieb.et Zucc.) Maxim., Ying-Yang-Huo in Chinese has been used as a traditional Chinese medicine and is deemed to "reinforce the kidney Yang". Previous studies showed that E. sagittatum could modulate the immune system and treat some chronic disease such as rheumatic arthritis, cardiovascular diseases and osteoporosis. The aim of this study is to evaluate the anti-inflammatory effects of ethyl acetate extracts (YYHs) of E. sagittatum and its mechanisms of action. METHODS: In order to explore the composition of YYHs, YYHs was analyzed using high performance liquid chromatography-mass spectrometry-mass spectrometry (HPLC-MS/MS) and in comparison with reference standards. Anti-inflammatory model was established in LPS-induced RAW264.7 cells. The levels of nitric oxide (NO) were measured with the Griess reagent. Production of tumor necrosis factor-alpha (TNF-α) and interleukin-2 (IL-2) were measured by enzyme-linked immunosorbent assays (ELISA). In addition, expression of p-p65 protein and TLR4/MD-2 complex was detected by western blots and flow cytometric, respectively. Nuclear factor kappa B (NF-κB) nuclear translocation was observed by fluorescence microscope. RESULTS: A total of eight compounds were identified, of which icariside II was the most abundant compound. YYHs (12.5-50 µg/mL) had no obvious cytotoxic effect on cells, and remarkably inhibited LPS-induced production of NO, TNF-α and IL-2 with a dose-dependent manner. Additionally, YYHs up-regulated expression of p-p65 and TLR4/MD-2 complex. Further research showed that YYHs significantly suppressed NF-κB p65 nuclear translocation. CONCLUSION: In brief, YYHs contributed to the inhibition of LPS-induced inflammatory response through the TLR4/MD-2-mediated NF-κB pathway and may be a potential choice to combat inflammation diseases. It includes a schema of pathways at the end of the paper.

Neuroprotective effects of Rhizoma Dioscoreae polysaccharides against neuronal apoptosis induced by in vitro hypoxia.

Rhizoma Dioscoreae polysaccharides (RDPS) are the primary active ingredient of Rhizoma Dioscoreae, which is a traditional Chinese medicine. RDPS have previously been shown to scavenge reactive oxygen species, and protect against D-galactose-induced mimetic aging. The present study aimed to investigate the neuroprotective effects of RDPS against hypoxia-induced neuronal cell apoptosis. Neuronal cells harvested from pregnant Sprague-Dawley rats were divided into groups, as follows: i) Normal control group; ii) hypoxia-induced apoptosis neuronal cell model; iii) 0.025 g/l RDPS-treated group; iv) 0.05 g/l RDPS-treated group; v) 0.1 g/l RDPS-treated group; and vi) 0.25 g/l RDPS treated group. Neuronal cell viability was investigated using an MTT assay, and neuronal cell apoptosis was analyzed using Annexin V-fluorescein isothiocyanate/propidium iodide double-staining, Hoechst 33342 fluorescent staining, Rhodamine 123 staining, polymerase chain reaction and immunocytochemical staining. The RDPS-treated neuronal cells exhibited improved viability, and decreased hypoxia-induced mitochondrial injury and apoptosis. In addition, the mRNA and protein expression levels of caspase-3 and B-cell lymphoma (Bcl)-2-associated X protein (Bax) were significantly downregulated, whereas the mRNA and protein expression levels of Bcl-2 were significantly upregulated, in the RDPS-treated hypoxic neurons, as compared with the apoptosis model (P<0.05). Furthermore, the ratio of Bcl-2 expression:Bax expression significantly increased following RDPS treatment, as compared with the apoptosis model (P<0.05). The results of the present study suggested that RDPS may attenuate hypoxia-induced neuronal cell apoptosis by altering the expression levels of key apoptosis-regulating proteins in hypoxic neurons.

Clinical Study on traditional Chinese medicine treatment of CSG and its impact on MTL, GAS and SS / 国际中医中药杂志

Objective To discuss of the therapeutic effects of treating three TCM types of Chronic superficial gastritis (CSG)disharmony of liver and stomach, weakness of the spleen and the stomach, and damp heat in the spleen and the stomach with chinese medicine, and to discuss its impact on the changes of three gastrointestinal hormones-- motilin (MTL), gastrin (GAS), and somatostatin (SS). Methods The data of 150 CSG patients in Shenzhen Bao’an Hospital from Feb, 2011 to Mar, 2013 were selected. These patients belonged to three kinds of syndrome, which was disharmony of liver and stomach, weakness of the spleen and the stomach, and damp heat in the spleen and the stomach, each syndrome had 50 cases. Different Chinese medicines were applied to different syndrome of patients. After one month treatment, the therapeutic effects and the changes of MTL, GAS, SS in these patients were detected by radioimmunoassay. Results After treatment, the total effective rate of the damp heat in the spleen and the stomach syndrome was 94%, the weakness of the spleen and the stomach type was 88%, disharmony of liver and stomach type was 90%, the difference was not statistically significant among the three types(χ2=3.84, P>0.05);after treatment, the serum MTL, SS level of disharmony of liver and stomach, weakness of the spleen and the stomach, and damp heat in the spleen and the stomach type were [damp heat in the spleen and the stomach type were (240.25±7.29)pg/ml, (68.23± 5.97)pg/ml;the weakness of the spleen and the stomach type were (260.16±8.84)pg/ml, (75.35±4.65)pg/ml;disharmony of liver and stomach type were(258.32±15.23)pg/ml, (80.12±7.88)pg/ml] higher than before treatment [damp heat in the spleen and the stomach type were (87.26±7.58)pg/ml, (49.34±4.93)pg/ml;the weakness of the spleen and the stomach type were (67.11±7.81)pg/ml, (57.15±5.11)pg/ml; disharmony of liver and stomach type were (89.26±7.60)pg/ml, (40.63±7.26)pg/ml], and the levels of serum GAS [(156.38±17.36)pg/ml, (146.15±15.56)pg/ml, (140.26±13.37)pg/ml] was lower than before treatment [(274.59 ± 17.82)pg/ml, (320.83 ± 15.02)pg/ml, (216.15 ± 13.77)pg/ml], the difference was statistically significant (P<0.05 or 0.01). Conclusions The changes of three hormones-MTL, GAS, SS can be used as an objective basis for the diagnosis and the judgment of healing efficacy of CSG treated by traditional Chinese medicine.

Development of innovative techniques for the endoscopic implantation and securing of a novel, wireless, miniature gastrostimulator (with videos).

BACKGROUND: Gastric stimulation via high-frequency, low-energy pulses can provide an effective treatment for gastric dysmotility; however, the current commercially available device requires surgical implantation for long-term stimulation and is powered by a nonrechargeable battery. OBJECTIVE: To test and describe endoscopic implantation techniques and testing of stimulation of a novel, wireless, batteryless, gastric electrical stimulation (GES) device. DESIGN: Endoscopic gastric implantation techniques were implemented, and in vivo gastric signals were recorded and measured in a non-survival swine model (n = 2; 50-kg animals). INTERVENTION: Five novel endoscopic gastric implantation techniques and stimulation of a novel, wireless, batteryless, GES device were tested on a non-survival swine model. MAIN OUTCOME MEASUREMENTS: Feasibility of 5 new endoscopic gastric implantation techniques of the novel, miniature, batteryless, wireless GES device while recording and measurement of in vivo gastric signals. RESULTS: All 5 of the novel endoscopic techniques permitted insertion and securing of the miniaturized gastrostimulator. By the help of these methods and miniaturization of the gastrostimulator, successful GES could be provided without any surgery. The metallic clip attachment was restricted to the mucosal surface, whereas the prototype tacks, prototype spring coils, percutaneous endoscopic gastrostomy wires/T-tag fasteners, and submucosal pocket endoscopic implantation methods attach the stimulator near transmurally or transmurally to the stomach. They allow more secure device attachment with optimal stimulation depth. LIMITATIONS: Non-survival pig studies. CONCLUSION: These 5 techniques have the potential to augment the utility of GES as a treatment alternative, to provide an important prototype for other dysmotility treatment paradigms, and to yield insights for new technological interfaces between non-invasiveness and surgery.

An endoscopic wireless gastrostimulator (with video).

BACKGROUND: Gastric electric stimulation (GES) at a high-frequency, low-energy setting is an option for treating refractory gastroparesis. The currently available commercial stimulator, the Enterra neurostimulator (Medtronic Inc, Minneapolis, MN), however, requires surgical implantation and is powered by a nonrechargeable battery. OBJECTIVE: To develop and test a miniature wireless GES device for endoscopic implantation in an experimental model. DESIGN: In-vivo gastric signals were recorded and measured in a nonsurvival swine model (n = 2; 110-lb animals). INTERVENTION: An endoscopically placed, wireless GES device was inserted into the stomach through an overtube; the two GES electrodes were endoscopically attached to the gastric mucosa and secured with endoclips to permit stimulation. MAIN OUTCOME MEASUREMENTS: Stable electrogastrogram measures were observed during GES stimulation. RESULTS: Electrogastrogram recordings demonstrated that gastric slow waves became more regular and of constant amplitudes when stomach tissues were stimulated, in comparison with no stimulation. The frequency-to-amplitude ratio also changed significantly with stimulation. LIMITATION: Nonsurvival pig studies. CONCLUSION: Gastric electric stimulation is feasible by our endoscopically implanted, wireless GES device.

Study on metabolites of ferulic acid and gallic acid compatible with danggui and chishao in rat plasma and urine / 中国中药杂志

<p><b>OBJECTIVE</b>To study the major metabolites of ferulic acid and gallic acid compatible with Danggui Chishaoyao in rat plasma and urine.</p><p><b>METHOD</b>The metabolites of ferulic acid and gallic acid in rat plasma and urine were analyzed after oral administration of compatible Danggui Chishaoyao using UPLC-Q-TOF-MS.</p><p><b>RESULT</b>On the basis of the mass information, it was inferred that in vivo metabolites of ferulic acid were be in the form of methylation products, sulfate conjugation products and glucuronidation conjugation products and so on; meanwhile, gallic acid was mainly transformed into eduction products and methylation products.</p><p><b>CONCLUSION</b>There are kinds of phase I and phase II metabolites of ferulic acid and gallic acid in rat plasma and urine, which provide a basis for its efficacious materials and action mechanism.</p>

Influence of different solutions on extracting of active components in Danggui Chishao drug pair / 中国中药杂志

<p><b>OBJECTIVE</b>To study the dissolution rate of active components of different extracting solvents of Danggui Chishao drug pair.</p><p><b>METHOD</b>The dissolution rates of phenolic acids (ferulic acid, vanillic acid and gallic acid), monoterpenes (gallic acid, peoniflorin, albiflorin, hydroxypeoniflorin and galloylpaeoniflorin) and phthalates (senkyunolide and ligustilide) contained in Danggui Chishao drug pair were determined by quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) and ultra high performance liquid chromatography in combination with triple-quadrupole mass spectrometry (UPLC-TQ-MS).</p><p><b>RESULT</b>The dissolution rates of phenolic acids and monoterpenes decreased with the increase in alcohol concentration, on the contrary the rates of phthalates increased. The relative dissolution rates of most active components were relatively high in water and low concentration alcohol than other solvents.</p><p><b>CONCLUSION</b>Liquid chromatography-mass spectrometry is practical for comprehensive multi-component assessment on traditional Chinese medicine preparation processes and can provide reference for optimization of processing parameters.</p>

Relationship between alcohol intake and TCM syndrome type in 1013 patients with cardio-cerebral diseases / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the relationship between alcohol intake and traditional Chinese medicine Syndrome type in patients with cardio-cerebral diseases.</p><p><b>METHODS</b>Study was conducted in 1013 selected inpatients with myocardial infarction (MI) or/and cerebral infarction (CI) by logistic multivariate regression analysis and comparison among groups.</p><p><b>RESULTS</b>The scores of blood stasis syndrome showed a decreasing tendency along the increasing of alcohol intake, but it cannot prove that taking alcohol could prevent blood stasis. The scores of dampness-heat syndrome increased in patients who had chronically taken alcohol in large dose.</p><p><b>CONCLUSION</b>Dampness-heat syndrome induced by alcohol is one of the basic mechanisms of cardio-cerebral diseases. Alcohol intake should be restricted.</p>