An integrated fecal microbiome and metabolome in rats reveal variations in gut microbiota and fecal metabolic phenotype of Semen Euphorbiae and Semen Euphorbiae Pulveratum.

Semen Euphorbiae (SE) is a toxic traditional Chinese medicine made from the dry or mature seed of Euphorbia lathyris L. Research demonstrates that the toxic side-effects from eating SE are associated with intestinal disturbance. By processing to produce Semen Euphorbiae Pulveratum (SEP), the toxicity is reduced, and diarrhea is attenuated. However, there are minimal studies on the differential effects between SE and SEP on microbiota and fecal metabolites. In this study, 16S rDNA sequencing and UPLC-Q-TOF/MS were interpreted with PCA and OPLS-DA multivariate analysis to understand the effect of SE and SEP on the gut microbiota and fecal metabolic phenotype in rats. Compared to the blank control group, the results showed that both SE and SEP were associated with increased microbes from the phylum Firmicutes and decreased Bacteroidetes, but the change was not as strong in the SEP administration group. Meanwhile, the fecal metabolism of rats also changed significantly, since 17 additional metabolites were detected in both groups, including amino acid metabolites, bacterial metabolites, and lipid metabolites. Our results indicate that the SEP administration group may reduce toxicity by differentially influencing intestinal metabolites and flora.

Construction of magnetic drug delivery system and its potential application in tumor theranostics.

Magnetic nanoparticles(NPs) are characterized by a rich variety of properties. Because of their excellent physical and chemical properties, they have come to the fore in biomedicine and other fields. The magnetic NPs were extensively studied in magnetic separation of cells, targeted drug delivery, tumor hyperthermia, chemo-photothermal therapy, magnetic resonance imaging (MRI) and other biomedical fields. Magnetic NPs are increasingly used in magnetic resonance imaging (MRI) based on their inherent magnetic targeting, superparamagnetic enzyme-like catalytic properties and nanoscale size. Poly(lactic-co-glycolic acid) (PLGA) is a promising biodegradable material approved by FDA and EU for drug delivery. Currently, PLGA-based magnetic nano-drug delivery systems have attracted the attention of researchers. Herein, we achieved the effective encapsulation of sized-controlled polyethylene glycol-3,4-dihydroxy benzyl-amine-coated superparamagnetic iron oxide nanoparticles (SPIO NPs) and euphorbiasteroid into PLGA nanospheres via a modified multiple emulsion solvent evaporation method (W1/O2/W2). NPs with narrow size distribution and acceptable magnetic properties were developed that are very useful for applications involving cancer therapy and MRI. Furthermore, SPIO-PLGA NPs enhanced the MRI T2 relaxation properties of tumor sites.The prepared SPIO NPs and magnetic PLGA nanospheres can be promising magnetic drug delivery systems for tumor theranostics. This study has successfully constructed a tumor-targeting and magnetic-targeting smart nanocarrier with enhanced permeability and retention, multimodal anti-cancer therapeutics and biodegradability, which could be a hopeful candidate for anti-tumor therapy in the future.

Characterization of diterpene metabolism in rats with ingestion of seed products from Euphorbia lathyris L. (Semen Euphorbiae and Semen Euphorbiae Pulveratum) using UHPLC-Q-Exactive MS.

Previous pharmacological studies have indicated that diterpenoids are the primary effective chemical cluster in the seeds of Euphorbia lathyris L. The seed products are used in traditional Chinese medicine in the forms of Semen Euphorbiae (SE) and Semen Euphorbiae Pulveratum (SEP). However, the metabolism of the plant's diterpenoids has not been well elucidated, which means that the in vivo metabolite products have not been identified. The current study screened the physiological metabolites of six diterpenes [Euphorbia factor L1 (L1), L2 (L2), L3 (L3), L7a (L7a), L7b (L7b), and L8 (L8)] in feces and urine of rats after oral administration of SE and SEP using UHPLC-Q-Exactive MS. A total of 22 metabolites were detected in feces and 8 in urine, indicating that the major elimination route of diterpenoids is via the colon. Hydrolysis, methylation, and glucuronidation served as the primary metabolic pathways of these diterpenoids. In sum, this study contributed to the elucidation of new metabolites and metabolic pathways of SE and SEP, and the new chemical identities can be used to guide further pharmacokinetic studies.

Metabonomics analysis of semen euphorbiae and semen Euphorbiae Pulveratum using UPLC-Q-TOF/MS.

Semen Euphorbiae (SE), the dry and mature seed of Euphorbia lathyris L., a common traditional Chinese medicine, has significant pharmacological activity. However, its toxicity limits its clinical application, and less toxic Semen Euphorbiae Pulveratum (SEP) is often used clinically. To explore the possible mechanism of SE frost-making and attenuation, this study used ultrahigh-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry to perform a comprehensive metabolomics analysis of serum and urine samples from rats treated with SE and SEP, and performed histopathological evaluation of liver, kidney and colon tissues. Meanwhile, the different metabolites were visualized through multivariate statistical analysis and the HMDB and KEGG databases were used to distinguish the differential metabolites of SE and SEP to reveal related metabolic pathways and their significance. In total, 32 potential biomarkers, 14 in serum and 18 in urine, were identified. The metabolic pathway analysis revealed that arachidonic acid metabolism, sphingolipid metabolism, tyrosine and tryptophan biosynthesis, the tricarboxylic acid cycle and seven other metabolic pathways were significantly altered. Importantly, compared with SE, SEP reduced the metabolic disorder related to endogenous components. The mechanism may be related to the regulation of lipid metabolism, intestinal flora metabolites, amino acid metabolism and energy metabolism. This study provided new insights into the possible mechanism of SE freezing and attenuation.

Comparison of Pharmacokinetics and Tissue Distribution Characteristics of Three Diterpenoid Esters in Crude and Prepared Semen Euphorbiae.

BACKGROUND: Semen Euphorbiae (SE) and Semen Euphorbiae Pulveratum (SEP) have a long history of medicinal use. SEP is the processed product of SE; both ancient and modern studies have shown that SEP has a lower toxicity compared to SE. To clarify the influence of processing on the pharmacological properties of SE and SEP, a study was carried out to compare the pharmacokinetics and distribution characteristics of three active compounds after oral administration of SE and SEP extracts. METHODS: A UPLC-MS/MS method was established to simultaneously determine the contents of Euphorbia factors L1, L2, and L3 in rat plasma and mouse tissues after an oral administration of crude and processed SE with approximately the same dosage. Plasma and heart, liver, spleen, lung, kidney, and colon tissue samples were treated with ethyl acetate and separated by gradient elution on a C18 column with a mobile phase of 0.1% formic acid and methanol. RESULTS: The established method had good selectivity, linear range, accuracy, precision, stability, matrix effect, and extraction recovery. The area under the concentration time curve, time to maximum concentration, maximum concentration, half-life of elimination, and mean retention time of plasma samples in SEP-treated group decreased, and the clearance in SEP-treated group increased. Moreover, the active component concentrations in colon, liver, and kidney tissues were more followed by those in the heart, lungs, and spleen. CONCLUSION: These results indicate that the processing could influence the pharmacokinetics and tissue distribution of Euphorbia factors L1, L2, and L3 after oral administration of crude and processed SE. The data obtained may lay a foundation for the clinical use of SE and for further study on the processing mechanism of SE.

Curcumin from Jianghuang (Rhizoma Curcumae Longae) protects against exposure to ultraviolet B by antioxidation and attenuating mitochondrion-dependent apoptosis.

OBJECTIVE: To investigate the protective effect of curcumin extracted from Jianghuang (Rhizoma Curcumae Longae) against ultraviolet B (UVB) and the possible mechanism. METHODS: Effects of curcumin were detected in vivo and in vitro. Morphological changes of white guinea pig skin were assessed by hematoxylin and eosin staining. HaCaT cell proliferation was measured by 3-[4,5-dimethylthylthiazol-2-yl]-2,5 diphenyltetrazolium broide (MTT) assays. The cell cycle distribution, apoptotic rate, level of reactive oxygen species (ROS), mitochondrial membrane potential, and intracellullar calcium ion concentration of HaCaT cells were detected by flow cytometry. Antioxidant levels in skin tissues and HaCat cells were measured by biochemical methods. RESULTS: UVB inhibited in vitro cell proliferation by inducing G2/M arrest, increasing ROS, apoptosis, and necrosis, and decreasing B-cell lymphoma-2, and increasing Bax, cytochrome c, and caspase-3 levels. CONCLUSION: Curcumin blocks the effects of UVB by reducing ROS and apoptosis, and reversing UVB-induced changes in the expression of apoptotic proteins. The mitochondrial pathway is involved in curcumin-regulated apoptosis.

Effects of Paeonol and Gastroretention Tablets of Paeonol on Experimental Gastric Ulcers and Intestinal Flora in Rats.

Paeonol, a major ingredient isolated from Moutan Cort, has various pharmacological effects. Our previous studies have shown that paeonol can exert antioxidant and anti-inflammatory therapeutic effects on ethanol-induced experimental gastric ulcer (GU). Therefore, in this study, we designed two GU models in rats induced by pyloric ligation (PL) and acetic acid and evaluated the protective effects of paeonol and gastroretention tablets of paeonol (GRT-Ps; 24, 48, and 96 mg/kg) on GU in rats and the effect of paeonol (48 mg/kg) on the intestinal flora. In vivo experiments showed that paeonol or GRT-Ps remarkably reduced gastric mucosal damage in a dose-dependent manner in the different types of models and improved the superoxide dismutase (SOD) activity and the malondialdehyde (MDA) content. And in fact, the sustained-release effect of GRT-Ps is more conducive to the improvement of GU compared with the rapid clearance of free drugs. In the PL-induced model, gastric secretion parameters, that is, pH and total acid, showed significant differences compared with the model group. In addition, paeonol treatment can improve the richness and diversity of the intestinal flora and increase the amount of beneficial bacteria, such as Lactobacillus. Paeonol and its stable sustained-release tablet GRT-Ps can promote ulcer healing by inhibiting oxidative stress and regulating the intestinal flora. This study can provide basis for the clinical treatment of GU with paeonol. Graphical Abstract.

A potential drug combination of omeprazole and patchouli alcohol significantly normalizes oxidative stress and inflammatory responses against gastric ulcer in ethanol-induced rat model.

Omeprazole (OME) is a representative of proton pump inhibitors and widely used in anti-ulcer treatment. However, OME may cause some inevitable side effects and the long-term consequences of OME could increase the risk of diarrhea. Patchouli Alcohol (PA), the main extract of Pogostemonis Herba, have demonstrated benefits in treating gastric ulcer (GU) with low toxicity. The present study aimed to investigate the synergistically protective effects of OME and PA against ethanol-induced GU in rats to study the involvement of antioxidant and anti-inflammatory activities. Moreover, the anti-apoptosis, anti-oxidant and anti-inflammatory effects in H2O2-induced gastric epithelial cells (GES-1) and LPS-induced RAW264.7 cells were determined, as well as the modulation of signaling proteins. The results demonstrated that PA alone or combined with OME provided remarkable benefits by reducing ulcer areas, modulating oxidant stress and inflammatory factors and the therapeutic efficacy was showed to be dose-dependent, which were partly superior to that of high-dose OME only. Additionally, co-treated regimen could superiorly down-regulate cell apoptosis and regulate the levels of oxidant activities and inflammatory cytokines on H2O2-induced GES-1 cells and LPS-induced RAW264.7 cells, which involved with cleaved caspase 3, Bcl-2 and BAX protein expressions and MAPK pathway. We provided a new understanding that the combination of OME and PA possessed gastroprotective effects on modulating cell apoptosis, antioxidant stress and anti-inflammatory responses against GU. Therefore, PA was inferred to take a potential and critic role in gastric mucosa protection.

Synthesis and anticancer activity of novel 9,13-disubstituted berberine derivatives.

Novel berberine derivatives with disubstituents on positions C9 and C13 were synthesized and evaluated for antiproliferative activities against human prostate cancer cell lines (PC3 and DU145), breast cancer cell line (MDA-MB-231) and human colon cancer cell lines (HT29 and HCT116). All compounds showed significantly enhanced antiproliferative activities compared with berberine. Notably, compound 18e exhibited the strongest cytotoxicity against PC3 cells with an IC50 value of 0.19 µM, and the highest selectivity index (SIPC3 > 20). Further studies showed that 18e could arrest the cell cycle at G1 phase, and significantly inhibit tumor cell colony forming and migration even at low concentrations. Interestingly, 18e could significantly induce cytoplasmic vacuolation, suggesting a different mode of action from berberine.

Resveratrol solid lipid nanoparticles to trigger credible inhibition of doxorubicin cardiotoxicity.

Background: Doxorubicin (DOX), a broad-spectrum chemotherapy drug, is clinically employed to treat cancers especially for breast cancer and lung cancer. But its clinical applications are limited by the dose-dependent cardiac toxicity. Resveratrol (Res), a polyphenolic antitoxin, has been proved to be capable of improving the cardiomyocyte calcium cycling by up-regulating SIRT-1-mediated deacetylation to inhibit DOX-induced cardiotoxicity. Purpose: The objective of this study was to develop a solid lipid nanoparticle (SLN) loaded with Res to trigger inhibition of DOX-induced cardiotoxicity. Methods: Res-SLN was prepared by emulsification-diffusion method followed by sonication and optimized using central composite design/response surface method. The Res-SLN was further evaluated by dynamic light scattering, transmission electron microscopy for morphology and high performance liquid chromatography for drug loading and release profile. And the Res distribution in vivo was determined on rats while the effect of inhibit DOX-induced cardiotoxicity was investigated on mice. Results: Res-SLN with homogeneous particle size of 271.13 nm was successfully formulated and optimized. The prepared Res-SLN showed stable under storage and sustained release profile, improving the poor solubility of Res. Heart rate, ejection fractions and fractional shortening of Res-SLN treating mice were found higher than those on mice with cardiac toxicity induced by single high-dose intraperitoneal injection of DOX. And the degree of myocardial ultrastructural lesions on mice was also observed. Conclusion: Res-SLN has a certain therapeutic effect for protecting the myocardium and reducing DOX-induced cardiotoxicity in mice.

Total Flavonoids Extracted from Oxytropis falcata Bunge Improve Insulin Resistance through Regulation on the IKKß/NF-κB Inflammatory Pathway.

Background. Insulin resistance (IR) is the main etiology of type 2 diabetes mellitus (T2DM). It has been known that total flavonoid extracts can markedly improve the hypoglycemic symptoms caused by IR. Nevertheless, the relevant molecular mechanism remains unclarified. Aim. This study aimed to investigate the antihyperglycemic effects and mechanism of the total flavonoid extract from Oxytropis falcata Bunge. Methods. STZ-induced T2DM rats (n = 35) were divided into 5 groups: model, low-, medium-, and high-dose total flavonoids, and pioglitazone groups. Ten healthy rats were used as controls. The serum insulin and inflammatory cytokines (MCP-1, TNF-α, and IL-6) level was measured by ELISA. The concentration of IRS-1, p-IRS-1, PKB p-PKB, PI3Kp85, and p-PI3K in skeletal muscles was determined by Western blot. The mRNA level of GLUT4, IκB, and NF-κB in skeletal muscle was detected by qRT-PCR. Results. The treatment of medium- and high-dose total flavonoids significantly reduced the FPG and P2hPG and enhanced insulin level in T2DM rats (P < 0.05). When compared with controls, the serum level of MCP-1, TNF-α, IL-6, IRS-1, and p-IRS-1 was significantly increased in T2DM rats, but the level of PKB, p-PKB, PI3Kp85, and p-PI3K expression was reduced (P < 0.05). The GLUT4 and IκB mRNA expression were significantly decreased, and NF-κB mRNA level was increased (P < 0.05). The treatment of low-, medium-, or high-dose total flavonoids markedly reversed the changes above (P < 0.05). Conclusion. Our study has confirmed the therapeutic effects of total flavonoids from Oxytropis falcata Bunge on IR. The flavonoids might reduce the production of inflammatory cytokines through downregulation of NF-κB expression in inflammatory pathway and regulate the IRS-1-PI3-K-PKB/Akt insulin pathway and thereby increased the GLUT4 expression.

Synergistic inhibitory effects of deferasirox in combination with decitabine on leukemia cell lines SKM-1, THP-1, and K-562.

A multi-center study from the French Myelodysplastic Syndrome (MDS) Group confirmed that iron chelation therapy is an independent prognostic factor that can increase the survival rate of patients who are suffering from transfusion-dependent low-risk MDS. In this study, we aimed to explore this clinical phenomena in vitro, by exploring the synergistic effect of the iron chelator Deferasirox (DFX) and the DNA methyl transferase inhibitor Decitabine (DAC) in the leukemia cell lines SKM-1, THP-1, and K-562. Treatment with both DFX or DAC promoted apoptosis, induced cell cycle arrest, and inhibited proliferation in all three of these cell lines. The combination of DFX and DAC was much greater than the effect of using either drug alone. DFX showed a synergistic effect with DAC on cell apoptosis in all three cell lines and on cell cycle arrest at the G0/G1 phase in K-562 cells. DFX decreased the ROS levels to varying degrees. In contrast, DAC increased ROS levels and an increase in ROS was also noted when the two drugs were used in combination. Treatment of cells with DAC induced re-expression of ABAT, APAF-1, FADD, HJV, and SMPD3, presumably through demethylation. However the combination of DAC and DFX just had strong synergistic effect on the re-expression of HJV.

Anti-tumor and immunomodulatory activities induced by an alkali-extracted polysaccharide BCAP-1 from Bupleurum chinense via NF-κB signaling pathway.

Bupleurum chinense is a well-known traditional Chinese medicine. Polysaccharides extracted from medical plants possess multiple healthy benefits. In the present study, an alkali-extracted polysaccharide (BCAP-1) was isolated from Bupleurum chinense, and evaluated its physicochemical features, anti-tumor activities and immunomodulatory effects. BCAP-1 was obtained by alkali-extraction, ethanol precipitation, and fractionation by DEAE-cellulose and Sepharose CL-6B columns. BCAP-1 markedly inhibited Sarcoma 180tumor growth in tumor-bearing mice, and increased the secretion of TNF-α in serum. MTT assay showed that BCAP-1 had no cytotoxicity against S-180 tumor cells. BCAP-1 enhanced the secretion of TNF-α and NO, and the transcripts of TNF-α and iNOS were increased. Meanwhile, BCAP-1 treatment induced the phosphorylation of p65 and decreased the expression of IκB in macrophages. These results suggest that BCAP-1 could activate macrophages through NF-κB signaling pathway, and the anti-tumor effects of BCAP-1 can be achieved by its immunostimulating features.

Effect of Yi Gong San Decoction on Iron Homeostasis in a Mouse Model of Acute Inflammation.

We investigated the effect of Yi Gong San (YGS) decoction on iron homeostasis and the possible underlying mechanisms in a mouse model of acute inflammation in this study. Our findings suggest that YGS regulates iron homeostasis by downregulating the level of HAMP mRNA, which may depend on regulation of the IL-6/STAT3 or BMP/HJV/SMAD pathway during acute inflammation.

Aß peptide secretion is reduced by Radix Polygalae-induced autophagy via activation of the AMPK/mTOR pathway.

Radix Polygalae is a traditional Chinese medicine that has been used as a sedative and to improve memory for a number of years. The impact of Radix Polygalae in patients with Alzheimer's disease has been investigated. However the mechanisms underlying its effects remain unclear. In the current study, the toxicity of various doses (100, 40, 20, 10, 5 and 0 µg/ml) of Radix Polygalae was measured in the human neuroblastoma cell line (SH-SY5Y) using an MTT assay. Changes in amyloid ß (Aß) levels in the supernatant of Chinese hamster ovary (CHO) cells overexpressing ß-amyloid pro-protein (APP) and BACE1 (CHO-APP/BACE1), were detected using an ELISA assay. In order to confirm that the Aß reduction was associated with autophagy, the autophagy marker protein, light chain 3 (LC3), was measured by western blot analysis and autophagosomes were assessed using MDC staining. In addition, the mechanism underlying the autophagy induced by Radix Polygalae was analyzed using western blotting to measure the protein expression of mammalian target of rapamycin (mTOR), p70s6k, Raptor, protein kinase B and adenosine monophosphate-activated protein kinase (AMPK), in addition to the phosphorylated forms of these proteins. The results demonstrated no significant toxicity of Radix Polygalae in SH-SY5Y cells, at a dose of 100 µg/ml. The secretion of Aß was markedly reduced following treatment with Radix Polygalae, and this reduction occurred in a dose-dependent manner. The autophagy levels were shown to be enhanced in the drug treatment group, using fluorescence microscopy. In addition, levels of LC3II/LC3I, the marker protein of autophagy, were also increased. The results of the current study suggest that Radix Polygalae may aid in the elimination of the Aß peptide, via the induction of autophagy, by the AMPK/mTOR signaling pathway. These results may provide a basis for further kin vivo investigation.

Dietary supplementation of Bacillus subtilis and fructooligosaccharide enhance the growth, non-specific immunity of juvenile ovate pompano, Trachinotus ovatus and its disease resistance against Vibrio vulnificus.

A feeding trial was conducted to investigate the effects of dietary administration of probiotic Bacillus subtilis and prebiotic fructooligosaccharide (FOS) on growth performance, immune responses and disease resistance of juvenile ovate pompano, Trachinotus ovatus. One thousand six hundred and twenty individuals (initial body weight: 10.32 ± 0.46 g, mean ± S.E) were fed nine practical diets according to a 3 × 3 factorial design: the basal diet as the control diet supplemented with three levels of B. subtilis (0, 1.05 × 10(7) or 5.62 × 10(7) CFU g(-1) diet), crossed with 0, 0.2% or 0.4% FOS. After an 8-week feeding experimental period, six fish per cage were sampled for immunity determination. Then 18 fish of each cage left were challenged by Vibrio vulnificus. The results showed that fish fed with 5.62 × 10(7) CFU B. subtilis g(-1) in combination with 0.2% FOS produced the highest specific growth rate, and were significantly higher than the groups fed with 0 and 0.2% FOS without B. subtilis supplementation (P < 0.05). Feed efficiency ratio significantly increased with the increasing doses of dietary FOS without B. subtilis added (P < 0.05). The immune assay showed that fish fed with the control diet produced the lowest respiratory burst activity and was significantly different from the groups fed the diets containing 0.2% FOS at each B. subtilis level and containing 0.4% FOS single (P < 0.05). Phagocytic activity was significantly decreased with the increasing doses of dietary B. subtilis at 0.4% FOS level (P < 0.05). Alternative complement pathway activity of the fish fed with 0.2% FOS single was significantly lower than those fed with 5.62 × 10(7) CFU B. subtilis g(-1) diet supplemented at each FOS level (P < 0.05). Fish fed with the control diet had the lowest lysozyme activity, and were significantly different from those fed with 0.2 or 0.4% FOS at 1.05 and 5.62 × 10(7) CFU B. subtilis g(-1) diet level. Moreover, fish fed with diets supplemented with 0.2% and 0.4% FOS at each B. subtilis level had notably lower cumulative mortality after 10 days following V. vulnificus infection (P < 0.05). Under the experimental conditions, dietary B. subtilis and FOS had a significant interaction on enhancing the immune responses and disease resistance of juvenile ovate pompano (P < 0.05).

The role of mitochondria-derived reactive oxygen species in hyperthermia-induced platelet apoptosis.

A combination of hyperthermia with radiotherapy and chemotherapy for various solid tumors has been practiced clinically. However, hyperthermic therapy has side effects, such as thrombocytopenia. Up to now, the pathogenesis of hyperthermia-induced thrombocytopenia remains unclear. Previous studies have shown that hyperthermia induces platelet apoptosis. However, the signaling pathways and molecular mechanisms involved in hyperthermia-induced platelet apoptosis have not been determined. Here we show that hyperthermia induced intracellular reactive oxygen species (ROS) production and mitochondrial ROS generation in a time-dependent manner in platelets. The mitochondria-targeted ROS scavenger Mito-TEMPO blocked intracellular ROS and mitochondrial ROS generation. By contrast, inhibitors of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, nitric oxide synthase, cyclooxygenase and lipoxygenase did not. Furthermore, Mito-TEMPO inhibited hyperthermia-induced malonyldialdehyde production and cardiolipin peroxidation. We also showed that hyperthermia-triggered platelet apoptosis was inhibited by Mito-TEMPO. Furthermore, Mito-TEMPO ameliorated hyperthermia-impaired platelet aggregation and adhesion function. Lastly, hyperthermia decreased platelet manganese superoxide dismutase (MnSOD) protein levels and enzyme activity. These data indicate that mitochondrial ROS play a pivotal role in hyperthermia-induced platelet apoptosis, and decreased of MnSOD activity might, at least partially account for the enhanced ROS levels in hyperthermia-treated platelets. Therefore, determining the role of mitochondrial ROS as contributory factors in platelet apoptosis, is critical in providing a rational design of novel drugs aimed at targeting mitochondrial ROS. Such therapeutic approaches would have potential clinical utility in platelet-associated disorders involving oxidative damage.

Antidiabetic effects of Tangnaikang on obese Zucker rats and the mechanism.

OBJECTIVE: To observe the effects of Tangnaikang (TNK), a compound traditional Chinese herbal medicine, on glucose metabolism and insulin resistance in obese Zucker rats. METHODS: Twelve male obese Zucker rats, 6 weeks old, were randomly divided into control group and TNK group (3.24 g/kg) after being fed for 2 weeks. All rats received high-fat diet and 4-week treatment. Body weight and blood glucose were tested every week. Oral glucose tolerance test (OGTT) was performed and fasting insulin level was tested on days 0, 14 and 28. Triglyceride, cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and free fatty acids (FFA) were tested on day 28. Glucose infusion rate (GIR) was tested by hyperinsulinemic-euglycemic clamp from day 29. The protein expressions of protein kinase B (Akt), phospho-Akt (p-Akt) (Thr308) and glucose transporter protein 4 (GLUT4) in skeletal muscle and GLUT4 in adipose tissue were measured after hyperinsulinemic-euglycemic clamp test. RESULTS: Compared with the control group, the fed blood glucose level and glucose level of OGTT at 120 min had a significant decline in TNK group on day 28, and TNK caused no alteration of the fasting serum insulin, and the GIR increased significantly in hyperinsulinemic-euglycemic clamp study. Furthermore, TNK increased Akt and p-Akt (Thr308) protein expressions in skeletal muscle and decreased the protein expression of GLUT4 in white adipose tissue. Body weight, and triglyceride, cholesterol, LDL-C and FFA contents were slightly decreased in the TNK group, but there were no statistically significant effects. CONCLUSION: TNK increases the protein expressions of Akt and p-Akt (Thr308) of the signal transduction pathway to influence the translocation of GLUT4 in skeletal muscle and improves glucose metabolism by reducing insulin resistance.

[Effects of Tangweian granule on 5-HT(2A)R in rat model with diabetic gastroparesis].

OBJECTIVE: To observe the effects of Tangweian granule on 5-HT(2A)R in rat model with diabetic gastroparesis (DGP). METHOD: The rats with diabetic gastroparesis induced by injecting alloxan and giving 200% Radix Rehmanniae preparata were divided into four groups randomly: Tangweian high dosage group, Tangweian low dosage group, motilium control group and the model control group, 10 rats each group. Each group was irrigated with drugs during establishing the model. Additionally, we chose 10 rats by way of normal control group. Further more, Tangweian high dosage group were irrigated stomach with gliclazide 20 mg x kg(-1) and Tangweian granule 31.75 g x kg(-1); Tangweian low dosage group were irrigated stomach with gliclazide 20 mg x kg(-1) and Tangweian granule 15.88 g x kg(-1); motilium control group were irrigated stomach with gliclazide 20 mg x kg(-1) and motilium 3.75 mg x kg(-1) and the model control group were irrigated stomach with distilled water. Then the effects of Tangweian granule on 5-HT(2A)R were observed. RESULT: The curative group had better effects than the control group in lowering the blood sugar and the level of 5-HT(2A)R content (P < 0.01). And there was significant difference between the curative group and control group (P < 0.05). CONCLUSION: It is verified that Tangweian granule has obvious effects on lowering the blood sugar and improving the level of 5-HT(2A)R.

[Cloning of distinguishing DNA sequences of Gastrodia elata Blume and application of them in identifying gastrodia tuber].

Gastrodia elata Bl. is a famous and costful traditional Chinese medicine. Their genomic DNA fingerprints were investigated using a modified Randomly Amplified Polymorphic DNA method. DNA fragments common to all or to fine populations were identified and recovered. Five DNA fragments were proven not to be reported through DNA cloning, PCR identifying, nucleotide sequencing and bioinformatics analyses and were received in and recorded by NCBI GenBank. Gastrodine contents of the Gastrodia tuber samples were determined using high performance liquid chromatography technique. The distribution of the five DNA fragments in 9 Gastrodia elata Blue populations and the correlation with gastromedicine content were studied. The results show the distribution of these DNA sequences varied greatly among the populations whereby DNA Sequence 1 was the common and distinguishing molecular marker for all the populations studied and DNA Sequence 2 may relate to higher gastrodine content. In conclusion, these DNA marker sequences can be employed to identify genuine gastrodia tubers, better varieties and optimize their selection and cultivating.